National Park Service

U.S. Department of the Interior

Natural Resource Stewardship and Science

Conservation Assessment for Triple-ribbed Milkvetch 

(Astragalus tricarinatus, Fabaceae) in Joshua Tree 

National Park

Natural Resource Report NPS/JOTR/NRR—2015/999
ON THE COVER

Triple-ribbed milkvetch, showing the cream-colored flowers and gray, well-spaced leaflets. Photo taken in Joshua Tree 

National Park, Upper East Deception Canyon, Little San Bernardino Mountains, Riverside County, California. 

Photograph by: Tasha La Doux.
Conservation Assessment for Triple-ribbed Milkvetch 

(Astragalus tricarinatus, Fabaceae) in Joshua Tree 

National Park

Natural Resource Report NPS/JOTR/NRR—2015/999

Naomi S. Fraga1

, Tasha La Doux2, Linda Prince1, Mitzi Harding2, and Josh Hoines2

1

Rancho Santa Ana Botanic Garden

1500 N. College Ave.

Claremont, CA 19711-3157

2 Joshua Tree National Park 

74485 National Park Drive 

Twentynine Palms, CA 92277

August 2015

U.S. Department of the Interior

National Park Service

Natural Resource Stewardship and Science 

Fort Collins, Colorado
ii

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This report is available in digital format from the Joshua Tree National Park website 

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format optimized for screen readers, please email irma@nps.gov.

Please cite this publication as:

Fraga, N., T. La Doux, L. Prince, M. Harding, and J. Hoines. 2015. Conservation assessment for 

triple-ribbed milkvetch (Astragalus tricarinatus, Fabaceae) in Joshua Tree National Park. Natural 

Resource Report NPS/JOTR/NRR—2015/999. National Park Service, Fort Collins, Colorado.

NPS 156/129380, August 2015 
iii

Contents 

Page

Figures....................................................................................................................................................v

Tables.....................................................................................................................................................v

Appendices............................................................................................................................................vi

Summary..............................................................................................................................................vii

Acknowledgments.................................................................................................................................ix

Acronyms and Initialisms.....................................................................................................................ix

Contacts.................................................................................................................................................ix

Introduction............................................................................................................................................1

Scope and Purpose..........................................................................................................................1

Background.....................................................................................................................................1

Species Description................................................................................................................................4

Taxonomic History ................................................................................................................................4

Phylogenetic studies.......................................................................................................................6

Biology and Ecology..............................................................................................................................7

Life History ....................................................................................................................................7

Reproductive Biology.....................................................................................................................9

Genetics........................................................................................................................................10

Habitat ..........................................................................................................................................11

Climate .........................................................................................................................................17

Vegetation and Associated species...............................................................................................17

Distribution and Abundance ................................................................................................................20

Status of Populations............................................................................................................................20

Threats..................................................................................................................................................22

Conservation Status .............................................................................................................................23

Ex-situ Conservation: Seed Bank Holdings.........................................................................................23

Research and Management Recommendations....................................................................................24

Research .......................................................................................................................................24

Population-level and phylogenetic studies..............................................................................24
iv

Contents (continued)

Page

Life-history, ecology, and reproductive biology studies.........................................................25

Park Management.........................................................................................................................26

Field Surveys and Habitat Modeling.......................................................................................26

Annual Monitoring ..................................................................................................................26

Protection.................................................................................................................................26

Literature Cited ....................................................................................................................................79
v

Figures 

Page

Figure 1. Joshua Tree National Park (shown in brown) is one of three large National Park 

Service units located in southern California. .........................................................................................2

Figure 2. The known global distribution of Astragalus tricarinatus is limited to 45 

occurrences (blue circles) in San Bernardino and Riverside counties of southern 

California. ..............................................................................................................................................3

Figure 3. Astragalus tricarinatus fruit, flowers, and habit ...................................................................5

Figure 4. A large tufted individual of Astragalus tricarinatus, erect inflorescences, 

flowers, and leaves.................................................................................................................................5

Figure 5. Astragalus bernardinus leaves, flowers, fruits, and weak, wiry stems often 

twining in shrubs....................................................................................................................................5

Figure 6. New shoots emerging from the base of Astragalus tricarinatus. ..........................................8

Figure 7. Habitat for Astragalus tricarinatus in the Little San Bernardino Mountains of 

Joshua Tree National Park. ..................................................................................................................14

Figure 8. Geologic map of southern California (USGS 2009), showing parent materials 

that Astragalus tricarinatus occurs on throughout its range, including within Joshua Tree 

National Park. ......................................................................................................................................15

Figure 9. Geologic units associated with Astragalus tricarinatus locations in Joshua Tree 

National Park. ......................................................................................................................................16

Figure 10. Frequency distribution graph for population size of Astragalus tricarinatus. ..................20

Tables 

Page

Table 1. Distinguishing morphological characters for Astragalus bernardinus and A. 

tricarinatus.............................................................................................................................................6

Table 2. Average number of seeds per fruit for Astragalus tricarinatus estimated from 

two studies. ............................................................................................................................................7

Table 3. Sampled populations for population genetic study of Astragalus tricarinatus.....................10

Table 4. Weather data from three RAWS stations (WRCC 2014) spanning the 

geographic and elevation range of Astragalus tricarinatus.................................................................17
vi

Appendices 

Page

Appendix A: Germination Study .........................................................................................................28

Appendix B: Genetic study..................................................................................................................31

Appendix C: GIS habitat model for Astragalus tricarinatus...............................................................56

Appendix D: Known occurrences for Astragalus tricarinatus............................................................64

Appendix E: Pilot study for Astragalus tricarinatus in Joshua Tree National Park............................68

Appendix F: Astragalus tricarinatus monitoring protocol ..................................................................74
vii

Summary 

Background

Triple-ribbed milkvetch (Astragalus tricarinatus A. Gray) is a perennial herb in the pea family 

(Fabaceae) endemic to southern California. It occurs in Riverside and San Bernardino counties in the 

transition zone between the Mojave and Sonoran deserts (CNDDB 2014, CCH 2014). Astragalus 

tricarinatus was listed by the federal government as endangered in 1998 due to its narrow 

distribution, limited number of individuals, and vulnerability to ground disturbance activities related 

to maintenance of an oil pipeline in Big Morongo Canyon (USFWS 1998, 2009). At the time of 

listing, A. tricarinatus had only been observed in four locations (Big Morongo Canyon, Whitewater 

Canyon, Mission Creek, and Aqua Alta Canyon) during the previous 20 years; however, more than 

one individual had only been observed in Big Morongo Canyon (USFWS 1998, 2009). Habitat and 

distribution of this species was thought to be limited to a few scattered individuals in canyons and 

sandy wash bottoms mostly of the southeastern San Bernardino Mountains and western Little San 

Bernardino Mountains. Fewer than 100 individuals had been documented by 1998. Since 2004,

surveys have yielded more information on the distribution, habitat preference, and abundance of A. 

tricarinatus (Amsberry and Meinke 2007, Fraga and Pilapil 2012, White 2004a, 2004b). We 

currently estimate 3100 individuals represented by 45 occurrences, and ten of these occurrences host 

more than 50 individuals.

The purpose of this report is to provide a comprehensive review of A. tricarinatus biology, ecology, 

distribution, taxonomic history, conservation status, and to provide management recommendations 

for populations that occur within Joshua Tree National Park (Park or JOTR) based on all known 

current information and recent studies conducted by Rancho Santa Ana Botanic Garden (RSABG) 

and JOTR. Here we present original research investigating seed germination, population genetics, 

and a habitat model for A. tricarinatus, with emphasis on plants that occur within JOTR. Finally, a 

monitoring protocol is provided for a pilot study initiated in the Park for assessing demographics and 

reproductive capacity. 

Original Research

Germination trials conducted between 2004 and 2013 indicate that seeds require some form of 

scarification to germinate. Clipping the seed coat at the hilum yielded 97–100% germination, 

whereas scarifying the seed coat using sandpaper yielded 68–96% germination. No treatment, a cold 

water soak prior to sowing, cold moist stratification, and boiling water followed by 24 hours of water 

soak had the lowest success rates with 0–4% germination rates. 

The population genetic study utilized ISSR (inter simple sequence repeat) markers and found 

individual populations were generally cohesive; however, the populations did not form reciprocally 

monophyletic population groups. This suggests either historic or contemporary gene flow among all 

three populations sampled. All three populations have similar levels of genetic diversity, however 

each population was found to have unique genetic diversity. 

The GIS habitat model utilized five parameters (elevation, slope, aspect, soil mapunit, and vegetation 

association) to assign probability values for potential habitat to the area within the Park. These 
viii

environmental datasets helped characterize occupied habitat for A. tricarinatus. The output from the 

model was used to prioritize areas for ground-truthing surveys, which yielded two new occurrences

for the species.

Recommendations

Conservation efforts for A. tricarinatus must focus on building upon baseline data, especially where 

gaps exist, by supporting research, field surveys, and long-term monitoring. Proposed research 

includes expanding the current population genetic study to determine levels of genetic diversity and 

gene flow across the range, among central and peripheral populations, and to evaluate source-sink 

dynamics among populations, including the occurrences characterized as “waifs.” Other research that 

would be particularly useful for future conservation efforts include a phylogenetic study to determine 

the closest relative of A. tricarinatus and its phylogenetic uniqueness, as well as an ecophysiological 

study to investigate drought tolerance across the range of the species. In addition, pollinator 

syndrome and breeding mechanism studies would aid tremendously in developing future 

management strategies. Current management practices in JOTR will include implementing a 

standardized monitoring protocol, as presented in this report, as well as continued efforts to survey 

and map known occurrences. Field surveys utilizing the existing habitat model will be used to search 

for new populations as well as improve the model. Finally, a detailed management plan that identifies 

a Conservation Area with Core Habitat will be developed in accordance with the Coachella Valley 

Multiple Species Habitat Conservation Plan and the USFWS 5-year Review. Management actions 

will be developed to ensure protection of essential ecological processes and protection of biological 

corridors and linkages critical for the long-term survival of A. tricarinatus.
ix

Acknowledgments 

The authors wish to thank the many people who participated in the development and implementation 

of the rare plant program at Joshua Tree National Park, and in particular to the efforts supporting the 

protection of Astragalus tricarinatus throughout its range. San Bernardino National Forest, The 

Wildlands Conservancy, and US Fish & Wildlife Service provided access and authorization to collect 

material as part of this study. We thank the RSA and UCR herbaria for access to specimens, and 

Duncan Bell for photos and information on recent surveys. Finally, we wish to thank Lise Grace, 

Nita Tallent, Scott White, and Alice Miller for helpful comments and careful review of this 

manuscript. This report is a collaborative effort between Joshua Tree National Park, Joshua Tree 

National Park Association, and Rancho Santa Ana Botanic Garden.

Acronyms and Initialisms

JOTR Joshua Tree National Park

RSABG Rancho Santa Ana Botanic Garden

USFWS United States Fish & Wildlife Service

CCH California Consortium of Herbaria 

CNDDB California Natural Diversity Database

CVMSHCP Coachella Valley Multiple Species Habitat Conservation Plan

EO Element Occurrence (location record from CNDDB)

Herbaria acronyms: C = Natural History Museum of Denmark; US = Smithsonian Institute; 

MO = Missouri Botanical Garden; UC = University of California Berkeley; 

RSA = Rancho Santa Ana Botanic Garden; UCR = University of California Riverside

RAWS Remote Automatic Weather Station

TWC The Wildlands Conservancy

Contacts (alphabetical order)

Naomi Fraga, Conservation Botanist, RSABG

Rancho Santa Ana Botanic Garden

1500 North College Avenue

Claremont, CA 91711

(909) 62508767 x231 

nfraga@rsabg.org

Tasha La Doux, Botanist JOTR

74485 National Park Dr.

Twentynine Palms, CA 92277

(909) 964-7304

ladouxtash@earthlink.net

1

Introduction 

Scope and Purpose 

Two federally listed plant species occur within the Joshua Tree National Park (JOTR) boundaries 

(Figure 1): Astragalus tricarinatus (triple-ribbed milkvetch, Fabaceae) and Erigeron parishii

(Parish’s fleabane, Asteraceae). Astragalus tricarinatus was federally listed as endangered in 1998 

primarily due to maintenance impacts from a crude oil pipeline running through occupied habitat in 

Big Morongo Canyon. Other factors contributing to the decision for federal protection included the 

restricted global distribution, limited number of individuals, and vulnerability to stochastic events 

(USFWS 1998). The purpose of this conservation assessment is to review and summarize all known 

background information on A. tricarinatus including taxonomic history, rarity and conservation 

status, habitat information, biology and ecology, known and potential distribution, and threats. We 

also report findings from germination trials and population genetic studies conducted at RSABG, in 

addition to results for a GIS-based habitat model developed at JOTR. Finally, we provide 

recommendations for management specific to JOTR, including a monitoring protocol to collect 

demographic and reproductive data, and suggestions for future research on A. tricarinatus. 

Background

Astragalus tricarinatus (Fabaceae) is endemic to southern California and is restricted to Riverside 

and San Bernardino counties; it primarily occurs in the transition zone between the Mojave and 

Sonoran Deserts (Figure 2). Currently, there are 45 documented occurrences (CCH 2014, CNDDB 

2014, see Appendix D). Ten of these are considered historic (not observed in over 20 years), whereas 

at the time of listing (November 1998), there were 15 occurrences, eight considered historic, and 

only one had more than one individual (USFWS 2009). Since 2004, a number of new populations 

have been discovered during field surveys; this, in addition to research conducted at known 

populations, has yielded valuable information on the distribution, habitat preference, and abundance 

of A. tricarinatus throughout its range (Amsberry and Meinke 2007, Fraga and Pilapil 2012, White 

2004a, 2004b). Astragalus tricarinatus is a perennial herb with white to cream flowers and a 

distinctive three keeled fruit that give the plant its common name, triple-ribbed milkvetch 

(Wojciechowski and Spellenberg 2012; Figure 3). It is a spring blooming species, generally between 

April and June, but can bloom as early as February. This species occurs between 390–1525 m (1285–

5005 ft) in elevation, primarily on steep slopes, but sometimes in rocky and gravelly washes (Fraga 

and Pilapil 2012). Threats to this species include the presence of exotic plant species, trail and road 

maintenance, illegal off-highway vehicle activity, residential development, wildland fire and 

suppression activities, climate change, pipeline maintenance, habitat fragmentation, reduced 

pollinator services, and small population sizes (Fraga and Pilapil 2012, USFWS 2009).
2

Figure 1. Joshua Tree National Park (shown in brown) is one of three large National Park Service units located in southern California. Two 

federally listed plants, Erigeron parishii (red triangles) and Astragalus tricarinatus (blue circles), are found within the boundaries of Joshua Tree 

National Park.
3

Figure 2. The known global distribution of Astragalus tricarinatus is limited to 45 occurrences (blue circles) in San Bernardino and Riverside 

counties of southern California. Three localities, Upper Covington Flat, Eureka Peak, and Whitewater Canyon, were sampled (yellow stars) for the 

genetic study presented in Appendix B. The weather stations (red triangles) used for a climate analysis across the range of the species are 

presented in Table 4.
4

Species Description

Astragalus tricarinatus is a relatively short-lived, herbaceous perennial (Figure 3) in the pea family 

(Fabaceae) that is loosely tufted from a woody taproot (Barneby 1964, Wojciechowski and 

Spellenberg 2012). Plants are 5–70 cm tall and finely strigose throughout, with straight, appressed 

hairs. Numerous stems arise from the base and are more or less erect to ascending in clumps 

(Barneby 1964). During dry seasons and/or after fruiting, the stems and leaves die back to ground 

level. The leaves are 7–20 cm long, with 17–27 leaflets that are 3–12 mm long, well-spaced, more or 

less narrowly obovate, elliptic and ovate, silvery canescent adaxially, and more or less green 

abaxially (Barneby 1964, Wojciechowski and Spellenberg 2012). The inflorescence is a loose 

raceme, 6-18 cm long with 5–15 flowers that are widely spreading to ascending; the peduncles are 

erect to ascending, 9–20 cm long (Figure 4). The calyx is 6.1–7.6 mm with mixed black and white 

hairs; the calyx tube is 4.1–5 mm; the flowers are cream colored, the banner is 12.6–15.7 mm long, 

recurved at a more or less 45° angle, the keel is 9.7–11 mm long. The fruit is a legume with two 

locules; at maturity the body is ascending and 24–42 mm long, 3.5–5.5 mm wide, more or less linear, 

three sided, glabrous, thin and papery; the stalk-like base is 1–2.5 mm long, stout, and jointed at top; 

the upper suture is a narrow ridge (Barneby 1964, Jones 1923, Wojciechowski and Spellenberg 

2012). There are 2–20 seeds per fruit. Chromosome counts are not reported for A. tricarinatus, 

however the new world Astragalus have chromosome numbers in an anueploid series with base 

numbers of x = 11, 12, 13, 14, 15 (Spellenberg 1976).

Taxonomic History 

Astragalus tricarinatus was first described by Asa Gray in 1876 from a herbarium specimen 

collected by Charles C. Parry in Whitewater Canyon, San Bernardino County, California (Gray 

1876). In his revision of the North American species of Astragalus, Marcus Jones (1923) placed A. 

tricarinatus in section Hamosi, however this placement has not been followed by subsequent authors 

because section Hamosi was based on A. hamosus L., which is an annual European species and 

thought to be unrelated (Rydberg 1927). Dr. Per Axel Rydberg (1927) transferred A. tricarinatus to 

the genus Hamosa Medik., then created a new section, Tricarinatae, within Hamosa with H. 

tricarinata (A. Gray) Rydb. serving as the type (Barneby 1964). Subsequent authors have followed 

Gray’s (1876) original treatment by placing A. tricarinatus in the genus Astragalus L. (Barneby 

1964, 1968, Isley 1986, Wojciechowski and Spellenberg 2012). Rupert Barneby (1964) transferred A. 

tricarinatus from its earlier placement in section Hamosi of Astragalus to section Leptocarpi

subsection Tricarinati, recognizing the section originally described by Rydberg in the genus Hamosa

at the rank of subsection in Astragalus. Section Tricarinatae of Hamosa included about a dozen 

species, but as a subsection is now reduced to only include two closely related species: A. 

tricarinatus and A. bernardinus M.E. Jones. 

Based on morphological similarities, A. bernardinus is considered the closest relative to A.

tricarinatus (Barneby 1964). Their close affinity was first described by Jones (1895) and Jepson 

found the species to be so similar that he included A. bernardinus in his circumscription of A. 

tricarinatus (1936). These two species have nearly overlapping geographic distributions, but differ in 
5

their habitat preference, growth habit, leaf morphology, flower color, and size of flower and fruit 

(Barneby 1964; Table 1, Figures 3, 4, and 5). Astragalus pachyphus has also been confused with A. 

tricarinatus, however the geographic range for these two species do not overlap and they are 

morphologically distinct. Specimens of A. pachyphus from the Central Valley in Kern County where 

previously misidentified as A. tricarinatus (Jones 1923, Jepson 1936). Astragalus pachyphus can be 

distinguished from A. tricarinatus by its narrower, linear, or linear-oblong leaflets and its laterally 

flattened fruit of almost woolly texture (Barneby 1964). 

Figure 3. Astragalus tricarinatus fruit (left), flowers (center), and habit (right). Photos by Mitzi Harding, 

Tasha La Doux, and Duncan Bell (left to right).

Figure 4. A large tufted individual of Astragalus tricarinatus (left), erect inflorescences (left and center), 

flowers (center), and leaves (right). Photos by Mitzi Harding.

Figure 5. Astragalus bernardinus leaves (left), flowers (left), fruits (center), and weak, wiry stems often 

twining in shrubs (right). Photos by Mitzi Harding.
6

Table 1. Distinguishing morphological characters for Astragalus bernardinus and A. tricarinatus.

