First Report of Wheat dwarf virus and Its Vector (Psammotettix provincialis) Affecting Wheat and Barley Crops in Syria

A field survey covering the major cereal-production areas of Syria was conducted during May 2009. A total of 938 wheat and 971 barley samples with typical symptoms of viral infection were collected from 45 wheat and 58 barley fields. All collected samples were tested by the tissue-blot immunoassay (1) at the Virology Laboratory of ICARDA, Syria using six specific cereal virus antisera, including polyclonal antibody AS-0216 for Wheat dwarf virus (WDV) provided by the German Collection of Microorganisms and Cell Cultures (DSMZ). Serological tests showed that WDV was detected in 16 wheat (cv. Cham 8) and five barley (cv. Arabic abiad) samples collected from Al-Hasskah governorate (eastern region of Syria) and showing dwarfing, yellowing, and reduced heading. Samples that reacted with WDV antiserum were transmitted from infected plants to healthy plants of oat (Avena sativa L.), barley (Hordeum vulgare L.), wheat (Triticum aestivum L.), and some grass species using four different leafhopper species, collected from Syrian wheat and barley fields, in a persistent manner. Leafhopper transmission tests indicated that only Psammotettix provincialis Ribaut was able to transmit Syrian barley WDV isolates (SB 1248-09 and SB 1249-09) from infected barley plants to healthy barley (48 plants became infected of 50 plants inoculated) and oats (45 of 50) under greenhouse conditions. The identity of P. provincialis was confirmed by the British Museum. Total DNA was extracted from six WDV-positive samples (three wheat and three barley) and tested by PCR using WDV primer set (WDV-F: 5′-TTGAGCCAATCTTCGTC-3′; WDV-R: 5′-GGAAAGACTTCCTGGGC-3′) described by Oluwafemi (2). All six Syrian WDV-positive samples generated amplicons around the expected size (~253 bp). The amplicons from one isolate from wheat (SW 2131-09, GenBank Accession No. HQ113095) and one isolate from barley (SB 1248-09, GenBank Accession No. HQ113096) showed they had 86% sequence identity with each other, suggesting that both isolates can be considered to belong to the same species (3). Barley isolate SB 1248-09 had 99% sequence identity to an Iranian isolate of Barley dwarf virus (FJ620684.1) and 92% identity to most European barley-WDV isolates (e.g., Germany [AM942044.1] and Hungary [FM999832.1]), whereas, the wheat isolate (SW 2131-09) had 98 to 100% identity with most European wheat-WDV isolates (e.g., Czech Rep [FJ546191.1] and Germany [AM296023.1]) and a Chinese isolate (EF536868.1). WDV has been reported to infect cereals in few countries in West Asia and North Africa (Turkey, Tunisia, and Morocco) and causes economic losses on wheat in many countries in Europe (e.g., Sweden). WDV has been reported to be transmitted in a persistent manner only by leafhoppers (P. alienus Dahlbom) (4) to a wide range of cereal and wild grasses. Two strains of WDV are known, one that primarily infects wheat and another that infects barley. To our knowledge, this is the first record of WDV (both strains) infecting wheat and barley crops in Syria and the first report of P. provincialis as a WDV vector worldwide. References: (1) K. M. Makkouk and A. Comeau. Eur. J. Plant Pathol. 100:71, 1994. (2) S. Oluwafemi. Afr. J. Biotechnol. 5:590, 2006. (3) J. Stanley et al. Page 301 in: The International Committee on the Taxonomy of Viruses. 8th Report. Elsevier/Academic Press, London, 2005. (4) J. Vacke. Biol. Plant. 3:228, 1961.

