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湖南省马铃薯主产区马铃薯病毒种类及流行分析 总被引：2，自引：0，他引：2
马铃薯是世界第四大粮食作物,其病毒病危害严重。2010年对湖南马铃薯主产区采集的66个病毒标样进行了RT-PCR检测,结果表明,检测出的马铃薯病毒有马铃薯Y病毒(PVY)、马铃薯卷叶病毒(PLRV)、马铃薯X病毒(PVX)、马铃薯S病毒(PVS)、马铃薯A病毒(PVA)和马铃薯纺锤块茎类病毒(PSTVd)。其中PVS的检出率最高,为54.5%,其次是PVX,检出率为45.5%,PVY的检出率为39.4%,PSTVd和PVA的检出率均为21.2%,PLRV的检出率为18.2%。2~4种病毒的复合侵染现象较为普遍。PVY中重组型PVY占85.7%。 相似文献
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马铃薯纺锤块茎类病毒(PSTVd)的检测与防治研究进展 总被引：1，自引：0，他引：1
详细阐述了马铃薯类病毒(PSTVd)鉴定技术的研究进展,比较了几种鉴定技术(指示植物、往返电泳、核酸斑点杂交及RT-PCR)的优缺点,并对类病毒的防治措施进行了论述。 相似文献
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延安地区马铃薯纺锤块茎类病毒的RT-PCR检测与序列分析 总被引：1，自引：0，他引：1
以陕西省延安地区8个乡镇种植2～3代的马铃薯叶片为材料,Trizol法提取马铃薯叶片总RNA,以已公布的马铃薯纺锤块茎类病毒（PSTVd）基因序列设计合成特异引物,反转录合成cDNA,RT-PCR扩增目的基因片段并电泳检测,回收纯化目的片段,然后进行克隆和测序,采用DNAstar软件分析序列一致性.结果显示：在马铃薯叶片中均扩增到与预期大小一致的目的片段,表明这些乡镇的马铃薯均感染了PSTVd,8个采样点PSTVd的目的片段大小为247～251bp,出现少量碱基的突变或缺失,说明PSTVd在延安地区部分乡镇发生了一定程度的变异,与国内外其他14个地区之间的序列一致性为804% ～992%. 相似文献
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马铃薯病毒病严重影响马铃薯产量和质量,因此,在马铃薯生产中防控马铃薯病毒病尤为重要。用马铃薯A病毒(Potato virus A,PVA)、马铃薯Y病毒(Potato virus Y,PVY)、马铃薯X病毒(Potato virus X,PVX)、马铃薯M病毒(Potato virus M,PVM)、马铃薯S病毒(Potato virus S,PVS)、PVY+PVA、PVY+PVX、PVY+PVS以及PVY+类病毒(Potato spindle tuber viroid,PSTVd)共9种病毒或病毒(类病毒)组合于苗期接种黑龙江省10个马铃薯主栽品种,调查这些品种的株高、产量、大薯率和植株症状的变化。结果表明,复合侵染中PVY和PSTVd和单一侵染中PVY对马铃薯品种高度影响最大。10个供试马铃薯品种对含有PVY侵染的5个处理(PVY、PVY+PVX、PVY+PVA、PVY+PVS、PVY+PSTVd)均表现出了明显的症状。PVY+PVX和PVY+PSTVd复合侵染对10个马铃薯品种的产量和大薯率影响较大。调查了生产中常见病毒病和类病毒在黑龙江省主栽品种上造成的症状和危害,有助于在生产中对病害的识别和防控,降低病害的危害,提高马铃薯的质量和产量。 相似文献
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《中国马铃薯》2016,(1):31-38
采用内蒙古自治区采集的感染马铃薯纺锤块茎类病毒(Potato spindle tuber viroid,PSTVd)的马铃薯材料,克隆了PSTVd的全长基因,命名为内蒙古分离株(PSTVd-IM),对其序列进行分析。结果表明,该序列大小为359 bp,其与俄罗斯的PSTVd分离株(EF044305.1)序列吻合。将该分离物与国内外已经报道的PSTVd株系进行了比较,构建了系统进化树,确定了其分类地位;通过序列比对和二级结构分析,发现PSTVd-IM与已报道的PSTVd三类致病类型株系:强株系(U23058.1)、中间株系(AY937179.1)和弱株系(M14814.1)的同源性分别为97.78%,98.61%和99.44%;且与这三种类型株系在中央保守区、致病区和可变区结构域存在差异,说明这些结构域可能与其致病性相关。PSTVd-IM的序列与二级结构分析有利于明确其来源及致病性的差异位点,对于PSTVd的致病机制的研究具有重要意义。 相似文献
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马铃薯X病毒的RT-PCR和IC-RT-PCR检测 总被引：1，自引：0，他引：1
马铃薯X病毒(Potato virus X,PVX)是侵染马铃薯重要病毒之一,通常引起花叶症状,在田间常与其他病毒混合感染导致马铃薯的毁灭性减产。PVX尚无有效的药剂可以防治,加强对PVX的快速检测是一个亟待解决的课题。本研究应用反转录聚合酶链式反应(RT-PCR)和免疫捕捉反转录聚合酶链式反应(IC-RT-PCR)技术检测马铃薯X病毒。结果表明:IC-RT-PCR方法可检测出稀释至1.0×10-3的粗汁液中的病毒;RT-PCR方法可从稀释至1.0×10-4的总RNA中扩增出特异的目的条带。这两种方法均具有较高的检测灵敏度,均可用于马铃薯X病毒的检测。 相似文献
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《中国马铃薯》2015,(3):162-166
马铃薯X病毒(PVX)、马铃薯M病毒(PVM)和马铃薯A病毒(PVA)是导致马铃薯种薯退化的重要病毒,有时复合侵染,因此建立快速、准确检测体系尤为重要。试验从中、英文文献中查找引物,通过筛选和综合评价,每种病毒确定1对特异性引物,再通过对PCR部分Mg2+、d NTPs、Taq DNA聚合酶浓度梯度优化,最终建立了稳定的三重RT-PCR反应体系,得到长度分别为711、520、273 bp的特异性条带。应用该体系和DAS-ELISA方法同时对田间150份马铃薯毒源样品进行检测,两种方法检测结果吻合,三重RT-PCR体系的灵敏度更高、特异性更强,可以用于马铃薯田间样品检测。 相似文献
