甘肃省马铃薯主要病毒病发生情况调查

2015年-2016年,在甘肃省10个地市24个马铃薯主栽县(区)146个生态区域(乡镇)采集了757份具有典型症状的马铃薯样品,应用DAS-ELISA法进行检测,筛查6种主要病毒(PVX、PVY、PLRV、PVA、PVS和PVM)。结果表明:631份样品检测到病毒,PVS的检出率最高,达47.03%,PVY次之,为33.82%,PVA最低,只有0.63%;发生复合侵染的病毒主要为PVY+PVS,复合侵染率达到10.13%,三种病毒复合侵染主要是PVY+PVS+PVM;病毒种类和感病程度与品种、地域有关。     

马铃薯纺锤块茎类病毒的检测和防治

马铃薯纺锤块茎类病毒病(potato spindle tuber viroid,PSTVd)是一种严重为害马铃薯生产的病害,降低产量20%—30%。防治的主要措施是应用无类病毒的种薯。由于目前还没有脱掉类病毒的有效措施,只能从未被饱和侵染的群体中鉴定筛选出未被侵染的个体,再脱掉其它病毒,作为核心繁殖材料。1987年以来,利用自制的电泳设备,以往复聚丙烯酰胺凝胶电泳法(return-polyacrylamide gel electrophoresis,R-PAGE)检测类病毒,筛选出未感病的个体,再用茎尖组织培养法脱掉其它病毒。经用马铃薯卷叶病毒等8种病毒酶标抗体鉴定筛选,获得既无类病毒也无主要马铃薯病毒的克新1、2、3和4号等主栽马铃薯品种的核心种。并已提供给省内外的良种场繁殖推广。1989和1990年抽样检测克山良种场繁殖的原种、一级和二级良种,未检测到类病毒。     

马铃薯主产区病毒病发生情况调查分析

在我国马铃薯主产区黑龙江、内蒙古、甘肃、贵州采集了752份具有典型病毒病症状的样品和疑似样品,应用DAS ELISA方法进行检测。主要筛查6种病毒：PVX、PVY、PVS、PLRV、PVM、PVA。结果显示: 270份样品检测到病毒,总体上,PVY的检出率最高,PVS次之,33份样品是由多种病毒混合侵染造成的,田间PVY+PLRV混合侵染率最高,PVS+PVY次之。试管苗PVS病毒发生较多,原原种和大田种薯PVY病毒发生比例最高。     

湖南省马铃薯病毒病发生情况调查

为了解湖南省马铃薯种薯质量和主要病毒病发生情况,2019年-2020年马铃薯秋作和冬作期间,对长沙、益阳、湘潭、澧临等马铃薯生产区的155个马铃薯样品,运用反转录-聚合酶链式反应(RT-PCR)和双抗体夹心酶联免疫吸附检测(DAS-ELISA)技术,筛查6种主要马铃薯病毒,包括马铃薯X病毒Potato virus X(PVX)、马铃薯Y病毒Potato virus Y(PVY)、马铃薯M病毒Potato virus M(PVM)、马铃薯S病毒Potato virus S(PVS)、马铃薯A病毒Potato virus A(PVA)、马铃薯卷叶病毒Potato leaf roll virus(PLRV)。检测结果表明:6种马铃薯病毒病在湖南均有不同程度的发生,单一和两种病毒复合感染植株占比最高,其次是3种病毒复合感染,存在极少数植株复合感染4～5种病毒病情况。在秋作马铃薯中,PVY检出率达到29.41%;PVS和PVA检出率均为27.94%;PVM、PVX、PLRV的检出率分别为20.59%、19.12%、17.65%。在冬作马铃薯中,PVX检出率最高,达到31.03%;其次是PLRV,...     

