我国东北部五省市SMV对大豆主栽品种的毒力测定

 自东北三省、山东及北京中国科学院遗传所农场(培育黄淮海豆区大豆品种)采集SMV毒株231个,用各地当前大豆主栽品种56个测定毒株的毒力。结果将SMV分为3个株系群,7个亚群,13个株系,即S1,S2-1M,S2-1N,S2-2M,S2-2N,S2-3M,S2-3N,S3-1M,S3-1N,S3-2M,S3-2N,S3-3M,S3-3N。除S1外,各亚群包括引起系统花叶(M)及系统坏死(N)2类株系。大豆品种按抗性归成8类。病毒群体毒力自北向南逐渐增强,黑龙江省的毒株群体毒力最弱,北京的最强,品种抗性也自北向南逐渐增高,黑龙江省大多数为高感品种,吉林多为中感品种,辽宁为中抗品种,北京全属高抗品种,品种抗性水平与病毒群体毒力相互适应。     

Characterization of two distinct Polish isolates of Pepino mosaic virus

In Poland in 2002 and 2005 two different isolates of Pepino mosaic virus signed PepMV-SW and PepMV-PK were obtained. Both isolates were compared on the basis of their symptomatology on a series of plant species. In addition, the isolates were characterized by the nucleotide sequence analysis of the triple gene block, coat protein and a part of the polymerase genes. The studies showed that the Polish isolates differ from each other and belong to two strains. PepMV-SW was highly similar to European isolates, showing extensive sequence identity, ca. 99%. Pairwise comparisons of PepMV-PK with other PepMV isolates from the GenBank database showed that the highest nucleotide sequence identity was with two isolates: Ch2 from Chile and US2 from the USA.     

Quantification of within-field spread of soybean mosaic virus in soybean using strain-specific monoclonal antibodies

ABSTRACT Strain-specific monoclonal antibodies were used to follow the temporal increase and spatial spread of soybean mosaic virus (SMV) strain G-5 released from a point source. The use of strain-specific monoclonal antibodies allowed discrimination of within-field temporal and spatial spread of SMV strain G-5 from non-G-5 SMV isolates that originated from exogenous field sources. SMV isolates originating from exogenous sources have potential to alter the temporal and spatial pattern of within-field virus spread, which could potentially affect the choice of models used to quantify within-field pathogen spread. Analysis of SMV epidemics in field-plot experiments indicated that the logistic model was the most appropriate model to describe and compare the temporal spread of SMV among years. On the basis of ordinary runs analyses, within-field spread of SMV strain G-5 was random in 1991 and 1994, but was mostly aggregated in 1992 and 1993. Non-G-5 SMV isolates arising from exogenous sources displayed a random spatial pattern over time. This is the first study in which pathogen incidence (detection of SMV using strain-specific monoclonal antibodies) as opposed to disease incidence (based on symptoms) was employed to monitor and model SMV spread in time and space.     

Differential necrotic lesion formation in soybean cultivars in response to soybean mosaic virus

In this study, we examined the necrosis phenotype on leaves of two cultivars of soybean (ZheA8901 and Nannong1138-2) that show varying level of resistance to soybean mosaic virus (SMV). The necrotic symptoms seen on inoculated and systemic leaves of soybean cultivar ZheA8901 were reminiscent of programmed cell death (PCD). The cell death phenotypes were evaluated using the TUNEL method, quantification of hydrogen peroxide (H₂O₂) and salicylic acid, callose production, as well as by monitoring expression of defence genes GmPR-1 and GmNPR1. Our results show that SMV inoculation induced PCD on ZheA8901 is associated with rapid increase in H₂O₂, increased SA and callose accumulation and higher defence gene expression.     

