富贵竹中杆状DNA病毒湖北分离物的分子鉴定

 【目的】证明湖北发病富贵竹上是否存在杆状DNA病毒属（Badnavirus）病毒,分析来自富贵竹及其它作物Badnavirus病毒不同分离物间的分子差异。【方法】通过分段克隆测序的方法获得湖北富贵竹中 Badnavirus病毒基因组全序列,利用BLAST工具及其它生物学软件进行序列分析。【结果】富贵竹Badnavirus病毒湖北分离物基因组为环状结构,全长7 522 bp;基因组正链包含7个ORFs,推测其编码蛋白的分子量分别为17.58、14.93、214.77、11.86、11.31、16.12和11.00 kD。获得的基因组与福建富贵竹斑驳病毒（Dracaena mottle virus , DrMV）大小仅相差9个核苷酸,两者基因组核苷酸序列一致率为99.7％,ORFs 1～3编码氨基酸序列的一致率分别为99.3%、100%、99.2%,而与其它已报道的14种Badnavirus病毒分离物间的核苷酸全序列一致率为32.0%～44.0%。【结论】叶片表现褪绿斑驳等症状的湖北富贵竹中存在Badnavirus病毒,该病毒与DrMV为同种病毒,已报道的富贵竹中Badnavirus病毒分离物间分子差异非常小,这与已报道的其它作物中Badnavirus病毒不同分离物间存在非常大的分子差异不同。     

Identification and transmission of Piper yellow mottle virus and Cucumber mosaic virus infecting black pepper (Piper nigrum) in Sri Lanka

Sri Lankan black pepper with symptoms of yellow mottle disease contained a mixture of viruses: Piper yellow mottle virus (PYMV) particles (30 × 130 nm), Cucumber mosaic virus (CMV, 30 nm diameter isometric particles), and unidentified, isometric virus-like particles (30 nm diameter). An effective purification procedure is described for PYMV. Immunosorbent and conventional electron microscopy successfully detected badnavirus particles only when at least partially purified extracts were used. PYMV was confirmed as the cause of the disease, with the other two viruses apparently playing no part in producing symptoms. PYMV was transmitted by grafting, by the insect vectors citrus mealy bug ( Planococcus citri ) and black pepper lace bug ( Diconocoris distanti ), but not by mechanical inoculation or through seeds. The CMV isolate was transmitted to indicator plants by mechanical inoculation and by the vector Aphis gossypii , but not by Myzus persicae ; but neither mechanical nor insect transmission of CMV to black pepper was successful. A sensitive polymerase chain reaction assay was developed to detect PYMV in black pepper.     

甘薯杆状病毒湖南分离物基因组全序列测定及比较分析

 利用Small RNA测序技术对采自湖南的甘薯杆状病毒进行鉴定,获得与SPBV-B序列具有相似性的contigs 161个。首先设计小片段引物,PCR扩增分离出706 bp的目的片段,与SPBV-B (FJ560944)相似性为78%。分段设计引物,进行基因全序列PCR扩增。引物组合SPBV-BF1/R1、SPBV-BF2/R4和SPBV-BF5/R5分别扩增出3 150、2 900和3 500 bp目的条带。序列拼接获得2个SPBV-B基因组全序列,大小分别为7 894 bp(MK052980)和7 981 bp(MK052981),包含完整编码阅读框。进化分析表明MK052980和MK052981与SPPV聚为同一分支,与SPPV相似性分别为81%和83%,与SPBV-B相似性分别为86%和94%。MK052980和MK052981具有89.5%的相似性。经编码阅读框氨基酸序列比对,MK052980序列的ORF3a氨基酸序列存在变异。这是国内首次报道SPBV-B全基因序列。     

Status of Banana Streak Disease in Africa: Problems and Future Research Needs

Streak disease of banana and plantain caused by banana streak virus (BSV) was first reported in the Ivory Coast in 1974 and occurs in at least 16 countries in Africa. Based on genomic characteristics, BSV has been shown to be a member of genus Badnavirus. Efficient and reliable diagnostic methods for BSV have recently become widely available. This paper summarizes the current knowledge on its causal agent, geographical distribution, symptomatology, transmission, host range, available diagnostic techniques and management options for the disease in Africa. Further research needs are identified in light of the widespread occurrence of BSV in most plantain/banana germplasms and the difficulties in obtaining BSV-free plantlets through tissue culture.     

