bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Cover Crop Residue and Preplant Herbicide on Early Leaf Spot of Peanut
Authors: Cantonwine E. G. , Culbreath A. K. , Stevenson K. L.
Published: June 2007

Epidemics of early leaf spot, caused by Cercospora arachidicola, of peanut (Arachis hypogaea) are delayed in strip-tilled compared to conventionally tilled fields. This effect may be due to applications of glyphosate used to kill the winter cover crop in strip-tilled fields and/or the presence of cover crop residue at the soil surface of strip-tilled fields. Preplant herbicide (no herbicide, glyphosate, and paraquat), reciprocal residue (plus residue in conventionally tilled plots and minus residue in strip-tilled plots), and added straw mulch were evaluated to determine their effects on early leaf spot epidemics (AUDPC based on incidence and severity, and final percent defoliation) in conventionally tilled and strip-tilled plots. Additional experiments were conducted to characterize the effects of mulch (straw, fumigated straw, and plastic straw [Textraw]) treatments on disease, and to study tillage effects on disease in nonrotated peanut fields. Glyphosate and paraquat had no effect on AUDPC values or defoliation. The addition of straw to conventionally tilled plots significantly reduced disease levels. Cover crop and straw treatments had no significant effect on disease in the strip-tilled plots. AUDPC values were highest in the bare soil plots, lowest in the straw and fumigated straw plots, and intermediate in the plots with Textraw. Fewer initial infections were detected in the Textraw plots compared to the bare soil plots based on results of a trap leaf experiment. Strip-tillage did not consistently suppress early leaf spot epidemics in nonrotated fields. These results show that the presence of cover crop residue is partly responsible for the early leaf spot suppression observed in strip-tilled fields. Cover crop residue may interfere with the dispersal of primary inoculum from overwintering stroma in the soil to the plant tissues.
Detection of Latent Infections of Ralstonia solanacearum Race 3 Biovar 2 in Geranium
Authors: Swanson Jill K. , Montes Luis , Mejia Luis , Allen Caitilyn
Published: June 2007

Ralstonia solanacearum race 3 biovar 2 is a regulated quarantine pathogen that infects solanaceous hosts such as potato as well as geranium, where it causes either bacterial wilt (also known as Southern Wilt) or a symptomless latent infection. Geranium growers and government regulators need reliable detection methods to identify infected plant material before it is exported. We previously found that R. solanacearum-infected geranium plants can shed millions of bacteria in effluent water that flows from pots. We tested a nondestructive sampling method wherein effluent water from infected plants grown under commercial conditions was both dilution plated and filter concentrated for real-time polymerase chain reaction (PCR). Under field conditions in Guatemala, effluent shedding of infected geranium plants was highly variable. Comprehensive growth chamber studies confirmed that latently infected and mildly symptomatic geranium plants often but not invariably shed detectable numbers of bacteria in their effluent. At the peak of bacterial shedding, just under 90% of infected plants shed detectable bacteria whereas, at the lowest point, 44% shed detectable numbers of pathogen cells. Bacterial shedding peaked several weeks after inoculation regardless of whether plants were symptomatic or latently infected. Bacterial stem population sizes did not correlate with either effluent population sizes or disease index rating. Finally, we found that the effluent from geranium plants grown in volcanic rock scoria medium contains inhibitors that reduce the effectiveness of real-time PCR detection methods.
Distribution and Molecular Characterization of Resistance-Breaking Isolates of Beet necrotic yellow vein virus in the United States
Authors: Liu H.-Y. , Lewellen R. T.
Published: June 2007

Beet necrotic yellow vein virus (BNYVV) is the causal agent of rhizomania in sugar beet (Beta vulgaris). The virus is transmitted by the plasmodiophorid Polymyxa betae. The disease is controlled primarily by the use of partially resistant cultivars. During 2003 and 2004 in the Imperial Valley of California, partially resistant sugar beet cultivars with Rz1 allele seemed to be compromised. Field trials at Salinas, CA have confirmed that Rz1 has been defeated by resistance-breaking isolates. Distinct BNYVV isolates have been identified from these plants. Rhizomania-infested sugar beet fields throughout the United States were surveyed in 2004–05. Soil surveys indicated that the resistance-breaking isolates not only existed in the Imperial Valley and San Joaquin Valley of California but also in Colorado, Idaho, Minnesota, Nebraska, and Oregon. Of the soil samples tested by baited plant technique, 92.5% produced infection with BNYVV in ‘Beta 6600’ (rz1rz1rz1), 77.5% in ‘Beta 4430R’ (Rz1rz1), 45.0% in ‘Beta G017R’ (Rz2rz2), and 15.0% in ‘KWS Angelina’ (Rz1rz1+Rz2rz2). Analyses of the deduced amino acid sequence of coat protein and P-25 protein of resistance-breaking BNYVV isolates revealed the high percentage of identity with non-resistance-breaking BNYVV isolates (99.9 and >98.0%, respectively). The variable amino acids in P-25 proteins were located at the residues of 67 and 68. In the United States, the two amino acids found in the non-resistance-breaking isolates were conserved (AC). The resistance-breaking isolates were variable including, AF, AL, SY, VC, VL, and AC. The change of these two amino acids cannot be depended upon to differentiate resistance-breaking and non-resistance-breaking isolates of BNYVV.
Host Resistance toMonilinia vaccinii-corymbosiin Flowers and Fruits of Highbush Blueberry
Authors: Lehman Jeffrey S. , Igarashi Suzu , Oudemans Peter V.
Published: June 2007