Infectivity analysis of a soybean isolate of Mungbean yellow mosaic India virus by agroinoculation

Yellow mosaic disease (YMD) of legumes endemic to South Asia are caused by begomoviruses transmitted by whiteflies. Based on molecular characterization, two distinct viruses – Mungbean yellow mosaic India virus (MYMIV) and Mungbean yellow mosaic virus (MYMV) – were found previously to be the etiological agents of YMD in legumes. Here, host range studies with a soybean isolate of MYMIV (MYMIV-[Sb]) were carried out by both whitefly transmission and agroinoculation. MYMIV-[Sb] was similar to a cowpea isolate of MYMIV (MYMIV-[Cp]) in its ability to infect cowpea, thus differing from blackgram (MYMIV) and mungbean (MYMIV-[Mg]) isolates, which do not infect cowpea. Genomic analysis of DNA A and DNA B components of these MYMIV isolates show characteristic differences in complete DNA B nucleotide sequence correlating with host range differences.     

Virus transmission by host-specific strains ofOlpidium bornovanus andOlpidium brassicae

Zoospores of 12 isolatesO. bornovanus from geographically diverse sites and representing the three host specific cucurbit strains were tested as vectors for seven viruses using watermelon bait plants and the in vitro acquisition method. All isolates of the cucumber, melon, and squash strains transmitted melon necrotic spot carmovirus (MNSV) and cucumber necrosis tombusvirus (CNV) but none transmitted petunia asteroid mosaic tombusvirus (PAMV) or tobacco necrosis necrovirus (TNV). The isolates varied as vectors of three other carmoviruses: cucumber leaf spot virus (CLSV); cucumber soil borne virus (CSBV); and squash necrosis virus (SqNV). All cucumber isolates transmitted CLSV and SqNV but not CSBV. Some of the melon isolates transmitted CLSV and SqNV but none transmitted CSBV. Two squash isolates transmitted CSBV and SqNV but not CLSV. Two isolates ofO. brassicae transmitted only TNV and a third did not transmit any of the viruses. The species of bait plant sometimes affected transmission. The most efficient vector strains ofO. bornovanus, as determined by reducing zoospores and virus in the inoculum, were the cucumber strain for CLSV; the cucumber strain for CNV if cucumber was the bait plant or melon strain if watermelon was the bait plant; and the squash strain for SqNV. The plurivorous strain ofO. brassicae was the most efficient vector of TNV.Olpidium bornovanus is the first vector reported for CSBV and is confirmed as a vector of SqNV. It is proposed that all carmoviruses may have fungal vectors.Ligniera sp. did not transmit any of the viruses in one attempt.Abbreviations CLSV cucumber leaf spot virus - CNV cucumber necrosis virus - CSBV cucumber soil borne virus - MNSV melon necrotic spot virus - PAMV petunia asteroid mosaic virus - SqNV squash necrosis virus - TNV tobacco necrosis virus - TBSV tomato bushy stunt virus     

马铃薯Y病毒属三种病毒通用型单克隆抗体的鉴定

构建了马铃薯Y病毒属的大豆花叶病毒(Soybean mosaic virus,SMV)、马铃薯Y病毒(Potato virus Y,PVY)、三叶草黄脉病毒(Clover yellow vein virus,CLYVV)的原核表达载体pET28-SMV CP、pET28-PVY CP和pET28-CLYVV CP,经IPTG诱导、表达和纯化,获得SMV、PVY和CLYVV的CP蛋白。采用ELISA方法,用9株实验室制备的单克隆抗体对含SMV、PVY、CLYVV的病毒汁液和纯化的重组CP蛋白进行检测分析。结果表明,9株单克隆抗体均识别SMV病毒,2D3、4F9、4G12和6E7单克隆抗体能够识别PVY病毒,4F9和4G12能够识别CLYVV病毒,但PVY、CLYVV病毒与抗体的亲和力低于SMV病毒;对于重组表达的衣壳蛋白:SMV、CLYVV与抗体2D3、4F9、4G12和6E7反应强烈,PVY与4F9和4G12抗体反应稍强。综上,本研究鉴定出能识别SMV、PVY、CLYVV的通用单克隆抗体4F9和4G12。     