Character Astragalus bernardinus Astragalus tricarinatus

Habitat stony areas among desert shrubs, juniper

woodlands

exposed rocky slopes, canyon walls 

along desert washes

Habit herbaceous perennial often twining in 

shrubs, inflorescence generally among 

branches 

loosely tufted herbaceous perennial with 

woody caudex and erect inflorescences

Stem weak, wiry, slender stems, 10–50 cm woody caudex with erect, stiff stems, 5–

70 cm

Leaflet number 7–19 17–27

leaflet shape narrow to broadly lanceolate narrowly obovate 

flower color Pale to dark lilac white to cream

banner 7.1–10.2 mm 12.6–15.7 mm

keel 6.8–9.4 mm 9.7–11 mm

calyx tube 2.7–4.1 mm 4.1–5 mm

Fruit three-sided, glabrous, 20–30 mm long, 4–5 

mm wide, lower surface often flat or 

shallowly channeled

three-sided, glabrous, 24–42 mm long, 

3.5–5.5 mm wide, narrow ridge on upper 

suture

Phylogenetic studies

Astragalus is one of the largest and most diverse genera of flowering plants in the world with over 

2,500 annual and perennial herbaceous plant species occurring throughout the northern hemisphere 

and South America. Western North America is one of the centers of diversity for the genus with 

about 400 species reported to occur here (Sanderson 1991). The genus has received considerable 

attention from plant taxonomists and several studies have examined systematics and phylogenetic 

relationships within the North American species (Gray 1964, Jones 1923, Rydberg 1929, Barneby 

1964, Isley 1986, Sanderson 1991, Liston 1992, Sanderson and Doyle 1993, Wojciechowski and 

Spellenberg 2012, Scherson et al. 2005). The aneuploid Astragalus species from the New World 

(chromosome numbers ranging from x=11–15) have been informally called Neo-Astragalus and form 

a monophyletic clade nested within old world Astragalus species which have euploid chromosome 

numbers (x =8, 16, 32; Sanderson and Doyle 1993). Astragalus tricarinatus likely occurs in the NeoAstragalus clade based on the placement of other species that are thought to be relatively closely 

related, although A. tricarinatus has not been sampled in any published phylogenetic studies so 

relationships within the Neo-Astragalus clade remain largely unresolved (Scherson et al. 2005).
7

Biology and Ecology

Life History

Astragalus tricarinatus is a relatively short-lived perennial that is thought to survive 3–5 years 

(Amsberry and Meinke 2007, Sanders 1999, USFWS 2009). There are no published demographic 

studies that have determined average lifespan of plants, although preliminary observations of plants 

in JOTR support a lifespan of less than 5 years (La Doux and Harding, unpublished data, also see 

Appendix E). A pilot study to assess occurrence viability was conducted by Amsberry and Meinke in 

2005 and 2006 at two locations in the Mission Creek watershed (Amsberry and Meinke 2007). They 

randomly selected 26 plants for a series of size parameter and reproductive capacity measurements. 

In 2005, they reported an overall mean stem length of 20.9 cm while the mean number of flowering 

stems per plant ranged from 18–46 throughout the season. They observed annual dieback at their 

study site with plants producing new shoots from a woody caudex (Figure 6). In addition, they 

reported that all reproductive plants appeared to be perennial, 38% had obvious woody bases

(Amsberry and Meinke 2007). 

Reports of seed production are relatively high for A. tricarinatus (estimated 2,759 seeds per plant) 

compared to reports from seed counts for other species of Astragalus (72–450 seeds; Amsberry and 

Meinke 2007, Gisler and Meinke 2001). An average of 12 viable seeds per fruit were estimated from

eight plants at Catclaw Flat (Amsberry and Meinke 2005). Similarly, an assessment of seed 

accessions at RSABG found 2–20 viable seeds per fruit with an average of 11 seeds per fruit (Table 

2). Based on these relatively high values of seed production per plant, it is likely that A. tricarinatus

maintains a large soil seed bank in-situ. Mechanisms for seed dispersal have not been documented; 

however given that A. tricarinatus has larger populations at the heads of canyons and smaller,

presumably waif, populations downstream, it is likely that some seeds are dispersed by water and/or 

gravity (USFWS 2009). Other vectors (e.g. ants, rodents, birds) for seed dispersal are of interest for 

future studies, as the upland sites appear to have abundant seed banks (White 2004a, 2004b,

Amsberry and Meinke 2007).

Table 2. Average number of seeds per fruit for Astragalus tricarinatus estimated from two studies. 

Study Date Location

# of sampled 

individuals Fruit #

Total # 

seeds

Avg seed

per fruit

RSABG 

accession 23184 05/04/2004

Wathier Landing 15 352 3808 10.82

Amsberry and 

Meinke 2007 05/05/2005

Catclaw Flat 8 59 714 12.10
8

Figure 6. New shoots emerging from the base of Astragalus tricarinatus (top photo). Dead branches are 

often present among new growth (bottom photo). Photos by Duncan Bell and Tasha La Doux, 

respectively.
9

Reproductive Biology 

Astragalus tricarinatus has specialized flowers that are characteristic of the legume subfamily

Papilionoidae (Fabaceae). The flowers are bilaterally symmetrical, the corolla being composed of 

five petals, including a generally broad banner that is held dorsally, two free wings (lateral to the 

banner), and a keel that consists of two fused petals that are opposite and positioned ventrally relative 

to the banner (Green and Bohart 1975). The flowers are borne on a raceme that is indeterminate and 

the inflorescence may bloom for several days to a week as individual flowers open along the axis 

(Barneby 1964, Green and Bohart 1975, Karron 1988, Figure 3). Sexual reproduction in the genus 

Astragalus is usually mediated by medium to large bees although pollinators for A. tricarinatus have 

not been observed. Astragalus flowers, like many papilionacous legumes, have a “tripping” 

mechanism for pollination (Karron 1988). Pollination of a flower is triggered when a bee lands on the 

keel and inserts its head under the banner; the keel is depressed (tripped) forcing the sexual column

(anthers and stigma) into contact with the bee. This action of tripping the flower allows pollen to be 

deposited on the bee, and at the same time the stigma can receive pollen brought by the bee from 

another plant. 

Astragalus tricarinatus is a narrow endemic restricted to southern California; its global distribution 

falls within a 518 sq km (200 sq mi) area (CNDDB 2014, CCH 2014). Such a narrow distribution

may have consequences for the reproductive biology of this perennial herb. Several studies have 

investigated reproductive strategies within the genus Astragalus, a genus with a large number of 

narrow endemics. These studies provided evidence that most narrowly distributed Astragalus species 

are self-compatible and by contrast, their more widely distributed congeners are often selfincompatible (Ralphs and Bagley 1988, Karron 1989, Kaye 1999). To the contrary for A. lentiginosus 

var. coachellae, a narrow endemic restricted to California’s Sonoran Desert, Meinke et al (2007) 

found that while bagged flowers were capable of setting fruits with viable seeds, the selfing rates 

were extremely low (<2%). These data support outcrossing as a primary means of seed production

for A. lentiginosus var. coachellae despite its narrow geographic range. Data from a population 

genetic analysis for A. tricarinatus (reported in Appendix B) is similar to the A. lentiginosus var. 

coachellae results in that the genetic diversity values favor a self-incompatible reproductive system 

for A. tricarinatus. Two of the values we calculated for genetic diversity, θ

II and GSTB, fall within a 

range more similar to outcrossing species than to selfing species (Appendix B). The breeding system 

of A. tricarinatus has not been studied; nonetheless A. tricarinatus is thought to be an outcrossing, 

perennial species of short to moderate life span. A study of the reproductive biology of A. 

tricarinatus involving bagging experiments, pollinator observations, and controlled pollinations 

could provide more information on pollination mechanisms, breeding system, and the ability to 

produce viable seed from self-pollination for this rare species.

Consistent with most species of Astragalus, A. tricarinatus does not show evidence of vegetative 

reproduction and relies on germination of seeds to recruit new individuals into populations (Barneby 

1964, Karron 1989, Kaye 1999). Plants described as seedlings have been observed in multiple years 

in large populations, including years with above (2004, 2005) and below (2006) average 

precipitation, suggesting that germination of seeds may occur frequently (White 2004a, 2004b,

Amsberry and Meinke 2007). However, this information is not sufficient to assess long-term viability 
10

of populations and more information is needed on the relative importance of life stages and their 

contribution to the species life cycle (i.e. seed bank, seedling success rates, and reproductive capacity 

of various life stages). 

Genetics

Long-term survival of any species is dependent upon its ability to endure adverse circumstances and 

adapt to environmental change. This ability is directly tied to the genetic diversity of the species. 

Knowledge of the breadth of genetic diversity and the distribution of that diversity is critical to 

management of populations. In ideal circumstances the entire breadth of genetic diversity would be 

preserved or protected, but this is rarely an option. Populations on the fringe of a distribution often 

experience more extreme environmental conditions than the species as a whole, and are more 

susceptible to random genetic drift and subsequent extirpation. Similarly, small populations are at 

greater risk of extinction due to stochastic events. Information on genetic diversity within and among 

populations can be used to infer historic gene flow and other evolutionary processes in addition to 

aiding in the management of individual populations. To better understand the genetic diversity of A. 

tricarinatus located within JOTR, a study of relative genetic diversity was undertaken using an 

analysis of ISSR genotype data. Two populations within JOTR, one near Eureka Peak (Population 1) 

and the other below Covington Flats (Population 2), were compared to a single population in 

Whitewater Canyon (Population 3) at Catclaw Flat on land managed by The Wildlands Conservancy

(Table 3, Figure 2). Details of the study are provided in Appendix B. 

Table 3. Sampled populations for population genetic study of Astragalus tricarinatus. EO= CNDDB 

Element Occurrence, TWC= The Wildlands Conservancy. 

Popn

ID Location EO# Latitude Longitude

Elevation

m (ft)

Land 

Manager

Population 

size

1 Eureka Peak 32 34.029 -116.356 1486 (4875) JOTR ~215

2 Covington Flats 28 33.989 -116.309 1435 (4710) JOTR ~70

3 Catclaw Flat 20 34.039 -116.678 1035 (3395) TWC ~100

No prior genetic work has been done on A. tricarinatus, though a few population genetic studies have 

been conducted on other species of Astragalus (Liston 1992, Karron et al. 1988, Travis et al. 1996, 

and Alexander et al. 2004). In general, the genus is notorious for low levels of genetic diversity 

(Liston 1992, Wojciechowski et al. 1999, Kazempour Osaloo et al. 2003, Wojciechowski et al. 

2004). For example, in allozyme-based studies, variation is generally low (GST <0.10, but see 

Astragalus clarianus and A. pauperculus; Liston 1990). However, recent population genetic studies 

using other fragment-based markers (ISSRs and AFLPs; Karron et al. 1988 and Alexander et al. 

2004) have detected more diversity in some Astragalus species. Based on the findings of our study, 

Astragalus tricarinatus is intermediate in diversity relative to those fragment-based studies (Table 

B10 in Appendix B).

Each population was characterized by a number of unique ISSR bands, and the number of unique 

bands was similar across the three populations (14-20 bands). Fourteen unique bands were 

characterized in the Eureka Peak population, 18 unique bands in the Covington Flats population, and 
11

20 unique bands in the Whitewater Canyon population. Population 2, from Covington Flats in JOTR, 

had the highest values for all population descriptors including the number of alleles (Na), number of 

effective alleles (Ne), Shannon’s Information Index (I), Nei’s gene diversity (h), Nei’s unbiased gene 

diversity (uh), and percentage polymorphic loci (%P) (see Tables B3-B5 in Appendix B). Individual

populations were generally cohesive, with the majority of samples from each population forming a 

clade or grade (see Figures B4-B6 in Appendix B). The populations did not form reciprocally 

monophyletic population groups however, suggesting either historic or contemporary gene flow 

among all three populations sampled. 

According to our data, the Eureka Peak and Covington Flats populations are more closely related to 

each other (NeiP = 0.034; uNeiP=0.028) than either are to the Whitewater Canyon population, but 

they are also much closer geographically. Interestingly, the Covington Flats population is slightly 

more closely related to the Whitewater Canyon population (NeiP=0.079–0.081; uNeiP=0.073–0.074) 

than is the Eureka Peak population (NeiP=0.082–0.086; uNeiP=0.076–0.080) despite being further 

away. This is graphically demonstrated in both the PCoA plots (Figures B1-B3 in Appendix B) and 

the UPGMA trees (Figures B4-B6 in Appendix B). All three populations have similar levels of 

genetic diversity. These analyses provide evidence for past or contemporary gene flow among all 

three populations, and that gene flow appears to be high based upon the low GST values.

Habitat 

Astragalus tricarinatus was thought to occur primarily on sandy or gravelly soils in washes and 

canyon bottoms, often at the base of scree slopes (Sanders 1999, USFWS 2009). However, since the 

discovery of a large population at the head of Whitewater Canyon in 2004, the preferred habitat for 

this species is now thought to be steep, rocky slopes and ridge tops (White 2004a, 2004b). This shift 

in understanding has led to several additional occurrences documented in similar upland habitats 

(White 2004a, 2004b, Fraga and Pilapil 2012). Because A. tricarinatus occurrences in washes and 

canyon bottoms are transitory, and usually consist of few plants, it is thought that plants in these 

habitats are waifs washed down from upland source populations. Documented occurrences of A. 

tricarinatus range in elevation between 390–1525 m (1285–5005 ft) (CCH 2014, CNDDB 2014).

Astragalus tricarinatus is found primarily on southern and westerly slopes with a gradient of 17° or 

more (Fraga and Pilapil 2012). In canyon bottoms the slopes are gentler with a gradient of 9° or less. 

Based on data collected from 50 plants in Upper East Deception Canyon (see Appendix E) of the 

Little San Bernardino Mountains, plants were found on SE- to SW-facing slopes with an average 

slope of 43° (median = 42°, minimum = 20°, maximum = 90°). Surveys and monitoring for this 

species is extremely difficult due to the steep slopes and unstable soils (Figure 7).

The A. tricarinatus substrate in Whitewater Canyon has been described as “unproductive looking 

gravelly soil” and “unusual metamorphic rock” (White 2004a, 2004b). According to Alice Miller 

(JOTR Vegetation Branch Chief between 2008-2010), the largest occurrences in JOTR “occur on 

shallow to deep, poorly developed, sandy or sandy-skeletal soils over weakly to moderately 

cemented bedrock derived from granite and gneiss. These soils are very friable and erosive, and 

somewhat excessively drained with medium to high runoff; they have low organic content, and low 

water holding capacity. Soil surfaces are dominated by gravel and fine gravel and have gravelly sand 
12

or gravelly loamy sand textures.” Although the soil at known population locations have not been 

analyzed, and therefore any superficially “unique” characteristic cannot be defined at this time, it has

a distinctive white to gray-green color that can be identified from a distance during field surveys 

(Figure 7). Soil analysis is needed to determine if soil properties are similar across the range of the 

species and if exceptional soil properties exist at locations with the largest occurrences. An effort to 

define which soil series is present at known populations across the range of the species would be very 

informative, as the current soil map used for the park only utilizes the soil “mapunit”, which 

describes an area of similar topography, climate, soil, and vegetation, but is quite coarse in scale.

According to the NRCS soil map of JOTR (Houdeshell et al 2014), there are three soil mapunits 

associated with typical A. tricarinatus habitat within JOTR: 

1) Xerric Torriorthents-Bigbernie association: This mapunit is very common throughout the 

Little San Bernardino Mountains (16,519 acres) and therefore captures the majority of the 

known occurrences in the park, (i.e. Upper East Deception Canyon). This mapunit is 

composed of 45% Xeric Torriorthents and similar soils, 25% Bigbernie and similar soils, and 

30% dissimilar minor components. The landform is described as steep mountain slopes (30-

75%) with east to southwest aspects. The parent material is colluvium derived from granitoid 

and/or gneiss over residuum weathered from granitoid and/or gneiss. The Xeric Torriorthents

are somewhat excessively drained soils, as are the other soils listed below, and are classified 

as Xeric Torriorthents. Bigbernie soils are classified as sandy-skeletal, mixed, thermic Typic 

Torriorthents.

2) Smithcanyon gravelly sand: This mapunit is very restricted, with only 2,806 acres in the 

Little San Bernardino Mountains. The Eureka Peak occurrences are found on this mapunit, 

which is composed of 80% Smithcanyon, dry and similar soils, as well as 20% dissimilar 

minor components. The landform is described as steep mountain slopes (30-75%) with east to 

southwest aspects. The parent material is colluvium derived from granitoid and/or gneiss 

over residuum weathered from granitoid and/or gneiss. Smithcanyon, dry soils are classified 

as being mixed, thermic, shallow Xeric Torripsamments.

3) Smithcanyon-Stubbespring-Rock outcrop complex: This mapunit is not very common in the 

park with only 4,075 acres designated in the Little San Bernardino Mountains. The two 

occurrences near Quail Mountain are found on this mapunit, which is composed of 40% 

Smithcanyon and similar soils, 25% Stubbespring and similar soils, 20% rock outcrop, and 

15% dissimilar minor components. The landform is defined as hilly backslopes or side slopes 

with southeast to northeast aspects and 15–50% slopes. Parent material is colluvium derived 

from granitoid over residuum weathered from granitoid. Stubbespring soils are classified as 

loamy, mixed, superactive, thermic, shallow Xeric Haplargids.

Astragalus tricarinatus is mapped as occurring on the following geologic substrates: maffic, felsic, 

gneiss, granodiorite, and alluvium (USGS 2009, NPS GRI 2014; Figures 8 and 9). According to the 

unpublished digital geologic map of JOTR (based upon USGS source maps), the majority of known 

localities are found on the geologic unit titled “Klmiff - Layered mafic, intermediate, and felsic rocks, 

foliated (Late Cretaceous)”, which is generally dark-colored with equal abundance of mafic and 
13

felsic rocks. This geologic unit is widespread throughout the Little San Bernardino Mountains (NPS

GRI 2014, Figure 9).

The largest occurrences known in JOTR occur on the Kmif geologic unit near Eureka Peak as well as 

the Kggif geologic unit near Quail Mountain. The latter is defined as “granitic, granodioritic, and 

intermediate rocks, foliated (Kggif)”. it is a light-colored substrate derived from granitoid rock 

material. Kmif is defined as “mafic and intermediate plutonic rocks.” It is fine- to coarse-grained and 

medium to dark gray in color. These rocks range in composition from granodiorite to tonalite and 

quartz diorite and contain biotite and biotite-hornblend, which could explain the slightly greenishgray hue of the substrate at the largest A. tricarinatus populations (Figure 7). Both Kggif and Kmif

are widespread throughout the Little San Bernardino Mountains. (NPS GRI 2014, Figure 9). 

There are a handful of localities in sandy wash corridors representing waif or transient occurrences. 

Although this habitat does not seem to support large populations, the waif or transient occurrences

may play a role in gene flow between occurrences in isolated canyons. These occurrences are found 

on the Qyaly geologic unit, which are present on piedmonts and pediment aprons, and defined as 

“young alluvium, light-sourced piedmont apron, younger unit.” The material consists of 

unconsolidated young sand and pebbly sand derived from quartz-rich light-colored granite (NPS GRI 

2014; Figure 9). The sandy wash soils are Carrizo, which are very deep sandy-skeletal hyperthermic 

Typic Torriorthents (Houdeshell et al 2014).

To better understand the habitat preferences of A. tricarinatus, and to be able to remotely identify 

probable habitat of this species, in 2012 a habitat model for JOTR (see Appendix C) was created in 

ArcGIS Desktop using a Python script (v2.6, © Copyright 1990-2014, Python Software Foundation). 

The model uses up to five environmental parameters (elevation, slope, aspect, soil mapunit, and/or 

vegetation association) to assign probability values of suitable habitat to the area within the park. A 

data layer representing occupied A. tricarinatus habitat within JOTR was created using a 15 m radius 

buffer around known point localities. The model uses this layer to narrow the values of the input 

parameters to only those that characterize at least 95% of the occupied habitat, and creates a 

weighted overlay of the results. Surveys in 2013 designed to ground truth the model yielded the 

discovery of 2 new populations in areas identified as highly probable habitat. The model was 

designed to be employed iteratively as more occurrences are discovered, and/or more detailed layers 

describing environmental parameters become available (such as soil series). 

An early attempt at creating the model involved manually identifying ranges of the environmental 

parameters which best characterize occupied habitat in JOTR. It was determined that there are four 

major vegetation associations (Single-leaf Pinyon Pine / Muller’s Oak Woodland Association, 

California Juniper / Blackbush Association, Muller Oak — California buckwheat — Narrowleaf 

goldenbush Association, and Catclaw Acacia — Desert Almond Association) and two soil mapunits 

(Xerric Torriorthents-Bigbernie association, and Smithcanyon gravelly sand) associated with >94% 

of known A. tricarinatus habitat in JOTR. The soil “mapunit” describes an area of similar 

topography, climate, soils and vegetation. It was also determined that more than 63% of the mapped 

occupied habitat is between 1370–1470 m (4495–4823 ft), 96% of occurrences have slopes ranging 

between 5–84% (3–40°), and 68% showed an aspect ranging between 155°–262° (SSE–W).
14

Figure 7. Habitat for Astragalus tricarinatus in the Little San Bernardino Mountains of Joshua Tree 

National Park. Top photo shows Erin Babich (red arrow) scrambling up a steep slope in Upper East 

Deception Canyon while surveying for A. tricarinatus. Bottom photo shows a closer view of the unique soil 

where these plants occur; a permanent plot was established in 2013 at this location near Eureka Peak. 