纳米磁珠结合实时荧光RT-PCR技术检测大丽花潜隐类病毒

正大丽花潜隐类病毒(Dahlia latent viroid,DLVd)是Verhoeven et al.(2013)用Return-PAGE和s-PAGE技术从大丽花中鉴定出的一种新类病毒。根据2015年国际病毒分类委员会分类报告,DLVd为马铃薯纺锤形块茎类病毒科啤酒花矮化类病毒属成员。该病毒单独侵染时,大丽花没有明显的症状,其与马铃薯纺锤块茎类病毒(Potato spindle tuber viroid,PSTVd)复合侵染时可使大丽花叶片卷曲(Tsu-     

福建省福清地区马铃薯病毒病病原的分子检测

2017年调查福建福清地区马铃薯病毒病的发生情况,以明确该地区马铃薯主要病毒病原。共采集了46份疑似感染病毒的马铃薯植株,提取总RNA,利用RT-PCR技术进行分子检测,结果表明,福清地区危害马铃薯的病毒有马铃薯Y病毒Potato virus Y(PVY)、马铃薯卷叶病毒Potato leaf roll virus(PLRV)、马铃薯S病毒Potato virus S(PVS),检出率分别为56.52%、17.39%和10.87%,以PVY检出率最高,说明PVY是危害该地区马铃薯样品的主要病毒病原。通过病毒复合侵染进行分析,发现该地区存在病毒复合侵染马铃薯现象。研究结果可为福清地区马铃薯种薯的引进和病毒病害防治提供参考依据。     

RT-PCR技术对宁夏马铃薯脱毒种薯病毒检测的研究

病毒病是影响马铃薯生产的重要病害,为了解宁夏地区马铃薯种薯生产中的病毒发生情况,在银川市、固原市和西吉县不同种植模式的脱毒种薯生产基地,对不同品种的原原种、原种、一级种在不同生育期随机采集样品,利用RT-PCR检测技术对4种马铃薯主要病毒PVX、PVY、PVS、PLRV和类病毒PSTVd进行检测。结果表明:PVX、PVY、PVS、PLRV在3个基地均有不同程度检出,成熟期病毒检出率分别为35%、28%、35%和34%,未检出PSTVd;4种病毒检出率在马铃薯现蕾期以后呈现显著性增长;病毒检出率随着种薯繁育级别增加差异显著;6个马铃薯品种均为感病品种;大田种植种薯的病毒检出率显著高于防虫网室。     

马铃薯病毒性退化及防止

马铃薯生长期间,出现植株变矮、分枝减少、叶片皱缩、花叶或卷叶、生长势衰退、块茎变小、产量一年不如一年,这种现象称为马铃薯病毒性退化。一、退化的原因马铃薯退化是由病毒侵染造成的。侵染危害马铃薯的病毒有二十几种,这些病毒一旦侵染马铃薯植株和块茎,就能造成各种不同的病态和不同程度的减产。病毒侵染块茎后,随块茎的种植使病毒代代相传。种植感病毒的马铃薯时间愈长,病毒侵入的机会就愈多（有的植株,常受２～３种病毒复合侵染）,病毒性退化就愈来愈重。最后患病的种薯在生产上失去了利用价值。二、如何防止马铃薯病毒性退…     

多重RT-PCR同时检测6种马铃薯病毒及1种类病毒

病毒病是限制我国马铃薯生产的重要因子之一。本研究通过引物设计和体系优化建立了一套多重RT-PCR检测方法，可以在一个反应体系中同时检测6种马铃薯病毒和1种马铃薯类病毒以及1个马铃薯内参基因，包括马铃薯Y病毒(potato virus Y, PVY)、马铃薯M病毒(potato virus M, PVM)、马铃薯S病毒(potato virus S, PVS)、马铃薯X病毒(potato virus X, PVX)、马铃薯A病毒(potato virus A, PVA)、马铃薯卷叶病毒(potato leaf curl virus, PLRV)、马铃薯纺锤块茎类病毒(potato spindle tuber viroid, PSTVd)和细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunitⅠ,CoxⅠ)基因的特异片段，产物大小依次为181、226、275、565、630、681、359、500 bp,符合理论预期。采用建立的方法以相应病毒和类病毒的质粒作模板进行检测，其检测灵敏度在1.6×10^-3～1.8×10^-1 ...     

