辣椒抗马铃薯Y病毒(PVY)研究进展

马铃薯Y病毒(Potato virus Y,PVY)是限制辣椒生产的主要病毒病害之一,该病毒是马铃薯Y病毒组(Potyvirus)的典型代表,能够侵染辣椒等茄科作物,造成重大的经济损失。应用抗病资源、选育抗病品种是克服PVY对辣椒危害最为经济有效的方法。文中综述了辣椒PVY病毒的发现、流行和检测与分类,并针对辣椒抗PVY育种工作描述了抗性鉴定方法、抗原筛选、抗性遗传、分子定位和标记开发的研究进展,介绍了PVY抗性基因与病毒的互作机理,并就辣椒抗PVY育种中存在的问题及未来研究方向进行了讨论。     

多抗马铃薯病毒及类病毒的amiRNA双元表达载体的构建及遗传转化

马铃薯病毒及类病毒病害严重影响了中国马铃薯的生产。为了培育抗病毒及类病毒的马铃薯新材料,本研究采用人工miRNA(artificial microRNA,amiRNA)策略,以马铃薯X病毒(potato virus X,PVX)与马铃薯Y病毒(potato virus Y,PVY)编码沉默抑制子P25蛋白和HC-pro蛋白的基因为靶标,利用5'RACE技术对P25基因、HC-Pro基因潜在的剪切热点进行预测。再采用overlapping PCR技术,以拟南芥mi R159a前体为骨架,设计特异性引物,构建分别靶向P25和HC-Pro基因的amiRNA表达载体。同时构建靶向马铃薯内源基因Virp1的amiRNA表达载体,将构建完成的三个amiRNA表达载体串联获得amiRNAs三联体,分别转入根癌农杆菌菌株EHA105,并用农杆菌介导法转化马铃薯品种克新13。经PCR鉴定,结果表明,目的基因片段已成功转入马铃薯中。     

RYSC3和Rxsp分子标记在马铃薯抗病育种中的应用

马铃薯Y病毒(PVY)和马铃薯X病毒(PVX)是对马铃薯的产量和品质影响最主要的两种病毒,马铃薯中含有对PVY和PVX具有极端抗性的基因Ryadg和Rx,聚合抗性基因是防治PVY、PVX对马铃薯生产影响最有效的策略。本研究利用与Ryadg和Rx紧密相连的分子标记RYSC3和Rxsp,对青海省主栽马铃薯品种青薯9号与大西洋品种及其杂交F1代进行了标记检测。结果显示,亲本青薯9号仅含RYSC3标记,亲本大西洋仅含Rxsp标记;在198个基因型的F1代分离群体中,含RYSC3标记的材料有91份,含Rxsp标记的材料有86份,同时含Rxsp和RYSC3标记的材料共41份;RYSC3和Rxsp标记基因型的分离比例在5%显著水平下经卡平方检验都符合孟德尔分离比例1:1,抗病亲本均属于单显性基因型,基因型符合Rrrr×rrrr。本研究以期为青海省马铃薯抗病毒新品种的选育和早期鉴定提供科学依据。     

抗马铃薯Y病毒的烟草种质资源抗性分析

为了解我国烟草种质资源对马铃薯Y病毒(Potato virus Y,PVY)的抗性状况,为烟草抗病育种的亲本选择提供信息,本研究通过苗期PVY汁液摩擦接种,从500多份烟草种质资源中筛选出假川烟等15份抗PVY资源。与va位点缺失的抗PVY品种NC55杂交F1的PVY抗性鉴定结果表明,假川烟、Havana K-2-24的PVY抗性与NC55的va位点等位。e IF4E-1基因特异分子标记检测表明,假川烟、Havana K-2-24、Yellow speicial和C151的PVY抗性与e IF4E-1基因缺失有关。本研究鉴定的抗PVY烟草种质资源,可为种质资源的育种利用提供参考。     

中国农科院烟草研究所在烟草马铃薯Y病毒抗性基因挖掘方面取得新进展

正日前,从中国农业科学院烟草研究所获悉,烟草马铃薯Y病毒病抗性基因挖掘研究取得新进展,揭示了烟草响应马铃薯Y病毒的生物学过程,锁定了6个重要抗性基因。烟草马铃薯Y病毒病是由马铃薯Y病毒(Potato virus Y,PVY)引起的一种烟草系统性侵染病害,该病毒是马铃薯Y病毒科(Potyviridae)马铃薯Y病毒属(Potyvirus)的典型成员,在生产中对烟草品质造成严重影响。近年来,     