Monilinia vaccinii-corymbosi, the causal agent of mummy berry disease, infects blueberry flowers via the gynoecial pathway. To describe the expression of host resistance in highbush blueberry (Vaccinium corymbosum), fungal growth in the styles and colonization of the locules were compared among five blueberry cultivars in a series of controlled greenhouse experiments. Styles were harvested 1 and 4 days postinoculation, and the length colonized by hyphae was determined using fluorescence microscopy. At 8 weeks after inoculation, fruit were harvested and scored for the presence of hyphae in the locules. The infection frequency of styles ranged from 0.33 to 0.71, and only cv. Weymouth had significantly lower infection frequency than the other cultivars. The mean length of the colonized portion of the stylar canal ranged from 0.126 to 0.434 mm after 1 day and 1.62 to 3.59 mm after 4 days. Hyphae in the styles of cv. Weymouth exhibited the least growth, whereas hyphae in the styles of cultivars Jersey and Rancocas were significantly longer. The distance of style penetrated for cultivars Bluecrop and Coville was intermediate. The mean disease incidence of locules differed significantly. Values for cultivars Weymouth and Jersey were the smallest (0.038 and 0.039) and largest (0.249 and 0.236), respectively. The results demonstrate that a component of resistance to infection by M. vaccinii-corymbosi is expressed during growth in the gynoecial pathway.
Identification and Quantification of Polymyxa graminis f. sp. temperata and P. graminis f. sp. tepida on Barley and Wheat
Authors: Vaïanopoulos C. , Bragard C. , Moreau V. , Maraite H. , Legrève A.
Published: June 2007

Polymyxa graminis f. sp. temperata and P. graminis f. sp. tepida are distinguished on the basis of their specific ribosomal DNA sequences. In order to evaluate whether or not host specialization is associated with the special form, the occurrence of infection of both forms on barley and wheat was studied. P. graminis inocula were obtained from soils collected in Belgium and France. Their ribotypes were characterized using molecular tools specific to P. graminis f. sp. temperata or P. graminis f. sp. tepida such as restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified rDNA, nested and multiplex PCR. Both special forms were found in each country and coexisted in some soils. The host specificity of P. graminis special forms for barley and wheat was studied from two soils collected at Gembloux (Belgium) and Chambon-sur-Cisse (France), each infested by bymo- and furoviruses. P. graminis f. sp. temperata is more frequent on barley and P. graminis f. sp. tepida on wheat. Furthermore, the quantification of each form on barley and wheat by two separated real-time quantitative PCR assays confirms the observations on the vector specialization. These results suggest a certain but not exclusive host specificity of P. graminis special forms.
Development of PCR Assays for the Identification of Species and Pathotypes of Elsinoë Causing Scab on Citrus
Authors: Hyun J. W. , Peres N. A. , Yi S.-Y. , Timmer L. W. , Kim K. S. , Kwon H.-M. , Lim H.-C.
Published: June 2007

Two scab pathogens of citrus, Elsinoë fawcettii and E. australis, cause citrus scab and sweet orange scab, respectively, and pathotypes of each species have been described. The two species cannot be readily distinguished by morphological or cultural characteristics and can be distinguished only by host range and the sequence of the internal transcribed spacer (ITS) region. In this study, random amplified polymorphic DNA (RAPD) assays clearly distinguished E. fawcettii and E. australis, and the sweet orange and natsudaidai pathotypes within E. australis also could be differentiated. We developed specific primer sets, Efaw-1 for E. fawcettii; Eaut-1, Eaut-2, Eaut-3, and Eaut-4 for E. australis; and EaNat-1 and EaNat-2 for the natsudaidai pathotype within E. australis using RAPD products unique to each species or pathotype. Other primer sets, Efaw-2 and Eaut-5, which were specific for E. fawcettii and E. australis, respectively, were designed from previously determined ITS sequences. The Efaw-1 and Efaw-2 primer sets successfully identified E. fawcettii isolates from Korea, Australia, and the United States (Florida) and the Eaut-1 to Eaut-5 primer sets identified both the sweet orange pathotype isolates of E. australis from Argentina and the natsudaidai pathotype isolates from Korea. The EaNat-1 and EaNat-2 primer sets were specific for isolates of the natsudaidai pathotype. The Efaw-1 and Efaw-2 primer sets successfully detected E. fawcettii from lesions on diseased leaves and fruit from Korea and primer pairs Eaut-1, Eaut-2, Eaut-3, Eaut-4, and Eaut-5 detected E. australis from lesions on sweet orange fruit from Brazil.
Management of Xanthomonas Leaf Blight of Onion with Bacteriophages and a Plant Activator
Authors: Lang Jillian M. , Gent David H. , Schwartz Howard F.
Published: June 2007