梨泡状溃疡类病毒的RT-PCR检测及序列分析

正梨泡状溃疡类病毒(Pear blister canker viroid,PBCVd)为马铃薯纺锤块茎类病毒科(Pospiviroidae)苹果锈果类病毒属(Apscaviroid)成员~([1])。该类病毒在自然条件下主要侵染野生和栽培的梨和苹果~([2~4]),梨和榅桲多数品种感染PBCVd后无明显症状,通过嫁接传染该类病毒可诱导梨指示植物     

一种快速简便检测马铃薯病毒的方法——病毒细菌协同凝集法(VBA)的研究

在国内、外首次选用金黄色葡萄球菌(Staphylococc aureus)的No.180菌株用于病毒细菌协同凝集试验(Virobacterial agglutination test简称VBA)检测了来自马铃薯的4种不同形态粒子的病毒:马铃薯X病毒(PVX)、马铃薯丫病毒(PVY)、烟草花叶病毒(TMV)和马铃薯卷叶病毒(PLRV),其灵敏度达2.7～6.1ng/ml,检出病汁液的最大稀释度达10~4～10~5(PLRV1:500),较国内、外采用的Cown I菌株的灵敏度提高5～10倍。与血清学方法相比,VBA在2～3分种内就可获得结果,无假阳性反应,其灵敏度显著高于间接酶联法和免疫电镜,而接近于A蛋白酶联法。经抗血清致敏的菌体在4℃下保存4个月,对VBA检测的灵敏度没有影响。用VBA对采自田间的93个马铃薯病样进行检测,它们大多受2～3种病毒(PVX、PVY及PLRV)的复合侵染;和用间接酶联法及鉴别寄主检测的结果趋向一致。室内和田间试验均表明VBA灵敏度高、特异性强、快速简便和经济,尤其适合在基层单位中推广应用。     

Characterization of potato potyvirus Y (PVY) isolates from seed potato batches. Situation of the NTN, Wilga and Z isolates

A collection of 38 PVY isolates from seed potato batches, originating from several Western European countries, was characterized by using current biological, serological and molecular tools differentiating PVY strains and groups. The correlation between the three kinds of tests was good but not absolute. No single serological or PCR method was able to discriminate among the five isolate groups found. Twenty-nine isolates belonged to the PVY^N strain and six to the PVY^O strain. No PVY^C was found. Two other isolates reacted serologically like PVY^O, but were unable to elicit a hypersensitive response from the Ny_tbr gene and probably represent the PVY^Z group. At the molecular level, these two isolates showed a combination of both PVY^O and PVY^N and could be recombinants of these strains. Another isolate reacted serologically like PVY^O, but induced vein necrosis in tobacco, like PVY^N-Wilga. Some PVY^N isolates caused tuber ring necrosis in glasshouse conditions. These might belong to the PVY^NTN group. The PVY^NTN, PVY^N-Wilga and PVY^Z groups probably represent pathotypes within strains PVY^N and PVY^O, respectively. The present study also confirms previous reports showing a high genetic variation at the 5 end within the PVYN strain.     

一种快速简便检测马铃薯病毒的方法——病毒细菌协同凝集法(VBA)的研究

 在国内、外首次选用金黄色葡萄球菌(Staphylococc aureus)的No.180菌株用于病毒细菌协同凝集试验(Virobacterial agglutination test简称VBA)检测了来自马铃薯的4种不同形态粒子的病毒:马铃薯X病毒(PVX)、马铃薯丫病毒(PVY)、烟草花叶病毒(TMV)和马铃薯卷叶病毒(PLRV),其灵敏度达2.7~6.1ng/ml,检出病汁液的最大稀释度达10⁴~10⁵(PLRV1:500),较国内、外采用的Cown I菌株的灵敏度提高5~10倍。与血清学方法相比,VBA在2~3分种内就可获得结果,无假阳性反应,其灵敏度显著高于间接酶联法和免疫电镜,而接近于A蛋白酶联法。经抗血清致敏的菌体在4℃下保存4个月,对VBA检测的灵敏度没有影响。用VBA对采自田间的93个马铃薯病样进行检测,它们大多受2~3种病毒(PVX、PVY及PLRV)的复合侵染;和用间接酶联法及鉴别寄主检测的结果趋向一致。室内和田间试验均表明VBA灵敏度高、特异性强、快速简便和经济,尤其适合在基层单位中推广应用。     

