沙漠研究 24-1, 33-36 (2014)
Ἃ₍◊✲ 24-1, 33 - 36 (2014)
Journal of Arid Land Studies
Journal of Arid Land Studies－ DT11 Refereed Paper －㸫DT11 Refereed Paper㸫

Rapid and Mass Propagation of the Economically Important Desert Plant
Capparis decidua for its Afforestation Program
Namita VIJAY1), Sarita ARYA*1) and I.D. ARYA2)
Abstract: Capparis decidua is an important constituent of the desert ecosystem and plays vital roles in the rural economy of western
Rajasthan. It provides people with food (vegetable and pickle), medicine, fodder, wood for carving, fuel etc. Natural populations ofCapparis decidua are being depleted, since its fruits are used as a food. Hence planting stock is not available in required quantities.
The present study was undertaken with an aim to set up a protocol for in vitro propagation. The season of explants showed direct
influence on bud break. Nodal segments of mature superior selected trees were collected. The nodal shoot explants harvested
during the months of March, April, September and October were found to be best for establishment of cultures of Capparis decidua.
Surface sterilized explants were inoculated onto MS medium supplemented with 17.76µM BAP + 0.54µM NAA + ascorbic acid (50mg/l) + adenine sulphate (50 mg/l) + citric acid (30 mg/l) for axillary bud break. The proliferated shoots were excised from motherexplants and further multiplied on MS medium supplemented with 8.88 µM BAP + 0.57 µM IAA + additives. These in vitro
regenerated shoots were elongated on MS medium + additives. Elongated in vitro shoots were in vitro rooted on one fourth strength
MS medium supplemented with 4.9µM IBA + additives and incubated at 31°C, where 94% of in vitro shoots rooted. Rooted
plantlets were hardened, acclimatized and established in soil, where they exhibited normal growth. The protocol developed is able toproduce superior planting stock for various afforestation and reforestation programmes.
Key Words: Afforestation, Capparis decidua, Reforestation

1. Introduction
Capparis decidua (Forsk.) Edgew, commonly known as
Kair, is an important indigenous shrub. In India, Kair can befound in the dry regions of Rajasthan, Gujarat, Punjab,Haryana, Madhya Pradesh and many parts of Tamil Nadu,
Karnataka and Andhra Pradesh. It is a small much branchedtree or shrub of the Thar desert and arid regions in Southern
Asia with a mass of slender, leafless branches, with small
caduceus leaves which are found only on young shoots. The
tree rarely exceeds a height of five meters. The mature fruits
serve as a valuable and integral source of nutrition for villagers
of arid and semiarid regions, and the immature fruits arecollected from natural stands and serve an additional source ofincome and nutrition for the rural poor (Singh and Singh, 2011).
The tree provides hard, heavy and termite resistant timber(Gupta et al., 1989). The seed contains 20% of high quality
edible oil. The stem and root bark extracts containisocodonocarpine and other alkaloids which are effective intreating asthma, inflammation, cough etc. (Ahmed et al.,1989). The immature fruits are used to cure stomach
problems. So diverse are the uses of every part of Capparis* Corresponding Author: aryaid@gmail.com

decidua that it is called as “Sandal Wood” of the desert in India(Harsh and Tiwari, 1998).
The plant is facing depletion of natural populations due tooverexploitation, change in climate condition, and the cuttingof plants in natural habitat for urbanization and agriculturalreasons (Anon, 2001; Khan et al., 2003). To maintain andsustain forest vegetation, conventional approaches like grafting,layering and cutting have been used for propagation.
Nonetheless, these conventional methods of plant propagation
have limited applicability (Yadav and Singh, 2011; Yadav et al.,2012). There is wide genetic variability in the field
population of Capparis decidua as it is an open pollinated plant.
The seeds have a short period of viability. Micropropagation
offers a rapid means of afforestation, multiplying woody
biomass, and conservation of elite and rare germplasm (Bajaj,1986; Karp, 1994). The technique of cell and tissue culture,under controlled and defined conditions, has contributed inraising new plants, manipulation of plant without conventional
breeding mechanism and methods, shortening germination anddevelopment phase of plants. Genetically poor gene pool is
only now available. Plant tissue culture offers an opportunity
for rapid multiplication of desired tree species.