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First Report of Chickpea chlorotic stunt virus Infecting Legume Crops in Tunisia

Plant Disease ◽

10.1094/pdis-10-10-0708 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1321-1321 ◽

Cited By ~ 3

Author(s):

A. Najar ◽

S. Kumari ◽

N. Attar ◽

S. Lababidi

Keyword(s):

Faba Bean ◽

Economic Losses ◽

Healthy Plant ◽

High Sequence Identity ◽

Serological Tests ◽

First Report ◽

Coat Protein Region ◽

Beet Western Yellows Virus ◽

Sequence Identity ◽

Legume Crops

During a survey of legume crops in the northeast and northwest regions of Tunisia in April 2010, plants showing yellowing, reddening, and stunting symptoms were observed. A total of 281 symptomatic samples were collected: 142 plants from 10 chickpea (Cicer arietinum L.) fields, 84 plants from six faba bean (Vicia faba L.) fields, and 55 plants from six pea (Pisum sativum L.) fields. All samples were tested by the tissue-blot immunoassay procedure with the following monoclonal antibodies (MAbs): a broad-spectrum legume-luteovirus MAb (5G4), Faba bean necrotic yellows virus (FBNYV; genus Nanovirus, family Nanovirudae) (3-2E9; provided by J. Vetten, BBA, Braunschweig, Germany), Beet western yellows virus (BWYV; genus Polerovirus, family Luteoviridae) (A5977; Agdia, Elkhart, IN), Bean leafroll virus (BLRV; genus Luteovirus, family Luteoviridae) (4B10), Soybean dwarf virus (SbDV; genus Luteovirus, family Luteoviridae) (ATCC PVAS-650; American Type Culture Collection ATCC, Rockville, MD,), and a mixture of three MAbs (5-2B8, -3D5, and -5B8) to a Syrian isolate of Chickpea chlorotic stunt virus (CpCSV) (1). Serological tests showed that CpCSV was detected in 121 samples (43.06%) (62 chickpea, 57 faba bean, and 2 pea), followed by FBNYV (detected in three faba bean and three pea), BWYV (detected in three chickpea and one faba bean), and BLRV (detected in one pea sample). FBNYV, BLRV, and BWYV have been previously detected in faba bean and chickpea in Tunisia (4), but to our knowledge, this is the first report of CpCSV affecting legumes in Tunisia, which was found in seven chickpea, seven faba bean, and two pea fields. CpCSV has been reported to naturally infect legume crops such as chickpea, lentil, field pea, and faba bean as well as some leguminous weeds and a few wild non-legume plants species in many countries in West Asia and North Africa and causes economic losses on chickpea in Eritrea, Ethiopia, and Syria (1–3). Serological results of CpCSV was confirmed in four (two pea, one faba bean, and one chickpea) samples by reverse transcription (RT)-PCR using CpCSV specific primers (F:5′-TAGGCGTACTGTTCAGCGGG-3′ and R:5′-TCCTTTGTCCATTCGAGGTGA-3′) (3), which produced an amplicon of expected size (413 bp). No amplification was observed from healthy plant extracts. Sequence analysis revealed that the four Tunisian isolates (TuV 258-201 collected from faba bean [GenBank Accession No. HQ199310], TuC 215-201 collected from chickpea [HQ199307], and TuP 163-201 [HQ199308] and TuP 166-201 collected from pea [HQ199309]) were most similar to each other with a high sequence identity (99%) and clustered with isolates of CpCSV from Syria (GenBank Accession No. EU541270), Egypt (EU541269), and Morocco (EU541267), to which they were most closely related (98%). The Tunisian isolates also showed high sequence identity (96%) in the coat protein region with Ethiopian (GenBank Accession No. EU541257) and Sudanese (EU541263) isolates. However, all isolates are distinct from BWYV, BLRV, and SbDV (less than 70% sequence identity). Since CpCSV is transmitted by aphids only, additional studies are needed to identify the host range of the virus and the efficient aphid vectors to better understand the epidemiology of this virus under Tunisian conditions References: (1) A. D. Abraham et al. Arch.Virol. 154:791; 2009. (2) N. Y. Asaad et al. J. Phytopathol. 157:756, 2009. (3) S. G. Kumari et al. Phytopathol. Mediterr. 47:42, 2008. (4) A. Najar et al. Phytopathol. Mediterr. 39:423, 2000.

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First Report of Aster Yellow Phytoplasmas (‘Candidatus Phytoplasma asteris’) in Canadian Grapevines

Plant Disease ◽

10.1094/pdis-93-6-0669a ◽

2009 ◽

Vol 93 (6) ◽

pp. 669-669 ◽

Cited By ~ 10

Author(s):

C. Y. Olivier ◽

D. T. Lowery ◽

L. W. Stobbs ◽

C. Vincent ◽

B. Galka ◽

...