Xanthomonas leaf blight of onion (Allium cepa), caused by Xanthomonas axonopodis pv. allii, continues to be a challenging and yield-threatening disease in Colorado and other regions of onion production worldwide. Studies were conducted to develop management strategies for this disease that are equally effective and more sustainable than the current practices of making multiple applications of copper bactericides. Mixtures of bacteriophages and the plant defense activator, acibenzolar-S-methyl, were evaluated under field and greenhouse conditions for their abilities to reduce Xanthomonas leaf blight severity. Bacteriophage populations in the phyllosphere of onion were monitored over time. Bacteriophage populations persisted on onion leaves for at least 72 to 96 h under field and greenhouse conditions, respectively. Under field conditions at one location, biweekly or weekly applications of bacteriophages reduced disease severity by 26 to 50%, which was equal to or better than weekly applications of copper hydroxide plus mancozeb. Acibenzolar-S-methyl also successfully reduced disease severity by up to 50% when used alone preventatively or followed by biweekly bacteriophage applications. Reductions in disease severity generally were not associated with improvements in onion bulb size or yield. Integration of bacteriophage mixtures with acibenzolar-S-methyl appears to be a promising strategy for managing Xanthomonas leaf blight of onion, and could reduce grower reliance on conventional copper bactericide applied with ethylenebisdithiocarbamate fungicides.
Inheritance of Tomato yellow leaf curl virus Resistance Derived from Solanum pimpinellifolium UPV16991
Authors: de Castro Ana Pérez , Díez María José , Nuez Fernando
Published: June 2007

Resistance to tomato yellow leaf curl disease (TYLCD) in accession UPV16991 Solanum pimpinellifolium has been previously reported by our group. A breeding program was developed from an initial S. lycopersicum × S. pimpinellifolium UPV16991 cross. This first cross was followed by several selfing generations. Selection for resistance to Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Sardinia virus (TYLCSV) was carried out for plants of each generation. One partially resistant F6 plant (L102) was chosen to form the family to study the genetic control of resistance to TYLCV. Crosses between four breeding lines susceptible to TYLCD and L102 were also performed to study the dominance of the resistance in S. lycopersicum genetic backgrounds. Response to TYLCV infection of P1, P2, F1, F2, BC1, and BC2 generations fitted, for this line, a monogenic control with partial recessiveness and incomplete penetrance. The percentage of homozygotic plants with partial resistance was 72.75. Among the four hybrids developed, the highest levels of resistance were found in the hybrid formed from the most vigorous S. lycopersicum line. These results must be considered for breeding purposes. Partial resistance derived from UPV16991 will be useful in homozygosis or combined with resistance genes from other sources.
Origin ofCeratocystis platanion NativePlatanus orientalisin Greece and Its Impact on Natural Forests
Authors: Ocasio-Morales Roberto G. , Tsopelas Panaghiotis , Harrington Thomas C.
Published: June 2007

Canker stain of plane tree recently was reported in a small area of southwestern Greece on natural populations of the important riparian species, oriental plane tree, Platanus orientalis. The fungus Ceratocystis platani (= C. fimbriata f. platani) was successfully isolated from infected, stained wood of 15 dead or dying trees on the Peloponnese Peninsula. Genetic analyses of these 15 isolates from Greece, using nuclear and mitochondrial DNA fingerprints, showed the fungus to be identical to the genotype reported from Italy, France, and Switzerland. A polymerase chain reaction-based microsatellite analysis of eight polymorphic loci discovered a new microsatellite allele in one of the isolates from Greece, but this may be due to a mutation after introduction of a single strain. Earlier studies indicated that the most common European genotype had been introduced from eastern North America to Italy during World War II. The recent introduction to Greece appears to have originated from Italy, France, or Switzerland, rather than from eastern North America, where the fungus is native. The pathogen is having a dramatic impact on the natural population of P. orientalis in southwestern Greece, and containment measures should be imposed before it spreads throughout the natural range of this ecologically and historically important host.
First Report of Anthracnose Caused by Elsinoe ampelina on Muscadine Grapes (Vitis rotundifolia) in Northern Florida
Authors: Yun H. K. , Louime C. , Lu J.
Published: June 2007

Anthracnose of grapes is an economically devastating disease caused by Elsinoe ampelina (2). Warm, humid weather favors disease development, and therefore in the United States, it is generally restricted to grape-growing areas east of the Rocky Mountains. Vitis vinifera is highly susceptible to the disease, which is one of the principal factors preventing the development of an industry with this high-quality grape in the southeastern United States. Growers in this area produce local species-such as muscadine grapes (V. rotundifolia Michx.) and hybrids. Muscadine grapes are known for their resistance or “immunity” to many diseases found in bunch (Euvitis spp. Planch.) grape species (1). As yet, there has been no formal report of anthracnose or its causal agent on muscadine grapes. E. ampelina was detected on muscadine leaves for the first time in the experimental vineyard at the Center for Viticulture and Small Fruit Research during the summer of 2006. Approximately 40% of the 52 muscadine cultivars in the collection showed circular or irregular black spots typical of anthracnose mainly on young leaves and tendrils. However, no symptoms were observed on fruits, shoot tips, or any other plant part. To confirm the causal agent, infected leaves were surface sterilized with 75% ethanol, dipped in 2% sodium hypochlorite for 15 s, rinsed in distilled water, dissected into small 0.5-cm leaf discs, and plated on potato dextrose agar (PDA) and incubated at 28°C. Single-spore isolates were grown on PDA. Colonies were slow growing and appeared as dark red mounds with some mycelia. Conidia were cylindrical and hyaline with pointed ends consistent with previous reports for E. ampelina (2). The identity was also confirmed by using the following PCR primers to the 18S RNA: left primer; TCCGTAGGTGAACCTGCGGA and right primer; TCCTACCTGAT CCGAGGTCA designed on the basis of the alignment of E. ampelina sequences deposited in NCBI database. To fulfill Koch's postulates, symptoms were reproduced by artificial inoculation onto young muscadines (cv. Carlos) and bunch (cv. Cabernet Sauvignon) grapevines. A conidial suspension was prepared from single-conidial cultures, and three experimental vines of each species were sprayed with 0.5 ml of suspension (2 × 105 conidia per ml), whereas three control plants were sprayed with distilled water. The plants were incubated in a moist chamber at 28°C with 16 h of light. The first typical symptoms appeared on V. vinifera 4 days postinoculation and on the muscadines 6 days postinoculation. To our knowledge, this is the first report confirming anthracnose disease on muscadine grapes.