Photos taken by Mitzi Harding (top) and Tasha La Doux (bottom). 
15

Figure 8. Geologic map of southern California (USGS 2009), showing parent materials that Astragalus tricarinatus occurs on throughout its range, 

including within Joshua Tree National Park.
16

Figure 9. Geologic units associated with Astragalus tricarinatus locations in Joshua Tree National Park. Geologic units, listed youngest to oldest: 

Qyaly - Young alluvium, light-sourced piedmont apron, younger unit (Holocene); Klmiff - Layered mafic, intermediate, and felsic rocks, foliated 

(Late Cretaceous); Kmif - Mafic and intermediate rocks, foliated (Late Cretaceous); Kggif - Granitic, granodioritic, and intermediate rocks, foliated 

(Cretaceous) (NPS GRI 2014).
17

Climate

Long-term persistence of A. tricarinatus within its native habitat and current distribution is of 

concern due to its highly restricted global distribution, small population sizes, and geographic 

isolation of known occurrences. The global distribution of A. tricarinatus is narrow, with the core 

populations (excluding Agua Alta Canyon and the Chuckwalla/ Orocopia Mountains) spanning 

approximately 40 km (25 mi) east to west and 20 km (12 mi) north to south. For this reason, it is not 

unexpected that the weather conditions would be similar in such a localized region, which might 

suggest that even minor shifts in temperature or rainfall patterns will impact the species. We report 

mean annual precipitation, average air temperature, minimum air temperature and maximum air 

temperature from data gathered at three Remote Automatic Weather Stations (RAWS) across the 

range of A. tricarinatus (Table 4, Figure 2). Based on the data from these RAWS, average 

precipitation ranges from 14.71–17.22 cm (5.79–6.78 in) and average annual temperature ranges 

from 16.04°–19.49°C (60.88–67.08°F). The White Water RAWS Station occurs at the lowest 

elevation and is only 1 km west of a known occurrence in the lower portion of Whitewater Canyon; 

this station reports the warmest maximum temperature based on averages across the last six years 

(Table 4). The Morongo Valley and White Water Stations report the widest temperature fluctuations; 

11–12°C (19–22°F) difference between maximum and minimum temperatures, compared to 3°C

(5°F) at the Lost Horse Station. The Lost Horse RAWS Station is 6.7 km northeast of the nearest 

known occurrence of A. tricarinatus in JOTR; however it is at a similar elevation to occurrences near 

Covington Flat. The Lost Horse weather station reports the coolest average air temperature, but is 

also the driest with 14.71 cm (5.79 in) average annual precipitation. Current and future climate 

conditions may impact the long-term survival of the species. Unfortunately, no studies have been 

done to determine thresholds for temperature extremes, nor do we know the impact of changes in 

precipitation patterns and/or amounts.

Table 4. Weather data from three RAWS stations (WRCC 2014) spanning the geographic and elevation 

range of Astragalus tricarinatus (Figure 2). National Weather Service identification number is in 

parentheses following the weather station name.

RAWS Weather station (NWS ID):

Lost Horse 

(45614)

Morongo Valley 

(45863)

White Water

(45628)

Mean annual precipitation cm (in) 14.71 (5.79) 17.22 (6.78) 15.40 (6.06)

Mean avg. air temp °C (°F) 16.04 (60.88) 15.78 (60.4) 19.49 (67.09)

Minimum avg. air temp °C (°F) 14.29 (57.72) 11.54 (52.78) 14.44 (58.26)

Maximum avg. air temp °C (°F) 17.07 (62.72) 23.73 (74.72) 25.11 (77.19)

Elevation m (ft) 1280 (4200) 780 (2561) 776 (2546)

# of years data collected 22 6 6

Vegetation and Associated species

The upland habitats most frequently associated with Astragalus tricarinatus include: Larrea 

tridentata Shrubland Alliance, Pinus monophylla Woodland Alliance, Quercus cornelius-mulleri 

Shrubland Alliance, and Juniperus californica Woodland Alliance; whereas the lowland wash 

habitats are generally classified as Ambrosia salsola Shrubland Alliance, Chilopsis linearis

Woodland Alliance, and Acacia greggii Shrubland Alliance (CNDDB 2014, Sawyer et al. 2009, 

USFWS 2009, Keeler-Wolf et al. 2005, Evens et al. 2012, La Doux et al. 2013). 
18

According to the vegetation classification for JOTR (Keeler-Wolf et al. 2005, Evens et al. 2012, La 

Doux et al. 2013), A. tricarinatus most commonly occurs in the following vegetation associations:

Pinus monophylla / Quercus cornelius-mulleri Woodland Association, Juniperus californica / 

Coleogyne ramosissima Association, Quercus cornelius-mulleri — Eriogonum fasciculatum —

Ericameria linearifolia Association, Acacia greggii — Prunus fasciculata Association. 

The following species are commonly associated with A. tricarinatus throughout its range: 

Acmispon glaber (Vogel) Brouillet deer weed

Ambrosia salsola (Torr. & A. Gray) Strother & B.G. Baldw. cheese bush

Bebbia juncea (Benth.) Greene var. aspera Greene sweet bush

Chilopsis linearis (Cav.) Sweet subsp. arcuata (Fosberg) Henrickson desert willow

Crossosoma bigelovii S. Watson ragged rock flower

Dudleya lanceolata (Nutt.) Britton & Rose lance-leaved live forever

Echinocereus engelmannii (Engelm.) Lem. hedgehog cactus

Encelia farinosa Torr. brittle bush

Eriodictyon trichocalyx A. Heller yerba santa

Ericameria linearifolia (DC.) Urbatsch & Wussow golden bush

Eriogonum fasciculatum Benth. California buckwheat

E. saxatile S. Watson hoary wild buckwheat

Juniperus californica Carrière California juniper

Hesperoyucca whipplei (Torr.) Trel. chaparral yucca

Hilaria rigida (Thurb.) Scribn. big galleta grass

Keckiella antirrhinoides (Benth.) Straw yellow bush snapdragon

Larrea tridentata (DC.) Coville creosote bush

Mirabilis laevis (Benth.) Curran desert wishbone bush

Opuntia basilaris Engelm. & J.M. Bigelow var. basilaris beavertail cactus

Peritoma arborea (Nutt.) H.H. Iltis bladder pod

Psorothamnus arborescens (A. Gray) Barneby Mojave indigo bush

Quercus cornelius-mulleri Nixon & K.P. Steele Muller’s oak

Rhus ovata S. Watson sugar bush

Salvia apiana Jeps. white sage

Senegalia greggii (A. Gray) Britton & Rose cat claw

Sphaeralcea ambigua A. Gray globe mallow

Stephanomeria pauciflora (Torr.) A. Nelson wire lettuce

Stipa coronata Thrub. crested needle grass

Yucca schidigera Ortgies Mojave yucca

Ziziphus parryi Torr. var. parryi Parry’s jujube
19

Within JOTR, A. tricarinatus is often associated with the following species (in addition to those 

listed above): 

Acmispon heermannii (Durand & Hilg.) Brouillet Heermann's lotus

Acmispon rigidus (Benth.) Brouillet shrubby deervetch

Arctostaphylos glauca Lindl. bigberry manzanita

Asclepias subulata Decne. rush milkweed

Cercocarpus betuloides Nutt. mountain mahogany

Chrysothamnus viscidiflorus (Hook.) Nutt. yellow rabbitbrush

Encelia actoni Elmer Acton's brittlebush

Ericameria paniculata (A. Gray) Rydb. black-banded rabbitbrush

Ericameria teretifolia (Durand & Hilg.) Jeps. round leaf rabbit brush

Eriogonum nudum Benth. var. pauciflorum S. Watson little-flower wild buckwheat

Eriogonum wrightii Benth. var. nodosum (Small) Reveal knot-stem bastard sage

Lepidospartum squamatum (A. Gray) A. Gray scale-broom

Linanthus pungens (Torr.) J.M. Porter & L.A. Johnson granite gilia

Lupinus andersonii S. Watson Anderson's lupine

Lupinus excubitus M.E. Jones grape soda lupine

Nolina parryi S. Watson Parry's beargrass

Pinus monophylla Torr. & Frem. singleleaf pinyon pine

Prunus fasciculata (Torr.) A. Gray desert almond

Psorothamnus schottii (Torr.) Barneby indigo bush

Quercus john-tuckeri Nixon & C.H. Mull. Tucker oak

Salvia mohavensis Greene Mojave sage

Stipa speciosa Trin. & Rupr. desert needle grass

Yucca brevifolia Engelm. Joshua tree
20

Distribution and Abundance 

The distribution of A. tricarinatus spans 126 km (78 mi) east to west, from the foothills of the San 

Bernardino Mountains, east to the Chuckwalla Mountains (Figures 1 and 2) and encompasses about a 

518 sq km (200 sq mi) area (CNDDB 2014, CCH 2014). We estimate that A. tricarinatus has a 

global population of nearly 3100 individuals distributed among 45 known occurrences in ten 

watersheds throughout its geographic range (CNDDB 2014, Fraga and Pilapil 2012). For 

comparison, at the time of listing A. tricarinatus was mostly known from historical occurrences (had 

not been seen in >20 years) within seven watersheds, representing seventeen occurrences and fewer 

than 100 individuals (USFWS 1998).

Status of Populations

Most occurrences (the CNDDB definition for an “occurrence” includes all individuals within 1⁄4 mile 

of each other) of A. tricarinatus are on public lands or private preserves including: Bureau of Land 

Management, Joshua Tree National Park, the San Bernardino National Forest, and The Wildlands 

Conservancy White Water and Mission Creek Preserves (CNDDB 2014). The two largest 

populations occur on land managed by The Wildlands Conservancy at Wathier Landing (estimated 

1300 individuals in 2010; EO# 14) and 1 mi WNW of Wathier Landing (estimated 900 individuals in 

2010; EO# 36). The area southwest of Eureka Peak in JOTR has the third largest occurrence with 

215 plants observed in 2011. Most known occurrences (70% or 30 occurrences) have ten or fewer 

individuals documented at the site or have an unknown number of individuals because the occurrence 

is “historic”, i.e. they have not been observed in over 20 years (CNDDB 2014). The remaining 11

occurrences have population sizes that range from 21 individuals to 96. A frequency distribution 

graph shows the disparity in population size at documented occurrences (Figure 10). Small 

populations likely represent transient or waif populations that are washed down from larger 

populations higher in the watershed. 

Figure 10. Frequency distribution graph for population size of Astragalus tricarinatus. Most occurrences

(70%) have 10 or fewer individuals and only two occurrences have more than 500 individuals.
21

Population trends for A. tricarinatus are difficult to discern for lack of systematic monitoring for the 

species across its range. Ten of the occurrences recorded in the CNDDB are historic and four of these 

have such vague locality information that they have been difficult to relocate (CNDDB 2014). 

According to the data in CNDDB (2014), only one known occurrence was recorded as having 

Excellent (A) habitat quality (Wathier Landing, EO# 36); it has no threats listed. Eight occurrences 

are rated as good (B), including the largest population at Catclaw Flat (EO# 14), as well as three 

occurrences in JOTR near Covington Spring (EO#s: 27, 28, 30). Seven occurrences are rated as fair 

(C), and nine occurrences are rated as poor (D). Twelve occurrences, including several historic 

occurrences, are ranked as unknown. Several of the low ranks are presumably due to small 

population size at those locations since few threats are listed in CNDDB (2014). 

In JOTR, there are 18 known A. tricarinatus occurrences, 12 of which have six or fewer individuals 

and are likely better categorized as waif occurrences (i.e. not likely to be effective populations). 

There is one occurrence with 24 individuals and five occurrences with ≥65 individuals recorded 

(only one of these exceeds 100 individuals). The long-term viability of A. tricarinatus in JOTR, 

therefore, is considered uncertain due to the small number of individuals and their isolation from one 

another. Within the Little San Bernardino Mountains in JOTR, there are approximately 12 acres of 

occupied habitat with an estimated 560 plants. The occurrence near Eureka Peak (EO#32) contains 

the highest number of plants, estimated at 215 individuals.

Prior to 2006, presence of Astragalus tricarinatus in JOTR was only known from three vouchers:

1. Patrick Leary and J. Hogan, 3 May 1975, Long Cyn, 1.5 mi N of Joshua Tree NM boundary; 

T2SR5ES22, 1800 ft.; Riverside County (UNLV9780). 

2. Harlan Lewis, 28 April 1940, Joshua Tree National Monument, Keys View; Riverside 

County (LA205753). Recently annotated to Astragalus bernardinus (Tom Huggins pers. 

comm.).

3. J. B. Feudge (1462), 9 May 1926, Keyes Ranch, Little San Bernardino Mts, 1220 m; 

Riverside County (POM147871).

In 2006, three new occurrences were discovered in JOTR; two along the southern boundary in East 

Deception Canyon and one in Long Canyon. All three occurrences were found in sandy washes and 

are considered waif occurrences (EO#s: 21, 22, 23). The discovery of these waifs prompted 

additional surveys upstream and eventually led to the discovery of a larger population (~80 

individuals total) in Upper East Deception Canyon in 2008 (EO#s: 28, 29). These occurrences grow 

on a unique light-colored substrate (as described earlier under the Habitat section), which provided a 

much stronger search image for field surveys. The Park was then able to launch a concerted effort for 

targeted surveys in similar habitats. Since 2008, eleven occurrences have been documented, 

including the third largest population known for the species (EO#32). 
22

Threats

Current threats to A. tricarinatus are not well documented, in part because accessing its habitat

makes it a challenging species to study. The greatest threat to this species is likely its small isolated 

populations with a limited global distribution - rendering it vulnerable to stochastic events (Holsinger 

2013). Threats associated with small population sizes (<500 individuals) are applicable to most 

(95%) of the occurrences (Figure 10). While it is likely that additional occurrences will be

documented with further field efforts, the chances seem low for discovering additional large 

populations (>500 individuals) simply based on the low number of large populations found to date, 

despite many years of concerted field efforts. In addition, the remoteness of suitable habitat makes 

new discoveries of large populations less likely.

At listing, threats to A. tricarinatus included maintenance activities for the crude oil pipeline in Big 

Morongo Canyon, vehicle use, and vulnerability due to small population sizes (USFWS 1998, 2009). 

The listing rule recorded past instances of plants destroyed during road grading activities and plants 

impacted in pipeline realignment procedures (USFWS 1998). These activities remain a threat in Big 

Morongo Canyon. Additional threats that have been listed for A. tricarinatus include mining activity, 

residential development, illegal off highway vehicle activity, the presence of exotic plant species, 

trail and road maintenance, wildland fire and suppression activities, and climate change (Fraga and 

Pilapil 2012, USFWS 2009). 

There is little significant mining activity within the range of A. tricarinatus, but one occurrence

(EO#18) is near an active quarry. There is a substantial amount of potentially minable gravel in 

Whitewater Canyon, therefore future mining is a concern (USFWS 2009).

Much of the known occupied habitat is extremely rugged and difficult to access; therefore most 

occurrences are not subject to direct impacts from development pressure (e.g. housing), however, the 

occurrences in Lower Mission Creek are currently threatened by development. There is concern for 

the indirect effects of residential or commercial development occurring nearby, especially along the 

southern boundary of JOTR, starting at state highway 62 all the way to Indio. With an increase in 

human population in this region, we can expect increased illegal OHV activity, spread of non-native 

or invasive species, disrupted pollinator services by native pollinators, and the potential for increased 

fire frequency. In addition, threats of urbanization and development into habitats downstream from 

large populations, such as in Lower Mission Creek, could prove quite pervasive. The role of waif or 

transient populations found along alluvial corridors remains unknown. It is possible that these 

transient occurrences, for example in East Deception Canyon, are in fact contributing to the flow of 

genes among populations that may otherwise be isolated from one another. 

Non-native species that are associated with and may pose a threat to A. tricarinatus include Bromus 

madritensis L. subsp. rubens (L.) Husn. (red brome) and Brassica tournefortii Gouan (Saharan 

mustard). However, direct impacts caused by the presence of exotic or invasive species have not been 

documented, nor are disease and predation known to be threats affecting A. tricarinatus (USFWS 

2009). In addition, the impact of local or global climatic changes on seed bank ecology, 
23

mortality/recruitment rates, longevity of individuals, reproductive success, or other life-history and 

ecological traits remain unknown for this species. 

Conservation Status

Astragalus tricarinatus was listed by the federal government as endangered in 1998 and is 

considered under the Coachella Valley Multiple Species Habitat Conservation Plan (CVMSHCP), a 

regional conservation plan developed under section 10(a)(1)(B) of the Endangered Species Act

(CVMSHCP 2007, 2014). Critical habitat for Astragalus tricarinatus has not been designated 

(USFWS 2009), and no recovery plan or outline has been published for A. tricarinatus (USFWS 

2014, species profile website).

The current rarity ranking of this species is provided below (CNPS 2014).

• Federal Listing Status: Endangered

• State Listing Status: None

• State Rank: None. S1

• California Native Plant Society: 1B.2 (Fairly endangered in California)

• Global Rank: G1

Ex-situ Conservation: Seed Bank Holdings

Conservation of plant species can be achieved by protection of habitats and populations in nature (insitu), or by the preservation of genetic diversity outside of an organism’s natural habitat (ex-situ), 

often in gene banks, botanic gardens, zoos, or translocation sites (Cohen et al. 1991). Ex-situ

conservation provides back-up for biological diversity that might otherwise be lost in nature due to 

human induced environmental change or natural extinction. Seed banks are considered one of the 

most economical forms of ex-situ conservation. It not only provides an excellent opportunity for 

maintaining genetic diversity in vulnerable populations, but they also provide a low impact means for 

research opportunities. In other words, without depleting or impacting the natural populations, seed 

bank material could be grown in a greenhouse setting to study some very important ecological 

questions, such as mating system, climate thresholds, seed viability, germination requirements, and 

fire ecology. Although there are no plans to augment native populations of A. tricarinatus at this 

point, this may be considered in the future if significant declines in population numbers are 

documented.

There are four seed accessions of A. tricarinatus stored in the seed bank at RSABG (Table A1 in 

Appendix A); two of these are from populations within JOTR. Germination studies of seeds for A. 

tricarinatus have been conducted to assess baseline seed viability and germination requirements. 

Seed germination trials conducted at RSABG report variable rates that range from 0 to 100% 

germination depending on treatment (RSABG internal data). Nicking or clipping the seeds using a 

knife or scalpel produced the highest germination rate consistently, with 97-100% germination rate. 

The lowest germination rates (i.e. no seeds germinating) were observed after “no treatment” or when 
24

seeds were soaked in water prior to sowing. Many Astragalus are well known for having hard 

impermeable seed coats that cause physical dormancy and prevent germination. The hard seed coats 

prevent water absorption, permeation of gases, or they may constrain the embryo, all of which can 

cause dormancy (Baskin and Quarterman 1969). Seeds with this dormancy mechanism require 

abrasion or scarification by chemical or mechanical means in order for germination to occur. Based 

on the trials presented here, it appears that clipping the seed coat provides the most reliable method 

for triggering germination in A. tricarinatus. The oldest A. tricarinatus seed accession currently in 

storage was collected in 2004. During germination trials in 2014, these seeds germinated at 97% (29 

of 30) after clipping, however 10 of the germinated seeds had poor cotyledon development and died, 

which lowered the post-germination survivorship to 63%. Whether the poor survivorship is due to the 

age of the seeds cannot be determined without further testing, therefore it is unknown if seeds will 

retain high viability in cold storage (-20 C) for long periods of time. Given that these seeds are likely 

to remain dormant in natural soil seed banks, we suspect that the seeds will remain viable in dry cold 

storage for many years. Additional details for the germination studies conducted at RSABG are 

summarized in Appendix A.

Research and Management Recommendations

Research

Many knowledge gaps still exist with regard to the life history, geographic distribution, habitat 

preference and ecology of A. tricarinatus. Research questions outlined here aim to advance our 

knowledge of this species to inform management strategies and long term conservation efforts. 

Population-level and phylogenetic studies

We recommend pursuing population genetic and phylogenetic research that would further our current 

knowledge of the genetic distinctiveness and diversity present within and among populations 

throughout the range. These studies could aid in prioritizing conservation efforts. Examples include: 

1. Expanding the population genetic study presented in this report to include additional 

populations across the range. The initial ISSR study (Appendix B) indicates that there are 

unique alleles present in the JOTR populations, i.e. alleles not present in the San Bernardino 

Mountains. However, due to the limited sampling used in the initial study it is possible these 

alleles are not limited to JOTR. Expanding this study will provide information regarding the 

uniqueness of the JOTR populations. 

2. There are no published phylogenetic studies that have sampled A. tricarinatus or its close 

relatives. It is hypothesized that A. bernardinus is the closest relative based on morphology, 

however additional information is needed regarding the phylogenetic position of A. 

tricarinatus relative to other species of Astragalus and its evolutionary history. 

3. Levels of gene flow among populations of A. tricarinatus are not well understood and the 

parameters that were analyzed in the current study are not well suited to study contemporary 

gene flow. An expanded paternity analysis utilizing the genotypes of the mother plants and 
25

their offspring compared with a pool of potential fathers within a population would provide 

information on contemporary gene flow and directionality. 

4. The role of transitory populations or waif individuals for gene flow among populations would 

be of great interest to long-term protection of the species. If these transitory gene pools play 

an important role in connecting populations, conservation of the supporting habitats might 

become a greater priority. 

Life-history, ecology, and reproductive biology studies

We recommend ecological studies addressing any gaps in current knowledge of basic biology for A. 

tricarinatus. In particular, studies that would significantly add to the long-term management and 

conservation of the species include, but are not limited to: 

1. Long-term population viability, including waifs and transient populations found in the wash 

corridors.

2. Determine rates of survivorship for individuals and determine meaningful demographic size 

classes (e.g. seedlings, juveniles, and adults).

3. Identify effective pollinators and learn more about pollinator biology, including timing of 

pollination, life cycles, pollen dispersal distances, host plants, etc.

4. Evaluate potential seed dispersal vectors (animals, wind, etc.)

5. Determine whether self-pollination is possible, or whether any reproductive barriers are 

present (e.g. self-incompatibility, inbreeding depression).

