Potato Virus Y from Petunia can cause Symptoms of Potato Tuber Necrotic Ringspot Disease (PTNRD)

A potyvirus known to be an important agent involved in causing a disease of trailing petunias, was identified as being a member of the necrotic strain of potato virus Y (PVY) using a number of monoclonal antibodies. The sequence of the coat protein gene for the PVY isolate was determined and when compared with sequences for other PVY strains it was shown to cluster closely with isolates of PVY^NTN and to have a recombination point present within the coat protein common with other isolates of PVY^NTN. When inoculated onto potato tuber necrotic ringspot disease (PTNRD) susceptible potato cultivars the petunia isolate was found to be capable of causing necrotic tuber symptoms, consistent with those caused by other isolates of PVY^NTN. Due to the number of similarities it is thought the petunia isolate belongs to the PVY^NTN group of isolates. Out of 24 species of bedding and pot plant crops tested, 19 were shown by mechanical inoculation to be susceptible to PVY, highlighting not only a clear risk to a number of commercially important plant species from PVY^NTN infected trailing petunias, but also other susceptible crops grown in these areas.     

侵染扶桑的烟草花叶病毒分离物鉴定

从表现叶斑驳症状的扶桑病株上获得一病毒分离物,电镜下可见约300 nm×18 nm的杆状粒子,其与烟草花叶病毒抗血清呈明显的阳性反应,dsRNA约为6.4 kbp。根据烟草花叶病毒(tobacco.mosaic virus,TMV)的RNA序列设计引物,进行RT-PCR检测,扩增出约800 bp的预期特异片段。将PCR产物连接pMD18-T载体,转化大肠杆菌DH5α,得到了含有目的片段的重组子。序列分析表明,与周雪平等报道的序列(GenBank AJ011933.1)同源性达99％。通过生物学、病毒粒子观察、血清学以及分子生物学实验结果,确定该病毒分离物为TMV。     

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.     

青蒿中番茄斑萎病毒和烟草花叶病毒的分子鉴定及相关序列分析

番茄斑萎病毒(tomato spotted wilt virus, TSWV)和烟草花叶病毒(tobacco mosaic virus, TMV)是2种重要的植物病原病毒, 对多种经济作物的产量和品质均造成严重影响。2021年-2022年, 在云南省丽江市烟草种植区不同烟区采集叶片黄化、皱缩以及无症状的青蒿Artemisia caruifolia样品共计14份, 利用免疫金标速测卡和RT-PCR对其病原病毒进行检测。利用免疫金标速测卡检测结果显示, 在所检样品中有9份样品检测出TSWV, 检出率为64.28%, 有3份样品检测出TMV, 检出率为21.43%, 2种病毒复合侵染的检出率同样为21.43%;利用RT-PCR对复合侵染的3份样品进行分子检测, 结果显示, 在3份复合侵染青蒿样品中获得3条TSWV N基因序列、3条TMV cp基因序列和2条TMV RdRp部分序列。TSWV青蒿分离物与分离自云南的TSWV-2分离物相似性最高, 为99.6%;TMV青蒿分离物与分离自辽宁的TMV-Shenyang分离物和分离自云南的TMV-Yongren-1相似性最高, 均大于99.4%。这是首次发现TSWV和TMV 2种不同属病毒复合侵染青蒿。     

番茄黄化曲叶病毒北京分离物侵染性克隆的构建与其致病性

为明确番茄黄化曲叶病毒北京分离物（Beijing isolate of tomato yellow leaf curl virus,TYLCV-BJ）致病性的强弱,以感染TYLCV-BJ的番茄叶片DNA为模板PCR扩增获得该分离物基因组全长序列,并构建该分离物的侵染性克隆,将其分别接种到番茄、烟草和拟南芥植株上,比较该分离物和TYLCV上海分离物2(TYLCV-Shanghai 2,TYLCV-SH2)致病性的差异。结果显示,该分离物基因组全长序列同TYLCV-SH2的相似度为99.03%,在番茄和烟草植株上TYLCV-BJ比TYLCV-SH2发病更早,症状更重,TYLCV DNA和外壳蛋白积累量更高。TYLCV-BJ可以通过农杆菌Agrobacterium tumefaciens注射法在拟南芥中复制和系统侵染,而TYLCV-SH2不能有效侵染拟南芥。表明TYLCV-BJ的致病性强于TYLCV-SH2,所建立的侵染性克隆有广泛的研究和应用价值。     