References: (1) J. Lu et al. Acta Hortic. 528:479, 2000. (2) R. C. Pearson and A. C. Gohen. Anthracnose. Pages 18–19 in: Compendium of Grape Diseases. The American Phytopathological Society. St. Paul, MN, 1994.
First Report of Subanguina radicicola, the Root-Gall Nematode Infecting Poa annua Putting Greens in Washington State
Authors: Mitkowski N. A.
Published: June 2007

In the fall of 2006, a golf course in Snoqualmie, WA renovated five putting greens with commercially produced Poa annua L. sod from British Columbia, Canada. Prior to the renovation, the greens had been planted with Agrostis stolonifera L. cv. Providence, which was removed during the renovation. In February of 2007, chlorotic patches were observed on the newly established P. annua greens. When the roots were examined, extensive galling was observed throughout plant roots. Galls were slender and twisted in appearance and less than one millimeter long. Upon dissection of washed galls, hundreds of eggs were exuded into the surrounding water droplet and both mature male and female nematodes were observed. Further morphometric examination of males, females, and juvenile nematodes demonstrated that they were Subanguina radicicola (Greef 1872) Paramanov 1967 (1). Amplification of nematode 18S, ITS1, and 5.8S regions, using previously published primers (2), resulted in a 100% sequence match with the publicly available sequence for S. radicicola, GenBank Accession No. AF396366. Each P. annua plant had an average of six galls (with a range of 1 to 8), primarily located within the top 2 cm of the soil. All five new P. annua putting greens at the golf course were infested with the nematode. Additionally, P. annua from two A. stolonifera cv. Providence greens that had not been renovated was infected, suggesting that the population occurred onsite and was not imported from the Canadian sod. S. radicicola has been identified as causing severe damage in New Brunswick, Canada on P. annua putting greens and in wild P. annua in the northwestern United States, but to our knowledge, this is the first report of the nematode affecting P. annua on a golf course in the United States.

References: (1) E. L. Krall. Wheat and grass nematodes: Anguina, Subanguina, and related genera. Pages 721–760 in: Manual of Agricultural Nematology. Marcel Dekker, New York, 1991. (2) N. A. Mitkowski et al. Plant Dis. 86:840, 2002.
First Report of Brown Rot Caused by Monilinia fructicola on Various Stone and Pome Fruits in the Czech Republic
Authors: Duchoslavová J. , Širučková I. , Zapletalová E. , Navrátil M. , Šafářová D.
Published: June 2007

Monilinia fructicola, a causal agent of brown rot, is one of the most important fungal pathogens of stone fruits. The disease causes major crop losses in peach, plum, prune, nectarine, and apricot. M. fructicola is commonly present in Asia, North and South America, and Australia. This is a quarantined pathogen in Europe; restricted occurrence has been observed in Austria and France. Recently, it was detected in Hungary and Switzerland on peach and nectarine fruits imported from Italy and Spain (1,4). During a survey in the summer of 2006, 56 samples were tested for the presence of Monilinia spp. M. fructicola was detected in 15 samples from 11 locations in the western area (Bohemia) of the Czech Republic, mainly on peaches (Prunus persica), apples (Malus × domestica), and sweet and sour cherries (Cerasus avium and C. vulgaris) and rarely on flowering plum (Prunus triloba) and Malus × moerlandsii cv. Liset. On the other hand, the pathogen was not detected on fruits of apricot (Prunus armeniaca) or pear (Pyrus communis). In all cases, M. fructicola was detected on fruits except for a single occurrence of the pathogen on a shoot of the Malus × domestica. The pathogen was always detected in mixed infections with M. fructigena and/or M. laxa. On both fruits and the shoot, symptoms appeared as brown, sunken lesions covered with grayish pustules. Many infected fruits became dry and mummified because rot progressed through the fruit surface. The infected shoot died back (3). M. fructicola was identified by means of colony and conidial morphology and molecular characteristics. The colonies cultivated on potato dextrose agar were entire and the colony surface was even. The color of the colony was gray, and sporulating colonies showed concentric rings that changed to a hazel color. Conidia were ellipsoid, hyaline, and 13.5 to 17.7 × 8.3 to 10.5 μm. Preliminary morphological identification was confirmed by PCR (2) on DNA isolated directly from mycelium on the examined fruits. A product that was 280 bp long was obtained in all cases. The BLAST analysis of our PCR product sequences showed 100% homology to sequences of M. fructicola (GenBank Accession Nos. DQ491506, AY2891185, Z73778, and AB125615). One sequence from our study was deposited in GenBank (Accession No. EF378628). To our knowledge, this is the first report of the quarantined fungus M. fructicola in the Czech Republic.

