Hohhs. H. :I.. Kedd}. L). V. R.. Kajerhui~r~. K.. ;rnd Kedd!.. 11. S. 1087. l ' s c ~I ' d~r c c ! ;~~l~i g c n coating ilnd proteln A coaling EI.ISA procedures lor detection 01' tllree pcamtt \ i~u a e a. I'l;~nt Lliacasc 7 1 : 747-749. Direct antigcn coating (UAC') and prolein h coutlng (I' C') l o~r n~ oi ~nd~rcci c n~~r n e-l~~~h c d immunosorhent assay (T I IS.4) urrc starrdsrdi/ed .ind cornpi~rcd NIIII lltc douhlc-iit~tihod! handuich (1)AS)torm cti~~rect EL.ISA Sorthe~r usel~~lnea In ~hcdctect~c~n ol tllrer pcilnut ttruscr: peanu! mottle tirus (I'MVI. tonlato sported *ill tlrua (ISWV), s~l d lrldtatl pci~nttt C~U I I I~ tirub (IPCV). I'MV wiisdetectahlc in peanut secdr and pea t~\suc.nt a I. 10.000dilutton In huttc~ uith the DAC and a! a 1:1.000 dilut~on wilh the I'GC' proccdurcr. WIIII the I)AS procedure. I'M\' %;I* detectable in pea tissuc at a 1: 1.000 dilutior~ and in peanul seed at it I:lOO dilution 1 SH'V u;lr dctcctahle in peanut lea\cs;~t a I:I.000d1lution ~t t h the I)AC method and a[ I:100ui1l1 the l'A(' and D A S methods. II'C'V uas dctectahle In peanut leave\ at n 1:100tlilttt1on h! the I):\('. I'At'. ant1 [)AS methods. Senrititity ol' DAC' ;~nd I'AC' nlcthad\ &; I S. therefore, r't>~llp,~ri~hlc to !II;II ti1 rhc 1)ASprocedurc under ~hcshorr incuhi~t~on pcrlt)d (I 2 hrat .lS(')vondilto~~\cniployed III lhc experimenlb.
A recombinant strain of Sclerotinia sclerotiorum hypovirus 2 (SsHV2) was identified from a North American Sclerotinia sclerotiorum isolate (328) from lettuce (Lactuca sativa L.) by high-throughput sequencing of total RNA. The 5=-and 3=-terminal regions of the genome were determined by rapid amplification of cDNA ends. The assembled nucleotide sequence was up to 92% identical to two recently reported SsHV2 strains but contained a deletion near its 5= terminus of more than 1.2 kb relative to the other SsHV2 strains and an insertion of 524 nucleotides (nt) that was distantly related to Valsa ceratosperma hypovirus 1. This suggests that the new isolate is a heterologous recombinant of SsHV2 with a yet-uncharacterized hypovirus. We named the new strain Sclerotinia sclerotiorum hypovirus 2 Lactuca (SsHV2L) and deposited the sequence in GenBank with accession number KF898354. Sclerotinia sclerotiorum isolate 328 was coinfected with a strain of Sclerotinia sclerotiorum endornavirus 1 and was debilitated compared to cultures of the same isolate that had been cured of virus infection by cycloheximide treatment and hyphal tipping. To determine whether SsHV2L alone could induce hypovirulence in S. sclerotiorum, a full-length cDNA of the 14,538-nt viral genome was cloned. Transcripts corresponding to the viral RNA were synthesized in vitro and transfected into a virus-free isolate of S. sclerotiorum, DK3. Isolate DK3 transfected with SsHV2L was hypovirulent on soybean and lettuce and exhibited delayed maturation of sclerotia relative to virus-free DK3, completing Koch's postulates for the association of hypovirulence with SsHV2L. IMPORTANCEA cosmopolitan fungus, Sclerotinia sclerotiorum infects more than 400 plant species and causes a plant disease known as white mold that produces significant yield losses in major crops annually. Mycoviruses have been used successfully to reduce losses caused by fungal plant pathogens, but definitive relationships between hypovirus infections and hypovirulence in S. sclerotiorum were lacking. By establishing a cause-and-effect relationship between Sclerotinia sclerotiorum hypovirus Lactuca (SsHV2L) infection and the reduction in host virulence, we showed direct evidence that hypoviruses have the potential to reduce the severity of white mold disease. In addition to intraspecific recombination, this study showed that recent interspecific recombination is an important factor shaping viral genomes. The construction of an infectious clone of SsHV2L allows future exploration of the interactions between SsHV2L and S. sclerotiorum, a widespread fungal pathogen of plants. Sclerotinia sclerotiorum (Lib.) de Bary is a cosmopolitan fungal plant pathogen that causes necrotic diseases (e.g., Sclerotinia stem rot) in more than 400 plant species, which result in yield losses in major crops each year (1-3). However, the diseases caused by S. sclerotiorum have not been adequately controlled by conventional technologies thus far (4, 5). A number of mycoviruses have been molecularly characterized and ...