Sequence comparison and transmission of <Emphasis Type="Italic">Blackcurrant reversion virus</Emphasis> isolates in black,red and white currants with black currant reversion disease and full blossom disease symptoms

We collected samples from black, red and white currants showing symptoms of blackcurrant reversion disease (BRD) and full blossom disease (FBD), cultivated in the Czech Republic. Blackcurrant reversion virus (BRV) was detected in all symptomatic plants. After amplification, a substantial part of the 3′ non-translated region (3′-NTR) of RNA2 of 15 new isolates of BRV was sequenced and compared with sequences available in the literature and GenBank. We did not find significant sequence diversity among isolates associated with either FBD or BRD. BRV was graft-transmitted from FBD infected red currant to black currant where symptoms of BRD were observed. Further sequence analysis of BRV isolates resulted in a phylogenetic tree with four branches, each consisting of six to nine isolates. No correlation with geographic origin was visible on the tree as isolates from various countries occurred in all four branches. We also found no correlation between the host and the topology of the tree: most of black currant isolates occurred in branches 3 and 4, but also occurred in branches 1 and 2. Only one white currant and one red currant isolate occurred in branches 3 and 4, respectively. The sequence identity of the Czech isolates in this region ranged from 91.9 to 99.8%. The 17 plant species growing within and in the close vicinity of the BRD-infested plantation were tested negative for BRV by RT-PCR as natural hosts of BRV. BRV was successfully transmitted by mechanical inoculation from black currant to Nicotiana occidentalis and N. tabacum cv. Xanthi, the latter being a new host for BRV. The infection was confirmed by PCR and sequencing.     

First report of tomato chlorotic dwarf viroid isolated from symptomless petunia plants (Petunia spp.) in Japan

A viroid was detected for the first time in symptomless petunia plants (Petunia spp.) and identified as Tomato chlorotic dwarf viroid (TCDVd) based on an analysis of the complete genomic sequence. These petunia plants are a likely source of inoculum for tomato or potato plants because TCDVd induces severe symptoms on these plants. The genomic sequence of this petunia isolate from Japan shared 100 % identity with petunia isolates from the Netherlands and United Kingdom and a tomato isolate from Japan. Phylogenetic analysis showed that all petunia isolates and the tomato isolate from Japan formed a monophyletic clade.     

Detection of “Candidatus Phytoplasma asteris” associated with henon bamboo witches' broom in Korea

Witches' broom disease in bamboo (Phyllostachys nigra Munro var. henonis) was found in Yeoungyang, Korea. In transmission electron micrographs, phytoplasma-like bodies were detected in the phloem cells of diseased plants but not in those of healthy plants. The presence of phytoplasmas was confirmed by amplification of a 1.8-kb DNA fragment using a primer pair specific for the region containing a 16S rRNA gene and an intergenic spacer region between the 16S and 23S rRNA genes. Comparision of the 16S rRNA gene sequences showed that the causal phytoplasma belongs to “Candidatus Phytoplasma asteris,” and shared the highest degree of similarity with the sequence of the onion yellows (OY) isolate in Japan. This is the first phylogenetic identification of phytoplasma infection of bamboo in Korea.     