贵州马铃薯Y病毒外壳蛋白基因的克隆与遗传分析

本研究从中国贵州不同海拔地区采集感染马铃薯Y病毒(PVY)病害叶片,利用试管捕捉RT-PCR(TC-RT-PCR)方法克隆到长度为804 bp的8个目的片段,序列分析表明为PVY外壳蛋白(PVY-CP)基因.通过与已经发表的PVY-CP基因序列进行比较和遗传分析,并根据PVY的CP基因全长和5'端序列构建系统发育树,结果表明,本研究分离的PVY,7株为O株系,1株为NTN株系.     

烟草PVY抗性的遗传分析与分子标记筛选

本文以抗马铃薯Y病毒(简称PVY)的烟草品种RY5,感病品种Coker176为亲本,构建F1、正反交F2和正反交BC1群体,苗期摩擦接种PVY的抗性遗传分析结果表明,接种后第21d群体PVY抗性数据符合孟德尔单基因隐性质量性状的遗传模型。接种后第28d群体PVY抗性数据偏离孟德尔单基因隐性质量性状的遗传模型。提取F2代群体中抗病和感病单株DNA,从多条RAPD引物和一对SCAR引物中,筛选出两个紧密连锁的分子标记。RAPD标记O12V3695与RY5的抗病基因对应的显性等位基因位点(Va)间的遗传距离为2.10cM,而SCAR标记与Va间的遗传距离为2.52cM,这两个分子标记可用于抗PVY抗性育种。     

烟草马铃薯Y病毒病的发生原因与防治对策

烟草马铃薯Y病毒病(PVY)近年来在安徽省的发生与危害日益严重,目前已成为安徽烟区最主要的病毒病.本文分析了安徽烟区近年来烟草马铃薯Y病毒病的发生特点及流行原因,并提出了相应的防治对策.     

PVY引起马铃薯块茎细胞坏死的相关基因研究

本研究采集健康马铃薯块茎和被PVY侵染的马铃薯块茎为材料,选取马铃薯RCCR基因和LSD1基因为目标基因。首先提取总RNA后通过反转录PCR(RT-PCR)、琼脂糖凝胶电泳,证明RCCR基因和LSD1基因成功获得,然后进行荧光定量PCR(qPCR)测定在PVY侵染的马铃薯块茎中RCCR基因和LSD1基因的相对表达量,最后又进行了目标基因编码蛋白的结构分析。结果表明,这2个基因的相对表达量均有所下降,推测这2个基因与马铃薯块茎细胞坏死相关,为之后研究PVY引起马铃薯块茎细胞坏死的相关分子机制做了前期探索,为今后的分子育种和抗病药物筛选提供了可选的靶点。     

脱毒马铃薯试管苗快繁及微型薯诱导技术

马铃薯是无性繁殖作物,在生产繁殖过程中极易感染病毒,并世代传递,病毒为害逐年加重,产量逐年下降。侵染马铃薯的主要病毒有马铃薯卷叶病毒（PLRV）、马铃薯Y病毒（PVY）、马铃薯X病毒（PVX）、马铃薯A病毒（PVA）、马铃薯S病毒（PVS）、     

Early selection for extreme resistance to potato virus Y and tobacco etch virus in potato using a β-glucuronidase-tagged virus

The Ry_sto gene from Solanum stoloniferum introduced into potato cultivars (Solanum tuberosum L. ssp. tuberosum) confers resistance to potato virus A, potato virus V and potato virus Y (PVY). In addition to PVY, tobacco etch virus (TEV) and a TEV construct that encodes β‐glucuronidase (TEV‐GUS) were inoculated to determine the inheritance of resistance to these viruses in progenies obtained from potato cultivars containing the Ry_sto gene. While cultivars ‘Karlena’ and ‘Delikat’ were susceptible, ‘Bettina’ and clone 927eY were resistant to PVY, TEV and TEV‐GUS, as determined by enzyme linked immunosorbent assay, biotest and GUS assay, respectively. The segregation ratios obtained from the progenies of ‘Bettina’בDelikat’ and 816eY בKarlena’ indicate that resistances to PVY and TEV are governed by one dominant gene or two genes tightly linked in coupling phase. Evidently, Ry_sto confers broad spectrum resistance to potyviruses. TEV resistance could be reliably detected 4 days after inoculation with the TEV‐GUS construct by GUS assay. Therefore, the GUS‐tagged TEV construct can be used for early selection for resistances based on the gene Ry_sto or closely linked genes.     