Keyword(s):

Prince Edward Island ◽

Taqman Probe ◽

First Report ◽

Sequence Identity ◽

Flower Abortion ◽

Freeze Dried ◽

Wide Range ◽

Canadian Food Inspection Agency ◽

Pcr Technology ◽

Elm Yellows

In North America, elm yellows, aster yellows (AY), and X-disease phytoplasmas have been detected in American grapevines (1), and recently, Bois noir was detected in Canadian vineyards from British Columbia (BC) and Ontario (ON) (2). Typical symptoms of grapevine yellows (GY) include leaf rolling and chlorosis, uneven or total lack of lignification of canes, flower abortion or berry withering, and stunting. In 2006 and 2007, independent surveys were conducted by the Canadian Food Inspection Agency (CFIA) and Agriculture and Agri-Food Canada (AAFC) to detect phytoplasmas in Canadian vineyards containing different cultivars in BC, ON, Québec (QC), Nova Scotia, New Brunswick, and Prince Edward Island. The CFIA collected and tested 651 fresh leaf samples from recently imported grapevines and older grapevines in the same or neighboring blocks displaying symptoms typical of those associated with disease caused by phytoplasmas. Many vineyards were surveyed only once. AAFC collected and tested 3,485 samples from symptomatic and asymptomatic grapevines from established vineyards in ON, BC, and QC. The same vineyards were sampled in ON and BC both years; QC vineyards were only sampled in 2007. AAFC-collected leaf samples were freeze dried and stored at –20°C before processing. CFIA samples were tested by a modified real-time PCR assay and TaqMan probe targeting the 16S ribosomal RNA gene that detects a wide range of known phytoplasmas (2). Positive samples were confirmed by conventional PCR using the phytoplasma-specific primers P1/P7 (3) and the resulting ~1,800-bp fragment was cloned and sequenced as previously described (2). DNA extracted by AAFC was amplified by nested PCR technology with universal phytoplasma specific primer pairs P1/P6 and R16R2/R16F2 (3) and the resulting 1,200-bp fragment was cloned and sequenced. Two plants, one located in ON in 2006 and the other in BC in 2007, were found to be infected with an AY-like phytoplasma by the CFIA. The phytoplasmas detected in both infected plants had a 99.9% nt sequence identity with AY phytoplasma sequences from GenBank (Accession Nos. AF222063 and AY665676, respectively), with the BC isolate also showing 100% identity to a strain of AY, ash witches'-broom phytoplasma (GenBank Accession No. AY566302). AAFC detected phytoplasma DNA in both years in a total of 17 symptomatic plants and 21 asymptomatic plants from different vine varieties in ON, BC, and QC. Positive samples were found to have a 99.0% nt sequence identity to AY subgroup 16SrI-A (GenBank Accession No. AY180956). Sequences were exchanged for confirmation of phytoplasma identity and were deposited in Genbank under Accession Nos. FJ659844 and FJ824597. Phytoplasma strains were identified for all plants in which phytoplasmas were detected. Results show that AY is present in vineyards in the provinces of ON, BC, and QC. To our knowledge, this is the first report of AY being detected in grapevines in Canada. References: (1) E. Boudon-Padieu. Bull. O I V, 79:299, 2003. (2) M. Rott et al. Plant Dis. 91:1682, 2007. (3) E. Tanne et al. Phytopathology 91:741, 2001.

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First Report of the Occurrence of Wheat dwarf virus in Wheat in China

Plant Disease ◽

10.1094/pd-91-0111b ◽

2007 ◽

Vol 91 (1) ◽

pp. 111-111 ◽

Cited By ~ 24

Author(s):

J. Xie ◽

X. Wang ◽

Y. Liu ◽

Y. Peng ◽

G. Zhou

Keyword(s):