References: (1) E. Bosshard et al. Plant Dis. 90:1554, 2006. (2) K. J. D. Hughes et al. EPPO Bull. 30:507, 2000. (3) J. M. Ogawa et al., eds. Compendium of Stone Fruit Diseases. The American Phytopathological Society, St. Paul, MN, 1995. (4) M. Petróczy and L. Palkovics. Plant Dis. 90:375, 2006.
First Report of Phytophthora citricola Occurring on Fagus sylvatica in Slovenia
Authors: Munda A. , Žerjav M. , Schroers H.-J.
Published: June 2007

During 2005 and 2006, eight declining, mature Fagus sylvatica trees with severe crown dieback were encountered at two stands near Ljubljana and Kamnik in central Slovenia. Bleeding cankers were seen mostly in the lower parts of the stem. Fresh cankers showed orange brown necroses of the inner bark and cambium. Older cankers were dark brown and surrounded by cracks and shedding bark. Small pieces of marginal tissue were excised from fresh necrotic lesions and incubated in the dark at 20°C on P5ARP and cornmeal (CMA) agar plates. Soil samples collected from around the base of these declining trees were submerged in sterile distilled water. Leaves of Rhododendron catawbiense were used as bait to test for the presence of Phytophthora spp. Several similar isolates were obtained from trees and soil from both stands. Colonies were narrowly petaloid, slow growing on P5ARP, and fast growing on CMA. The colonies produced abundant sporangia when submerged in pond water. Semipapillate, noncaducous sporangia were ovoid to obpyriform, but were occasionally distorted, and 30 to 69 (49) μm long and 23 to 44 (34) μm wide. Neither hyphal swellings nor chlamydospores developed. Isolates were homothallic showing paragynous antheridia, spherical oogonia of 22 to 31 (25) μm diameter, and plerotic oospores of 20 to 27 (23) μm diameter. The morphological characters resembled those described for Phytophthora citricola (1). ITS rDNA, spanning ITS 1 and 2, plus the 5.8S rDNA were generated using primers ITS4 and ITS5. Isolates from symptomatic trees and the soil yielded identical sequences and were the same as various sequences deposited for P. citricola at GenBank. The ITS rDNA of one representative strain was deposited at GenBank (Accession No. EF423556). Mycelial plugs from one of the P. citricola strains grown on CMA were used to inoculate stem wounds of seven potted seedlings and wounds made on four freshly cut, healthy branches of F. sylvatica. Sterile agar plugs were used as controls. The test was carried out over 4 weeks at 20°C. Extensive necrotic lesions developed around inoculation points on seedlings and branches, whereas the controls showed no symptoms. P. citricola could also be reisolated from margins of these lesions. During the past decade, declining F. sylvatica trees were observed in an increasing number of stands in Germany (2). Several Phytophthora species appeared to be involved in this decline, but P. citricola was the most frequently recovered species (2,3). To our knowledge, this it the first report of P. citricola found associated with a decline of European beech in Slovenia.

References: (1) D. C. Erwin and O. K. Ribeiro. Pages 282–287 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) T. Jung. Forst Holz. 60:131, 2005. (3) T. Jung et al. Mycologist 19:159, 2005.
First Report of Ascochyta rabiei Causing Ascochyta Blight of Cicer pinnatifidum
Authors: Can C. , Ozkilinc H. , Kahraman A. , Ozkan H.
Published: June 2007

In July 2005, small (2 to 5 mm), elongated, dark brown spots on the stems of Cicer pinnatifidum Jaub. & Spach. were observed on plants grown in the rocky hills of the Kahramanmaras Province. To understand this phenomenon, field trips to Kahramanmaras, Adiyaman, and Sanliurfa provinces were conducted in the summer of 2006. C. pinnatifidum plants exhibiting symptoms similar to Ascochyta rabiei (Pass.) Lab. were collected during May and June. The plants had flowers and pods with seeds at the time of collection. Ascochyta blight symptoms on stems were not extensive. None of the plants had leaf symptoms, but one plant had lesions on its pods. Twelve plants exhibiting Ascochyta blight symptoms were taken to the laboratory, and necrotic parts were used for isolation of the fungi on potato dextrose agar (PDA). After 3 to 5 days of culturing on PDA, characteristic beige-to-dark brown colony development of A. rabiei from explants was observed and five isolates from different locations were recovered. The fungal colony growth was slow and limited conidia formed on PDA. The isolates were also cultured on chickpea meal agar (CMA) and Czapek Dox Agar (CDA) media. Abundant conidia formation occurred only on CMA, 10 to 12 days after culturing. Conidia were one-celled similar to that of A. rabiei of chickpea and single-spore isolations were done. C. pinnatifidum and chickpea cv. Gokce (C. arietinum L.) were inoculated with spore suspensions of 5 × 105 spores per ml (2). Ten- to twelve-day-old seedlings were used for inoculation in the experiments. Brown-black lesions at the crown region on C. pinnatifidum seedlings were observed 9 to 10 days after inoculation, and characteristic Ascochyta blight symptoms on stems developed on chickpea cv. Gokce. The fungus was reisolated from the infected seedlings. For molecular characterization, mating type of the isolates was determined by PCR using A. rabiei specific Tail1, Com1, and Sp21 primers (1). A single band of Mat 1.2 specific 500- bp product was amplified by PCR from five of the A. rabiei isolates of C. pinnatifidum. This confirmed that the isolates from C. pinnatifidum are A. rabiei.