6. Conduct a more detailed soil analysis. For example, define which soil series are present at 

populations and determine unique chemical or physical properties across the range of the 

species. Also, determine if exceptional soil properties exist at the locations that hold the 

largest populations.

7. Document physiological response to various rainfall patterns (amount/timing of rains, 

monsoonal rains), as well as changing environmental conditions such as shifting seasons or 

higher/lower temperature extremes.

8. Determine germination rates after fire (smoke treatment on seeds), as well as the ability to 

resprout from the base post-fire.

9. Determine germination and survivorship rates for seeds of varying ages, under various

storage methods, and further testing on scarification methods to mimic natural conditions 

(both physical and chemical treatments).

10. Test for changing environmental conditions in a controlled environment. For example, the 

effect of extended periods of drought or increased competition from non-native species on 

survivorship.
26

11. Improve our understanding of preferred habitat parameters by conducting additional surveys 

and analyzing with habitat models. 

12. Determine whether rhizobium and mycorrhizae relationships are necessary for or enhance 

growth for this species through cultivation experiments.

Park Management

Field Surveys and Habitat Modeling

Continued efforts to locate new populations utilizing the habitat model will remain a priority, as well 

as improving upon the habitat model. Field surveys will focus on high probability areas according to 

the maps produced by the current model (see Appendix C). Because the model is meant to be 

iterative, any new data points must be uploaded immediately, and the model updated before another 

field survey takes place. According to the current habitat model, geospatial data is most effective if

taken for each individual more than 15 m apart, so time permitting, additional geospatial data will be 

collected. Future habitat models, if possible, will incorporate absence points, as well as providing a 

means to weight the parameters differently. In addition, we hope to include habitat information from 

the entire geographic range in future habitat models.

Finally, future efforts will prioritize then revisit all known occurrences in order to document 

population status and changes over time. Revisits will likely need to occur every 3–5 years, however 

the revisitation rate will need to be adapted according to any observed trends. 

Annual Monitoring

As recommended in the CVMSHCP (2007, 2014) and by the USFWS (2009) 5-year Review, annual 

monitoring will occur in JOTR. We propose following the protocol presented in Appendix F at the 

two permanent plots for a minimum of five years, or until better data can be used to adapt improved 

protocols. The frequency of monitoring could be reassessed after the first five-year period. With 

consistent annual data collection, results from statistical analyses on demographics and reproductive 

biology for this species will be much more effective at predicting any correlative relationships. In 

particular, long-term viability of the two A. tricarinatus occurrences being monitored, as well as 

longevity of individuals, will be addressed. Adding waif or transient populations found in wash 

corridors to the annual monitoring would provide valuable information about their role in the longterm viability of the species. Finally, there are a number of recommendations presented in Appendix 

E pertinent to annual monitoring that will be followed or addressed.

Protection

Complete protection should be attempted for all individuals and occupied habitat of A. tricarinatus,

in accordance with the Endangered Species Act (USFWS 2014). Collecting of specimens can be 

allowed for the purpose of research or recovery efforts, but must be permitted by NPS and reported 

to USFWS. 

Areas within JOTR with known populations or individuals will be designated as a strict “no 

disturbance” zone, which would exclude any kind of ground disturbance, vegetation pulling, 

trimming, or removal. In particular, fire suppression activities (e.g. hand lines) should not be allowed 
27

in and around the known populations. Any newly proposed trails or access routes will require field 

surveys and GIS analysis prior to plan approval in order to ensure the route avoids all known 

individuals and potential habitat. 

Joshua Tree National Park was identified in the Coachella Valley Multiple Species Habitat 

Conservation Plan (CVMSHCP) as a potential Conservation Area (CVMSHCP 2007, 2014). Now 

with the current information in place, a detailed management plan adopting the goals and objectives 

of the CVMSHCP will be developed. Specifically, the plan will need to specify a Conservation Area

with Core Habitat in accordance with the CVMSHCP and the USFWS 5-year Review, as well as 

develop management actions that will ensure protection of essential ecological processes and protect 

biological corridors and linkages (CVMSHCP 2014, USFWS 2009). 

Finally, as a measure to protect the genetic diversity found within JOTR, an ex-situ seed bank will be 

established. Seeds will be collected from all (or most) known populations and placed in long-term 

storage at RSABG. All seed collection, viability testing, and seed processing will follow ethical and 

current protocols, such as those described in the Seeds of Success (2014) technical report.
28

Appendix A: Germination Study

By Naomi Fraga, Michael Wall, and Evan Meyer, Rancho Santa Ana Botanic Garden

Seeds in the legume family (Fabaceae), including Astragalus, are well known for having hard 

impermeable seed coats that prevent germination and cause physical dormancy. Hard seed coats can 

cause dormancy because they prevent absorption of water, permeation of gases, or they may 

constrain the embryo (Baskin and Quarterman 1969). Hard seed coats generally become permeable 

and seeds often germinate after the seed coat is abraded or scarified by chemical or mechanical 

scarification. In nature, seeds of Astragalus tricarinatus are likely scarified through a heating/cooling 

cycle, which may include some type of chemical interaction from the soil, and/or the physical action 

of sand or gravel abrading the seed coat during rain events. Regardless of how the seed coat is 

broken, doing so allows germination to occur when seeds are moistened. 

Currently, there are four seed accessions of Astragalus tricarinatus held at RSABG (Table A1), all 

localities are from the Little San Bernardino Mountains. Two accessions are from within Joshua Tree 

National Park (JOTR), collected in 2011 from the Eureka Peak and Upper Covington Flat areas. The 

other two accessions are from Wathier Landing and Whitewater Canyon area, collected in 2004 and 

2011, respectively.

Amsberry and Meinke (2007) conducted a pilot study on germination requirements for A. 

tricarinatus. Although the methods used for scarification were not included in the report, they found 

that 80% of viable seeds germinated within 72 hours after “scarification” and wetting. Between 2004 

and 2013 a variety of treatments were tested at RSABG, success rates varied from 0–100%, 

depending on the pre-treatment of seeds (Table A2). All germination tests at RSABG were conducted 

on 0.5% agar solution on clear plastic examination plates maintained at 11 hrs. light at 20°C and 13 

hrs. dark at 12°C. Prior to germination, seeds were sterilized using a 20% bleach and 1% tween 

solution. Unsuccessful treatments included: “no-treatment” and “water soak”, both yielding 0% 

germination; boiling water followed by a 24-hour soak yielded 2% germination; and cold moist 

stratification yielded 4% germination. More successful treatments included: scarification of seed coat 

using abrasion (sandpaper rubbed for 3 min), which yielded 68-96% germination; and clipping the 

seed coat at the hilum using a scalpel, which yielded 97–100%.

Additional germination trials could focus on longevity of seeds in the seed bank and analyzing the 

potential contributors to a chemical scarification process.
29

Table A1. Current seed accession information for Astragalus tricarinatus at the RSABG seed bank. 

ACC # Date Collector State County

Land 

Manager Location

23439 14-Jun-11 Mitzi Harding California Riverside JOTR Little San Bernardino 

Mountains; 

approximately 1 mile 

west of Eureka Peak

23438 14-Jun-11 Mitzi Harding California Riverside JOTR Little San Bernardino 

Mountains; 

approximately 1.5 

miles south of Upper 

Covington Flats 

parking 

area/backcountry 

trailhead

21384 29-Apr-04 Michael Wall California San 

Bernardino

The 

Wildlands 

Conservancy

Little San Bernardino 

Mountains; ESE of 

Mount San Gorgonio, 

headwater region of 

N fork of Whitewater 

River; E end of 

Catclaw Flat at 

Wathier Landing, ca. 

6.0 mi. WNW along 

trail from stone house

23425 3-May-11 Duncan S. Bell California Riverside The 

Wildlands 

Conservancy

Little San Bernardino 

Mountains; just off 

the Pacific Crest Trail 

between the West 

Fork of Mission Creek 

and Whitewater 

Canyon. USGS 

Quad: Catclaw Flat
30

Table A2. Germination trial information for seeds stored at the RSABG seed bank.

ACC #

Date 

tested Treatment # Tested Total Germ. % Observations

23425 16-Apr-12 CL 15 15 100 all seedlings with very healthy root and 

cotyledon development

23439 19-Jul-11 CL 20 20 100 healthy root and cotyledon 

development

23425 05-Jul-11 CL 24 24 100 all seedlings with healthy vigorous root 

and cotyledon development

21384 21-Jun-04 CL 30 29 97 19 of 30 with healthy root/ cotyledon; 

10 of 30 with stunted, deformed root 

and/or shoot or had cotyledons detach 

from root

23425 18-Mar-13 SC 50 48 96 48 with healthy root and cotyledon. 1 

with stunted root, normal cotyledon. 1 

ungerminated seed hard, probable that 

seed coat was insufficiently scarified.

23425 02-Aug-11 SC 50 34 68 healthy root and cotyledon 

development; severe mold on plate 

rapidly occurred killing seedlings 

23425 02-Aug-11 CS1 50 2 4 healthy root and cotyledon 

development; ungerminated seeds 

hard

23425 02-Aug-11 HW1 50 1 2 healthy root and cotyledon 

development; ungerminated seeds 

hard

23425 02-Aug-11 WS 50 0 0 all seeds hard, no imbibition

23425 02-Aug-11 NT 50 0 0 all seeds hard, no imbibition

CL Clip, rupture seed coat using a pin knife or scalpel. Seeds punctured at hilum

NT No Treatment

CS1 Cold Moist stratification; up to 14 days

WS Water soak prior to sowing

HW1 Boiling water; soak cooling 24 hours

SC Scarification of seed coat using abrasion (sandpaper rubbed for ca. 3 min)
31

Appendix B: Genetic study

By Linda Prince and Naomi Fraga, Rancho Santa Ana Botanic Garden

Astragalus tricarinatus occurs primarily on mafic, felsic, gneiss, granodiorite, and alluvium

substrates in the Little San Bernardino Mountains. Historically, waif populations were thought to be 

the only known locations for this rare endemic of southern California. However, since 2004 a number 

of new populations have been discovered during field surveys, including large populations in Joshua 

Tree National Park (JOTR), yielding valuable information on the distribution, habitat preference, and 

abundance of A. tricarinatus (Amsberry and Meinke 2007, Fraga and Pilapil 2012, White 2004a, 

2004b). No prior genetic work has been done on Astragalus tricarinatus, and the genus is notorious 

for low levels of genetic diversity in general (Liston 1992, Wojciechowski et al. 1999, Kazempour 

Osaloo et al. 2003, Wojciechowski et al. 2004). A few population genetic studies have been 

conducted on other Astragalus species (Liston 1992, Karron et al. 1988, Travis et al. 1996, and 

Alexander et al. 2004), but no prior information is available for this taxon. Numerous studies have 

compared the variability of data generated by allozymes and repeat based markers (AFLPs, RAPDs, 

and ISSRs in particular), and shown that higher genetic diversity can be detected by repeat based 

markers, especially within populations (see the recent reviews by Nybom and Bartish 2000, and 

Nybom 2004). Based on prior studies showing low levels of diversity within the genus, we employed 

ISSR markers to gather baseline genetic diversity estimated for two populations located within 

JOTR, and compared them to a single population from outside the park. Populations sampled include 

Population 1 from Eureka Peak (-116.3557 / 34.0290, elev. 1486 m), Population 2 near Covington 

Flats (-116.3089 / 33.9889, elev. 1435 m), and Population 3 in Whitewater Canyon (-116.67905 /

34.03819, elev. 1033 m). For a comprehensive list of all known populations, see Table D1 in 

Appendix D; also see Figure 2 in the main report to see localities for sampled populations.

Fragment based methods such as ISSRs have some limitations relative to allozymes. Because they 

are primarily dominant data, it is not possible to determine heterozygosity, thus the statistical tests 

that may be applied are more limited. It is possible to assess relative levels of diversity. Dominant 

markers such as ISSRs, RAPDs, and AFLPs cannot be analyzed using the same measures as codominant markers such as microsatellites and allozymes. Some scientific publications frequently 

report heterozygosity, gene diversity, and other indices that cannot be calculated from dominant data 

in which heterozygosity and homozygosity are unknown. The only valid data that can be reported 

include the percentage of polymorphic loci (%P), ΦST (an analog of FST based on genetic distance), 

genetic distance (GD), and Shannon’s information index (I) (Culley 2014.).

Inappropriate measures include number of alleles per locus (A), number of alleles per polymorphic 

locus (AP), proportion of observed heterozygotes (HO), proportion of expected heterozygotes under 

Hardy-Weinberg equilibrium (HE), proportion of the total diversity that is partitioned among 

populations (FST), analog of FST that incorporates effects of small and unequal population/sample 

sizes (Θ), and an analog of FST but calculated differently under the assumption of Hardy-Weinberg 

equilibrium (GST). These measures cannot be used because they rely on knowing the number of 

homozygous and heterozygous individuals. With dominant data it is impossible to calculate the 
32

frequency of dominant alleles (Culley 2014). Nonetheless, these measures are often provided within 

peer-reviewed publications, therefore we provide them here for relative comparison purposes.

Experimental Design

Three populations of Astragalus tricarinatus were sampled; two from JOTR (Eureka Peak and 

Covington Flats) and a single population from Whitewater Canyon (see Figure 2 in the main report). 

For each population, 2 to 4 leaves from a minimum of 30 individuals were collected and immediately 

placed in silica gel desiccant. Plants were sampled haphazardly, including extreme edges of the 

population. Sampled individuals were also geo-referenced using hand-held GPS units accurate to 

approximately 3 m. Voucher specimens for all populations are housed in the herbarium collection of 

RSABG and JOTR. All laboratory work was conducted in the Laboratory of Molecular Evolution 

and Systematics (molecular lab) at RSABG. Any remaining leaf material was saved and stored in the 

molecular lab at RSABG.

DNA was extracted following a minor modification of the Doyle and Doyle (1987) method 

(inclusion of a 50oC incubation with Proteinase K (Life Technologies, Grand Island, New York, 

U.S.A.) for 20 min prior to the 65oC incubation step). DNA was resuspended in 100μL 1X TE buffer 

(10 mM Tris, 1 mM EDTA, pH 7.5). No secondary cleaning methods were employed. Quantification 

was carried out using 2μL of the stock DNA extraction with a NanoVue spectrophotometer (GE 

Healthcare Biosciences, Pittsburgh, Pennsylvania, U.S.A.). All DNA was diluted in water, to a 

working concentration of 10 ng/μL. Remaining DNAs are stored in the -80oC freezer collection of 

the molecular lab at RSABG.

Data collection

DNA for three to four individuals per population were selected, based upon quantity of DNA 

available, for ISSR marker screening. Approximately 30 different fluorescently tagged ISSR primers 

were amplified using Phusion high fidelity polymerase (with the 5X GC Buffer; New England 

Biolabs, Ipswich, Massachusetts, U.S.A.). Because ISSR primers are anchored microsatellite 

primers, polymerase stutter or slipping is expected to be a problem. When the polymerase attempts to 

copy the template DNA, it can slip, thereby adding an additional repeat, or skip a repeat, leading to 

the generation of incorrect fragment sizes. The use of Phusion polymerase minimizes the likelihood 

of these fragments. Annealing temperature varied based upon primer composition, as indicated in 

Table B1. Fluorescently tagged fragments (1-3 μL) were diluted in 10 μL Hi-Di formamide and coloaded with 0.5-0.75μL Liz 1200 internal size standard, electrophoresed and visualized on an 

Applied Biosystems Inc. (Carlsbad, CA, U.S.A.) 3130xl Genetic Analyzer (50 cm array, POP7 

polymer) following the manufacturer’s directions. Applied Biosystem’s GeneMapper v. 4.0 software

was used to score the data using the built-in AFLP module, with automatic binning.

Data were reviewed for each sample and the number and variability of bands per sample per primer 

noted. A number of primers either failed to produce any fragments or were virtually invariant across 

the samples tested. The results of the initial screening are summarized in Table B1. From the primers 

screened, the three primers with the most variability and larger number of fragments were selected 

for population wide analysis (ISSR 813FAM, 815VIC, 880VIC). Only peaks with heights greater 

than 100 in at least one individual were scored, and only peaks with height >50 were scored as 
33

present. All samples were amplified and run in triplicate. Only peaks that reproducibly produced 

scorable peaks (as described above) were analyzed. Background noise ranged from 5-7 with a few up 

to 10. A (primarily) binary data matrix (0, 1, ?) was generated and analyzed under several different 

criteria. The matrix included 6 monomorphic loci: 26, 30, 50, 144, 145, and 146. In the first set, all 

loci and all samples were analyzed, including 3 samples with large amounts of missing data (due to 

single primer failure OR no scorable fragments). Missing data were scored as ?. In the second set, the 

3 samples that contained large amounts of missing data had the missing data changed from ? to 0. 

The third set excluded those 3 samples from the analyses. These different analyses were conducted to 

assess the impact of missing data on the overall population diversity and structure estimates.

The phylogenetic software package PAUP*4.0 (test version a129, Swofford 2002) was used to create 

a pairwise distance matrix under the “RFLP/AFLP” option [Nei-Li (fragments); L=17]. Distance 

dendrograms were also generated using both Neighbor Joining (NJ) and the unweighted pair group 

method with arithmetic mean (UPGMA; under both average distance and total distance). Both NJ 

and UPGMA are simple agglomerative or bottom-up data clustering methods used in bioinformatics

for the creation of phylogenetic trees. Branch support was estimated using 1000 bootstrap replicates 

(NJ or UPGMA as appropriate).

The number of different alleles (Na), number of effective alleles (Ne), Shannon's information index 

(I, Lewontin 1972), Nei’s (1973) gene diversity (h) and unbiased gene diversity (uh), and percentage 

polymorphic loci (%P) were calculated in GenAlEx v. 6.5 (Peakall and Smouse 2006). Estimates of 

genetic diversity (h), genetic structure (θ

I

, an FST analog; θ

II

, an estimate most similar to Nei’s GST), 

and GSTB (a Bayesian estimate of GST) were estimated in HICKORY v. 1.1 (Holsinger and Lewis 

2007). HICKORY values were estimated using the “f-free” model since estimates of f from dominant 

data can be unreliable (Holsinger et al. 2002). Each HICKORY analysis was run in triplicate to 

ensure the Bayesian estimation had reached stationarity. Genetic diversity measures were estimated 

in two different programs as the numbers differ slightly, with HICKORY consistently providing 

lower estimates of diversity. A genetic distance matrix was generated in GenAlEx (for use in 

Principal Coordinates Analyses (PCoA or PCO; also conducted in GenAlEx) to identify groups of 

samples with the highest allelic similarity. Plots (based on the first two axes) for each analysis are 

provided in Figures B1-B3. 

Results

Effect of missing data

Population 2, from Covington Flats in JOTR, had highest values for all population descriptors 

including the number of alleles (Na), number of effective alleles (Ne), Shannon’s Information Index 

(I), Nei’s gene diversity (h), Nei’s unbiased gene diversity (uh), and percentage polymorphic loci 

(%P), regardless of how missing data were treated (see Tables B3-B5). On the other hand, population 

1, from Eureka Peak in JOTR, had the lowest values for all population descriptors.

Population genetic distance (see Table B6) and genetic structure (see Tables B7-9), as measured by 

θ

I

, θ

II, and GSTB, values ranged from 0.163-0.164 for GSTB, 0.258-0.260 for θII, and 0.437-0.438 for 
34

θ

I

. The values were similar regardless of how missing data were treated and the variation did not 

change with replicate analyses.

Similar topologies were found for NJ (data not shown) and UPGMA analyses (see Figures B4-B6 for 

UPGMA dendrograms) regardless of the treatment of missing data. Most individuals from Eureka 

Peak and Whitewater Canyon group together within their respective populations, with the Covington 

Flats population forming an intermediate grade between the other two. There is very little branch 

support for the topology. Analysis 2 (missing data = 0) resulted in a topology with the most 

intermingling of individuals from different populations, but those differences were not supported by 

bootstrap values ≥50%. The effect of missing data was most obvious in the PCO graph of Analysis 1 

(Figure B1), where individuals with large amounts of missing data (scored as ?) formed an outlying 

cluster.

Population Diversity

Each population was characterized by a number of unique ISSR bands not found in the other 

populations sampled, with 14 unique bands in the Eureka Peak population, 18 unique bands in the 

Covington Flats population, and 20 unique bands in the Whitewater Canyon population. 

Additionally, the Covington Flats-JOTR population had a high frequency (20-22 individuals out of 

the 31 sampled) of two bands (#131 and 132) that were either absent from the Eureka Peak 

population or present in very low frequency (1-2 out of the 31 individuals sampled) in the 

Whitewater Canyon population. The same was true for the Whitewater Canyon population, with 28 

out of 31 individuals exhibiting band #54 and #59, which were absent in the other two populations.

