Artichoke yellow ringspot virus infecting vetch (Vicia sativa) in Greece

A seed-transmitted virus was isolated from vetch seedlings showing severe stunting, reduced lamina and leaf mottling. Identification was based on host range, seed transmission, coat protein and nucleic acid analysis, electron microscopy and serology. This is the first record of vetch as a natural host of artichoke yellow ringspot virus (AYRSV). http://www.phytoparasitica.org posting Jan. 13, 2002.     

Evidence for Lettuce big‐vein associated virus as the causal agent of a syndrome of necrotic rings and spots in lettuce

Lettuce big‐vein associated virus (LBVaV, genus Varicosavirus) was shown to be responsible for characteristic necrotic symptoms observed in combination with big‐vein symptoms in lettuce breeding lines when tested for their susceptibility to lettuce big‐vein disease (BVD) using viruliferous Olpidium virulentus spores in a nutrient film technique (NFT) system. Lettuce plants showing BVD are generally infected by two viruses: Mirafiori lettuce big‐vein virus (MiLBVV, genus Ophiovirus) and LBVaV. New mechanical inoculation methods were developed to separate the two viruses from each other and to transfer both viruses to indicator plants and lettuce. After mechanical inoculation onto lettuce plants MiLBVV induced vein‐band chlorosis, which is the characteristic symptom of BVD. LBVaV caused a syndrome of necrotic spots and rings which was also observed earlier in lettuce plants inoculated in the NFT system, resembling symptoms described for lettuce ring necrosis disease (RND). This observation is in contrast with the idea that LBVaV only causes latent infections in lettuce. De novo next‐generation sequencing demonstrated that LBVaV was the only pathogen present in a mechanically inoculated lettuce plant with symptoms, providing evidence that LBVaV was the causal agent of the observed necrotic syndrome and thus fulfilling Koch’s postulates for this virus. The necrotic syndrome caused by LBVaV in lettuce is referred to as LBVaV‐associated necrosis (LAN).     

Host range comparison of the causal agents of pepper yellow vein and lettuce big vein

A number of solanaceous and composite plant species were tested as hosts for the causal agents of pepper yellow vein (PYVA) and lettuce big vein (LBVA), transmitted by a pepper and a lettuce isolate ofOlpidium brassicae, respectively. The agents had the following artificial hosts in common:Lycopersicon esculentum, Solanum melongena, Physalis floridana, Nicandra physaloides, Lactuca sativa, Sonchus oleraceus andL. virosa. Capsicum annuum, S. villosum, S. nigrum, Crepis vesicaria andSenecio vulgaris were infected by PYVA, but not by LBVA.Cichorium endivia andTaraxacum officinalis were not infected by any of the two agents.N. physaloides, although not colonized by the pepper isolate ofOlpidium, still became infected by PYVA.     

Characteristics of a resistance-breaking isolate of potato virus Y causing potato tuber necrotic ringspot disease

An Austrian isolate of potato virus Y^NTN, the causal agent of potato tuber necrotic ringspot disease (PTNRD), was serologically compared with seven Dutch PVY^N isolates. Using polyclonal and monoclonal antibodies, it was found indistinguishable from PVY^N. Determination of the nucleotide sequence of the coat protein cistron and comparison of the deduced amino acid sequence with coat protein sequences of other potyviruses revealed a high level of homology with PVY^N coat protein sequences. This confirmed the close taxonomic relationship of PVY^NTN with the PVY^N subgroup of potato virus Y. PVY^NTN is able to overcome all resistance genes known so far in commercial potato cultivars. Remarkably, transgenic PVY-protected tobacco plants are also resistant to PVY^NTN infection upon mechanical and aphid-mediated inoculation. These experiments indicate that genetically engineered resistance offers great potential in protection of potato to new aggressive strains of PVY^N.     

A new Fusarium lineage within the Gibberella fujikuroi species complex is the main causal agent of mango malformation disease in Brazil