References: (1) M. P. Barve et al. Fungal Genet. Biol. 39:151, 2003. (2) M. S. A. Khan et al. Plant Pathol. 48:230, 1999.
First Report of Leaf Spot and Twig Blight of Rhododendron spp. Caused by Phytophthora hibernalis in Spain
Authors: Álvarez L. A. , Pérez-Sierra A. , García-Jiménez J. , Abad-Campos P. , Landeras E. , Alzugaray R.
Published: June 2007

During the early spring of 2004, an estimated 20% of containerized nursery stocks of Rhododendron spp. in Asturias (northern Spain) were affected by a foliar disease that has reoccurred annually. Leaf spots were dark brown to almost black, generally oval to round, visible from both sides of the leaf, and expanded to affect the entire leaf including the petiole. Affected leaves abscised from the plant. A Phytophthora sp. was consistently isolated from symptomatic leaf tissues on PARBH medium (3) and hyphal tips were transferred onto potato dextrose agar (PDA). Colonies grown on PDA at 20°C were submerged, had a growth rate of 2.2 mm/day, and had lobes of compact mycelium. Sporangia were semipapillate and caducous with a pedicel (20.0-) 37.7 (-52.5) μm long. Sporangia were asymmetrical in shape with the broadest point near the apex: 25.2 to 40.4 μm long × 10.2 to 15.8 μm wide (average 33.1 × 12.6 μm), and length/width ratio was 2.8:1. Chlamydospores were not observed. Isolates were homothallic and oogonia ranged from 26.5 to 27.5 μm in diameter. Antheridia were mostly amphigynous but occasionally paragynous. Oospores were plerotic and 23.1 to 25.5 μm in diameter. These characteristics conformed to those of Phytophthora hibernalis Carne (2). Sequences of the internal transcribed spacer regions on the isolates and comparison with other sequences in GenBank showed that they were identical to P. hibernalis (Accession No. AY827556.1 from Citrus sp.). For pathogenicity tests, four isolates of P. hibernalis were used to inoculate detached leaves of Rhododendron hybrid Brigitte. The underside of five detached leaves was inoculated with a drop of 40 μL of a suspension of 104 zoospores/ml. Controls were inoculated with a 40-μL drop of sterile distilled water. Leaves were incubated in a moist chamber at 20°C in the dark. A quantification of the lesion area was made 8 days after inoculation using the software Assess-APS. All inoculated leaves developed necrotic lesions that ranged from 0.246 to 1.512 cm2. P. hibernalis was reisolated from infected tissue. Symptoms were not detected on the controls. The test was repeated twice and similar results were obtained each time. P. hibernalis has been described previously as causing brown rot on citrus in Spain (4) and was isolated from rhododendron plants in California and Oregon (1). To our knowledge, this is the first record of P. hibernalis causing foliar blight on Rhododendron species in Spain as well as in Europe.

References: (1) C. Blomquist et al. Online publication. doi:10.1094/PHP-2005-0728- 01-HN. Plant Health Progress, 2005. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul MN. 1996. (3) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (4) J. J. Tuset. An. Inst. Nac. Investig. Agrar. Ser. Prot. Veg. N.7, 1977.
First Report of Cucumber vein yellowing virus in Melon in France
Authors: Lecoq H. , Dufour O. , Wipf-Scheibel C. , Girard M. , Cotillon A. C. , Desbiez C.
Published: June 2007

During the fall of 2003, mild mosaic symptoms were observed in melon (Cucumis melo L.) plants grown in glasshouses near Eyragues (southeastern France) resembling those caused by the Bemisia tabaci transmitted Cucumber vein yellowing virus (CVYV, genus Ipomovirus, family Potyviridae). In addition, large numbers of B. tabaci were observed to be colonizing these crops. The identification of CVYV was established through differential host range reaction, immunosorbent electron microscopy (IEM), and reverse transcription (RT)-PCR experiments. Crude sap from symptomatic leaves was used to inoculate differential host plants. Mild mosaic symptoms were observed on melon, and cucumber developed vein-clearing symptoms typical of CVYV. No symptoms were observed in Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana, N. tabacum, and Vigna sinensis. Numerous, slightly flexuous, elongated virus particles were observed in infected plant extracts; these particles were decorated by a polyclonal antiserum raised against a Sudanese CVYV isolate. To confirm CVYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St. Louis, MO) were obtained from the original symptomatic melon tissues. RT-PCR was carried out using CVYV-specific primers CVYV-CP-5′: 5′-GCTTCTGGTTCTCAAGTGGA-3′ and CVYV-CP- 3′: 5′-GATGCATCAGTTGTCAGATG-3′ designed according to the partial sequence of the coat protein gene of CVYV-Isr (GenBank Accession No. AF233429) (2). A 540-bp fragment corresponding to the central region of CVYV coat protein was amplified from total RNA extracted from symptomatic but not from asymptomatic melon tissue. Direct sequencing was done on RT-PCR products (GenBank Accession No. EF441272). The sequence was 95 and 99% identical to that reported for CVYV isolates from Israel and Spain, respectively. CVYV was first described in Israel and has recently emerged as the cause of important diseases in Spain and Portugal (1,3). Shortly after detecting CVYV during 2003, efforts were made to eradicate the virus in susceptible crops. CVYV was not detected again during intensive surveys conducted in southeastern France during 2004, 2005, and 2006, suggesting that the CVYV detected during 2003 resulted from an accidental introduction and that the virus has not become established in France.