Elucidation of virus-host interactions to enhance resistance breeding for control of virus diseases in potato

Potato virus Y (PVY) and Potato mop-top virus (PMTV) are viruses whose geographical distribution is expanding and economic losses are increasing, in contrast to most of other viruses infecting potato crops. Most potato cultivars lack broad-spectrum resistance to the new, genetically complex strains of PVY, and no efficient resistance to PMTV is known in potato. Control of the vectors of these viruses is not an efficient or possible strategy to prevent infections. Studies on molecular virus-host interactions can discover plant genes that are important to viral infection or antiviral defence. Both types of genes may be utilized in resistance breeding, which is discussed in this paper. The advanced gene technologies provide means to fortify potato cultivars with effective virus resistance genes or mutated, non-functional host factors that interfere with virus infection.     

Natural Genes and Mechanisms for Resistance to Viruses in Cultivated and Wild Potato Species (Solanum spp.)

The types of resistance to viruses in potato are immunity, extreme resistance coded for by the R genes and hypersensitivity coded for by the N genes. Immunity prevents the establishment of virus infection, whereas extreme resistance is expressed as an extremely low virus titre resulting from restricted virus multiplication or localization of virus infection in the plant without obvious necrotic reactions. Expression of the N genes can also lead to localization of the virus infection followed by necrosis at the infection site. Tolerance to virus infection and resistance to inoculation and virus vectors are useful components of field resistance. Virus or virus strain specific resistance genes have been identified in cultivated and wild potato species and transferred to cultivated potato. However, combining resistance to the three globally most important viruses PLRV, PVY and PVX has proved difficult, and new sources of broad spectrum resistance to viruses are required for potato breeding. New molecular techniques will enhance the isolation of the natural resistance genes and studies of gene regulation.     

马铃薯Y病毒株系分类及中国大田马铃薯感染的马铃薯Y病毒株系谱研究进展

为了更科学、合理地检测、检疫马铃薯Y病毒(PVY)株系,和全面掌握侵染中国大田马铃薯的PVY株系情况,总结了PVY株系的分类方法,分析了中国大田马铃薯感染的PVY株系谱。认为目前广泛使用的血清型划分法、基因组重组与否划分法、PTNRD效应划分法、过敏反应互作基因划分法,以及Singh等提出的株系群划分法都无法对PVY株系予以完全合理的划分。提出有必要鉴定PVY基因组中决定马铃薯病理效应的基序并通过对基序测序来鉴定PVY株系,通过对株系间基因组差异区段的短序列测序来鉴定PVY株系是目前最接近基序测序分类法的思路。指出感染中国大田马铃薯的PVY目前有PVY^O、PVY^N、PVY^NTN、PVY^N:O、PVY^N-Wi和PVY^NW这6种株系,隶属于2个株系群PVY^O和PVY^N,其中PVY^N株系和PVY^N株系群病样占绝大多数,以非重组株系为主,但重组株系已占一定比例,其发展趋势值得关注。     

Quantity production of anther-derived haploids from a multiple disease resistant tobacco hybrid. I. Frequency of plants with resistance or susceptibility to tobacco mosaic virus (TMV), potato virus Y (PVY), and root knot (RK)

Summary Methods are described for producing large numbers of haploid plantlets from anthers of a flue-cured tobacco hybrid with monogenic resistance to tobacco mosaic virus, (TMV), potato virus Y (PVY) and root knot (RK), respectively. Additional details are given on colchicine treatment for converting haploids to doubled haploids (DH's) and on the frequency of spontaneous DH's among untreated plantlets. Disparate genetic ratios of TMV-resistant to TMV-susceptible plants were obtained among colchicine-treated haploid plantlets, induced DH's and untreated haploids when compared with F₂ and BC₁ progenies. Haploids (gametes) with the gene for TMV resistance occurred more frequently than expected and plantlets with the gene for RK resistance occurred less frequenctly than expected. Transmission of the gene for PVY resistance differed only slightly from Mendelian expectations. These unexpected ratios, in addition to the frequent occurrence of plastid chimeras among anther-derived plantlets, strengthened our conviction that haploidy is somehow associated with mutation.Joint contribution from the Departments of Genetics, Crop Science and Plant Pathology, North Carolina State University, and the Tobacco Research Laboratory, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, Oxford, North Carolina. Paper No. 5576 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, North Carolina.     