Triticum Aestivum L ◽

Coli Strain ◽

Gansu Province ◽

Dwarf Virus ◽

Shanxi Province ◽

Wheat Dwarf Virus ◽

Alkaline Lysis ◽

First Report ◽

Pcr Products ◽

Dideoxynucleotide Chain Termination Method

In May of 2004 and 2005, several diseased wheat (Triticum aestivum L.) plants showing extreme dwarfing, various types of yellowing, and reduced or no heading were found in the breeding fields of the Institute of Crop Science, Shanxi Academy of Agricultural Sciences, Taiyuan, Shanxi Province, China. On the basis of these symptoms, infection with Wheat dwarf virus (WDV) was suspected. Total DNA was extracted from diseased plants with the DNeasy Plant Mini kit (Qiagen, Hilden, Germany). Primers were designed based on the WDV-Enkoping1 genome sequence (NC_003326) (1), including: 245f, 5′-CGACTACGCCTGGCGAACATTTG-3′ (residues 245–267); 806r, 5′-TCTGGCATTGCCTGTTTCGG-3′ (complementary to residues 787–806); 1381f, 5′-CAGTGACATCTTCGCCGGAG-3′ (residues 1381–1400); and, 1886r, 5′-ACTCCGTAAGCCTCGAATCC-3′ (complementary to residues 1867–1886). With primer pairs 245f/806r, 1381f/1886r, 245f/1886r, and 1381f/806r, PCR products of 560, 506, 1642, and 2,275 bp were expected, respectively. After amplification, fragments of the expected sizes were seen on 1% (w/v) agarose gels. The fragments were purified by using a DNA gel extraction kit (TaKaRa, Dalian, China) and cloned into the pGEM-T vector (Promega, Madison, WI). The plasmids were transformed into E. coli strain DH5α and plasmid DNA was isolated from overnight cultures by alkaline lysis. Insert sequences were determined using the dideoxynucleotide chain termination method with an automated sequencer (ABI BigDye 3.1, Applied Biosystems, Foster City, CA). At least three independently isolated clones were analyzed for each PCR product. The compiled 2,750 nt sequence (GenBank Accession No. DQ868525) was 98.1, 98.5, 97.8, and 97.9% identical to WDV-Enkoping1 (NC_003326), WDV-SE (X02869), WDV-B (AM040732), and WDV-F (AM040733), respectively. Therefore, the virus isolate (WDV-TY) was identified as WDV (genus Mastrevirus, family Geminividae). Wheat samples collected from different provinces from 2004-2006 were also infected with WDV as indicated by PCR using the same primer pairs. For Shijiazhuang (Hebei Province), Yangling (Shanaxi Province), Kunming (Yunnan Province), Yuncheng (Shanxi Province), Tianshui (Gansu Province), Gangu (Gansu Province), and Zhenzhou (Henan Province), 13 of 14, 6 of 6, 5 of 5, 4 of 4, 2 of 3, 1 of 2, and 1 of 1 samples were positive, respectively, indicating a broad distribution of WDV in China. To our knowledge, this is the first report of WDV in wheat in China. Reference: (1) A. Kvarnheden et al. Arch Virol. 147:205, 2002.

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First Report of Tomato chlorosis virus in China

Plant Disease ◽

10.1094/pdis-12-12-1163-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1123-1123 ◽

Cited By ~ 17

Author(s):

R. N. Zhao ◽

R. Wang ◽

N. Wang ◽

Z. F. Fan ◽

T. Zhou ◽

...

Keyword(s):