References: (1) I. M. Cuadrado et al. Plant Dis. 85:336, 2001. (2) H. Lecoq et al. J. Gen. Virol. 81:2289, 2000. (3) D. Louro et al. Plant Pathol. 53:241, 2004.
The Course of Colonization of Two Different Vitis Genotypes by Plasmopara viticola Indicates Compatible and Incompatible Host-Pathogen Interactions
Authors: Unger Sabine , Büche Claudia , Boso Susana , Kassemeyer Hanns-Heinz
Published: June 2007

The course of colonization of leaf mesophyll by the causal agent of grapevine downy mildew, Plasmopara viticola, in a susceptible and a resistant grapevine genotype was examined in order to characterize the development of the pathogen in compatible and incompatible host-pathogen interactions. Within a few hours after inoculation, the pathogen was established in the susceptible Vitis vinifera cv. Müller-Thurgau and formed primary hyphae with a first haustorium. No further development occurred in the following 10 to 18 h. The next step, in which the hyphae grew and branched to colonize the intercellular space of the host tissue, was observed 1.5 days after inoculation. After 3 days, the intercostal fields were entirely filled with mycelium and sporulation was abundant under favorable environmental conditions. The first infection steps were essentially the same in the resistant V. rupestris. However, the invasive growth of P. viticola was delayed, and further development ceased before the intercostal fields were filled with mycelium.
Real-time PCR Quantification and Mycotoxin Production of Fusarium graminearum in Wheat Inoculated with Isolates Collected from Potato, Sugar Beet, and Wheat
Authors: Burlakoti Rishi R. , Estrada Rolando , Rivera Viviana V. , Boddeda Anuradha , Secor Gary A. , Adhikari Tika B.
Published: June 2007

Fusarium graminearum causes Fusarium head blight (FHB) in small grains worldwide. Although primarily a pathogen of cereals, it also can infect noncereal crops such as potato and sugar beet in the United States. We used a real-time polymerase chain reaction (PCR) method based on intergenic sequences specific to the trichodiene synthase gene (Tri5) from F. graminearum. TaqMan probe and primers were designed and used to estimate DNA content of the pathogen (FgDNA) in the susceptible wheat cv. Grandin after inoculation with the 21 isolates of F. graminearum collected from potato, sugar beet, and wheat. The presence of nine mycotoxins was analyzed in the inoculated wheat heads by gas chromatography and mass spectrometry. All isolates contained the Tri5 gene and were virulent to cv. Grandin. Isolates of F. graminearum differed significantly in virulence (expressed as disease severity), FgDNA content, and mycotoxin accumulation. Potato isolates showed greater variability in producing different mycotoxins than sugar beet and wheat isolates. Correlation analysis showed a significant (P < 0.001) positive relationship between FgDNA content and FHB severity or deoxynivalenol (DON) production. Moreover, a significant (P < 0.001) positive correlation between FHB severity and DON content was observed. Our findings revealed that F. graminearum causing potato dry rot and sugar beet decay could be potential sources of inoculum for FHB epidemics in wheat. Real-time PCR assay provides sensitive and accurate quantification of F. graminearum in wheat and can be useful for monitoring the colonization of wheat grains by F. graminearum in controlled environments, and evaluating wheat germplasms for resistance to FHB.
Photosynthetic Declines in Phytophthora ramorum-Infected Plants Develop Prior to Water Stress and in Response to Exogenous Application of Elicitins
Authors: Manter Daniel K. , Kelsey Rick G. , Karchesy Joseph J.
Published: June 2007

Phytophthora ramorum, causal agent of sudden oak death, is responsible for widespread oak mortality in California and Oregon, and has the potential to infect 100 or more species. Symptoms range from stem girdling and shoot blight to leaf spotting. In this study, we examined the physiological impacts of P. ramorum infection on Rhododendron macrophyllum. In stem-inoculated plants, photosynthetic capacity (Vcmax) significantly declined by ≈21% 3 weeks after inoculation in visibly asymptomatic leaves. By 4 weeks, after the development of significant stem lesions and loss in water transport capacity, water stress led to stomatal closure and additional declines in photosynthetic capacity. We also report the isolation, characterization, and biological activity of two P. ramorum elicitins. Both elicitins were capable of inducing a hypersensitive-like response in one incompatible (Nicotiana tabacum SR1) and three compatible hosts (R. macrophyllum, Lithocarpus densiflorus, and Umbellularia californica). Infiltration of leaves from all three compatible hosts with both P. ramorum elicitins caused significant declines in chlorophyll fluorescence (Fv /Fm). For all four species, the loss of photosynthetic capacity was directly proportional to H+ uptake and ethylene production, two common components of the hypersensitive response. This is the first report of elicitins causing photosynthetic declines in compatible hosts independent of plant water stress.
Phylogenetic Relationships of Xylella fastidiosa Strains Isolated from Landscape Ornamentals in Southern California
Authors: Hernandez-Martinez Rufina , de la Cerda Karla A. , Costa Heather S. , Cooksey Donald A. , Wong Francis P.
Published: June 2007