Mango malformation is a serious disease in tropical and subtropical areas of the world and has been attributed to various Fusarium spp., including F. mangiferae , F. proliferatum , F. sacchari , F. sterilihyphosum and F. subglutinans . Isolates of Fusarium associated with mango malformation from Brazil, Egypt, India, South Africa and the United States were evaluated through amplified fragment length polymorphisms (AFLPs) and partial DNA sequences of the genes encoding β-tubulin ( tub2 ) and translation elongation factor 1-α ( tef1 ). These techniques were used to delimit species and to estimate the genetic and phylogenetic relatedness of the isolates. In the AFLP analysis, most of the Brazilian isolates formed a unique cluster. Additionally, one small cluster was formed by isolates of F. sterilihyphosum from Brazil and South Africa, and another by isolates of F. mangiferae from Egypt, India, South Africa and the United States. In the phylogenetic analysis, most of the Brazilian isolates represented a new phylogenetic lineage in the Gibberella fujikuroi species complex, where they formed a sister clade to F. sterilihyphosum. Representatives of both clades were pathogenic to mango (cv. Tommy Atkins) and Koch's postulates were completed for isolates belonging to the new lineage and to F. sterilihyphosum . Thus, most of the mango malformation disease in Brazil is due to a distinct phylogenetic lineage of Fusarium , and to a lesser extent by F. sterilihyphosum. The new phylogenetic lineage identified in this study, together with F. mangiferae and F. sterilihyphosum , are the only known taxa of Fusarium proven to be capable of causing mango malformation.     

Molecular,biological and serological variability of Zucchini yellow mosaic virus in Tunisia

A study was conducted to better understand the population structure of Zucchini yellow mosaic virus (ZYMV), a severe virus affecting cucurbit crops worldwide, in Tunisia and to estimate whether the use of resistant cultivars may provide durable control. Analysis of the polymerase and coat protein (NIb‐CP) partial sequences of 83 isolates collected in the three main cucurbit‐growing areas in Tunisia showed that ZYMV grouped into two distinct clusters within ZYMV molecular group A. An important variability was observed in the MREK motif of the P3 protein, a motif associated with tolerance breaking in ZYMV‐tolerant zucchini squash cultivars. Interestingly, significant differences were found in the distribution of the MREK variants in the two clusters defined by the partial NIb‐CP sequences, MREK and MKEK sequences being more common in cluster 1 and cluster 2, respectively. When combining NIb‐CP and P3 sequence information, ZYMV molecular variability was shown to be significantly higher in the Cap Bon region than in the Bizerte area. An important biological variability was observed in a subset of 23 isolates regarding symptomatology in susceptible or resistant cucurbits. Some isolates overcame ZYMV tolerance or resistance in zucchini squash and melon, but not in cucumber. Three serotypes were differentiated using a set of 13 monoclonal antibodies (MAbs). Seven parameters characterizing the 23 isolates, including molecular, serological and biological properties, were used for a multiple component analysis (MCA). This analysis revealed that symptom intensity of a given isolate was similar in different susceptible cucurbit hosts, suggesting similar degrees of aggressiveness in different hosts.     

Parsley latent virus,a new and prevalent seed-transmitted,but possibly harmless virus of Petroselinum crispum