㸦Received, September 13th, 2013; Accepted, January 17th, 2014㸧

Forest Genetics and Tree Breeding Division, Arid Forest Research Institute, Jodhpur 342005, Rajasthan, India
1) Arid Forest Research Institute, Jodhpur 342005, Rajasthan, India

2) Arid Forest Research Institute, Jodhpur 342005, Rajasthan, India

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2. Material and Methods
2.1. In vitro shoot induction
2.1.1. Plant materialExplants in the form of nodal segments (2.0-3.5 cm) werecollected from healthy, disease free mature field grown plantaof Capparis decidua for axillary bud induction andproliferation from places near Sarnau village (Rajasthan).
2.1.2. Surface sterilization
Explants were washed with Tween-80 and followed bybavistin and streptomycin. Further, surface sterilization wascarried out by 0.1% HgCl2 for 3-10 min and finally they werewashed thrice with autoclaved distilled water.
2.1.3. Effect of plant growth regulators on shoot induction
Nodal segments were inoculated on MS (Murashige andSkoog, 1962) medium supplemented with 3% (w/v) sucrose(Himedia, India) and solidified with 0.8% Agar (Himedia,India). Medium was enriched with ascorbic acid (50 mg/l),
citric acid (30 mg/l), and adenine sulphate (50 mg/l). The
medium was then incorporated with different plant growth
regulators. Cytokinins BAP and Kn were tested either alone
or in combination with auxin NAA. The pH of the medium
was adjusted to 5.8 and autoclaved at 15 lb pressure for 20
minutes at 121°C.

2.2. In vitro shoot multiplication
Multiplication of the shoots starts after bud break initiationfrom in vitro cultured plants. After 3-4 weeks in vitro
regenerated shoots of the cultures attain the height of 2-3 cm
which were cut into shoot segments containing at least two
nodes 1.5 to 2 cm in length and carefully subcultured on MS
medium supplemented with various concentration of cytokinin
BAP and Kn alone and in combination with auxin IAA andNAA. Additives such as ascorbic acid, citric acid, andadenine sulphate for shoot multiplication were also used in themedium. The multiple shoots proliferated from cultures wererepeatedly transferred to fresh cultured medium. Repeatedtransfer was done after 25-30 days. Shoot produced were
either used for further multiplication of shoots or forelongation.
2.3. In vitro rootingWell developed in vitro shoots of 5-6 cm length were usedfor various in vitro rooting experiments. These in vitro shoots
were transferred to MS medium supplemented with auxins inorder to obtain in vitro rooting. Different auxins (IBA, NAAand IAA) were tested at different concentration in the medium
for in vitro rooting. Effect of strength of MS medium (1/4×,1/3×, 1/2×, 3/4×, 1×) was investigated for in vitro rooting of invitro raised shoots.