Xylella fastidiosa is an insect-borne, xylem-limited pathogenic bacterium that has been associated with a rise in incidence of diseased landscape ornamentals in southern California. The objective of this study was to genetically characterize strains isolated from ornamental hosts to understand their distribution and identity. Strains of X. fastidiosa isolated from ornamentals were characterized using a multiprimer polymerase chain reaction (PCR) system, random amplified polymorphic DNA (RAPD)-PCR, and sequence analysis of the 16S-23S rDNA intergenic spacer region (ISR). Based on RAPD-PCR and 16S-23S rDNA ISR, strains isolated from daylily, jacaranda, and magnolia clustered with members of X. fastidiosa subsp. sandyi and caused oleander leaf scorch but not Pierce's disease symptoms in glasshouse assays on oleander and grape, respectively. This demonstrated both that our groupings based on genetic characterization were valid and that strains of X. fastidiosa subsp. sandyi are present in hosts other than oleander. Strains isolated from Spanish broom, cherry, and one strain isolated from western redbud clustered with X. fastidiosa subsp. fastidiosa members. Strains isolated from purple-leafed plum, olive, peach, plum, sweetgum, maidenhair tree, crape myrtle, and another western redbud strain clustered with members of X. fastidiosa subsp. multiplex. All strains isolated from mulberry and one from heavenly bamboo formed a separate cluster that has not yet been defined as a subspecies.
Multiplex Real-Time Quantitative PCR to Detect and Quantify Verticillium dahliae Colonization in Potato Lines that Differ in Response to Verticillium Wilt
Authors: Atallah Z. K. , Bae J. , Jansky S. H. , Rouse D. I. , Stevenson W. R.
Published: June 2007

Potato early dying (PED), also known as Verticillium wilt, caused by Verticillium dahliae, is a seasonal yield-limiting disease of potato worldwide, and PED-resistant cultivars currently represent only a small percentage of potato production. In this study, we developed a real-time quantitative polymerase chain reaction (Q-PCR) approach to detect and quantify V. dahliae. The efficiency of the designed primer pair VertBt-F/VertBt-R, derived from the sequence of the β-tubulin gene, was greater than 95% in monoplex Q-PCR and duplex (using Plexor technology) procedures with primers PotAct-F/PotAct-R, obtained from the sequence of the actin gene, designed for potato. As few as 148 fg of V. dahliae DNA were detected and quantified, which is equivalent to five nuclei. Q-PCR detected V. dahliae in naturally infected air-dried potato stems and fresh stems of inoculated plants. Spearman correlations indicated a high correlation (upward of 80%) between V. dahliae quantifications using Q-PCR and the currently used plating assays. Moreover, Q-PCR substantially reduced the variability compared with that observed in the plating assay, and allowed for the detection of V. dahliae in 10% of stem samples found to be pathogen free on the culture medium. The described Q-PCR approach should provide breeders with a more sensitive and less variable alternative to time-consuming plating assays to distinguish response of breeding lines to colonization by V. dahliae.
Asexual Reproduction of Phytophthora capsici as Affected by Extracts from Agricultural and Nonagricultural Soils
Authors: Sanogo S.
Published: June 2007

Formation of sporangia and zoospores in species of Phytophthora is known to be influenced by soil microbial and chemical composition. In Phytophthora capsici, the study of the relationship of soil chemical composition to production of sporangia and zoospores has been limited. P. capsici is a soilborne pathogen of a wide array of vegetable crops, including chile pepper (Capsicum annuum) in New Mexico. Production of sporangia and zoospores by P. capsici was evaluated in extracts of soils from three different environments in New Mexico: (i) agricultural environments with a long history of chile pepper cropping and occurrence of P. capsici (CP), (ii) agricultural environments with no history of chile pepper cropping and no occurrence of P. capsici (Non-CP), and (iii) nonagricultural environments consisting of forests and rangelands (Non-Ag). There was a significant difference in production of P. capsici asexual propagules, expressed as natural log (number of sporangia × number of zoospores), among the three environments (P = 0.0298). Production of propagules was 9 to 13% greater in Non-Ag than in CP or Non-CP environments. Stepwise multiple discriminant analysis and canonical discriminant analysis identified the edaphic variables Na, pH, P, organic matter content, and asexual propagule production as contributing the most to the separation of the three environments. Two significant (P < 0.0001) canonical discriminant functions were derived with the first function, accounting for ≈75% of the explained variance. Based on the two discriminant functions, ≈93, 86, and 89% of observations in CP, Non-CP, and Non-Ag environments, respectively, were classified correctly. Soils from agricultural and nonagricultural environments differentially influence production of sporangia and zoospores in P. capsici, and soil samples could be effectively classified into agricultural and nonagricultural environments based on soil chemical properties and the production of asexual propagules by P. capsici in soil extracts.
Microsatellite and Minisatellite Analysis of Leptosphaeria maculans in Australia Reveals Regional Genetic Differentiation
Authors: Hayden Helen L. , Cozijnsen Anton J. , Howlett Barbara J.
Published: June 2007

The population genetic structure of the fungal pathogen Leptosphaeria maculans was determined in Australia using six microsatellite and two minisatellite markers. Ascospores were sampled from Brassica napus stubble in disease nurseries and commercial fields in different sites over 2 years. The 13 subpopulations of L. maculans exhibited high gene (H = 0.393 to 0.563) and genotypic diversity, with 357 haplotypes identified among 513 isolates. Although the majority of genetic variation was distributed within subpopulations (85%), 10% occurred between the regions of eastern and Western Australia, and 5% within regions. FST analysis of subpopulation pairs also showed the east-west genetic differentiation, whereas factorial correspondence analysis separated Western Australian subpopulations from eastern ones. Bayesian model-based population structure analyses of multilocus haplotypes inferred three distinct populations, one in Western Australia and an admixture of two in eastern Australia. These two regions are separated by 1,200 km of arid desert that may act as a natural barrier to gene flow, resulting in differentiation by random genetic drift. The genetic differentiation of L. maculans isolates between eastern and Western Australia means that these regions can be treated as different management units, and reinforces the need for widespread disease nurseries in each region to screen breeding lines against a range of genetic and pathogenic populations of L. maculans.