Disease Incidence ◽

Mainland China ◽

Economic Losses ◽

Degenerate Primers ◽

First Report ◽

Sequence Identity ◽

Cp Gene ◽

Specific Fragment ◽

Tomato Chlorosis Virus ◽

Beijing China

In October 2012, a severe yellowing disease was found on greenhouse and plastic house tomato (Solanum lycopersicum) plants in Beijing, China. The disease incidence varied from 5 to 80% in each of six fields across Haidian and Daxing districts. The lower leaves showed symptoms of interveinal chlorosis, leaf brittleness, and limited brown necrotic flecks, similar to symptoms induced by Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) (two members of genus Crinivirus, family Closteroviridae) (4). A large number of whiteflies (Bemisia tabaci) were also observed. Leaf samples were taken from eight symptomatic and two asymptomatic tomato plants in two plastic houses in the Haidian district and total RNA was isolated from the 10 samples using TRIzol reagent (Tiangen, Beijing, China). Nested reverse transcription (RT)-PCR was performed to test the presence of ToCV and TICV with degenerate primers HS-11 and HS-12 and specific primers ToC-5/ToC-6 or TIC-3/TIC-4 for ToCV or TICV, respectively (1). With ToCV primers, a 463-bp specific fragment was amplified from eight symptomatic samples but not from two asymptomatic samples, and there was no amplification with TICV primers from any sample. Sequence analysis of the amplified fragment showed 99% nucleotide sequence identity with the heat shock protein 70 homolog (HSP70h) gene of ToCV isolates from Japan (GenBank Accession No. AB513442), Spain (DQ136146), Florida (AY903448), and Greece (EU284744). The presence of ToCV was confirmed by amplification of a 848-bp fragment covering the coat protein (CP) gene of ToCV with primers CP-F (5′-GAATCTTTTAGAAGCTTTGGTTTAAGG-3′) and CP-R (5′-GATCCTCTTGATCCTCATAGATTTC-3′) (3). The CP had 97 to 99% amino acid sequence identity to the above-mentioned four ToCV isolates. A sequence of the CP gene obtained from one isolate was deposited at GenBank (KC311375). Additionally, virions were isolated from 25 g of symptomatic samples followed Klaassen's method (2) and their lengths were estimated to be about 800 to 850 nm by transmission electronic microscopy To our knowledge, this is the first report of ToCV on tomato in mainland China. Tomato is one of the most widely cultivated crops in China and the spread of ToCV in China may cause significant economic losses. Further information on the prevalence and incidence of ToCV is required to assess the potential impact of this virus. References: (1) C. I. Dovas et al. Plant Dis. 86:1345, 2002. (2) V. A. Klaassen et al. J. Gen. Virol. 75:1525, 1994. (3) H. Tomoki et al. J. Gen. Plant Pathol. 76:168, 2010. (4) G. C. Wisler et al. Phytopathology 88:402, 1998.

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First Report of the Occurrence of Wheat dwarf virus Infecting Wheat in Slovenia

Plant Disease ◽

10.1094/pdis-11-16-1642-pdn ◽

2017 ◽

Vol 101 (7) ◽

pp. 1336 ◽

Cited By ~ 2

Author(s):

M. Viršček Marn ◽

I. Mavrič Pleško

Keyword(s):

Dwarf Virus ◽

Wheat Dwarf Virus ◽

First Report

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First Report of Dimethachlon Resistance in Field Isolates of Sclerotinia sclerotiorum on Oilseed Rape in Shaanxi Province of Northwestern China

Plant Disease ◽

10.1094/pdis-07-13-0730-pdn ◽

2014 ◽

Vol 98 (4) ◽

pp. 568-568 ◽

Cited By ~ 15

Author(s):

F. Zhou ◽

F. X. Zhu ◽

X. L. Zhang ◽

A. S. Zhang

Keyword(s):