Parsley latent virus, a hitherto undescribed virus, was isolated from 38 out of 54 samples of seed of parsley (Petroselinum crispum) of 17 out of 24 cultivars and from all five European countries tested, but not from some samples from the USA. It could easily be detected in seedlings and also in seeds germinated on moist filter paper, but not in dry seeds or in seeds soaked in water. Strawberry latent ringspot virus was detected in five samples. The parsley virus is symptomless in parsley and caused latent systemic infection inGomphrena globosa, three cultivars ofSpinacia oleracea and weak and often transient systemic symptoms inChenopodium amaranticolor, C. giganteum, C. glaucum andC. quinoa, but did not infect any other species out of all 32 species of seven plant families tested in total.The virus could easily be transmitted mechanically but not by seven aphid species in the non-persistent manner. Dilution end-point was between 100 and 1000, thermal inactivation between 55 and 60°C and ageing in vitro between 7 and 10 days.Purification yielded a single infectious component. The particles were spherical, ca. 27 nm in diameter, with a sedimentation coefficient of 127.5 S, a buoyant density of 1.449 g/ml, an RNA content of 36% and one type of protein with a relative molecular mass of 22×10³. Purificition without Triton and urea resulted in preparations with aggregates each consisting of 12 particles in icosahedral array.The virus differs from all viruses described so far and did not show clear serological affinity with antisera to any of 34 widely differing viruses tested. It does not seem of direct practical importance and may be easily overlooked.Samenvatting In zaailingen van peterselie (Petroselinum crispum) werd een nog niet eerder beschreven virus aangetroffen. Het virus kon niet worden aangetoond door toetsing van droge of in water geweekte zaden opChenopodium quinoa maar wel in op filtreerpapier gekiemde zaden en vooral in zaailingen. Het werd aangetroffen in 38 van de 54 getoetste herkomsten, in 17 van de 24 getoetste rassen en in zaad vermeerderd in alle zes hierop onderzochte Europese landen maar niet in enkele zaadmonsters uit de USA. In sommige monsters bevatten nagenoeg alle zaden het virus. In vijf herkomsten werd eveneens het nog niet eerder in peterselie gerapporteerde latente aardbeikringvlekkenvirus geconstateerd. Dit virus kan bij toetsing gemakkelijk worden herkend door systemische symptomen inC. amaranticolor en komkommer.In geïnfecteerde peterselieplanten zijn geen afwijkingen waargenomen. Het virus kon niet op non-persistente wijze worden overgebracht met zeven bladluissoorten maar wel gemakkelijk met sap. Van 32 getoetste plantesoorten van zeven families, waaronder vier schermbloemigen, kon het virus slechts worden overgebracht op vierChenopodium-soorten,Gomphrena globosa en alle drie getoetste spinazierassen. AllenC. quinoa (Fig. 1),C. giganteum, C. glaucum en soms ookC. amaranticolor (Fig. 2) reageerden met vaak voorbijgaande systemische symptomen. Een lokalelesietoetsplant werd niet gevonden. Zaadovergang bijC. quinoa kon niet worden aangetoond.Voor de houdbaarheid van het infectievermogen werden de volgende waarden gevonden: verdunningseindpunt 100–1000, thermaal inactiveringspunt 55–60°C en houdbaarheid in vitro 7–10 dagen.Zuivering door homogenisatie in fosfaatcitroenzuurbuffer, behandeling met Triton X-100 en ureum en differentiële en daarna dichtheidsgradiëntultracentrifugering leverde preparaten op met uniforme deeltjes van ca. 27 nm diameter (Fig. 3B), een sedimentatiecoëfficiënt van 127,5 S, een zweefdichtheid van 1,449 g/ml, een RNA-gehalte van 36% en een relatieve moleculaire massa van de eiwitondereenheid van 22×10³. Bij zuivering zonder toepassing van Triton en ureum werd een extra zone verkregen met aggregaten van 12 deeltjes in icosaëdrische rangschikking (Fig. 4). In ruw plantesap waren slechts met grote moeite enkele deeltjes met behulp van de elektronenmicroscoop te vinden.Het virus reageerde niet met antisera tegen 33 bolvormige virussen en luzernemozaïekvirus (Tabel 1). Of de zwakke reactie verkregen met één antiserum tegen het tomate-aspermievirus een verre serologische verwantschap inhoudt, dan wel het gevolg is van een verontreiniging, werd niet vastgesteld.Het virus wordt beschouwd als een geheel nieuw virus waarvoor de naamlatent peterselievirus wordt voorgesteld. Het lijkt door zijn beperkte waardplantenreeks en symptoomloosheid in de vatbaar bevonden soorten, behalve in enkele als toetsplant te gebruikenChenopodium-soorten, nauwelijks van praktische betekenis.     

水稻新病害叶鞘黑斑病的病原鉴定

 2011年7月,在湖南省花垣县水稻上发现一种新的病害,症状表现为叶鞘上出现长椭圆形黑斑,病健交界模糊。分离到的4隔孢弯孢霉(HNHY001)经离体和活体人工接种均可产生黑斑;另一种黑孢霉菌株(HNHY002)不引致叶鞘黑斑。从湖南省栽培水稻150份种子样本中共分离到弯孢霉菌株27个,其中4个菌株具有4隔孢子。这4个菌株中1个菌株不产生子座,其余3个均产生分枝子座。经人工接种除不产生子座的外,其余3个菌株均能产生典型的叶鞘黑斑。HNHY001的rDNA-ITS序列(GenBank登录号JQ360963)经BLAST搜索,与之最接近的几个序列为膝曲旋孢腔菌(Cochliobolus geniculatus)及其有丝分裂产孢种—4隔孢弯孢霉。鉴于单独以及与其它4隔孢弯孢霉菌株进行对峙培养均未产生有性阶段,建议根据此菌无性阶段特征即分生孢子较直、4横隔、子座分枝等特征,鉴定为膝曲旋孢腔菌有丝分裂产孢种之一的假弯孢(C.fallax)。水稻叶鞘黑斑病系国内外首次报道。     

湖南水稻上1种新矮缩病的病原研究

 Summer rice was suffered extensive damage from a new dwarf disease in Hunan Province in year 2009. In this study, the causal agent of this disease was confirmed as Southern rice black-streaked dwarf virus (SRBSDV) by nested RT-PCR, RT-PCR and PCR product sequencing.     