4种病毒抑制剂对昭通烟草番茄斑萎病毒病和马铃薯Y病毒病的防治效果

番茄斑萎病毒病(Tomato spotted wilt virus, TSWV)和马铃薯Y病毒(Potato virus Y,PVY)是云南烟草生产上的主要病毒病,本研究旨在筛选有效的药剂防控烟草番茄斑萎病毒病和马铃薯Y病毒病,从而减少烟草经济损失。通过小区试验,分别研究了1%香菇多糖水剂、3%超敏蛋白微粒剂、5%氨基寡糖素水剂和2%嘧肽霉素水剂对田间烟草番茄斑萎病毒病和马铃薯Y病毒病的防治效果。结果表明：试验所选的4种病毒抑制剂对烟草番茄斑萎病毒病和马铃薯Y病毒病均具有一定的防控作用,1%香菇多糖、3%超敏蛋白、5%氨基寡糖素和2%嘧肽霉素对烟草TSWV的防治效果分别为：31.11%~39.90%、54.34%~59.91%、8.69%~38.21%和22.75%~43.32%,4种药剂对烟草PVY的防治效果分别为55.08%~71.06%、68.10%~82.24%、37.76%~55.36%和58.07%~68.86%,其中,3%超敏蛋白的防效最好,其烟草TSWV和PVY防治效果分别高于50%和60%,其次是2%嘧肽霉素,对烟草TSWV和PVY防治效果分别高于22%和58%。防治烟草番茄斑萎病毒病和马铃薯Y病毒病以3%超敏蛋白微粒剂的防治效果最佳,其次是2%嘧肽霉素水剂。     

马铃薯抗病毒遗传及与产量性状的关系

本文研究了马铃薯对卷叶和花叶病毒抗性的遗传及与产量性状的关系.结果表明,抗病性与群体产量没有显著的遗传相关性,抗病性主要通过保证马铃薯植株的正常生长发育而间接保持群体的高产稳产性能.马铃薯对卷叶病毒抗性、株高、茎粗、有效株率和主要产量性状的遗传主要由亲本的累加基因效应决定,且同一亲本在不同性状上的一般配合力效应差异较大.因此,抗病高产育种中各目标性状的协调选择是十分重要的.群体有效株率和茎粗的选择可使抗病性和丰产性同步提高.马铃薯对花叶病毒的抗性、加性和非加性作用同时存在.群体抗病性、有效株率和产量的遗传进度高,选择潜力大.     

一步法RT-PCR和两步法RT-PCR对马铃薯病毒诊断研究的比较

本研究采用试剂盒和Total试剂两种方法从马铃薯叶片和茎秆中提取总RNA,同时设计了一步法RT-PCR和两步法RT-PCR的反应体系,依据马铃薯X病毒、马铃薯Y病毒、马铃薯S病毒、马铃薯A病毒以及马铃薯卷叶病毒的基因组RNA保守序列设计特异性引物对,并运用两种RT-PCR反应体系对染病的马铃薯进行病毒检测,结果与报道的这五种马铃薯病毒核苷酸序列进行BLAST比对, 表明扩增的五种马铃薯病毒靶带的序列与靶序列完全一致。两种诊断方法对马铃薯病毒核酸进行浓度检测,结果证明,两步法RT-PCR具有较高灵敏性。但这两种方法均可快速、简便、有效地诊断马铃薯病毒。     

马铃薯Y病毒的株系分化及检测方法研究进展

马铃薯Y病毒是烟草上的主要病毒之一。目前中国关于PVY分子变异的报道还较少,但变异株系的出现应该引起科研工作者的高度重视。笔者简述PVY的株系分化进程,并从检测技术的优缺点及应用等方面总结PVY检测的指示植物法、酶联免疫法、聚合酶链式反应、核酸杂交等几种方法。指出随着植物种质资源的引进和生态条件的改变,PVY变异株系将不断出现,这对株系检测方法提出了更高的要求,因此在实际应用中要综合运用各种检测方法提高检测的准确性。