Oilseed Rape ◽

Sclerotinia Sclerotiorum ◽

The United States ◽

Northwestern China ◽

Shaanxi Province ◽

Resistant Isolate ◽

Economic Losses ◽

First Report ◽

Field Isolates ◽

Wide Range

Sclerotinia sclerotiorum (Lib.) de Bary is a necrotrophic fungal pathogen causing diseases in a wide range of plants, including oilseed rape (3). Substantial economic losses caused by S. sclerotiorum have been reported in the United States, Canada, Brazil, South Africa, Hungary, India, Nepal, and Japan (1). Application of fungicides is the principal tool for controlling S. sclerotiorum because of lack of high level of host resistance. Dicarboximide fungicides such as dimethachlon have been widely used to control S. sclerotiorum in recent years in China and field isolates with reduced sensitivity to dimethachlon have been reported in Jiangsu Province of eastern China (2). In order to understand the current status of dimethachlon resistance in S. sclerotiorum isolates of northwestern China, 196 and 344 isolates of S. sclerotiorum collected from oilseed rape fields in 10 counties throughout Shaanxi Province in 2011 and 2012, respectively, were assayed for sensitivity to dimethachlon using 5 μg ml–1 dimethachlon as a discriminatory dose. Mycelial plugs (6 mm in diameter) cut from the margin of a 48-h-old colony were placed in the center of petri dishes containing potato dextrose agar (PDA) amended with 5 μg ml–1 dimethachlon; PDA without fungicide served as the control. Cultures were incubated at 26°C and colony growth was measured after 72 h of incubation. Isolates that showed growth on PDA amended with fungicide were tentatively considered resistant to dimethachlon, whereas the completely inhibited isolates were considered sensitive. Results showed that 1.02% or 2 isolates of the 196 isolates collected in 2011 and 3.78% or 13 isolates of the 344 isolates collected in 2012 were resistant to dimethachlon. For all the isolates considered resistant and 42 randomly selected sensitive isolates, 50% effective concentrations (EC50) were determined on PDA amended with a series of dimethachlon concentrations. The average EC50 value of dimethachlon for sensitive isolates was 0.29 ± 0.02 μg ml–1 Resistance ratios (EC50 of resistant isolate / average EC50 of sensitive isolates) for the two resistant isolates detected in 2011 were 10.28 and 23.83, respectively, whereas resistance ratios for the 13 resistant isolates detected in 2012 ranged from 24.90 to 101.97. The average EC50 value of dimethachlon for the 13 resistant isolates detected in 2012 was 19.05 μg ml–1, and EC50 values for the two resistant isolates detected in 2011 were 2.98 and 6.91 μg ml–1, respectively. These results indicated that both resistance frequency and resistance level increased from 2011 to 2012. Bioassay results of three resistant isolates indicated that there was positive cross-resistance between dimethachlon and other dicarboximide fungicides such as iprodione and procymidone. To our knowledge, this is the first report of dimethachlon resistance in S. sclerotiorum in Shaanxi Province of northwestern China. The molecular mechanism of dimethachlon resistance in field isolates of S. sclerotiorum remains to be studied. Although resistance frequency is low at present, dimethachlon resistance should be kept in mind and fungicide resistance management tactics such as use of biological control agents, fungicide tank-mixing, or alternating dimethachlon with other fungicides having different modes of action is recommended in controlling S. sclerotiorum. References: (1) M. D. Bolton et al. Mol Plant Pathol. 7:1, 2006. (2) H. X. Ma et al. Plant Dis. 93:36, 2009. (3) L. H. Prudy. Phytopathology 69:875, 1979.

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First Report of Zonate Leaf Spot of Glycine max Caused by Cristulariella moricola in Korea

Plant Disease ◽

10.1094/pdis-01-12-0028-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 906-906 ◽

Cited By ~ 1

Author(s):

S. E. Cho ◽

J. H. Park ◽

J. K. Choi ◽

H. D. Shin

Keyword(s):

Glycine Max ◽

Its Region ◽

Economic Losses ◽

Mountainous Area ◽

Potential Threat ◽

Voucher Specimen ◽

First Report ◽

Leaf Spots ◽

Initial Symptoms ◽

Wide Range

Soybean (Glycine max (L.) Merr.) is native to East Asia including Korea and is widely grown and consumed as an edible seed. In August 2011, following a prolonged period of cool and moist weather, zonate leaf spots were observed in local soybean (cultivar unknown) planted in a mountainous area of Goseong, central Korea. A voucher specimen was collected and entered at the Korea University herbarium (KUS-F26049). Initial symptoms included grayish green-to-grayish brown spots without border lines. As the lesions enlarged, they coalesced, leading to leaf blight and premature defoliation. Sporophores on the leaf lesions were dominantly hypophyllous, rarely epiphyllous, solitary, erect, easily detachable, and as long as 750 μm. The upper portion of the sporophores consisted of a pyramidal head that was ventricose, 275 to 500 μm long, and 80 to 160 μm wide. The fungus was isolated from leaf lesions and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 18°C without light, but conidia were not observed in culture. The morphological and cultural characteristics were consistent with those of Cristulariella moricola (Hino) Redhead (2,3). An isolate was preserved in the Korean Agricultural Culture Collection (KACC46401). Genomic DNA was extracted with the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 453 bp was deposited in GenBank (Accession No. JQ036182). A BLAST search in GenBank revealed that the sequence showed an exact match with that of C. moricola from Acer negundo (JQ036181) and >99% similarity with that of Grovesinia pyramidalis, teleomorph of C. moricola from Juglans sp. (Z81433). To determine the pathogenicity of the fungus, sporophores with the pyramidal head were carefully detached from a lesion on the naturally infected leaflet with fine needles. Each sporophore was transferred individually onto four places of six detached healthy soybean leaflets. The leaflets were placed in humid chambers at 100% relative humidity and incubated at 16 to 20°C (4). Symptoms were observed after 2 days on all inoculated leaflets (one to four lesions/leaflet). The lesions enlarged rapidly and reached ~20 mm diameter in a week. A number of sporulating structures and immature sclerotia were formed on the abaxial surface of the leaf 2 weeks after inoculation. The pathogen was reisolated from lesions on the inoculated leaflets, confirming Koch's postulates. No symptoms were observed on the control leaflets kept in humid chambers for 2 weeks. C. moricola was known to cause zonate leaf spots and defoliation on a wide range of woody and annual plants (1), but not on G. max. To our knowledge, this is the first report of Cristulariella infection in cultivated soybeans. Since the infections may be limited to the mountainous area with low night temperature and high humidity, economic losses seem to be negligible. However, the disease could be a potential threat to the safe production of soybeans in areas with prolonged periods of cool and moist weather. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.arsgrin.gov/fungaldatabases/ , January 7, 2012. (2) H. B. Lee and C. J. Kim. Plant Dis. 86:440, 2002. (3) S. A. Redhead. Can. J. Bot. 53:700, 1975. (4) H. J. Su and S. C. Leu. Plant Dis. 67:915, 1983.