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马铃薯类病毒病是威胁世界马铃薯生产的主要病害之一,更影响着发展中国家马铃薯产业的发展。2008~2010年,黑龙江省农业科学院植物脱毒苗木研究所连续3年举办了马铃薯病害检测技术国际培训班,对部分发展中国家从事马铃薯工作的人员进行培训,在培训过程中进行了马铃薯类病毒方面的问卷调查,调查结果显示:这些发展中国家对马铃薯纺锤块茎类病毒比较了解,发病较重的国家不多,但总体检测水平不高,甚至不进行马铃薯类病毒的检测,多数国家不重视马铃薯纺锤块茎类病毒病的防治工作。因此,马铃薯纺锤块茎类病毒病是发展中国家的隐患。 相似文献
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Summary A test procedure for PSTVd is described based on immobilisation of plant sap on filter paper, by dotting or tissue printing
followed by RT-PCR. Tests were carried out using primarily and secondarily infected potato plants, primarily infected in vitro
plants, and potato tubers. Print PCR was shown to be suitable for testing large samples of potato plants whereas dot PCR is
recommended for in vitro plantlets and tuber tissue. Bulking one infected plant to 4 or 9 healthy plants gave reliable results
with secondarily infected potato plants, but sometimes the test failed to detect PSTVd in primarily infected in vitro plants.
Dotted and printed paper squares could be stored at 4°C for at least 2 weeks in Triton X-100 solution or under dry conditions.
Storing at room temperature can lead to unreliable results. 相似文献
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Return-polyacrylamide gel electrophoresis (R-PAGE) and tomato bioassay followed by R-PAGE were compared for the detection of potato spindle tuber viroid (PSTVd) from individual true potato seed (TPS). Both methods detected PSTVd from single TPS. TPS extract formulated as sap or nucleic acids in two different buffers did not affect the percentage of viroid detection on tomato plants. There was some evidence of viroid inhibitor in TPS extracts but not in nucleic acid extracts of TPS. Because R-PAGE is more rapid than the tomato bioassay followed by R-PAGE, the former was used to determine the extent of PSTVd in TPS of China’s Keshan Potato Research Institute breeding material. Over 1700 individual TPS were tested. Twenty-four of the 46 seedlots tested (inbred and outcrossed) contained PSTVd. The viroid was detected in 70% of lots from inbred lines compared to 38% of lots from outcrosses. TPS (20 lots) stored in paper bags at room temperature as far back as 1965 were also tested, and PSTVd was detected in TPS stored for 21 years. 相似文献
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Summary The potential use of a simple, sensitive and non-radioactive method for detecting potato spindle tuber viroid (PSTVd) in germinated
true potato seeds, based on nucleic acid hybridization with a PSTVd-specific DNA probe labelled with digoxigenin, was investigated.