Occurrence of the A2 mating type and oospores ofPhytophthora Infestans in potato crops in Israel

Seventeen metalaxyl-sensitive and 21 metalaxyl-resistant isolates ofPhytophthora infestans collected from blighted potato fields during the years 1983–1988 were tested for mating type on rye seed agar medium. All isolates except one (MS3, collected in 1986 at Sufa, in the western Negev) were found to belong to the A2 mating type. A2 isolates produced oospores within 2 weeks when cultured together with isolate 163 (A₁ from the U.S.A.) or with the A₁ isolate MS3 from Israel. When cultured singly, A₂ isolates produced some oospores within 4–8 weeks. Blighted potato tubers harvested from potato crops artificially inoculated with a mixture of A1+A2 sporangia were found to contain some oospores. No oospores were detected in blighted tubers harvested from A₂ + A₂ inoculated crops. It was concluded that the A2 mating type ofP. infestans has occurred in Israel since 1983 or even earlier. The rare occurrence of the A1 mating type was unexpected and indicated that sexual reproduction of the fungus in the country might be limited.     

Effect of Phenylpyrrole-resistance Mutations on Ecological Fitness of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and their Genetical Basis in <Emphasis Type="Italic">Ustilago maydis</Emphasis>

Mutants of Botrytis cinerea and Ustilago maydis highly resistant to fludioxonil were isolated at a high frequency, after nitrosoguanidine or UV mutagenesis, respectively, and selection on media containing fludioxonil. Tests on the response of mutant strains to high osmotic pressure resulted in the identification of two fludioxonil-resistant phenotypes (FLD_osm/s and FLD_osm/r), regarding the sensitivity to high osmolarity. Approximately 95% of fludioxonil-resistant mutants were found to be more sensitive to high osmotic pressure than the wild-type parent strains. Genetic analysis of phenylpyrrole-resistance in the phytopathogenic basidiomycete U. maydis, showed that fludioxonil-resistance was coded by three unlinked chromosomal loci (U/fld-1, U/fld-2 and U/fld-3), from which only the U/fld-1 mutation coded an osmotic sensitivity similar to that of the wild-types. Cross-resistance studies with fungicides from other chemical groups showed that the mutations for resistance to phenylpyrroles affect the sensitivity of mutant strains to the aromatic hydrocarbon and dicarboximide fungicides, but not to the benzimidazoles, anilinopyrimidines, phenylpyridinamines, hydroxyanilides or the sterol biosynthesis inhibiting fungicides. A study of fitness parameters in the wild-type and fludioxonil-resistant mutants of B. cinerea, showed that all osmotic sensitive (B/FLD_osm/s) isolates had significant reductions in the characteristics determining saprophytic fitness such as mycelial growth, sporulation, conidial germination and sclerotial production. Contrary to that, with the exception of mycelial growth, the fitness parameters were unaffected or only slightly affected in most of the osmotic resistant (B/FLD_osm/r) isolates. Tests on cucumber seedlings showed that the osmotic-sensitive strains were significantly less pathogenic compared with the wild-type and B/FLD_osm/r strains of B. cinerea. Preventative applications of the commercial products Saphire 50 WP (fludioxonil) and Rovral 50 WP (iprodione) were effective against lesion development on cotyledons by the wild-type and the mutant strains of B. cinerea that were resistant to the anilinopyrimidine cyprodinil (B/CPL-27) and to the hydroxyanilide fenhexamid (B/FNH-21), but ineffective, even at high concentrations, against disease caused by the fludioxonil-resistant isolates (B/FLD) and a mutant strain resistant to the dicarboximide iprodione (B/IPR-1). Experiments on the stability of the fludioxonil-resistant phenotype showed a reduction of resistance, mainly in osmotic-sensitive isolates, when the mutants were grown on inhibitor-free medium. A rapid recovery of the high resistance was observed after mutants were returned to the selection medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain of B. cinerea, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of osmotic-resistant mutants was higher than the osmotic-sensitive ones. Furthermore, competition tests, in planta, showed a significant reduction of the frequency of both phenylpyrrole-resistant phenotypes, with a respective increase in the population of the wild-type strain of the pathogen.     