Rice seedbeds as a source of primary infection by Rice yellow mottle virus

The effect of contamination of rice seedlings by Rice yellow mottle virus (RYMV) in seedbeds on the onset and spread of rice yellow mottle in the field was investigated. Rice seedlings were artificially contaminated in seedbeds at different rates (0.1, 0.5, and 2.5%) and pooled in bundles before transplantation, as done by farmers. RYMV was successfully transmitted through contaminated hands and bundling healthy and diseased seedlings together. Hand contamination was responsible for 4.5% infection. Disease incidence in the field after secondary spread reached 32% for 2.5% seedbed contamination rate but remained limited (less than 10%) for all other rates. Eradicating infected plants from seedbeds lessened disease incidence in the field. This technique may be used in conjunction with other prophylactic measures to efficiently control rice yellow mottle disease.     

稳定性肥料配施微生物菌剂对莴笋生长及品质的影响研究

通过以蔬菜硫基稳定性复合肥替代普通化肥作为微生物菌剂的增效载体,研究了蔬菜硫基稳定性复合肥减量对灌漠土地区露地莴笋生长及品质的影响.采用田间试验方法,设置6个处理:不施肥料(CK)、当地习惯施肥处理(CF)、100％稳定性复合肥化肥(T100)、80％稳定性复合肥化肥+菌剂(T80)、60％稳定性复合肥化肥+菌剂(T6...     

Enzyme-linked immunosorbent assay (ELISA) and dispersedye immuno assay (DIA): Comparison of simultaneous and separate incubation of sample and conjugate for the routine detection of lettuce mosaic virus and pea early-browning virus in seeds

Two modifications of ELISA and DIA were compared in relation to sensitivity of detection of two plant viruses and suitability for large-scale routine testing. DIA is a solid phase immuno assay like ELISA, in which the enzyme conjugate is replaced by a dye sol conjugate and substrate incubation is replaced by immediate dissolving of the dye molecules from the conjugate with an organic solvent. Sample and conjugate were incubated separately (ELISA 1, DIA 1) or simultaneously (ELISA 2, DIA 2). The seed-borne viruses viz. lettuce mosaic virus (LMV) and pea early-browning virus (PEBV) were subjected to the assays. DIA detected LMV in a purified extract ofNicotiana benthamiana. However, compounds present in crude virus-free and virus-containing plant extracts strongly interfered with DIA, necessitating adaptation of DIA to plant viruses in crude extracts.With the extracts of lettuce and pea seeds ELISA 2, in comparison with ELISA 1, resulted in equal (LMV) or slightly higher (PEBV) extinction values and in a higher ratio between extinction values of virus-containing and virus-free samples. The higher sensitivity of ELISA 2 in combination with its higher efficiency as a result of simultaneous sample and conjugate incubation, indicates the potential of this method for large-scale indexing.Samenvatting Twee modificaties van ELISA en DIA werden vergeleken met betrekking tot hun gevoeligheid voor het aantonen van twee plantevirussen en hun geschiktheid voor routinematige toepassing. DIA is een serologische toetsmethode die veel overeenkomst vertoont met ELISA, maar waarin het enzymconjugaat is vervangen door een conjugaat met gedispergeerde kleurstofdeeltjes en de incubatie met enzym-substraat door het direct oplossen van de kleurstofmoleculen van het conjugaat met een organisch oplosmiddel. Incubatie van monster en conjugaat vond zowel gescheiden (ELISA 1, DIA 1) als gelijktijdig (ELISA 2, DIA 2) plaats. Twee met zaad overgaande virussen, te weten slamozaïekvirus (LMV) en vroege-verbruiningsvirus van erwt (PEBV) werden bij het onderzoek betrokken. Met DIA kon LMV worden aangetoond in een gezuiverd extract vanNicotiana benthamiana. In ruwe planteëxtracten bleken echter stoffen voor te komen die in DIA sterke niet-specifieke reacties tot gevolg hadden. Verder onderzoek is dan ook noodzakelijk om DIA geschikt te maken voor het aantonen van plantevirussen in ruwe extracten van planten. Betere resultaten werden verkregen met de beide ELISA-modificaties. Met de extracten uit slazaad en erwtezaad gaf ELISA 2 vergelijkbare (LMV) of iets hogere (PEBV) extinctiewaarden dan ELISA 1. Bovendien was de verhouding tussen de extinctiewaarden van virusziek materiaal en die van virusvrij materiaal, bij ELISA 2 hoger dan bij ELISA 1. De grotere gevoeligheid van ELISA 2 en de grotere doelmatigheid ten gevolge van de gelijktijdige incubatie van monster en conjugaat duiden op de bijzondere geschiktheid van deze methode voor routinematige toepassing op grote schaal.     