水蜡A病毒在呼和浩特市和哈尔滨市紫丁香上的发生及其基因组分子特征分析

为明确侵染紫丁香Syringa oblata并引起褪绿花叶症状的病毒种类及其基因组分子特征,利用透射电子显微镜对分离自呼和浩特市和哈尔滨市的紫丁香病样中的病毒粒子进行观察,并通过小RNA高通量测序和RT-PCR技术对其进行检测分析。结果表明,在紫丁香显症叶片的病毒粗提液中观察到长约600 nm、宽约13 nm的线状病毒粒子。利用小RNA高通量测序和RT-PCR技术从病样中检测到水蜡A病毒（Ligustrum virus A,LVA）,发病率为3.7%。呼和浩特市紫丁香分离物LVA-Sob的基因组序列全长8 525 nt,包含6个开放阅读框,分别编码Rep（1 968 aa）、TGB1（229 aa）、TGB2（107 aa）、TGB3（60 aa）、CP（294 aa）和NABP（119 aa）共6个蛋白。序列一致性分析表明,分离物LVA-Sob与韩国水蜡树分离物LVA-SK的基因组序列一致率高达97.9%,而与我国辽宁省暴马丁香分离物LVA-DX的基因组序列一致率仅为73.6%。在这3个LVA分离物基因组中没有检测到重组事件;基于基因组和cp基因序列的系统发育树显示这3个LVA分离物形成一个分支,并与瑞香S病毒（daphne virus S,DVS）有较近的亲缘关系。     

Identification of a new pathotype of Puccinia hordei with virulence for the resistance gene Rph7

Barley leaf rust resistance gene Rph7, derived from barley accession Cebada Capa, is the most effective R-gene for resistance to Puccinia hordei. Virulence for this gene was known in the USA, Israel and Morocco but not yet in Europe. We found an unexpected leaf rust infection in the field at Córdoba, Spain in 2004 on Rph7 carrying lines. This virulence for Rph7 was confirmed in growth chamber experiments, being the first report of Rph7 virulence in European populations of P. hordei. A collection of 680 barley accessions was screened for resistance against this new isolate. Twelve accessions showed segregation with individual plants showing resistance based on hypersensitivity (low infection type). These individual resistant plants were selected and grown in the greenhouse to obtain seeds.     

A Cucumber mosaic virus isolated from Momordica charantia L.

Momordica charantia L. plants systemically infected with Cucumber mosaic virus (CMV) were found in Oita Prefecture. The virus isolated from the host plant was characterized by biological, serological, and molecular biological methods. The purified virus was used to mechanically inoculate the host and produced green mottle, green mosaic, and/or chlorotic spots in the noninoculated upper leaves of the host. The virus was identified as an isolate of CMV containing genomic RNA3 derived from subgroup IA by several lines of evidence based on electron microscopy, serological detection, host range, symptoms, and the entire nucleotide sequence of RNA3.     

Efficacy of olive mill waste water and its derivatives in the suppression of crown gall disease of bitter almond

Olive mill waste water (OMW) and some of its indigenous bacterial strains were tested in vitro and in planta for their efficacy against crown gall disease caused by Agrobacterium tumefaciens. OMW and polyphenols displayed a high level of antibacterial activity, however the volatile fraction was less efficient as only a bacteriostatic effect was observed. In pot experiments, the percentage of bitter almond rootstock showing symptoms of crown gall was significantly reduced with the dosage rate of OMW 1% as compared to the control (highly natural infected soil treated with water). Five indigenous bacterial strains isolated from OMW exhibited an antagonistic effect against the bacterium. Based on the gene 16S rRNA sequence analysis, one isolate showed 99.2% similarity to known sequences of Bacillus subtilis, one isolate demonstrated high percentage similarities (99.3%) to the genera Bacillus pumilis, and two isolates were associated with Stenotrophomonas maltophilia and Pseudomonas putida 100% and 99.6% similarities respectively. Among these bacteria, the strain B1 proved efficient against the soil borne pathogen in vitro and pot experiments. Our study in controlled conditions suggested that the addition of OMW to soil exerts significant disease suppressiveness against A. tumefaciens. Thabet Yangui and Ali Rhouma contributed equally to this work and are regarded as joint first authors.     