浙江省马铃薯病毒病检测分析

马铃薯病毒病是影响马铃薯产量和品质的主要因素之一,其症状表现为花叶、黄化和卷曲等。2019年4月-5月,在浙江省湖州、杭州、绍兴、宁波、金华、台州和丽水等7个地市的主要马铃薯产区采集了具有典型病毒病症状的马铃薯样品,采用转录组测序和RT-PCR验证的方法进行病毒检测,共筛查并验证出6种马铃薯病毒,包括马铃薯X病毒Potato virus X(PVX)、马铃薯Y病毒Potato virus Y(PVY)、马铃薯S病毒Potato virus S(PVS)、马铃薯H病毒Potato virus H(PVH)、马铃薯M病毒Potato virus M(PVM)以及马铃薯奥古巴花叶病毒Potato aucuba mosaic virus(PAMV)。检测结果表明,在调查的这7个地市中每个地市都具有2种以上的病毒发生,其中PVH作为一种新鉴定的麝香石竹潜隐病毒属成员在除宁波以外的各地区都有发生;此外杭州市、湖州市以及绍兴市均检测出了PAMV。对检测到的PVS、PVH、PVM和PAMV外壳蛋白(coat protein,CP)序列进行系统发育分析表明,这些病毒均存在地域差异性。本研究为浙江省马铃薯病毒的防控奠定了重要基础。     

Spread of potato virus Y in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity

The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus Y^NTN (PVY^NTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVY^NTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses.     

Systemic Potato virus YNTN infection and levels of salicylic and gentisic acids in different potato genotypes

Endogenous levels of free and conjugated salicylic (SA) and gentisic (GA) acids, both putative signal molecules in plant defence, were analysed in order to investigate their involvement in the resistance of four potato ( Solanum tuberosum ) genotypes with different susceptibilities to Potato virus Y^NTN (PVY^NTN) infection: the highly susceptible cv. Igor and its extremely resistant transgenic line, the extremely resistant cv. Sante and the tolerant cv. Pentland Squire. The lowest levels of free and conjugated SA were observed in the extremely resistant cv. Sante, while free GA, which was detected in all the other varieties, was absent. The extremely resistant transgenic cv. Igor contained the highest basal total SA level and the lowest level of total GA of all four cultivars. In susceptible cv. Igor, but not in resistant transgenic cv. Igor, a systemic increase of free SA was measured 1 day postinfection (dpi). Even more significant increases of free and conjugated SA and GA were detected 11 dpi when systemic symptoms appeared. In inoculated but not in upper noninoculated leaves of resistant transgenic cv. Igor, significant increase of SA conjugates occurred, but not before 11 dpi. The increase of SA and GA in susceptible cv. Igor could contribute to the general elevated levels of phenolic compounds as a response to stress caused by virus infection. It appears that basal levels of SA and GA do not correlate with resistance to PVY^NTN in potato plants.     

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.     

一株PVYNTN-NW黑龙江马铃薯分离物的检测鉴定

 马铃薯Y病毒(Potato virus Y,PVY)是马铃薯、烟草等茄科作物上的重要病毒,在与寄主共同进化过程中产生了许多株系。本文从黑龙江马铃薯样品中得到PVY分离物A12。ELISA结果表明A12被PVY^O的单克隆抗体特异性识别。A12开放阅读框为9 186个核苷酸,编码3 061个氨基酸,与SYR-II-Be1分离物的核苷酸和氨基酸序列一致率均最高,分别为98.3%和99.2%。系统发育分析发现A12与PVY^NTN-NW株系SYR-II型的分离物聚类到一起;重组分析表明,A12是N-605和O_z的重组体,重组类型与SYR-II-Be1相同。综合以上结果表明,A12属于PVY^NTN-NW株系SYR-II型。但与常见PVY^NTN-NW株系分离物在珊西烟引起叶脉坏死不同,A12产生花叶症状。A12辅助成分-蛋白酶在182位和245位的氨基酸均为精氨酸,而其它PVY^NTN-NW株系分离物为赖氨酸。本研究结果可为黑龙江马铃薯PVY的早期检测和有效防控提供理论指导。