2.4. Hardening and acclimatization of in vitro propagatedplantlets
The tissue culture raised plants are heterotrophic in theirmode of nutrition and cannot withstand harsh environmental
conditions without proper hardening and acclimatization.
The rooted plantlets were taken out from the flasks, washedthoroughly with water to remove all traces of medium attachedto the roots and then transferred to glass bottles containing 1/3
volume of soilrite. These plantlets were nurtured with half
strength MS medium (without organics) twice a week for three
weeks and were kept in a tissue culture room. After three
weeks, these bottle were shifted to a mist chamber having
relative humidity 80-90% with a temperature of 30 ± 2°C.
The caps of bottles were removed and plantlets were allowed
to remain in the bottle for 3-4 days before they were transferred
to polybags containing a mixture of sand, soil and farmyard
manure in ratio of 1:1:1. In the mist chamber, the plants were
kept for four weeks and were irrigated with half strength MS
medium. Finally, hardened plants were transferred to bigger
pots and transferred to a net house.
All cultures were incubated at 28 ± 2°C temperature andrelative humidity of 70 ± 2%, maintained in culture rooms by
air conditioners. All cultures were kept under illumination of
16 hrs photoperiod with light intensity of 2500 lux, obtained bywhite cool fluorescent tubes of 40 watts (Philips, India).
Data collected were analyzed using CRD design ofexperiments because it is one of the most widely used andacceptable designs in the case of tissue culture experiments(controlled set of conditions and for homogenous materials)
which gives the best analysis. Experiments were repeatedthrice and data represent the mean of three experiments. Eachtreatment consisted of a minimum of twelve replicates.
Degree of variation was represented by mean, standard
deviation and standard error.

3. Results and Discussion
In this report we describe a tissue culture proceduredeveloped for rapid propagation of mature plants of C. decidua.
The nodal shoot explants harvested during the month ofMarch-April and September and October were found to be bestfor establishment of cultures of Capparis decidua. Duringsterilization, the shoots were sensitive to HgCl2 treatment.
The cut ends of all the explants exhibited browning in cultureand subsequently the entire explant necrosed and died. Thus
the brown portion was removed within 4-5 days ofsubculturing and explants were cultured on the same fresh
medium to avoid further browning. Deora and Shekhawat(1995) and Chahar et al. (2010) have also reported thatbrowning could be avoided by removing the necrotic portion

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Table 1. Effect of BAP + 0.54 µM NAA concentration in MS
medium supplemented with additives on in vitro axillary
bud induction. Data recorded after 4 weeks.
BAP+NAA(0.54)(µM /l)Response %M ean shootnumberM ean ShootLength (cm)Control

15.53 ± 2.22e

1.00 ± 0.00d

1.00 ± 0.03edcd

1.44 ± 0.18

1.26± 0.12d

4.44

22.22 ± 2.22

8.88

62.00 ± 2.00b

1.79± 0.63bc

1.99 ± 0.54bc

13.32

66.66 ± 0.00ab

2.13 ± 0.08b

2.09 ± 0.04b

17.76

75.55 ± 2.22
52.22 ± 4.44ba

3.03 ± 0.14
3.10± 0.16aa

2.27 ± 0.03
2.10 ± 0.03bcac

22.2
26.66

51.11 ± 2.22
F= 85.27**

2.78 ± 0.20
F= 20.66**

a1.89 ± 0.05
F=57.41**** = P<0.01, *= P<0.05. Values with common superscript are notsignificantly different at P<0.05, Duncan multiple range test.

Table 2. Effect of BAP concentration in MS medium supplemented
with 0.57 µM IAA + additives on in vitro shootmultiplication of C. decidua. Data recorded after 4weeks.
BAP +IAA(0.57)(µM /l)M ean shootnumberM ean shootlength (cm)

M ultiplication
FoldControl

7.31 ± 0.14e

1.62 ± 0.04d

1.83 ± 0.04edc

1.89 ± 0.03

2.44 ± 0.06dbb

2.10 ± 0.04

4.44 ± 0.05ba

3.02 ± 0.04
1.93 ± 0.12ca6.47 ± 0.04
6.55 ± 0.09a

4.44

9.78 ± 0.11

6.66

17.75 ± 0.87

8.88

25.86 ± 0.22
26.14 ± 0.24a

11.1
13.32

a

15.19 ± 0.16c 1.65 ± 0.04d
3.81 ± 0.04c
F = 2025.88** F = 188.42** F = 2048.35**** = P<0.01, *= P<0.05.Values with common superscript are notsignificantly different at P<0.05, Duncan multiple range test.
Table 3. Effect of IBA on in vitro rooting of Capparis decidua. One
fourth strength MS medium supplemented with additives wasused.
IBA(µM /l)Response %M ean root
numberM ean root
length (cm)Control
0.49