Production of discernible disease phenotypes in Canna by five plant viruses belonging to the genera Potyvirus,Cucumovirus and Badnavirus

Cannas are tropical and subtropical flowering perennial plants. The genus contains many species but most commercially grown cultivars are interspecific hybrids selected for their attractive foliage and flowers. Canna production is so lucrative that there are farmers and nurseries dedicated solely to its production. The specific issue that the canna industry faces is virus diseases. In this study, rhizomes of 24 canna cultivars were gathered and diagnostics conducted to detect Bean yellow mosaic virus (BYMV, Potyvirus), Canna yellow mottle virus (CaYMV, Badnavirus), Canna yellow streak virus (CaYSV, Potyvirus), Cucumber mosaic virus (CMV, Cucumovirus) and Tomato aspermy virus (TAV, Cucumovirus). Visual assessment of disease symptoms and diagnostic tests were carried out to identify the prevalent diseases and describe the symptoms that are associated with virus infection. BYMV, CaYMV and CaYSV caused severe mosaic and necrosis either in the leaf lamina or veins of infected leaves. Potyvirus infection suppressed red colouration in the foliage of some varieties. CaYMV and CaYSV often appeared in the same plant, suggesting they might represent a viral complex. CMV and TAV were rarely seen in these populations. Interestingly, CaYMV but not CaYSV could be mechanically inoculated to Phaseolus vulgaris plants.     

甘薯杆状病毒湖南分离物基因组全序列测定及比较分析

 利用Small RNA测序技术对采自湖南的甘薯杆状病毒进行鉴定,获得与SPBV-B序列具有相似性的contigs 161个。首先设计小片段引物,PCR扩增分离出706 bp的目的片段,与SPBV-B (FJ560944)相似性为78%。分段设计引物,进行基因全序列PCR扩增。引物组合SPBV-BF1/R1、SPBV-BF2/R4和SPBV-BF5/R5分别扩增出3 150、2 900和3 500 bp目的条带。序列拼接获得2个SPBV-B基因组全序列,大小分别为7 894 bp(MK052980)和7 981 bp(MK052981),包含完整编码阅读框。进化分析表明MK052980和MK052981与SPPV聚为同一分支,与SPPV相似性分别为81%和83%,与SPBV-B相似性分别为86%和94%。MK052980和MK052981具有89.5%的相似性。经编码阅读框氨基酸序列比对,MK052980序列的ORF3a氨基酸序列存在变异。这是国内首次报道SPBV-B全基因序列。     

侵染湖南甘薯种质资源的曲叶病毒和杆状病毒的检测及其亲缘关系分析

采用PCR技术对2015—2016年从湖南省多个市县采集的180份甘薯种质资源进行两种DNA病毒（sweet potato leaf curl virus,SPLCV和sweet potato badnavirus,SPBV）检测,并以PCR扩增产物序列为基础,对这两种病毒的亲缘关系进行分析。结果表明：（1）180份甘薯种质资源中,22份检测出SPLCV,占总数的12.2%;32份检测出SPBV,占总数的17.8%;8份同时检出SPLCV和SPBV这两种病毒,占总数的3.9%。（2）检出SPLCV的市县有长沙、永州、怀化、邵阳、郴州、常德、娄底、益阳、岳阳和株洲;检出SPBV的市县有长沙、永州、怀化、衡阳和邵阳;检出两病毒复合侵染的市县有怀化、长沙、永州和邵阳。（3）基于病毒全基因组序列分析显示,全球已报道的36个SPLCV分离物可分为3组,本研究获得的12个湖南分离物可分别归于3个组中,其中6个归于组Ⅰ,1个归于组Ⅱ,5个归于组Ⅲ,表明湖南SPLCV具有高度多样性。（4）基于病毒运动蛋白和外壳蛋白编码基因部分序列分析显示,全球已报道的36个SPBV分离物可分为4组,绝大部分分离物属于组Ⅰ,我国1个安徽分离物单独成组（组Ⅱ）,我国2个山东分离物和1个海南分离物构成1个组（组Ⅲ）,本研究获得的2个湖南分离物其中1个归于组Ⅰ,而另1个与已报道各分离物的核苷酸序列均存在显著差异,相似性仅为74.92%,单独成组（组Ⅳ）,表明我国SPBV具有高度的变异性,存在多个变异类型。本研究结果可为湖南地区这两种病毒病防控提供依据。