Population Structure

Individual populations were generally cohesive, with the majority of samples from each population 

forming a clade or grade (see Figures B4-B6), regardless of how missing data were treated. The 

populations did not form reciprocally monophyletic population groups however, suggesting either 

historic or contemporary gene flow between all three populations sampled. Principal Coordinates 

Analysis (PCoA) showed a similar pattern (Figures B2-B3), with the bulk of the samples from any 

given population clustering together and small areas of overlap among the populations. As noted 

above, this pattern was slightly obscured under Analysis 1, where individual samples with large 

amounts of missing data were included in the analysis as “?” (Figure B1). In this case, the two 

outlying clusters correspond to data from two markers: ISSR-813 or ISSR-815. In all three PCoA 

analyses, coordinates 1-3 explained approximately 70% of the variation. 

A variety of measures for population structure (θ

I

, θ

II, and GSTB) were examined to assess the 

distribution of individuals and gene flow within Astragalus tricarinatus. The Bayesian estimate of 

GST (GSTB) is considered the most conservative measure, followed by θ

II, and then θI

. Values, based 

on three independent Bayesian runs, ranged from 0.163-0.164 for GSTB, 0.258-0.260 for θ

II, and 

0.437-0.438 for θ

I

. Different methods of coding missing data had little effect on values obtained. θII

is the most relevant statistic as it provides information on the extent of genetic differentiation among 

contemporaneous populations and is less sensitive to incomplete population sampling.
35

The Eureka Peak and Covington Flats populations are more closely related to each other (NeiP = 

0.034; uNeiP=0.028) than either are to the Whitewater Canyon population, but they are also much 

closer geographically. Interestingly, the Covington Flats population is slightly more closely related to 

the Whitewater Canyon population (NeiP=0.079-0.081; uNeiP=0.073-0.074) than is the Eureka Peak 

population (NeiP=0.082-0.086; uNeiP=0.076-0.080) despite being slightly further away. This is 

graphically demonstrated in both the PCoA plots (Figures B1-B3) and the UPGMA trees (Figures 

B4-B6).

Discussion

Prior genetic work in Astragalus supported the claim that the genus is notorious for low levels of 

genetic diversity (Liston 1992, Wojciechowski and Hu 1999, Kazempour Osaloo et al. 2003, 

Wojciechowski et al. 2004). However, contrary to earlier genetic studies using comparative DNA 

sequences, allozyme studies have successfully investigated population level variation in a number of 

North American species (Table B10). The variation is still generally low (GST <0.10, with two 

exceptions: A. claranus and A. pauperculus). Recent population genetic studies using other fragmentbased markers (ISSRs and AFLPs; Karron1988 and Alexander et al. 2004) have detected 

significantly more diversity. Astragalus tricarinatus is intermediate in diversity relative to those 

fragment-based studies (Table B10).

The Bayesian estimates of population structure are typical of those found in other studies (Nybom 

2004), but are not conclusive in terms of their predictive value, with GSTB=0.163–0.164, θ

II=0.258–

0.260, and θ

I

=0.437–0.438. As a reminder, θ

I

is an FST analog, θ

II is an estimate most similar to 

Nei’s GST, and GSTB, a Bayesian estimate of GST. The latter is the most conservative of the three. The 

GSTB values are consistent with long-lived perennials of narrow geographic range that are 

outcrossers, animal-dispersed (via consumption), and considered of late successional status. In other 

words, based on these data A. tricarinatus appears to have enough gene flow (past or present) that the 

populations have not greatly differentiated genetically. The θ

I

values however, fall within the 

expected range for selfing species with gravity induced seed dispersal that are early successional 

annuals with a wide geographic range. As stated above, GSTB is very conservative, and θ

I

very 

generous, especially if the study fails to sample all extant populations. θ

II values are intermediate 

between reported values for short- and long-lived perennials and species with narrow to regional 

geographic distributions. They are also closer in value to outcrossing species than to selfing species. 

Astragalus tricarinatus is thought to be an outcrossing perennial species of short to moderate life 

span. It occurs in unstable or disturbed habitats (due to natural erosional processes) and appears to be 

restricted to a specific substrate. 

As stated above, each population was characterized by a number of unique ISSR bands, and the 

number of unique bands was similar across the three populations (14-20 bands). All three populations 

have similar levels of genetic diversity as well. There is evidence for past or contemporary gene flow 

among all three populations and that gene flow appears to be high based upon the low GST values.
36

Table B1. ISSR primers screened for population genetic analysis of Astragalus tricarinatus, including

annealing temperature employed and approximate number of major bands observed. Bold typeface 

indicates primers used in this study.

Primer

Number Base Composition Dye

Anneal

Temp.

(oC)

Number

of Bands

807 AGA GAG AGA GAG AGA GT FAM 50.0 1-9

808 AGA GAG AGA GAG AGA GC VIC 52.0 0

809 AGA GAG AGA GAG AGA GG FAM 52.0 8+

811 GAG AGA GAG AGA GAG AC VIC 52.0 7

812 GAG AGA GAG AGA GAG AA VIC 50.0 0

813 CTC TCT CTC TCT CTC TT FAM 50.0 13+

814 CTC TCT CTC TCT CTC TA NED 50.0 5

815 CTC TCT CTC TCT CTC TG FAM 52.0 10-16

817 CAC ACA CAC ACA CAC AA VIC 50.0 0

818 CAC ACA CAC ACA CAC AG FAM 52.0 4

820 GTG TGT GTG TGT GTG TC VIC 52.0 0

821 GTG TGT GTG TGT GTG TT PET 50.0 0

822 TCT CTC TCT CTC TCT CA VIC 50.0 4

823 TCT CTC TCT CTC TCT CC NED 50.0 4-10

825 ACA CAC ACA CAC ACA CT VIC 50.0 1

826 ACA CAC ACA CAC ACA CC PET 54.0 0

828 TGT GTG TGT GTG TGT GA FAM 50.0 0

830 TGT GTG TGT GTG TGT GG VIC 54.0 0

861 ACC ACC ACC ACC ACC ACC FAM 60.0 0

863 AGT AGT AGT AGT AGT AGT FAM 48.0 2

866 CTC CTC CTC CTC CTC CTC FAM 60.0 >20

868 GAA GAA GAA GAA GAA GAA VIC 48.0 3

869 GTT GTT GTT GTT GTT GTT PET 48.0 0

873 GAC AGA CAG ACA GAC A NED 48.0 0

874 CCC TCC CTC CCT CCC T FAM 54.0 10-15

880 GGA GAG GAG AGG AGA VIC 50.0 >15

881 GGG TGG GGT GGG GTG VIC 54.0 0
37

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JOTR – Eureka Peak; Population 2 

(ML678-ML708) = JOTR – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ?.

Locus

Individ. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35

ML646 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML647 0 0 1 0 1 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML648 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML649 0 0 1 0 1 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML650 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML651 0 0 1 0 1 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML652 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML653 0 0 1 0 1 0 1 0 0 0 0 0 1 0 1 1 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML654 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML655 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML656 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML657 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML658 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML659 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML660 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML661 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML662 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML663 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML664 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML665 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML666 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML667 0 0 1 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML668 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML669 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML670 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML671 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML672 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML673 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML674 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML675 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML676 0 0 1 0 1 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML678 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 0 1 0 0 1 1 0

ML679 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML680 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 1 1 0 0 0 0 0

ML681 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML682 0 1 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML683 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML684 0 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML685 0 1 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML686 0 1 1 1 1 1 1 1 0 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML687 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML688 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML689 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML690 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1 0 0 0 0 0

ML691 0 0 1 0 1 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML692 0 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 1
38

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35

ML693 0 0 1 0 1 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 1 0 1 1 1 0 0 1 1 1 0 0 0 0 0

ML694 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML695 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML696 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML697 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML698 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 0 0 0

ML699 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML700 0 0 1 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML701 0 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML702 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0

ML703 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML704 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML705 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML706 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML707 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML708 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML734 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 0 1 1 1 0 0 0

ML735 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0

ML736 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 1 1 1 0 1 0 0 0

ML737 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 1 0 1 0 0 0 0 0

ML738 0 0 1 0 1 1 1 0 0 0 0 0 0 0 1 1 1 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML739 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML740 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 0 0 0

ML741 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML742 0 1 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 1 1 0 0 0 0 0

ML743 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML744 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0

ML745 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 1 0 0 0 0 0 1 1 0 0 0 0 0

ML746 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 1 0 1 0 0 0

ML747 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 1 1 1 0 1 0 0 0

ML748 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 0 0 0

ML749 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML750 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 1 0 1 0 0 0 0 0 0 1 1 1 1 0 0 0

ML751 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML752 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML753 0 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML754 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 1 1 1 0 0 0 0 0

ML755 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML756 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 1 1 0 0 1 1 0

ML757 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 1 1 1 0 0 0

ML758 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 1 0 0 1 1 1 0 0 0 0 0

ML759 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 1 1 0 0 0

ML760 0 0 1 0 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 1 0 1 1 1 0 1 1 1 1 0 0 0 0 0

ML761 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML762 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML763 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0

ML764 1 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0
39

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68

ML646 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML647 0 0 1 0 1 0 0 0 0 0 0 1 0 0 1 1 0 1 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML648 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML649 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML650 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 1 0 1 1 1 1 0 0 0 0 0

ML651 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML652 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 0 1 0 0 0 1 1 0 0 0 0 0

ML653 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0

ML654 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML655 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML656 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML657 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML658 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML659 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML660 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML661 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML662 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML663 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML664 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML665 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML666 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML667 0 0 0 0 1 1 0 0 0 0 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML668 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0

ML669 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML670 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML671 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML672 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML673 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 1 0 0

ML674 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML675 0 0 0 0 0 0 0 0 0 0 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML676 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML678 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML679 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML680 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML681 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML682 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML683 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML684 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML685 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML686 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML687 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML688 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML689 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML690 1 1 0 0 0 1 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML691 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 1 0 0 0 0 0

ML692 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0
40

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68

ML693 0 0 0 0 1 0 0 1 0 1 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML694 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML695 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0

ML696 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0

ML697 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML698 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML699 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML700 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML701 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML702 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML703 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 1 0 1 0 1 1 0 1 0 0 0

ML704 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0

ML705 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML706 0 0 0 0 1 0 0 1 0 1 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0

ML707 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML708 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 1 1 0 0 0 0 0

ML734 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 1 1 0 0 1 1 0 0 1 0 0 0 0 0 0

ML735 0 0 1 0 0 0 0 0 0 0 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML736 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML737 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML738 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML739 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML740 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 1 0 0 0 1 0 0 0 0 0 ? 0 0 0

ML741 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 1 0 0 1 0 1 0 0 1 1

ML742 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML743 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML744 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML745 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML746 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML747 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML748 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML749 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML750 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML751 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML752 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML753 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML754 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML755 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 ? 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML756 0 0 0 0 0 0 0 0 0 0 1 ? 0 0 1 1 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML757 1 1 1 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 1 0 0 0 1 1 0 0 1 0 0 0 0 0 0

ML758 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0

ML759 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0

ML760 0 0 0 0 1 0 1 0 1 0 0 1 0 0 1 1 0 0 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0

ML761 1 1 0 0 1 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 1 0 1 0 1 1 0 0 0 0 0

ML762 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0

ML763 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 1 0 0 1 0 0 0 1 0 0 0 0 0 0

ML764 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0 1 1 0 0 1 1 0 0 1 0 0 0 0 0 0
41

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101

ML646 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 1 1 0 0 0 1 1

ML647 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 0 1 0 0 0 1 1 0 0 1 1

ML648 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 1 1 0 0 0 1 1

ML649 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 0 1 0 0 0 1 1

ML650 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 1 1 0 0 1 1

ML651 0 0 1 1 1 0 0 0 0 0 1 1 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML652 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 1 1 0 0 1 1

ML653 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 1 1 0 0 1 1

ML654 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 1 1 0 0 1 1

ML655 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 1 1

ML656 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 1 1 0 1 1 1

ML657 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 0 1 0 0 1 1 1 0 0 1 1

ML658 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 1 1

ML659 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 1 1 0 0 0 1 0

ML660 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 0 0 1 1 0 0 1 1

ML661 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 1 1 0 0 0 1 1

ML662 0 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 0 0

ML663 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 1 1 1 0 0 1 1

ML664 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 1 1 0 0 0 0 0

ML665 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 1 0 0 0 1 1 0 0 1 1

ML666 0 0 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 0 0 0 1 1 0 0 1 1

ML667 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML668 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 1 0 0 0 1 1

ML669 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML670 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 1 0 0 1 1 1

ML671 0 0 0 1 1 0 0 0 0 0 0 0 0 0 ? 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML672 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 1 0 0 0 1 1 0 0 1 1

ML673 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 1 1 1

ML674 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0

ML675 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML676 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML678 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 1 0 0 0 1 1

ML679 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 0 0

ML680 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 ? 1 0 0 0 1 1

ML681 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 0 1 1 0 0 1 0

ML682 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 1 0 0 0 1 1 0 0 0 0

ML683 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 1 1 0 0 0 0

ML684 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 1 0 0 0 1 1 0 0 0 0

ML685 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 1 1 1 0 1 1 0 1 0 0 1 1 1 0 0 0 0

ML686 0 0 0 1 1 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML687 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 1 0 0 0 1 1 0 0 0 0

ML688 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 0 0 0 1 1 0 0 0 1

ML689 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 0 0 0 1 0 0 0 1 0

ML690 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 0 0 0 0

ML691 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 0 0 1 0

ML692 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1 0 0 0 1 1 0 0 0 0
42

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101

ML693 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0 1

ML694 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 1 0 0 0 0 0

ML695 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML696 0 0 0 1 1 1 1 0 0 0 0 0 1 1 0 0 0 1 1 0 1 1 0 0 0 1 0 1 0 0 0 0 0

ML697 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1

ML698 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 1 1

ML699 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 1 0

ML700 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 1 1 1

ML701 0 0 0 1 1 1 1 0 0 0 0 0 0 1 1 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML702 0 0 0 1 0 1 1 0 0 0 0 0 0 1 1 0 0 0 1 1 1 1 0 0 0 0 1 1 0 0 0 1 1

ML703 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 1 0

ML704 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 1 1

ML705 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0

ML706 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0

ML707 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 1 0 0 0 1 1 0 0 1 0

ML708 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 1 0 0 0 0 0 0 0 1 1 1

ML734 0 0 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML735 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?

ML736 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML737 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML738 0 0 0 1 1 1 1 0 ? 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML739 0 0 ? 1 1 1 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML740 0 0 1 1 1 1 1 1 ? 0 0 0 0 0 0 0 0 0 0 0 ? 0 ? 0 0 0 0 0 0 ? 0 1 1

ML741 1 1 0 1 1 1 1 0 0 0 1 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML742 0 0 1 1 1 1 1 1 1 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML743 0 0 0 1 1 1 1 0 0 ? 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML744 0 0 0 1 1 1 1 1 1 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML745 0 0 ? 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML746 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML747 0 0 0 1 1 1 1 1 1 0 0 0 0 1 1 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML748 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 1 1

ML749 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML750 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML751 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML752 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML753 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML754 0 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML755 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 1 1

ML756 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML757 0 0 1 1 1 1 1 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 1 1

ML758 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML759 0 0 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML760 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML761 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1 1

ML762 0 0 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1

ML763 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 1 1

ML764 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0
43

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128

ML646 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 1 0 0 0 0 0 0 1

ML647 1 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0 0 1 0 0 0 1

ML648 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML649 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML650 0 1 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML651 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML652 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 1 0 1 0 1

ML653 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML654 1 0 0 0 1 0 0 0 0 0 1 1 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML655 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 1 1

ML656 1 1 0 0 1 1 0 0 1 1 1 1 1 1 0 1 1 0 0 0 0 0 1 0 1 0 1

ML657 1 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML658 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML659 1 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML660 0 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML661 1 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML662 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0 0 1 0 1 0 1

ML663 0 1 0 0 0 0 0 0 0 1 1 1 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML664 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 0 0 0 0 0 0 0 1 0 1

ML665 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML666 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 1 1 0 1

ML667 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 1 0 0 0 0 0 1 0 1 0 1

ML668 1 0 1 0 0 1 1 1 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML669 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 1 0 0 0 0 0 1 0 1

ML670 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 0 0 0 0 1 0 1 0 1

ML671 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML672 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML673 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 0 0 0 0 1 0 1 0 1

ML674 0 0 0 0 1 0 0 0 1 1 0 0 1 1 1 1 1 0 0 0 0 0 1 0 1 0 1

ML675 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 1 0 0 0 0 0 1 1 1 0 1

ML676 1 0 0 0 1 0 0 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 1 0 1 0 1

ML678 1 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 0 0 0 0 0 0 0 1 0 1

ML679 0 0 0 ? 0 0 0 0 0 0 1 1 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1

ML680 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 1 1 1 0 1

ML681 0 0 0 1 0 0 0 0 0 1 1 1 1 1 0 1 1 0 0 0 0 0 0 1 1 0 1

ML682 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 1 0 0 0 0 0 0 1 0 0 0 1

ML683 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 1 0 0 1

ML684 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 1 1 1 0 1

ML685 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML686 1 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML687 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 0 1

ML688 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 1 0 0 1 1 0 1

ML689 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1 0 0 0 1 0 0 0 1 0 1

ML690 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 1 1 0 0 1 0 1

ML691 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 0 0 0 0 1 0 0

ML692 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 1 1
44

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128

ML693 1 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1 1 0 0 0 0 0 1 0 1

ML694 0 0 0 0 1 0 0 0 0 1 0 0 1 1 0 1 0 1 1 0 1 0 1 0 1 0 1

ML695 1 0 0 0 1 0 0 0 1 1 0 0 1 1 0 1 1 0 0 0 0 0 1 0 1 1 1

ML696 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0 0 0 1

ML697 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 1 0 0 0 0 0 0 0 0 0 1 1

ML698 1 0 0 0 1 0 0 0 1 1 0 0 1 1 0 1 1 0 0 0 0 0 0 0 1 0 1

ML699 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1 0 0 0 1 0 1 1 1

ML700 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 0 1

ML701 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 1 1 0 1 0 1

ML702 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 1 0 1 0 0

ML703 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 1 0 0 0 1 0 1 0 0

ML704 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 1 1 0 0 0 1 0 1 0 1

ML705 0 0 0 0 1 0 0 0 0 1 0 0 1 1 0 1 0 1 1 0 1 1 1 0 1 1 0

ML706 0 0 0 0 1 0 0 0 1 1 0 0 1 1 1 1 1 0 0 0 1 1 1 0 0 1 1

ML707 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 1 1 1 0 1 0 1

ML708 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0

ML734 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 1 0

ML735 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 1 0 0 0 0 0 0 0 1 0 0

ML736 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 0 1

ML737 0 1 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 1 1 0 0 0 0 0 0 0 0

ML738 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML739 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML740 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0

ML741 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 1 0 0 0 0 0 0 0 0

ML742 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 1 0 0 0 0 0 0 0 0

ML743 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 1 1 0 0 0 0 0 0 1 0

ML744 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 0 1 0

ML745 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML746 1 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 0 0 1 0 0 0 0

ML747 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 0 0 0 0 0 0 0

ML748 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 0 0 0 1 0 1 0 0

ML749 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 0 0 0

ML750 1 0 0 0 1 0 0 0 1 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 0

ML751 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML752 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0

ML753 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 1 0 1

ML754 1 1 0 0 0 0 0 0 1 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML755 1 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML756 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0

ML757 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 1

ML758 1 1 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 0 0 0 0 0 0 0 0 0

ML759 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0

ML760 1 1 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 0 1 0 0 0 0 0 0 0 0

ML761 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0

ML762 1 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 1

ML763 1 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 1 1 0 0 0 0 0 1 0 1

ML764 0 0 0 0 1 0 0 0 0 1 0 0 1 1 0 0 1 1 1 0 0 0 0 0 1 0 0
45

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146

ML646 0 0 0 0 0 0 0 1 1 0 0 1 1 0 1 1 1 1

ML647 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1

ML648 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1

ML649 1 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 1 1

ML650 1 0 0 0 0 0 0 1 1 0 0 1 1 0 0 1 1 1

ML651 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 1 1 1

ML652 0 1 0 0 0 0 0 1 0 0 0 1 1 0 1 1 1 1

ML653 1 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1

ML654 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 1 1 1

ML655 0 1 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML656 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1

ML657 1 0 0 0 0 0 0 1 0 0 0 1 1 0 1 1 1 1

ML658 0 0 0 0 0 0 0 1 1 0 0 1 1 0 1 1 1 1

ML659 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 1 1 1

ML660 1 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1

ML661 1 0 0 0 0 0 0 1 0 1 0 1 1 1 1 1 1 1

ML662 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 1 1 1

ML663 1 0 0 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1

ML664 1 0 0 0 0 0 0 1 1 0 0 1 1 0 1 1 1 1

ML665 1 0 0 0 0 0 0 1 1 0 0 1 1 0 1 1 1 1

ML666 0 0 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1 1

ML667 1 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML668 1 0 0 0 0 0 0 0 0 1 0 1 0 1 1 1 1 1

ML669 1 0 0 0 0 0 0 1 1 1 1 1 1 0 1 1 1 1

ML670 1 0 0 0 0 0 0 1 1 1 0 1 1 0 1 1 1 1

ML671 1 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML672 1 0 0 1 0 0 1 1 0 0 0 1 0 1 1 1 1 1

ML673 1 0 0 0 0 0 0 1 1 1 0 1 1 0 1 1 1 1

ML674 1 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML675 1 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML676 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1 1 1 1

ML678 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1

ML679 1 0 0 0 0 0 0 0 1 1 1 0 1 0 1 1 1 1

ML680 0 0 0 0 0 0 0 1 0 1 0 1 0 0 1 1 1 1

ML681 1 0 1 1 0 0 0 0 1 1 1 0 1 0 1 1 1 1

ML682 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1

ML683 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 1 1 1

ML684 1 0 1 1 1 1 0 1 0 1 0 1 0 0 1 1 1 1

ML685 1 0 1 1 1 1 0 1 1 1 0 1 1 0 1 1 1 1

ML686 1 0 1 1 1 0 0 1 1 0 0 1 1 0 1 1 1 1

ML687 0 0 1 0 0 0 0 0 1 1 1 0 1 1 1 1 1 1

ML688 0 0 1 1 1 1 0 1 1 0 0 1 1 0 1 1 1 1

ML689 1 0 1 1 1 0 0 0 0 0 1 1 0 1 1 1 1 1

ML690 1 0 1 1 1 1 0 1 1 0 1 1 1 0 1 1 1 1

ML691 1 0 1 1 0 0 0 1 1 0 0 1 0 0 1 1 1 1

ML692 1 0 1 1 1 0 0 0 0 1 1 0 1 1 1 1 1 1
46

Table B2. ISSR data matrix (analysis 1 version) for Astragalus tricarinatus. Population 1 (ML646-ML676) = JTNP – Eureka Peak; Population 2 

(ML678-ML708) = JTNP – Covington Flats; Population 3 (ML734-ML764) = Whitewater Canyon. Character states = 0, 1, ? (continued).