Virulence and double-stranded RNA (dsRNA) profiles of 44 isolates of Fusarium virguliforme were compared. When grouped according to dsRNA profiles, isolates with large dsRNAs were significantly (P≤0.05) less virulent than isolates without dsRNAs. High-throughput sequence analysis of total RNA prepared from cultures with large dsRNAs identified two novel RNA viruses with genome sequences of approximately 9.3 kbp, which were named Fusarium virguliforme dsRNA mycovirus 1 and Fusarium virguliforme dsRNA mycovirus 2. The new viruses were most closely related to a group of unclassified viruses that included viruses of F. graminearum and Phlebiopsis gigantea and are related to members of the family Totiviridae.
Fifty‐two North American (NA) ancestral soybean [Glycine max (L.) Merr.] lines were screened for resistance to Bean pod mottle virus (BPMV), Soybean mosaic virus (SMV) strains G1 and G5, Tobacco ringspot virus (TRSV), and Tobacco streak virus (TSV). Seven ancestors, ‘CNS’, ‘Haberlandt’, ‘Ogden’, ‘Peking’, PI 71506, PI 88788, and ‘Tokyo’, were resistant to SMV‐G1. Sixteen entries, ‘A.K. (Harrow)’, ‘Capital’, CNS, FC 33243, Haberlandt, ‘Illini’, ‘Improved Pelican’, ‘Laredo’, ‘Lincoln’, ‘Mandarin’, ‘Mandarin (Ottawa)’, Ogden, ‘Palmetto’, Peking, PI 88788, and Tokyo were resistant to SMV‐G5. All ancestral lines tested were susceptible to BPMV and TRSV. Only one ancestor, ‘Tanner’, was resistant to TSV. On the basis of cultivar registration articles through 2002, there were 15 public soybean cultivars with reported resistance to SMV. The possible donors of resistance for each were identified. Two soybean ancestors, CNS and Ogden, were the most important possible sources of SMV resistance genes in U.S. commercial soybean cultivars, as the pedigree of 75 and 56% of the reported resistant cultivars contained CNS and Ogden, respectively. In most of the cultivar registration articles, reactions to SMV were not reported. With the relatively high frequency of SMV resistance in major ancestral lines, SMV resistance in U.S. cultivars may be more common than expected.
Soybean seed coat mottling often has been a problematic symptom for soybean growers and the soybean industry. The percentages of seed in eight soybean lines with seed coat mottling were evaluated at harvest after inoculating plants during the growing season with Bean pod mottle virus (BPMV), Soybean mosaic virus (SMV), and both viruses inside an insect-proof cage in the field. Results from experiments conducted over 2 years indicated that plants infected with BPMV and SMV, alone or in combination, produced seed coat mottling, whereas noninoculated plants produced little or no mottled seed. BPMV and SMV inoculated on the same plants did not always result in higher percentages of mottled seed compared with BPMV or SMV alone. There was significant virus, line, and virus-line interaction for seed coat mottling. The non-seed-coat-mottling gene (Im) in Williams isoline L77-5632 provided limited, if any, protection against mottling caused by SMV and none against BPMV. The Peanut mottle virus resistance gene Rpv1 in Williams isoline L85-2308 did not give any protection against mottling caused by SMV, whereas the SMV resistance gene Rsv1 in Williams isoline L78-379 and the resistance gene or genes in the small-seeded line L97-946 gave high levels of protection against mottling caused by SMV. The correlations (r = 0.77 for year 2000 and r = 0.89 for year 2001) between virus infection of the parent plant and seed coat mottling were significant (P = 0.01), indicating that virus infection of plants caused seed coat mottling.
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