Molecular Identification of Oat Mosaic Virus as a Bymovirus

A partial sequence of Oat mosaic virus (OMV) has been obtained for four isolates of the virus from four European countries. This represents the first available sequence data for this important disease of winter-sown oats. The longest clone of 1699 nucleotides was obtained from infected English oats using a degenerate primer, designed to members of the Potyviridae family. Alignment of the predicted amino acid sequence with members of the Potyviridae showed closest identity with viruses of the Bymovirus genus. The predicted amino acid sequence has one open reading frame corresponding to part of the NIb and capsid protein, with a 3 untranslated region of 351 nucleotides, followed by a poly(A) tail. PCR primers were designed to the coat protein and NIb gene of members of the Bymovirus genus and used to obtain partial sequences of 1441 nucleotides at the 3 end of infected oats from both Wales and France. A specific primer set designed to the English isolate was used to generate a product of 701 nucleotides from OMV-infected oat leaves from Ireland. All four isolates are highly conserved at the amino acid level.The first two authors contributed equally to the work     

传播木尔坦棉花曲叶病毒的烟粉虱隐种鉴定

为明确广东地区传播木尔坦棉花曲叶病毒(Cotton leaf curl Multan virus,CLCu Mu V)的烟粉虱隐种,采用分子鉴定、烟粉虱传毒试验与分子检测的方法,对传播CLCu Mu V的烟粉虱隐种进行了鉴定。结果表明:在广东棉花、红麻和黄秋葵曲叶病发生的田间,2014年采集的30头烟粉虱mt COI与Asia II 7隐种序列同源性为98.3%~99.6%,2015年采集的10头烟粉虱中,8头烟粉虱mt COI与MEAM1隐种序列同源性为99.2%~99.5%,2头烟粉虱mt COI与Asia II 7隐种序列同源性为99.8%~99.9%,说明烟粉虱种群包括入侵隐种MEAM1和土著隐种Asia II 7。在利用烟粉虱人工传毒试验中,MEAM1、Asia II 7和Asia II 1这3个隐种均可在棉花曲叶病株上饲毒获得CLCu Mu V及β卫星分子;除MEAM1隐种外,Asia II 7、Asia II 1隐种可传播CLCu Mu V,侵染红麻及黄秋葵植株引起曲叶病,前者传毒效率分别为50%和100%;而后者为33%和100%。以上3个烟粉虱隐种传毒接种棉花未见成功。     

A survey of tobacco viruses in tobacco crops and native flora in Greece

The most important tobacco producing areas in Greece were surveyed for virus presence, from 1997 to 2000. Tobacco seedlings or plants showing virus-like symptoms were randomly collected from seedbeds or fields, respectively, and tested by ELISA, and/or mechanical inoculation onto indicator plants. Potato virus Y (PVY), Cucumber mosaic virus (CMV) and Tobacco mosaic virus (TMV) were detected in all sampling areas, with TMV mainly found in oriental varieties. Tomato spotted wilt virus (TSWV) consisted a serious endemic virus in Northern Greece (Thrace, Central and Eastern Macedonia), whereas Alfalfa mosaic virus (AMV) was mainly found in regions, where alfalfa was cultivated in the vicinity of tobacco crops. Eggplant mottled dwarf virus (EMDV) was detected in several areas but always in very low incidence (<0.01%). Surveys were also conducted to assess the potential reservoir hosts of PVY, CMV and AMV among weeds collected from highly infected tobacco fields from 1998 to 2000. Among 3450 samples tested for PVY, plants from 17 species in 10 families were found infected. For CMV, 2891 weed samples were tested and 19 species in 12 families were positive. Assays for AMV infection were made on 961 samples and 12 species in 9 families were identified as hosts of this virus.