Locus

Individ. 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146

ML693 0 0 1 1 1 1 0 0 1 0 1 0 0 0 1 1 1 1

ML694 1 0 1 1 1 1 0 0 1 0 0 0 1 0 1 1 1 1

ML695 1 0 1 1 0 0 0 0 0 1 1 0 1 1 1 1 1 1

ML696 0 0 1 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1

ML697 0 1 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1

ML698 1 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1

ML699 1 0 1 1 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML700 1 0 1 1 1 0 0 1 0 1 1 1 0 1 1 1 1 1

ML701 1 0 1 1 1 1 0 1 0 1 0 1 0 0 1 1 1 1

ML702 1 0 1 1 0 0 0 1 1 0 0 1 1 0 1 1 1 1

ML703 1 0 0 1 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML704 1 0 0 0 0 0 0 0 1 0 0 0 1 0 1 1 1 1

ML705 1 1 1 1 1 1 0 0 0 0 1 0 0 0 1 1 1 1

ML706 0 1 1 1 1 1 0 0 0 0 0 0 0 0 1 1 1 1

ML707 1 0 1 1 1 1 0 0 1 0 1 0 1 0 1 1 1 1

ML708 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1 1 1

ML734 0 1 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1 1

ML735 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML736 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1

ML737 0 1 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 1

ML738 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML739 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML740 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 1 1

ML741 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1

ML742 0 1 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 1

ML743 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

ML744 0 1 1 1 1 0 0 0 0 1 0 0 1 1 1 1 1 1

ML745 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML746 1 1 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 1

ML747 0 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 1

ML748 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1

ML749 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1 1

ML750 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML751 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 1 1

ML752 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML753 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1

ML754 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML755 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML756 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML757 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1 1

ML758 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

ML759 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 1 1 1

ML760 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

ML761 0 0 1 0 1 1 0 0 1 1 0 0 1 1 1 1 1 1

ML762 0 0 0 0 0 0 0 0 1 0 0 1 1 0 1 1 1 1

ML763 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1

ML764 1 0 0 0 0 0 0 1 0 0 0 1 0 0 1 1 1 1
47

Tables B3-B5. Various population statistics and descriptors, under Bayesian allele frequency (BAFP; 

Lynch and Milligan 1994) criteria, for Astragalus tricarinatus based on ISSR data. Results based on 

GenAlEx analyses. Highest values are in bold typeface, lowest values underlined. Number of individuals 

sampled (SS), number of alleles (Na), number of effective alleles (Ne), Shannon’s Information Index (I), 

Nei’s gene diversity (h), Nei’s unbiased gene diversity (uh), percentage polymorphic loci (%P). h*1-3 are 

values of h from three, independent HICKORY analyses.

B3. Analysis #1 with missing data scored as ?.

Population SS Na Ne I h h*1 h*2 h*3 uh %P

1. JOTR-EP 31 Mean 1.164 1.197 0.199 0.124 0.138 0.138 0.138 0.129 52.74

SE 0.077 0.025 0.020 0.014 0.005 0.005 0.005 0.014

2. JOTR-CF 31 Mean 1.432 1.313 0.289 0.187 0.189 0.189 0.189 0.193 68.49

SE 0.072 0.030 0.022 0.016 0.007 0.007 0.007 0.016

3. WC 31 Mean 1.288 1.227 0.231 0.144 0.150 0.150 0.151 0.149 60.27

SE 0.076 0.025 0.020 0.014 0.007 0.007 0.007 0.014

Total Mean 1.295 1.246 0.239 0.152 0.159 0.159 0.159 0.157 60.50

SE 0.044 0.016 0.012 0.008 0.005 0.005 0.005 0.009 4.55

B4. Analysis #2 with missing data scored as 0.

Population SS Na Ne I h h*1 h*2 h*3 uh %P

1. JOTR-EP 31 Mean 1.185 1.207 0.208 0.130 0.138 0.138 0.138 0.135 54.79

SE 0.078 0.025 0.020 0.014 0.005 0.005 0.005 0.014

2. JOTR-CF 31 Mean 1.432 1.313 0.289 0.187 0.189 0.189 0.189 0.193 68.49

SE 0.072 0.030 0.022 0.016 0.007 0.007 0.007 0.016

3. WC 31 Mean 1.329 1.232 0.239 0.149 0.151 0.151 0.151 0.153 64.38

SE 0.077 0.025 0.020 0.014 0.007 0.007 0.005 0.014

Total Mean 1.315 1.251 0.245 0.155 0.159 0.159 0.159 0.160 62.56

SE 0.044 0.016 0.012 0.008 0.005 0.005 0.005 0.009 4.06

B5. Analysis #3 with taxa for which large amounts of data are missing excluded from the analyses.

Population SS Na Ne I h h*1 h*2 h*3 uh %P

1. JOTR-EP 29 Mean 1.144 1.196 0.196 0.123 0.138 0.138 0.138 0.128 51.37

SE 0.077 0.025 0.020 0.014 0.005 0.005 0.005 0.014

2. JOTR-CF 31 Mean 1.432 1.313 0.289 0.187 0.189 0.189 0.189 0.193 68.49

SE 0.072 0.030 0.022 0.016 0.007 0.007 0.007 0.016

3. WC 30 Mean 1.288 1.229 0.232 0.145 0.151 0.151 0.151 0.150 60.27

SE 0.076 0.025 0.020 0.014 0.007 0.007 0.007 0.014

Total Mean 1.288 1.246 0.239 0.152 0.159 0.159 0.159 0.157 60.05

SE 0.044 0.016 0.012 0.008 0.005 0.005 0.005 0.009 4.94
48

Table B6. Pairwise population genetic distances for Astragalus tricarinatus based on analyses of ISSR 

data. Analysis 1: missing data = ?; analysis 2: missing data = 0; analysis 3: taxa with large amounts of 

missing data excluded. Highest values are in bold typeface, lowest values underlined. NeiP = Nei’s 

genetic distance; uNeiP = Nei’s unbiased genetic distance. Pop1 = JOTR – Eureka Peak; Pop2 = JOTR –

Covington Flats; Pop3 = Whitewater Canyon.

Pairwise

Comparison

Analysis 1 Analysis 2 Analysis 3

NeiP uNeiP NeiP uNeiP NeiP uNeiP

Pop1-Pop2 0.034 0.028 0.034 0.028 0.034 0.028

Pop1-Pop3 0.086 0.080 0.082 0.076 0.085 0.080

Pop2-Pop3 0.081 0.074 0.079 0.073 0.081 0.074

Tables B7-B9. Astragalus tricarinatus estimates of population genetic structure based on HICKORY 

analysis results of ISSR data analyses. θ

I

is an FST analog, θ

II is an estimate most similar to Nei’s GST, 

and GSTB is a Bayesian estimate of GST.

B7. Analysis #1 with missing data scored as ?.

Run 1 Run 2 Run 3

θ

I

θ

II

GSTB θ

I

θ

II

GSTB θ

I

θ

II

GSTB

Mean 0.437 0.260 0.164 0.437 0.259 0.164 0.438 0.259 0.164

SE 0.026 0.029 0.012 0.025 0.029 0.012 0.025 0.029 0.012

B8. Analysis #2 with missing data scored as 0.

Run 1 Run 2 Run 3

θ

I

θ

II

GSTB θ

I

θ

II

GSTB θ

I

θ

II

GSTB

Mean 0.438 0.259 0.164 0.437 0.260 0.164 0.437 0.260 0.164

SE 0.025 0.029 0.012 0.025 0.029 0.012 0.025 0.029 0.012

B9. Analysis #3 with taxa (for which large amounts of data are missing) excluded from the analyses.

Run 1 Run 2 Run 3

θ

I

θ

II

GSTB θ

I

θ

II

GSTB θ

I

θ

II

GSTB

Mean 0.438 0.258 0.163 0.438 0.259 0.163 0.438 0.259 0.164

SE 0.025 0.029 0.012 0.026 0.029 0.012 0.025 0.028 0.012
49

Table B10. Comparison of Astragalus species with population level genetic studies. Habit = Annual (A) or 

Perennial (P).

Taxon Habit Distribution Marker FST or GST Ref

A. brewerii A Narrow Allozyme 0.000 (GST) 1

A. claranus A Narrow Allozyme 0.331 (GST) 1

A. nyensis A Narrow Allozyme 0.000 (GST) 1

A. pauperculus A Narrow Allozyme 0.725 (GST) 1

A. rattanii var. jepsonianus A Narrow Allozyme 0.053 (GST) 1

A. rattanii var. rattanii A Narrow Allozyme 0.058 (GST) 1

A. tener var. tener A Narrow Allozyme 0.059 (GST) 1

A. tener var. titi A Narrow Allozyme 0.000 (GST) 1

A. acutirostris A Widespread Allozyme 0.254 (GST) 1

A. gambelianus A Widespread Allozyme 0.211 (GST) 1

A. nuttallianus A Widespread Allozyme 0.000 (GST) 1

A. cremnophylax P Narrow AFLP 0.41 (FST) 2

A. linifolius P Narrow Allozyme 0.11 (FST*) 3

A. oniciformis P Narrow ISSR 0.1130-0.1134 (GST) 4

A. osterhouti P Narrow Allozyme 0.14 (FST*) 3

A. tricarinatus P Narrow ISSR 0.164 (GST) here

A. nothoxys P Widespread Allozyme 0.000 (GST) 1

A. pattersoni P Widespread Allozyme 0.01 (FST*) 3

A. pectinatus P Widespread Allozyme 0.07 (FST*) 3

1=Liston 1992, 2=Travis et al. 1996, 3=Karron et al. 1988, 4=Alexander et al. 2004. FST* = sum of 

CE+GOT values.
50

Figure B1. Principal coordinates analysis of ISSR data for Astragalus tricarinatus (analysis 1: missing 

data = ?). Coordinates 1 + 2 explain 60% of the variation, + coordinate 3 = 72%. Population 1 = JOTR –

Eureka Peak (red); Population 2 = JOTR – Covington Flats (green); Population 3 = Whitewater Canyon

(blue). Note the circled, outlying cluster. These are samples with large amounts of missing data. Circle = 

missing data for marker ISSR-815.
51

Figure B2. Principal coordinates analysis of ISSR data for Astragalus tricarinatus (analysis 2: missing 

data = 0). Coordinates 1 + 2 explain 54% of the variation, + coordinate 3 = 67%. Population 1 = JOTR –

Eureka Peak (red); Population 2 = JOTR – Covington Flats (green); Population 3 = Whitewater Canyon

(blue).
52

Figure B3. Principal coordinates analysis of ISSR data for Astragalus tricarinatus (analysis 3: excluding 

taxa with missing data). Coordinates 1 + 2 explain 56% of the variation, + coordinate 3 = 68%. Population

1 = JOTR – Eureka Peak (red); Population 2 = JOTR – Covington Flats (green); Population 3 = 

Whitewater Canyon (blue).
53

Figure B4. UPGMA analysis of ISSR data for Astragalus tricarinatus (analysis 1: missing data = ?). 

Population color code: JOTR – Eureka Peak (red); Pop 2 = JOTR – Covington Flats (green); Pop 3 = 

Whitewater Canyon (black). Bold lines indicate branches with bootstrap support ≥50%.
54

Figure B5. UPGMA analysis of ISSR data for Astragalus tricarinatus (analysis 2: missing data = 0). 

Population color code: JOTR – Eureka Peak (red); Pop 2 = JOTR – Covington Flats (green); Pop 3 = 

Whitewater Canyon (black). Bold lines indicate branches with bootstrap support ≥50%.
55

Figure B6. UPGMA analysis of ISSR data for Astragalus tricarinatus (analysis 3: excluding taxa with 

missing data). Population color code: JOTR – Eureka Peak (red); Pop 2 = JOTR – Covington Flats 

(green); Pop 3 = Whitewater Canyon (black). Bold lines indicate branches with bootstrap support ≥50%.
56

Appendix C: GIS habitat model for Astragalus tricarinatus

By Sean Murphy, Mitzi Harding, and Tasha La Doux, Joshua Tree National Park

Summary 

A habitat model was developed to identify potential habitat for Astragalus tricarinatus in Joshua 

Tree National Park (JOTR). The model is designed to highlight probable habitat based on 

quantitative characteristics associated with known localities. Parameters used in developing the 

probabilities include: slope, aspect, elevation, soil mapunit, and vegetation association. Geographic 

locations are based on voucher specimens, field surveys, and CNDDB occurrences. After a 

preliminary model was developed, high probability areas lacking presence points were targeted for a 

ground truthing exercise. The final model can be accessed as an ArcGIS Toolbox, labeled “Rare 

Plant Model” and is available on the JOTR Resource Division network shared drive.

Methods

The habitat model was developed in a python environment using Python v2.6 (© Copyright 1990-

2014, Python Software Foundation) because the tools available in ArcToolbox and capabilities in 

ModelBuilder (ArcGIS v10.1) were not sufficient to complete the task. The python script provided 

the flexibility to look at raster files, query properties of those raster files, and then implement 

dynamic statistics based on those properties. A script tool interface was created in ArcGIS Desktop, 

an environment familiar to GIS users, for the user to specify input parameters. The Astragalus

tricarinatus Habitat Model script can run one, some, or all of the following parameters based on user 

preferences: elevation, slope, aspect, soil mapunit, and/or vegetation association. A “presence point 

buffer” shapefile was produced in ArcGIS by creating circular polygons centered on each known 

locality with a 15-meter radius. The area within each polygon is then used to collect data for each of 

the parameters (e.g. averages, minimum/maximum). Also, the script was developed with the idea that 

it would be implemented iteratively with a ground truthing process, as it is based on presence of 

known locations. Absence points were not informative for the model and therefore are not 

incorporated into the script. Please note that the script requires an ArcInfo level of licensing and the 

Spatial Analyst extension.

A preliminary model was produced in ArcGIS by manually performing the functions now automated 

by the A. tricariniatus Habitat Model script. Range limits were manually calculated for each 

parameter using the buffered presence point layer. The ranges of these values were then used to 

create a limited raster for each parameter. Finally, we performed a weighted overlay of those limited 

rasters to create the final habitat model. The ranges of values used in creating the intermediate rasters 

were determined by narrowing the range for each parameter to include a minimum threshold of 60% 

of the area within the known habitat layer (buffered presence points). This threshold was determined 

to create range limits that best captured our understanding of the known habitat for the species, more 

specifically:

Four vegetation associations were included in the first model: Single-leaf Pinyon Pine / Muller’s Oak 

(Pinus monophylla / Quercus cornelius-mulleri) Woodland Association, California Juniper / 
57

Blackbush (Juniperus californica / Coleogyne ramosissima) Association, Muller Oak — California 

buckwheat — Narrowleaf goldenbush (Quercus cornelius-mulleri — Eriogonum fasciculatum —

Ericameria linearifolia) Association, Catclaw Acacia — Desert Almond (Acacia greggii — Prunus 

fasciculata) Association. These vegetation associations accounted for 95.2% of the area within the 

presence point buffer layer. 

Two soil mapunits were used for the calculations: Xerric Torriorthents-Bigbernie association, 

Smithcanyon gravelly sand. These two soil mapunits account for 94% of the area within the presence 

point buffer layer.

Elevation range was limited to 1370–1470 m, which accounted for 63.4% of the area in the presence 

point buffer layer.

Slope range was limited to 3°–40°, which accounted for 95.8% of the area in the presence point 

buffer layer.

Aspect range was limited to 155°–262°, which accounted for 68% of the area in the presence point 

buffer layer.

The output from this model was then used to prioritize areas for ground truthing surveys, which 

yielded two new occurrences for the species (see Figure C1). Originally, we had thought that using 

absence points would be helpful in producing the model, however, in the end this was not the case. It 

is possible that absence points could be useful in a model that focuses on a narrower range of field 

values for the various parameters (i.e. limit the analysis to one watershed). The first model was 

informative for the development of a more user-friendly and automated script that is less timeintensive and reduces user error. 

The final model is set up so that the user is not required to manually establish the range limits for the 

various parameters, as this method is tedious and inconsistent. The user only needs to select the 

various source data layers to be used in the analysis through a user-friendly interface, after which the 

model will perform calculations according to the specific scripts for each parameter (read below for 

details). Essentially, the model will create range limits for raster data such as slope, aspect, and 

elevation, capturing 95% of the area within the presence point buffer layer. For vector data such as 

soil and vegetation, only the type that represents the most area within the presence point buffer layer 

will be selected for. These limitations can be overcome either by adjusting the script (only to be done 

by advanced GIS/modeling specialists) or by creating source data layers that somehow combine 

relevant data. For example, by assigning a common code to all vegetation associations represented by 

the presence point buffer layer, the analysis could better represent the range of vegetation 

associations associated with the occurrence of the species. This type of manipulation to the source 

data layers can also offset any bias associated with an unequal number of data points in any given 

area. Because the model is based on weighted averages, it will always be prone to bias the results 

according to the habitat associated with the highest number of presence points. While this type of 

limitation may be appropriate for narrow habitat specialists, it can be misleading for plants that occur 

in a variety of habitats. For this reason, it may be reasonable to exclude certain presence points, if 
58

they represent anomalous habitat types (for example, a waif found in the wash below the main 

population on the slope). Conversely, it is important to continue to add points to the presence point 

buffer layer, as each additional point will hopefully increase the accuracy of the output. In particular, 

it is important to make sure that GPS points are recorded in the field for each individual plant, or a 

group of plants isolated by a 15-meter radius.

The model is meant to be iterative. In other words, the output of each model will guide field surveys 

to target high probability habitat, then as new populations/individuals are added to the database, the 

model will increase in accuracy. Also, since a date field is collected with each point, users can select 

data in a date range and rerun the model based on the subset. Doing so will allow the user to see how 

the model accuracy changes over time as the amount of presence points are increased.

Scripts

Elevation: When the user selects elevation as an analysis parameter and specifies a digital elevation 

model to use in the analysis, the script applies the corresponding elevation statistic calculations. First, 

it extracts elevation values that are within the presence point buffer. Second, it calculates the 

elevation standard deviation and average of the extracted values. Third, it assigns a negative standard 

deviation elevation value and a positive standard deviation elevation value. This is done by taking the 

calculated standard deviation and subtracting or adding two standard deviations from the average 

elevation value, accordingly. Based on the standard deviation range, the script’s last step takes the 

original digital elevation model and selects the elevation values within the standard deviation range 

to create a new raster. This raster is used in the last step of the script during the weighted overlay.

Slope: When the user selects slope as an analysis parameter and specifies a slope surface analysis 

product to use in the analysis, the script applies the corresponding slope statistic calculations. First, it 

extracts slope values that are within the presence point buffer. Second, it calculates the slope standard 

deviation and average of the extracted values. Third, it assigns a negative standard deviation slope 

value and a positive standard deviation slope value. This is done by taking the calculated standard 

deviation and subtracting or adding two standard deviations from the average slope value, 

accordingly. Based on the standard deviation range, the script’s last step takes the original slope 

raster and selects the slope values within the standard deviation range to create a new raster. This 

raster is used in the last step of the script during the weighted overlay.