4.44 ± 2.22e
50.55 ±2.78d

1.50 ± 0.50c
3.45 ± 0.43b

0.20 ± 0.00d
2.07 ± 0.64a

2.46
4.92

69.44 ± 2.78b

2.29 ± 0.24b2.12 ± 0.04a

94.44 ± 2.78
55.55 ± 2.78bc

a5.82 ± 0.31
3.14 ± 0.16ba

2.35 ± 0.07
1.70 ± 0.07b

44.44 ± 2.78c
F= 134.19**

1.75 ± 0.18c
F= 23.88**

1.30 ± 0.07c
F=41.34**

7.439.84a** = P<0.01, *= P<0.05.Values with common superscript are notsignificantly different at P<0.05, Duncan multiple range test.
Fig. 1. A-H: a) Axillary bud induction using nodal explants collectedfrom mature mother plants, b) Axillary bud proliferation
after first subculturing (after 3-4 weeks), c) In vitro shootmultiplication after second subculturing (after 3-4 weeks), d)Elongated shoots before rooting, e) In vitro rooted plantlets, f)Hardened and acclimatized plantlets, g) Micropropagatedplants transferred to poly-bags (after 3 months), h) Plants
transferred to pots (after 6 months).

within 3-4 days and re-culturing the explants on the same fresh
medium. In the present investigation, MS basal medium
supplemented with 17.76 µM BAP + 0.54 µM NAA andascorbic acid (50 mg/l), citric acid (30 mg/l), adenine sulphate(30 mg/l) was the most suitable for in vitro shoot induction(Table 1). Seventy five percent of the nodal explants
responded with 3-4 shoots/explant within 3-4 weeks (Fig. 1a).
Deora and Shekhawat, (1995) and Tyagi and Kothari, (1997)
also observed best shoot proliferation from nodal explants in
Capparis decidua on MS medium supplemented with BAPand NAA. The increased concentration of BAP showed
increased number of shoots and shoot length in the presence ofadditives. However, in the absence of the additives adecrease in shoot regeneration response was observed from
nodal explants. In the present study, axillary bud induction
was more in number on BAP supplemented medium ascompared to medium replaced with Kn. The efficiency of
BAP for shoot initiation is also documented for a number ofplant species (Kumar et al., 2010; Singh and Tiwari, 2012).
The shoots proliferated from axillary buds were established onMS medium and were used for in vitro shoot multiplication(Fig. 1b). Best shoot multiplication was obtained on MS
medium supplemented with 8.88 µM BAP + 0.57 µM IAA
and ascorbic acid (50 mg/l), citric acid (30mg/l), and adenine
sulphate (50 mg/l). A shoot multiplication rate of 6-7 fold
was obtained in a period of four weeks (Table 2 and Fig. 1c).
Similar results have been reported earlier by Deora andShekhawat (1995), Komalavalli and Rao (2000), andPhulwaria et al. (2012).
Elongated in vitro shoots of 5-6 cm length (Fig. 1d) wereused for various in vitro rooting experiments. In vitro rooting
was obtained when in vitro grown shoots were transferred on

36MS medium supplemented with auxins (IBA, NAA and IAA).
One fourth strength MS medium supplemented with 4.92 µM
IBA and ascorbic acid (50 mg/l), citric acid (30 mg/l) andincubated at 31°C gave the maximum rooting response of
94.44% with an average of 5-6 roots per shoot (Table 3 andFig. 1e). Compared to NAA and IAA, IBA was found to be abetter auxin for in vitro root induction. The present in vitro
rooting percentage appears to be superior compared withprevious reports published for the same plant species (Deoraand Shekhawat, 1995; Tyagi and Kothari, 1997; Chahar et al.,2010). Plants were hardened and acclimatized and put into
soil with 80% survival rate. (Figs. 1f-h)

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