Biological and molecular characterization of Potato yellow blotch virus,a new species of the genus Potyvirus

A new species of the genus Potyvirus infecting potatoes, with the proposed name Potato yellow blotch virus (PYBV), was discovered in a breeding line 99m-022-026 in Scotland. The infected plants show isolated yellow blotches on the leaves. The genome of PYBV contains a large open reading frame encoding a single polyprotein of 3054 amino acids. Sequence analysis shows that PYBV is closely related to Potato virus A (PVA), with an overall 72% identity at the nucleotide level for the whole genome. The least conserved P1 protease gene shares only 50% nucleotide identity with PVA. The host range of PYBV was comparable to PVA on solanaceaous and non-solanaceous indicator plant species with the exception of Solanum demissum A and Y. Different symptoms were also observed for PYBV and PVA in Nicotiana benthamiana, Nicotiana hesperis and Nicotiana occidentalis P1. The susceptibility of potato (Solanum tuberosum) cultivars to PYBV and PVA was similar. In over 5 years of investigation, PYBV has not been found in commercial seed and ware potato crops in Scotland.     

A new selective medium for Burkholderia caryophylli, the causal agent of carnation bacterial wilt

A new selective medium (APCA medium) was developed for the isolation of Burkholderia caryophylli , the causal agent of carnation bacterial wilt, from both plants and soil. The optimal concentration and combination of antibiotics was investigated to determine the most selective condition for growing B .  caryophylli . The resultant composition of the medium per litre was: 0·79 g (NH₄)₂SO₄, 1·0 g KH₂PO₄, 0·5 g MgSO₄ · 7H₂O, 0·2 g KCl, 2·0 g D-arabinose, 5 mg crystal violet, 50 mg cycloheximide, 50 mg polymyxin B sulphate, 50 mg ampicillin sodium, 10 mg chloramphenicol, 25 mg blue tetrazolium, and 15 g agar. Plating efficiency ranged from 119 to 174% with an average of 141% compared to that of nutrient agar. The bacterium was successfully isolated from contaminated soil and plant tissues with this medium. Moreover, the medium almost completely inhibited the growth of other plant pathogenic bacteria and soil saprophytes. This selectivity was high enough to detect B . caryophylli in contaminated soil.     

小麦苗枯病菌的ITS分析及PCR检测

 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。     

Description of a new disease on Erythrina sp. in Martinique (French West Indies) and preliminary characterization of the causal agent as a novel Erwinia species

In spring 1995, symptoms of partial defoliation were observed on Erythrina indica var. fastigiata trees, commonly used as windbreaks in banana plantations in Martinique (French West Indies). Browning of the bark surface was consistently observed at the base of defoliated branches. Bacteria were isolated as nearly pure cultures from typical necrotic lesions on the bark. Results of Gram stain, staining of flagella and biochemical tests indicated that all isolates belonged to the Enterobacteriaceae, and to the genus Erwinia . Numerical analysis of the results of 48 biochemical properties showed that Erwinia isolates from diseased erythrina trees constituted a homogenous phenotypic cluster, and that, although weakly pectolytic, these isolates were closely related to pectolytic Erwinia species: E. carotovora ssp. betavasculorum and ssp. wasabiae , and E. cacticida . However, they could be clearly distinguished by different biochemical characteristics from type strains of known plant pathogenic species of Erwinia , Pantoea and Enterobacter . Under greenhouse conditions, the isolates were pathogenic to Erythrina indica var. fastigiata cuttings, but not to banana and pineapple, which are major crops in Martinique.     

Brazilian Potato virus Y isolates identified as members of a new clade facilitate the reconstruction of evolutionary traits within this species

Potato virus Y (PVY) is a plant virus distributed worldwide that causes damage to several species of the Solanaceae family. It was established long ago that groups of PVY isolates defined by phylogenetic analyses correlate strongly with those demarcated by differential biological properties. Consequently, life‐history traits of this viral species can be inferred by phylogenetic analysis. In this study, characteristics of PVY isolates sampled in different tobacco fields in Brazil were analysed and most of the tested Brazilian PVY isolates were assigned to the recently described unconventional serogroup Y^U. The analysis of molecular diversity of the coat protein (CP) cistron from some Y^U isolates made it possible (i) to identify specific amino acid residues in the N‐terminal of the CP protein and (ii) to assign some Y^U isolates to a new PVY clade. The symptoms caused by isolates belonging to this new PVY ‘Brazilian’ clade and their ability to infect selected susceptible hosts led to the conclusion that neither veinal necrosis symptoms expressed on infected tobacco plants nor adaptation to potato or pepper hosts are ancestral characteristics of PVY. These observations suggest that PVY has gained a remarkable new biological property and broadened its host range over time.