Aspect: When the user selects aspect as an analysis parameter and specifies an aspect surface 

analysis product to use in the analysis, the script applies the corresponding aspect statistic 

calculations. Aspect values are cyclic values, meaning 0° and 360° are the same value, and not a 

range that starts with a low value and stops at the high value. Because aspect values are cyclic, a few 

more steps needed to be incorporated into the script. First, it extracts aspect values that are within the 

presence point buffer. Second, it isolates the values ranging from zero to 180° by setting values less 

than one and greater than 180° to null. Third, it calculates the aspect standard deviation and average 

extracted values subset. Fourth, it assigns a negative standard deviation aspect value and a positive 

standard deviation aspect value. This is done by taking the calculated standard deviation and 

subtracting or adding two standard deviations from the average aspect value, accordingly. Next, the 

third and fourth step are repeated for aspect values ranging from 180 to 360 degrees – a subset is 
59

created, average and standard deviation are calculated, and the low and high value in the standard 

deviation range are calculated. Based on the standard deviation range, the script’s last step takes the 

original aspect raster and selects the aspect values within both standard deviation ranges and creates a 

new raster. This raster is used in the last step of the script during the weighted overlay.

Soil: When the user selects soil as an analysis parameter and specifies soil polygons to use in the 

analysis, the script applies the corresponding soil statistic calculations. First, it clips soil mapunit to 

the presence point buffer. Second, it calculates coverage area for each soil mapunit. Third, it sorts 

through the area totals and isolates the soil mapunit with the most coverage (the maximum). Lastly, 

the maximum soil mapunit is selected out from the original soil polygons and converted into a raster 

format. This raster is used in the last step of the script during the weighted overlay.

Vegetation

When the user selects vegetation as an analysis parameter and specifies vegetation polygons to use in 

the analysis, the script applies the corresponding vegetation statistic calculations. First, it clips 

vegetation associations to the presence point buffer. Second, it calculates coverage area for each 

vegetation association. Third, it sorts through the area totals and isolates the vegetation association

with the most coverage (the maximum). Lastly, the maximum vegetation association is selected out 

from the original vegetation polygons and converted into a raster format. This raster is used in the 

last step of the script during the weighted overlay.

Weighted Overlay

The last part of the script takes the five parameters, or less if a parameter was excluded from the 

analysis, and overlays them using the weighted overlay tool. The tool was set to give each parameter 

equal weight. The result is an overlay that has values ranging from zero to five, or zero to the number 

of parameters being analyzed; five representing where the habitat is most likely located and zero 

representing where the habitat is least likely located.

Results and Discussion

The final model can be accessed as an ArcGIS Toolbox, labeled “Rare Plant Model” and is available 

on the JOTR Resource Division share drive. The model can be used for any species, provided

geospatial data layers with known locations and at least one of the corresponding parameters are 

available. 

Below are two examples of the automated model output. The first attempt (Figure C2) utilized all 

five data layers (elevation, aspect, slope, vegetation, and soil) without any modifications to the 

source data. As discussed above, the highest probability habitat is biased toward the single vegetation 

association and soil mapunit with the highest number of occurrences. For comparison, in order to 

represent all of the vegetation associations and soil mapunits with known localities, we modified the 

vegetation and soil source data layers for a second version of the model. This second version (Figure 

C3) also utilized all five parameters, but with the following modifications:

1) There are four vegetation associations represented by the presence point buffer layer. These 

were discussed above and were used to develop the preliminary habitat model. In order to 
60

capture and equally weight these vegetation associations, we decided to combine all four of 

them into one common name and code, therefore forcing the script to recognize them all as 

one vegetation association. 

2) We re-coded the three most common soil mapunits found within the presence point buffer 

layer, so that all would be included in the final overlay. Two of these soil mapunits were 

discussed above and were included in the preliminary model, and a third (SmithcanyonStubbespring-Rock outcrop complex) was added due to newly discovered populations found 

during the 2013 ground truthing surveys on that soil mapunit. 

As expected, the second version of the automated model significantly expands the area of “highly 

probable habitat” within the Little San Bernardino Mountains (Figure C3). The second version seems 

to reflect probable habitat more accurately according to the authors experience and understanding for 

preferred habitat of A. tricarinatus in JOTR. In this second model, areas with newly discovered 

robust populations (for example, northwest of Eureka Peak) are highlighted as “very high 

probability” habitat, whereas in the first model (Figure C2) they are not.

As discussed earlier, the model will only select for the most frequent category of each parameter, 

such that adjusting the input layers to best reflect the known habitat preferences will likely improve 

results. Finally, it is important to keep in mind that some areas of JOTR have been surveyed more 

thoroughly than others and efforts to record data points have not been driven by the 15-m radius 

requirement of this model.

Future Suggestions

Future efforts will focus on the following items. 

1) Field efforts to locate new populations will focus on high probability areas according to the 

map produced by version 2 of the current model (Figure C3).

2) Time permitting, geospatial data will be taken for each individual, unless there are multiple 

individuals within a 15-meter radius of the point.

3) New data will be added to the presence point buffer layer in a timely manner and the model 

will be updated accordingly.

4) Future models will consider the idea of utilizing absence points, as well as providing a means 

to weight the parameters differently.

5) Expand the geospatial scope of the model to include the entire range of the species; or at the 

very least incorporate habitat information from known localities outside the park boundary.

6) Additional trials will be done with the current model, to see if other modifications to the 

available data sources increase the accuracy of the output. For example, combining fields or 

removing parameters altogether. In addition, reducing the area being used in the model (e.g. 

to one watershed), may produce a more accurate prediction on a finer spatial scale.
61

Figure C1. Preliminary Astragalus tricarinatus habitat probability model. This model reflects a weighted overlay of a manually defined range of 

values for elevation, slope, aspect, soil mapunits, and vegetation associations that are most commonly associated with the known A. tricarinatus

localities within Joshua Tree National Park. Dark green represents the lowest probability habitat, while red represents areas of highest probability 

habitat. Ground truthing surveys were performed in spring and summer of 2013, yielding two new occurrences; one population of approximately 50 

individuals south of Quail Mountain, and another estimated at over 50 individuals on the steep slopes of the Little San Bernardino Mountains.
62

Figure C2. Astragalus tricarinatus habitat probability model output produced using the ArcGIS Toolbox automated script, Rare Plant Model. The 

model output reflects a weighted overlay of a statistically defined range of values for elevation, slope, and aspect representing values associated 

with 95% of the area in which the species occurs in Joshua Tree National Park. Soil mapunits and vegetation associations were restricted to the 

one type containing the highest frequency of occurrences. Areas falling within the target ranges of each parameter are assigned a value of “1”, 

and areas falling beyond the range limitations are assigned a value of “0”. Overlapping ranges are summed accordingly; five representing where 

the habitat is most likely located, and zero representing where the habitat is least likely located.
63

Figure C3. Astragalus tricarinatus habitat probability model output produced using the ArcGIS Toolbox automated script, Rare Plant Model. The 

model output reflects a weighted overlay of a statistically defined range of values for elevation, slope, and aspect representing values associated 

with 95% of the area in which the species occurs in Joshua Tree National Park. Soil mapunit and vegetation association source data were edited 

to combine all types that contain occurrences of the species, resulting in four vegetation associations and three soil mapunits included in the 

analysis. Areas falling within the target ranges of each parameter are assigned a value of “1”, and areas falling beyond the range limitations are 

assigned a value of “0”. Overlapping ranges are summed accordingly; five representing where the habitat is most likely located, and zero 

representing where the habitat is least likely located. 
64

Appendix D: Known occurrences for Astragalus tricarinatus

Table D1. List of 45 known occurrences of Astragalus tricarinatus. Sources CNDDB, CCH 2014, JOTR. EO = Element Occurrence; ElmDate = 

Year species was last seen at occurrence; County = Riverside (RIV), San Bernardino (SBD); KeyQuad = USGS 7.5-minute quadrangle map.

NOTE: EO#4 is now considered part of EO#5; EO#7 is now considered part of EO#3; EO#10 was deleted after specimen (Parish JUN 15 1895, 

UC1601949) was annotated to A. bernardinus; NA = EO# not yet assigned.

EO ElmDate County KeyQuad Elev (ft) Location General OwnerMgt

1 2011 SBD Morongo 

Valley 

2500 BIG MORONGO CANYON, NEAR 

COVINGTON PARK IN TOWN OF 

MORONGO VALLEY, LITTLE SAN 

BERNARDINO MOUNTAINS.

2 PLANTS SEEN IN 1983. NO PLANTS WERE FOUND 

IN 1987, 1992, 1994, OR 2005. 1 PLANT OBSERVED IN 

EACH POLYGON IN 2011.

SBD COUNTY, 

BLM

2 2011 RIV, 

SBD

Morongo 

Valley 

2200 WHERE BIG MORONGO CANYON 

CROSSES THE SAN 

BERNARDINO/RIVERSIDE COUNTY 

LINE, LITTLE SAN BERNARDINO MTNS

2 PLANTS OBSERVED IN 1986, 1 IN 1987, 40 IN 1992, 

13 IN 1993, 5 IN 1994, 1 SEEN IN 2004 IN WESTERN 

POLYGON, NONE FOUND IN MAIN CANYON IN 2005. 

2 PLANTS SEEN IN 2011 IN EASTERN POLYGON. 

INCLUDES FORMER OCCURRENCE #7.

BLM-BIG 

MORONGO CYN 

PRESERVE

3 1995 RIV White 

Water 

1800 WHITEWATER CANYON, NORTH OF 

WHITEWATER.

TYPE LOCALITY. NOT SEEN IN AREA IN 1987 (LOW 

RAINFALL). 1 IMMATURE PLANT FOUND IN 1995. 

COLLECTIONS FROM "WHITEWATER CANYON", 

"ATTRIBUTED TO THIS SITE. NEEDS FIELDWORK.

UNKNOWN

5 1946 RIV, 

SBD

Morongo 

Valley 

1700 DRY MORONGO CANYON AND WASH. INCLUDES FORMER OCCURRENCE #4. NEEDS 

FIELDWORK.

UNKNOWN

6 1994 RIV Morongo 

Valley 

2000 BIG MORONGO WASH, 1.9 MILES 

NORTH OF MOUTH, LITTLE SAN 

BERNARDINO MOUNTAINS.

8 PLANTS OBSERVED IN 1986, NONE FOUND IN 

1987, SEVERAL OBSERVED IN 1993, 1 IN 1994, NONE 

FOUND IN 2005.

BLM-BIG 

MORONGO CYN 

PRESERVE

8 1985 RIV Martinez 

Mtn. 

1500 AGUA ALTA CANYON, ABOUT 1.6 

MILES UPSTREAM FROM MARTINEZ 

CANYON, SANTA ROSA MTNS.

1 PLANT OBSERVED IN 1985. BLM

9 2011 RIV Catclaw 

Flat 

2600 MISSION CREEK, CA. 0.5 MILE N OF 

CONFLUENCE WITH W. FORK 

MISSION CREEK, SAN BERNARDINO 

MTNS.

2 PLANTS OBSERVED IN 1997, 8 PLANTS SEEN IN 

1998, AND 1 PLANT FOUND IN 2011.

BLM

11 1995 RIV Desert 

Hot 

Springs

2100 MISSION CREEK CANYON, NEAR 

GATE ON MISSION CREEK ROAD, 

EAST OF THE RANCH.

ONLY SOURCE OF INFO FOR THIS OCCURRENCE IS 

A 1995 COLLECTION BY BALLMER. BALLMER LISTED 

THE PRESENCE OF THIS PLANT AS "SCARCE 

ALONG THE ROAD".

UNKNOWN

12 2010 RIV White 

Water 

2400 WHITEWATER CANYON, NEAR 

WHITEWATER TROUT FARM AND 

NNW OF TROUT FARM FOR ABOUT 

1.2 MILES, SAN BERNARDINO MTNS

SITE BASED ON A 1987 COLLECTION BY PITZER AND 

A 2010 MISTRETTA ET AL. COLLECTION. NEED MAP 

DETAIL FOR THIS SITE.

BLM, UNKNOWN

13 1994 RIV Morongo 

Valley 

1800 BIG MORONGO CANYON, 4 ROAD MI 

BELOW THE CARETAKER'S HOUSE 

AT THE BIG MORONGO PRESERVE, 

LITTLE SAN BERNARDINO MTNS.

2 PLANTS OBSERVED IN 1994. NO PLANTS SEEN IN 

A 2005 SEARCH OF THE CANYON. 

BLM-BIG 

MORONGO CYN 

PRESERVE
65

EO ElmDate County KeyQuad Elev (ft) Location General OwnerMgt

14 2010 SBD Catclaw 

Flat 

3800 FROM WATHIER LANDING TO 1 MILE 

E. OF WATHIER LANDING, SAN 

BERNARDINO MTNS.

THOUSANDS OF PLANTS OBSERVED IN THE TWO 

WESTERNMOST POLYGONS IN 2010.

WILDLANDS 

CONSERVANCY, 

BLM

15 2011 RIV Catclaw 

Flat 

2500 MISSION CREEK CANYON, JUST S OF 

THE MOUTH OF W FORK MISSION 

CREEK, SAN BERNARDINO MTNS

ONE PLANT OBSERVED IN EACH OF THE 4 E. MOST 

POLYGONS IN 1998. 21 PLANTS OBSERVED IN THE 

W. MOST POLYGON BUT NO PLANTS SEEN IN WASH 

IN E. PART OF OCCURRENCE IN 2011.

THE 

WILDLANDS 

CONSERVANCY, 

BLM

16 UNK RIV Orocopia 

Canyon 

OROCOPIA RANGE. ONLY SOURCE FOR THIS SITE IS A SITE NAME 

MENTIONED IN BARNEBY (1964). NEEDS 

FIELDWORK.

UNKNOWN

17 1921 RIV Myoma THOUSAND PALMS CANYON, 

COLORADO DESERT.

ONLY SOURCES ARE A 1921 JAEGER COLLECTION 

FROM "THOUSAND PALMS" AND A 1921 SPENCER 

COLLECTION FROM "1000 PALMS CANYON". NEEDS 

FIELDWORK.

UNKNOWN

18 2009 RIV White 

Water 

2300 RIDGE E. OF WHITEWATER CANYON; 

~2 MI N OF TOWN OF WHITE WATER 

(I-10), NEAR SUPER CREEK QUARRY, 

SAN BERNARDINO MTNS.

2 PLANTS WERE SEEN IN 2009. BLM

19 2005 SBD Morongo 

Valley 

2400 BIG MORONGO CANYON, 0.8 MI S OF 

SE EDGE OF MORONGO VALLEY, 

LITTLE SAN BERNARDINO MTNS.

A SINGLE LARGE, REPRODUCTIVE PLANT WITH 

OVER 100 FLOWERING STEMS WAS OBSERVED IN 

2005. NO PLANTS COULD BE RELOCATED IN 2011.

BLM-BIG 

MORONGO CYN 

PRESERVE

20 2011 SBD Catclaw 

Flat 

3400 E END OF CATCLAW FLAT, 2 MI E OF 

WATHIER LANDING, WEST OF 

MORONGO VALLEY.

>100 PLANTS (INCLUDING SEEDLINGS) WERE 

OBSERVED IN 2005. 96 PLANTS OBSERVED IN 2011.

THE 

WILDLANDS 

CONSERVANCY

21 2006 RIV East 

Deception 

Canyon 

1725 EAST DECEPTION CANYON, NEAR 

PARK BOUNDARY, ~0.3 MI N OF END 

OF E DECEPTION RD, LITTLE SAN 

BERNARDINO MTNS.

3 PLANTS WERE SEEN AND DESCRIBED AS WAIFS 

IN 2006. VOUCHERED BY T. LA DOUX (#1454).

NPS-JOSHUA 

TREE NP

22 2006 RIV East 

Deception 

Canyon 

1804 EAST DECEPTION CANYON, NEAR 

PARK BOUNDARY, ~0.7 MI N OF END 

OF E DECEPTION RD, LITTLE SAN 

BERNARDINO MTNS.

1 PLANT WAS SEEN IN 2006. VOUCHERED BY T. LA 

DOUX (#1452).

NPS-JOSHUA 

TREE NP

23 2006 SBD Yucca 

Valley 

South

2748 LONG CANYON; STOP 2 IN SIDE 

CANYON EAST OF MAIN WASH, 

LITTLE SAN BERNARDINO MTNS.

1 PLANT, PROBABLY A WAIF, WAS FOUND IN 2006. 

VOUCHERED BY T. LA DOUX (#1446).

NPS-JOSHUA 

TREE NP

24 2011 SBD Catclaw 

Flat 

3200 MISSION CREEK; JUST NE OF THE 

PACIFIC CREST TRAIL, ABOUT 0.9 AIR 

MILE NORTH OF SBD / RIV COUNTY 

LINE.

IN 2011, THERE WERE 2 PLANTS GROWING ON THE 

WEST SIDE OF THE SIDE CANYON JUST BEFORE 

THE FIRST COTTONWOODS. EXTENSIVE SURVEY 

WAS NOT CONDUCTED; POSSIBLY MORE PLANTS IN 

THE AREA.

BLM

25 2011 RIV, 

SBD

Catclaw 

Flat 

3000 CA. 1.5 MI N OF CONFLUENCE OF 

MISSION CREEK & W FORK MISSION 

CREEK, NEAR RIV/SBD CO LINE, SAN 

BERNARDINO MTNS.

1 PLANT OBSERVED IN 1998 IN CENTER POLYGON. 

31 SEEN IN NORTHERN POLYGON AND 6 OBSERVED 

IN SOUTHERN POLYGON IN 2011.

BLM, UNKNOWN
66

EO ElmDate County KeyQuad Elev (ft) Location General OwnerMgt

26 2010 RIV East 

Deception 

Canyon 

4600 2.2 AIR MILES SW OF COVINGTON 

SPRING, W OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

IN NORTHERN POLYGON: 20 PLANTS OBSERVED IN 

2009, 21 ADDITIONAL PLANTS OBSERVED IN 2010. IN 

SOUTHERN POLYGON: 24 PLANTS OBSERVED IN 

2010.

NPS-JOSHUA 

TREE NP

27 2009 RIV East 

Deception 

Canyon 

3650 3.3 AIR MI SW OF COVINGTON 

SPRING, W OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

5 PLANTS OBSERVED IN 2009, WAIFS GROWING IN 

WASH

NPS-JOSHUA 

TREE NP

28 2008 RIV East 

Deception 

Canyon 

4400 2.2-2.9 AIR MI SSW OF COVINGTON 

SPRING, S OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

2008 POPULATION NUMBERS OBSERVED IN 

POLYGONS FROM NORTH TO SOUTH: 54, 1, 3, AND 5 

PLANTS. IN 2010, REVISITS TO THE TOP TWO 

POLYGONS OBSERVED 60 AND 3 PLANTS, 

RESPECTIVELY. 

NPS-JOSHUA 

TREE NP

29 2008 RIV East 

Deception 

Canyon 

4700 2.3 AIR MI SSW OF COVINGTON 

SPRING, S OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

4 PLANTS OBSERVED IN NE POLYGON AND 2 

PLANTS SEEN IN SW POLYGON IN 2008. 

NPS-JOSHUA 

TREE NP

30 2010 RIV East 

Deception 

Canyon 

3600 3.7 AIR MI SW OF COVINGTON 

SPRING, SW OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

4 PLANTS OBSERVED IN 2010. NPS-JOSHUA 

TREE NP

31 2006 RIV East 

Deception 

Canyon 

2100 BOTTOM OF DECEPTION CANYON, 

ABOUT 1.5 MI N OF CANYON MOUTH, 

LITTLE SAN BERNARDINO MTNS.

3 PLANTS OBSERVED IN 2006. NPS-JOSHUA 

TREE NP

32 2010 RIV Joshua 

Tree 

South 

5000 0.4 AIR MILES SW OF EUREKA PEAK; 

NW OF UPPER COVINGTON FLAT, 

LITTLE SAN BERNARDINO MTS.

175 PLANTS OBSERVED IN SE POLYGON IN 2010. 40 

PLANTS OBSERVED IN NW POLYGON IN 2011.

NPS-JOSHUA 

TREE NP

33 2009 RIV Joshua 

Tree 

South 

4000 2.9 AIR MI WSW OF COVINGTON 

SPRING, W OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

1 PLANT OBSERVED IN 2009. NPS-JOSHUA 

TREE NP

34 2009 RIV Joshua 

Tree 

South 

4500 2.4 AIR MI WSW OF COVINGTON 

SPRING, W OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

4 PLANTS OBSERVED IN 2009. NPS-JOSHUA 

TREE NP

35 2009 RIV Joshua 

Tree 

South 

3800 3.1 AIR MI WSW OF COVINGTON 

SPRING, W OF UPPER COVINGTON 

FLAT, LITTLE SAN BERNARDINO 

MTNS.

3 PLANTS OBSERVED IN 2009. VOUCHERED BY T. LA 

DOUX (#1953). 

NPS-JOSHUA 

TREE NP

36 2010 SBD Catclaw 

Flat 

4000 1 AIR MILE WNW OF WATHIER 

LANDING, SAN BERNARDINO MTNS.

300+ PLANTS OBSERVED IN EASTERN POLYGON 

AND 600+ OBSERVED IN WESTERN POLYGON IN 

2010.