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First Report of Soybean dwarf virus on Soybean in Wisconsin

Plant Disease ◽

10.1094/pdis.2004.88.11.1285a ◽

2004 ◽

Vol 88 (11) ◽

pp. 1285-1285 ◽

Cited By ~ 7

Author(s):

A. Phibbs ◽

A. Barta ◽

L. L. Domier

Keyword(s):

Leaf Tissue ◽

Dwarf Virus ◽

Economic Losses ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

First Report ◽

Elisa Testing ◽

Soybean Plants ◽

Soybean Dwarf Virus ◽

Das Elisa

Soybean dwarf virus (SbDV) causes widespread economic losses on soybean (Glycine max (L.) Merr.) in Japan (4), and has been reported on soybean in Virginia (2), in various legumes in the southeastern United States (1), and in peas in California (3). During late July and early August of 2003, soybean plants in Wisconsin were surveyed for SbDV. In 286 soybean fields at the R2-R4 growth stage, the uppermost fully unfurled leaf was collected from 10 plants at each of five sites. Samples were collected at random without regard to symptoms. SbDV symptom information was not recorded. Samples were stored on ice until frozen at -80°C. Five fields in four Wisconsin counties (Columbia, Lafayette, Sauk, and Waushara) tested positive for SbDV using double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). DAS-ELISA testing was conducted with reagents from Agdia, Inc (Elkhart, IN) following the manufacturer's protocol. Absorbance was read at 405 nm with a Stat Fax 2100 microplate reader (Awareness Technology, Inc., Palm City, FL) or visually evaluated. DAS-ELISA did not discriminate between strains of SbDV. The presence of SbDV was confirmed, and strain identity was inferred as dwarfing strain using reverse transcription-polymerase chain reaction (RT-PCR). Total RNA was extracted from homogenized leaf tissue, reverse transcribed, and amplified with the SuperScript One Step RT-PCR System (Invitrogen, Carlsbad, CA) and SbDV-specific primers (5′-CTGCTTCTGGTGATTACACTGCCG-3′ and 5′-CGCTTTCATTTAACGYCATCAAAGGG-3′). Size of the RT-PCR products (110 bp) was consistent with the dwarfing strain, SbDV-D. All locations that tested positive for SbDV showed soybean aphids, Aphis glycines Matsumura (Homoptera: Aphididae), on 100% of soybean plants. Several aphid species have been reported to vector SbDV, but at this time, vector relations in the Wisconsin infections are unknown. To our knowledge, this is the first report of SbDV infecting soybean in Wisconsin. References: (1) V. D. Damsteegt et al. Plant Dis. 79:48, 1995. (2) A. Fayad et al. Phytopathology (Abstr.) 90(Suppl.):S132, 2000. (3) G. R. Johnstone et al. Phytopathology (Abstr.) 74:795(A43), 1984. (4) T. Tamada et al. Ann. Phytopathol. Soc. Jpn. 35:282, 1969.

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