Characterization and detection ofCucumber green mottle mosaic virus in Greece

The host range specificity of Greek isolates ofCucumber green mottle mosaic virus (CGMMV) was determined and the ability of the virus to retain its infectivity in naturally contaminated soil, after storage at 4°C for at least 10 months, was established. An immunocapture RT-PCR protocol was developed for the detection of the virus and proved to be 10⁵ times more sensitive than DAS-ELISA and 10² times more sensitive than F(ab’)₂-ELISA using F(ab’)₂ fragments that were prepared from an antiserum, which was raised against a local virus isolate. In order to clarify dissimilarities that were revelaed between the coat protein (CP) genes of the Greek isolates after restriction mapping of their respective PCR products, the CP genes of three isolates were sequenced and found to be very similar but not identical to the CP gene of CGMMV-SH. http://www.phytoparasitica.org posting Aug. 27, 2002.     

Molecular analysis and virus transmission tests place Olpidium virulentus, a vector of Mirafiori lettuce big-vein virus and tobacco stunt virus, as a distinct species rather than a strain of Olpidium brassicae

The rDNA-ITS sequences of ten single-sporangium isolates of Olpidium virulentus (a noncrucifer strain of Olpidium brassicae), which transmits Mirafiori lettuce big-vein virus (MLBVV) and tobacco stunt virus (TStV), were compared with those of six single-sporangium isolates of O. brassicae. The sequence similarity within isolates of O. virulentus or O. brassicae was almost identical (98.5%–100.0%), but was low between the two species (79.7%–81.8%). In a phylogenetic analysis of the rDNA-ITS region, O. virulentus and O. brassicae fell into two distinct clusters, indicating that O. virulentus, a vector of MLBVV and TStV, is a distinct species rather than a strain of O. brassicae.     

Differentiation of Bean pod mottle virus isolates based upon host symptoms

Accessions from Glycine, Phaseolus, and Vigna genera were screened for their reactions to different subgroups of isolates of Bean pod mottle virus (BPMV) in order to establish a differential host system. Screening results indicated that the BPMV isolates differed in pathogenic aggressiveness but not in virulence. No major resistance genes were found in soybean (Glycine max) or G. soja since all screened accessions showed mosaic or necrotic symptoms to BPMV inoculation. However, these accessions expressed differences in severity of symptoms when challenged by various BPMV isolates. The inoculation of G. tomentella accessions did not result in mosaic symptoms, and some accessions did not support systemic infection of some of the isolates. Resistance, presented as a hypersensitive reaction, was observed in some of Phaseolus and Vigna genotypes, and resistant response or susceptibility was stable to all the isolates used in the screening. In conclusion, the selected G. soja genotypes PI 407019, PI 464889A, and PI 464928, and ‘Amsoy 71’ soybean may help to separate severe (reassortant) from mild isolates of BPMV based upon their phenotypic reactions.     

Incidence of viruses in <Emphasis Type="Italic">Alstroemeria</Emphasis> plants cultivated in Japan and characterization of <Emphasis Type="Italic">Broad bean wilt virus-2, Cucumber mosaic virus</Emphasis> and <Emphasis Type="Italic">Youcai mosaic virus</Emphasis>

Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings.     