UNKNOWN

37 2011 SBD Catclaw 

Flat 

3400 RIDGELINE BETWEEN MISSION 

CREEK & W. FORK MISSION CREEK, 

CA. 2 MILES E. OF CATCLAW FLAT, 

SAN BERNARDINO MTNS

62 PLANTS OBSERVED IN 2011. BLM
67

EO ElmDate County KeyQuad Elev (ft) Location General OwnerMgt

38 2011 RIV Catclaw 

Flat 

2930 RIDGE AT SE END OF CATCLAW 

FLAT, 1.1 AIR MILES NNW OF RED 

DOME, NE OF WHITEWATER RIVER, 

SAN BERNARDINO MTNS

1 PLANT OBSERVED IN 2011. BLM

39 2011 RIV Catclaw 

Flat 

3000 RIDGE BETWEEN W. FORK MISSION 

CREEK AND WHITEWATER RIVER, 

ABOUT 1 MI NE OF RED DOME, SAN 

BERNARDINO MTNS

51 PLANTS OBSERVED IN WESTERN POLYGON AND 

26 SEEN IN EASTERN POLYGON IN 2011.

BLM

40 2011 RIV Catclaw 

Flat 

2700 0.6 AIR MILE NNW OF RED DOME, 0.3 

MILE WEST OF PACIFIC CREST TRAIL, 

ALONG WHITEWATER RIVER, SAN 

BERNARDINO MTNS

1 PLANT OBSERVED IN 2010, 6 PLANTS SEEN IN 

2011.

BLM

NA 1921 RIV UNK UNK CHUCKWALLA MOUNTAINS ONLY KNOWN FROM A VOUCHER COLLECTED BY 

SPENCER (#1770, GH366431) 

BLM

NA 1975 RIV UNK 1800 LONG CYN, 1.5 MI N OF JOSHUA 

TREE NM BOUNDARY; T2SR5ES22; 

LITTLE SAN BERNARDINO MTNS

COMMUNITY TYPE: ENCELIA, HYMENOCLEA, DALEA 

CALIFORNICA; ONLY KNOWN FROM VOUCHER 

COLLECTED BY PATRICK LEARY 5/3/1975 

(UNLV9780). LIKELY A WAIF IN WASH.

NPS-JOSHUA 

TREE NP

NA 2011 RIV Joshua 

Tree 

South

5000 APPROX. 0.75 AIR MILES SSE OF 

EUREKA PEAK, LITTLE SAN 

BERNARDINO MOUNTAINS

OBSERVED BY M. HARDING, ESTIMATED 70 

INDIVIDUALS ON STEEP LOOSE SLOPES ABOVE 

WASH

NPS-JOSHUA 

TREE NP

NA 2010 RIV Joshua 

Tree 

South

3800 3.2 AIRMILES SW OF COVINGTON 

SPRING, WEST OF UPPER 

COVINGTON FLAT, LITTLE SB MTNS 

2 PLANTS OBSERVED IN 2010 NPS-JOSHUA 

TREE NP

NA 2013 RIV East 

Deception 

Canyon

5000 1.3 AIRMILES SW OF QUAIL PEAK, 

WEST OF JUNIPER FLAT, LITTLE SB 

MTNS

APPROX. 80 PLANTS OBSERVED BY M. HARDING IN 

STEEP DRAINAGES AND SLOPES 

NPS-JOSHUA 

TREE NP

NA 2013 RIV East 

Deception 

Canyon

4700 2.2 AIRMILES W OF STUBBE SPRING, 

LITTLE SB MTNS

24 PLANTS OBSERVED ON STEEP, LOOSE SLOPES 

BELOW RIDGE, POP’N ESTIMATED AT >100

NPS-JOSHUA 

TREE NP
68

Appendix E: Pilot study for Astragalus tricarinatus in Joshua 

Tree National Park

By Tasha La Doux and Mitzi Harding, Joshua Tree National Park

Introduction and Methods

In April of 2010, fifty Astragalus tricarinatus individuals were tagged in the Little San Bernardino 

Mountains (Figure E1) with the intention of tracking these individuals over time for life expectancy 

and to assess a variety of life history and reproductive traits. It became very clear during the first visit 

that frequent observations of the plants would result in extreme destruction to their habitat and likely 

lead to high mortality rates, therefore data collection was kept to a minimum until better protocols 

and study sites could be established. Revisits to these plants in 2013 were conducted as a means to 

glean what we could from the data, however we do not recommend revisiting these plants again.

In 2010, all 50 plants were in flower with green vegetative growth. The following parameters were 

recorded for each plant: height and two widths, GPS location, percent slope, aspect, reproductive 

phenology, habitat, associated species, and elevation. We did revisit these plants in 2013, however 

due to the dry conditions that year, no reproductive activity was observed. In addition, very few 

plants had any new growth, so height and two widths were recorded for the dead material only. 

Height was measured perpendicular to the slope of the ground. The longest axis of any plant material 

was recorded (dead or alive). During the 2010 season, new growth consistently exceeded the old 

growth so only one measurement (total growth) was recorded. Width measurements were taken 

parallel to the slope. The longest axis was measured first, then the second axis was measured 

perpendicular to the first axis (see Figures F2 and F3 in Appendix F). 

Reproductive phenology for each individual was recorded as one of the following flowering stages: 

• Vegetative = no reproductive activity

• Bud = No open flowers, head with green phyllaries and/or fresh buds

• Early Flowering = Flowers are open, no open flowers are senescing 

• Late Flowering = some or all flowers are senescing or going to fruit

• Fruiting = fruit/seed development with no open flowers

At the time of initial tagging (April 2010), most known locations of A. tricarinatus in the Park 

consisted of 1 to 5 individuals growing sporadically along steep canyon walls, therefore establishing 

a permanent plot with >10 individuals was not thought to be possible. In addition we were attempting 

to minimize our impact to any one site by tagging individuals throughout the canyon. 

After 2010, several new, and quite large, populations of A. tricarinatus were discovered in the Park 

(see discussion in main report), providing an opportunity to reconsider the monitoring protocol. In 

2013, we revisited the 50 plants tagged in 2010 and established two permanent plots, one of which 

incorporates seven of the plants tagged in 2010. A new monitoring protocol was drafted and 
69

employed on the new plots (see Appendix F), although we were not able to fully test efficiency and 

utility of the protocol due to the lack of vegetative growth and reproductive activity in 2013. 
70

Figure E1. In 2010, fifty Astragalus tricarinatus individuals were tagged (blue circles) and two permanent plots were established in 2013 (red 

squares) for the purpose of monitoring demographics and reproductive capacity.
71

The data collected each year will need to be tested and analyzed to assess whether adaptations need 

to be made before the following season. Ultimately, the objectives of the long-term monitoring 

protocol presented in this report are as follows:

1. To establish expected life span for individuals.

2. To assess longevity and long term viability of populations. 

3. To monitor reproductive capacity.

4. To establish mortality rates and reproductive capacity for size classes.

In addition to these objectives, there are a number of questions (see also the Recommendations 

section below) that can be built into the monitoring efforts as time and staffing allow. At a minimum, 

we recommend calculating the ovule/seed ratio as well as collecting/observing any pollinators.

It is impossible to express in this document the difficulties encountered while attempting to establish 

monitoring plots for this species. Due to the steep slopes, loose soils, and shallow rooting system 

exhibited by this species, accessing the individual plants is precarious at best. Any person responsible 

for performing data collection for this species must be forewarned that they need to be prepared for 

extremely unstable conditions on the slope. This is not to say that the person is necessarily unsafe, 

but rather their presence on the slope places a high risk for mortality on a federally listed plant. For 

this reason, the value of the data should always far outweigh the negative impacts of collecting the 

data to the long-term viability of the species. And perhaps more importantly, only the most agile and 

experienced hikers will be asked to perform these tasks.

Results and Discussion

All 50 plants tagged in 2010 were flowering, 88% had fruit in addition to flowers (though six of these 

plants were almost entirely in fruit). All plants were found on SE- to SW-facing slopes with an 

average grade of 43° (median = 42°, minimum = 20°, maximum = 90°). Elevation ranged from 

1339–1443 m (4393–4734 ft). Black aphids and ladybugs were observed on five separate individuals, 

a “honeybee” was observed visiting flowers on one individual. Insects were not captured. 

Based on data from 2010, plants varied in height from 10–70 cm, whereas total volume (HxW1xW2) 

varied from 7,500–2,361,528 cm3 (Median = 147,808 cm3

). Plants were measured following the 

protocol described in Appendix F. Measurements were taken on live material, which exceeded any 

dead material present.

In 2013, two of the 50 tagged individuals were found dead in the wash below the slope; eight plants 

were not relocated and are therefore assumed to be dead (mortality rate = 20%). The 40 plants remeasured varied in height from 18–53 cm, and total volume varied 12,312–433,964 cm3 (median = 

69,296 cm3

). Total volume is based on measurements taken of dead material, as none of the plants 

exhibited any new growth. It is possible that many of the individuals measured were in fact dead, but 

until we have conditions that allow for comparing normal growth in healthy individuals to dead 

material, we felt it premature to make these calls. In particular, there were two individuals that 
72

appeared to have a slightly grayer look to the stems (i.e. appeared to be dead), as compared to a 

yellow-brown present in the others, but because they were still rooted in the ground we took a 

conservative approach and assumed they were still alive.

Based on the experience from these two years of monitoring, it is clear that annual visits to the plants 

will be necessary to glean meaningful data. Tracking the response to seasonal precipitation will be 

very helpful in discerning mortality rates due to drought versus normal senescence. It will also allow 

for a better understanding of how well these plants can resprout from the base, how quickly they can 

respond to a rain, and how varying rainfall patterns/amounts affect reproductive capacity. Moreover, 

if, as indicated in previous reports (Amsberry and Meinke 2007, Sanders 1999, USFWS 2009), the 

lifespan is three to five years, capturing data annually for that time span will be essential. 

Relocating plants in 2013 was extremely difficult and time consuming, in part because the tags did 

not always stay in the ground. The notes and photos from the previous monitoring season were 

helpful in trying to positively identify some plants. For this reason, we recommend taking photos and 

including as much detail in the monitoring notes as time permits. 

Future monitoring

In 2013 two permanent belt transects were established in the Little San Bernardino Mountains, near 

Eureka Peak and Upper Covington Flats (Figure E1). Each is different in size and bearing (Table 

E1). The goal was to keep the survey area within each transect narrow enough that data could be 

collected with minimal impact to the site and still capture >10 individuals. More importantly, the plot 

area should be easily surveyed for new seedlings. The plot near Eureka Peak (ASTR-1) is 25 m long 

and 12 m wide; the X-axis starts at the top of the slope and runs exactly south (180) for 25 m; 18 

individuals were tagged and measured in May 2013. The plot below Upper Covington Flats (ASTR2) is 15 m long and 12 m wide; the X-axis starts at the bottom of the slope and runs NNE for 15 m; 

14 individuals were tagged and measured in June 2013. The endpoints for each belt transect are 

permanently marked with rebar. 

Table E1. Detailed location and orientation of the two permanent Astragalus tricarinatus monitoring plots 

located in the Little San Bernardino Mountains. All GPS data is recorded in NAD83 UTM, and bearings 

are recorded with a 12° east declination. 

Plot ID

Origin Location # of 

plants

Plot Size and Orientation

Zero point UTM E UTM N X-axis Y-axis 

ASTR-1 North end is 0,

runs downhill

559493 3765469 18 25m @ 180° ±6m on either side of line 

ASTR-2 South end is 0,

Runs uphill

~563753 ~376129

2

14 15m @ 30° ±6m on either side of line 

It was the intention of the authors to establish four permanent plots in 2013, however for a number of 

reasons this was not accomplished. The primary reasons for the shortcoming are due to a lack of sites 

with >10 plants easily accessible from a transect line, plus the poor conditions of that season (most 

plants were dormant). Although the plot west of Eureka Peak (ASTR-1) is fairly accessible without 

causing excessive soil damage while sampling, the other plot is much more difficult to traverse 
73

without causing soil disturbance. If possible, additional sites will be established, though it might be 

wise to wait a year or two to see how these two plots withstand data collection as described in the 

current protocol (provided in Appendix F). It is recommended that data collection occur annually, 

accordingly. Each year the data needs be assessed as to whether the value of the data exceeds any 

risk or damage to the plants, as well as to the time and resources required to obtain that data. After 

careful evaluation any suggested changes to the protocol can be adopted. 

Recommendations

• Conducting annual surveys is essential for building a robust long-term dataset that will allow 

meaningful conclusions about reproductive output and demographic trends. We recommend 

following the monitoring protocol provided in Appendix F annually for a minimum of 5 

years, on the two permanent plots.

• Additional permanent plots will be established. Careful consideration must be given in 

choosing a site, as the slopes can often be much more difficult to traverse than expected upon 

first observation. Ideally the plot will be small enough that new seedlings will be easily 

observed, plants can be accessed without excessive soil disturbance, and a minimum of 10 

individuals are found within 5 to 6 m of the transect line.

• We do not recommend revisiting the 50 tagged individuals from 2010.

• Establish a better understanding for dormancy in this species. Through annual monitoring of

the conditions under which they can resprout from the base will become clearer. 

• Determine meaningful demographic age and size classes.

• Determine rates of survivorship and reproductive output for age and size classes (if 

appropriate).

• Conduct analyses on how weather conditions affect demographics (survival rates for age/size 

classes) and reproductive biology (flowering period, seed production/viability) over time. 

Suggestions for important climate variables include: the amount and timing of rainfall, 

minimum precipitation per event or season, effect of summer rainfall, temperature extremes, 

and number of days under a certain temperature. 

• Tracking the same 25 inflorescences (five stems on 5 individuals) with the goal of capturing 

the very beginning of flowering (buds) to when the plant has gone completely to seed will 

establish an expected duration for each reproductive stage, as well as the timing of anthesis 

and stigma receptivity.

• Study the waifs and transient populations for longevity, survivorship, and reproductive output 

to determine if they differ from individuals found in larger populations.
74

Appendix F: Astragalus tricarinatus monitoring protocol

Ideally, each plot will be visited twice per season, targeting full flower and full fruit phenological 

stages (April to June). Two people are required for data collection and one day per plot per visit will

be required for data collection. All measurements will be recorded in metric.

Things to bring with you in the field:

1. Data sheets from previous season and several blank ones

2. Measuring tape (minimum 25 m)

3. Additional stakes and tags

4. Compass

5. Foldable measuring stick (2 m minimum)

6. Clip boards, Sharpie pens, pencils, 

7. GPS unit, extra batteries

8. Seed envelopes

9. Insect vials, net

10. Pin flags (~10 to 20)

Data collection:

1. The belt transects are 25 or 15 m in length (X-axis) and 12 m wide (Y-axis). The X-axis runs 

down the center of the belt creating a west and east side, 6 m on either side (Figure F1). 

2. Establish transect by laying down one measuring tape with the 0 point starting from the

origin and running to the established endpoint. One person walks to the endpoint with the 

tape measure reel, while the other person stands at the origin and guides them with the proper 

bearing. This will serve as the “X-axis” of the belt transect. The Y-axis is defined by the 

distance of a plant away from the X-axis, at a perpendicular angle (90°), in both the west and 

east directions within 6 m.

3. If conditions allow, pin flags can be placed along the x-axis every 2 m, or can be used to 

mark y-axis measurements. Remember that you will have to go back for them once you walk 

away, so it is best if they are not used unless they are easily retrieved. 

4. One person will be the recorder, and a second person will be the observer calling out data for 

the recorder. The recorder should stay out of the plot as much as possible.

5. For each plant the following information will be recorded/verified at each visit:

o Location of plant within transect. The location represents the center of the plant, 

to the nearest 0.1 m. The Y coordinate will be accompanied by “E” or “W”,

indicating which side of the X-axis the plant is located (Figure F1). 

o Plant ID (metal tag should be in ground next to plant, or attached to plant)

o Height1, width1, width2 of the new growth or live material only, measurements 

will be to the nearest cm (see Figures F3 and F4).

o Height2, width3, width4 of total cover, including old or dead material, only if 

greater than the above measurements. Measurements will be to the nearest cm 

(see Figures F3 and F4).

o Number of inflorescences

o Flowers or Pods/Stem. 
75

o Reproductive status

o Tag location

o Notes

6. If the tag is missing, attempt to use the past coordinates to deduce the plant ID. Write down 

the old ID number in the notes and make sure this information stays on the datasheet. Assign 

a new ID and attach new tag to the plant or stake it to the ground (preferred). Always record 

the location of the tag on the datasheet.

7. Be sure to look for and record any new seedlings or juvenile plants that were not previously 

recorded. Assign a new ID and tag, accordingly. 

8. On the first visit, randomly select five stems with flowers, use plastic tape to mark the stem 

by tying it to the base of the inflorescence. Count the number of flowers and circle “Flws” on 

the datasheet. If both flowers and pods are present on the first visit, count them separately 

and indicate how many of each by using the prefix “F“ or “P“; for example F-8, P-6 would 

mean there are eight flowers and six pods on that stem.

9. On the second visit, relocate the five stems with tape, count the number of pods on each stem, 

circle “Pod” on the datasheet. 

10. Collect one mature (but not dehisced) fruit from two different stems, place them in separate 

envelopes, then circle the appropriate box on the datasheet (for example, if you collected a 

pod from the 1st and 3rd stem on your datasheet, circle the numbers in R1 and R3). Envelopes 

will be labeled with following: date, collector initials, plot ID, and Plant ID. Remove tape 

from stem.

Explanation of Fields:

Height: measure perpendicular to ground, from base of plant to highest point (see Figure F2)

Width1 or 3: measure longest distance, parallel to ground (see Figures F2 and F3)

Width2 or 4: measure longest distance perpendicular to width 1 or 3 (see Figures F2 and F3)

Number of inflorescences: count the number of stems with flowers or pods from the current growing 

season only

# of Flws/stem: randomly choose five inflorescences, tie some tape around the stem at the base of 

inflorescence, then count the number of flowers on each inflorescence. Circle “Flws” on datasheet, or 

if both flws and pods are present apply a “F“ or “P“ as a prefix to the numbers, accordingly.

# of Pods/stem: locate the five inflorescences with tape and count the number of pods on each 

inflorescence. 

Reproductive Status: If there are no inflorescences from this season then call it vegetative (VEG), 

otherwise categorize as buds only (BUD), flowering (FLW), flowering and fruiting (FLW/FRT), 

fruiting (FRT), or post-fruiting (POST). Be sure to base this on the current season inflorescences 

only.

Tag location: Describe where the tag is located relative to the center of the plant, as well as whether 

it is attached to the plant or staked into the ground (e.g. on plant, SW)
76

Notes: Indicate anything about the plant that may be helpful or useful. For example, if it appears to 

be a juvenile or dying, location of the plant relative to other landmark, herbivory, etc. 

Important Notes:

 X- and Y-axis readings are to the nearest 0.1 m.

 All plant measurements will be to the nearest cm.

 Never leave a field blank (except in Notes).

 Timing of first monitoring should target when the plants are in full flower, though if they 

have some early stage fruit that is fine. The idea for the first visit is to capture the maximum 

number of flowers per stem. 

 Timing of second monitoring should target when the plants are in full fruit, if flowers are 

present, they are in late stages. The idea for the second visit is to capture the maximum 

number of mature fruits and therefore the maximum contribution to the seed bank.

 It is very important to minimize your impact to the area. For this reason, only one person 

should walk around inside the plot. Avoid stepping near, under, or above the base of the 

plant.

 Bring a copy of the previous years’ data with you to ensure all individuals are revisited. This 

will ensure that any new seedlings or juveniles are recognized. Data can be found in the 

Botany Program folder within the JOTR Resource Division share drive.

 Blank datasheets (Figure F4) can also be found in the Botany Program folder within the 

JOTR Resource Division share drive, be sure to print out several copies for each field day. 

 Data will be transcribed into an electronic version within a week of data collection. Hard 

copies of the datasheets will be archived.

 Do not remove the tape from a stem if doing so will damage the plant.

 If you cannot relocate the tape, pick a new stem.

Figure F1. Plot diagram for belt transects, showing two examples of how to measure the x, y axis 

readings for the location of a plant within the plot. Measurements are taken to the nearest 0.1 m. The yaxis reading is given as the distance from the x-axis with a cardinal direction (e.g. west or east) from the 

x-axis. Blue star location: x-axis = 5.1 m, y-axis = 4.2 m East. Red Star location: x-axis = 17.8 m, y-axis = 

2.7 m West.
77

Figure F2. Side view of a hill slope with two plants. Solid black lines demonstrate proper method for 

measuring width and height.

Figure F3. An aerial view of a plant; solid lines demonstrate where to take proper width measurements. 

Old or dead plant material is shown in brown with red outline, whereas new growth or live material is 

represented by green with blue outline. Width 1 and 2 represent live (green) material only (blue lines); 

width 3 and 4 represent total cover (red lines), which includes dead and live material. Width 1 and 3 

represent the longest axis, width 2 and 4 are always perpendicular (90°) to their respective axis.
78

Figure F4. Example datasheet for Astragalus tricarinatus monitoring.
79

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