Two simple procedures for the clarification of plant extracts suitable for a non-radioactive detection system were also investigated.
The nucleic acid hybridization technique could detect one PSTVd-infected seed in more than 150 healthy seeds. The benefits
of this non-radioactive detection system are discussed. 相似文献
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Mike Sun Susan Siemsen William Campbell Pablo Guzman Robert Davidson Jonathan L. Whitworth Terry Bourgoin Jeff Axford Willem Schrage Gary Leever Alan Westra Steve Marquardt Hossien El-Nashaar Jeff McMorran Oscar Gutbrod Tom Wessels Robert Coltman 《American Journal of Potato Research》2004,81(3):227-231
Fourteen United States (U.S.) seed potato certification agencies surveyed all U.S. seed potato growing areas for presence of the potato spindle tuber viroid (PSTVd). The survey included general surveillance, which involved searching for the occurrence of PSTVd in state seed potato certification records from 1990 through 2000, and a field survey, which involved testing selected crops for PSTVd infection by nucleic acid dot blot hybridization during 1999 through 2001. No PSTVd incident was documented in any of the state certification records, nor was PSTVd detected in the field surveys. All U.S. seed-growing areas were determined to be free of PSTVd. It is concluded that PSTVd has been eradicated and freedom from potato spindle tuber viroid has been successfully maintained in all of the seed potato growing areas in the United States. 相似文献
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R. P. Singh A. Boucher T. H. Somerville S. Coleman 《American Journal of Potato Research》1996,73(3):101-112
Using enzyme-linked immunosorbent assay (ELISA) and dotimmunobinding assay (DIBA) for potato viruses A (PVA), M (PVM), S (PVS), X (PVX), YN (PVYN), YO (PVYO) and leafroll (PLRV) and nucleic acid spot hybridization (NASH) for potato spindle tuber viroid (PSTVd), virus and viroid were detected reliably from single leaf discs (6 mm) of tissue-culture plantlets. Leaf discs taken from leaf positions (1 to 8) (bottom to top) can be used for detection of all viruses except PLRV where the lower leaves had higher concentrations of virus than the leaves from the upper part of the plantlet. Virus cultures were maintained for 1 to 4 years in several potato cultivars. The levels of virus remained reproducible except for PVM concentration, which was found to be very low in cv. Green Mountain. Using densitometry software, the DIBA spots were quantified and results were comparable to A405 values obtained by ELISA. PSTVd concentration as measured by densitometry from spots of NASH indicated no loss of viroid over 1–4 years in tissue culture in two potato cultivars. 相似文献
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马铃薯Y病毒一步RT-PCR检测试剂盒的研制 总被引：1，自引：0，他引：1
马铃薯Y病毒(Potato virus Y,PVY)对马铃薯的危害最大,可导致马铃薯退化,降低马铃薯产量。解决这一问题的重要途径就是培养脱毒种薯,但是否完全脱毒需要经过检测才能证实。本研究依据PVY CP基因序列设计合成了一对引物PY1、PY2,以带毒样品植物总RNA为模板,在同一个反应中同时加入反转录和PCR反应所需试剂,反应程序中包括反转录和PCR反应所需条件,进行反应扩增,带毒样品扩增得到340 bp的目的条带,而健康对照无此目的条带,从而建立了PVY的一步RT-PCR检测技术,并组装成试剂盒。该试剂盒具有良好的稳定性和特异性,灵敏度可以检测到带毒植物组织下限的6.25μg,高于ELISA(100μg)和NASH(15μg)的灵敏度,虽然和常规方法的灵敏度相同,但更为快速、简便、易于操作,适合脱毒苗和脱毒种薯生产单位做大量样品的检测。 相似文献