接种大豆花叶病毒条件下大豆上位叶离体培养物的生长与形态发生

 本研究从接种大豆花叶病毒(SMV)江苏株系Sa、Sg的大豆植株上取上位叶为外植体,研究不同株系对愈伤组织产生及芽与根分化的影响。结果发现,愈伤组织的诱导率及其形态特点在接种与未接种间无明显差异;接种Sa的其芽分化率低于未接种的,但接种Sg的其芽分化率反略高于未接种Sg的;接种Sa、Sg后,根的发生能力减弱,但对抗病品种的影响小于感病品种。     

Oospores associated with tomato seed may lead to seedborne transmission of<Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Tomato fruits at the green mature stage were inoculated with a mixed sporangial suspension of A₁ and A₂ isolates ofPhytophthora infestans. Other fruits were inoculated with either A₁ or A₂ sporangia. Seeds were extracted from the blighted fruits and sown in soil or on agar media to test for the transmission of late blight to the emerging seedlings. Only 23 (0.09%) of approximately 25,000 seedlings developed symptoms. All blighted seedlings originated from fruits inoculated with mixed A₁ + A₂ sporangia. Isolates ofP. infestans recovered from the emerging blighted seedlings were seemingly of oosporic origin, as they differed phenotypically (mating type, virulence, sensitivity to metalaxyl) from the parent isolates used to inoculate the fruits. The results suggest that transmission ofP. infestans might occur by seeds extracted from fruits carrying oospores and less probably by seeds extracted from fruits having no oospores. http://www.phytoparasitica.org posting April 30, 2004.     

Characterization of Grapevine Algerian latent virus isolated from nipplefruit (Solanum mammosum) in Japan

Alfalfa mosaic virus (AMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), Tomato bushy stunt virus nipplefruit strain (TBSV-Nf), and an unknown spherical virus were isolated from nipplefruit (Solanum mammosum) cultivated in Chiba Prefecture, Japan. The spherical virus was identified as Grapevine Algerian latent virus nipplefruit strain (GALV-Nf) from the genus Tombusvirus, based on its physical properties, serological relationships, and analysis of genomic RNA. The genomic RNA of GALV-Nf is 4731 nucleotides long and encodes five open reading frames as well as those of other tombusviruses. Nipplefruit infected with GALV-Nf had severe stunting, leaf deformation, and clear mosaic symptoms. This is the first report of an isolation of GALV in Japan. An erratum to this article is available at .     

我国部分地区常见农作物上黄瓜花叶病毒分离物核酸多样性分析

为明确我国黄瓜花叶病毒株系分化及系统进化基本情况,从湖南、新疆、青海和海南4省区采集1 367个样品对其进行酶联免疫和RT-PCR检测,并对分离获得的15个黄瓜花叶病毒(Cucumber mosaic virus,CMV)纯化分离物CP、MP、2b核苷酸序列进行相似性和进化树分析及生物学性状比较。结果表明,辣椒、龙葵和黄瓜的CMV阳性检出率较高,分别为54.13%、29.19%和18.46%。进化树分析显示CMV-Q5与CMV亚组II的亲缘性较高;CMV-N7为新发现的重组株系,其CP、2b基因属于CMV亚组IB,MP基因却属于CMV亚组II;其余13个分离物均属于CMV亚组IB。CMV-N7和CMV-Q5在系统寄主心叶烟和枯斑寄主苋色藜上引发的症状相似,但比对照株系CMV-P3613(IB)的发病时间要晚1~2 d,系统花叶较温和,枯斑较小。表明在以上4省区常见农作物上广泛流行的CMV存在分子变异。     

Similarity of clover yellow vein virus and pea necrosis virus

There still is confusion concerning the relationships between clover yellow vein virus (ClYVV), pea necrosis virus (PNV) and bean yellow mosaic virus (BYMV). Therefore, three Swedish isolates of ClYVV and its type strain have now been compared with three isolates of PNV. A bean mosaic isolate and three pea necrosis isolates of BYMV have been used for reference. Based on host range tests, serology, and light microscope studies of inclusion bodies, ClYVV and PNV isolates are now considered to be strains of one virus, with the first name having priority. ClYVV (including the original PNV) especially differs from BYMV in its ability to infect white clover, to produce local lesions on cucumber cotyledons (at least two cultivars), to go systemic inChenopodium quinoa (the two local selections used at Wageningen and at Uppsala), to be rather virulent onNicotiana clevelandii, and to provoke extensive nucleolar enlargements in its host cells. Serologically the two viruses are more or less distinct.