马铃薯纺锤块茎病

马铃薯纺锤块茎病吕文河（东北农业大学作物遗传育种系哈尔滨１５００３０马铃薯纺锤块茎病也叫马铃薯纤块茎病，它是由马铃薯纺锤块茎类病毒（Ｐｏａｔｏｓｐｉｎｄｌｅｔｕｂｅｒｖｉｒｏｉｄ，简称ＰＳＴＶ）引起的一种传染性病害。该病害发现于１９２２年，人们一直认...     

我国北方一季作区马铃薯纺锤块茎类病毒病的发生与防治

2003年的马铃薯生育期间,农业部薯类产品质量监督检验测试中心(张家口)的全体工作人员在农业部的统一安排布置下,对我国华北地区的内蒙古、山西和河北省的脱毒马铃薯主栽区域进行了田间实地抽样调查.从调查的结果看,华北地区马铃薯罹感病毒病的类型主要有普通花叶病毒病(PVX)、重花叶病毒病又称条斑坏死(PVY)、卷叶病毒病(PLRV)和纺锤块茎类病毒病(PSTVd).但对马铃薯产量影响较大,世代传递力强,茎尖分生组织培养难以脱除的是马铃薯纺锤块茎类病毒.这对我国马铃薯病毒病研究提出了新的课题.     

马铃薯生产中马铃薯纺锤块茎类病毒的综合防治

马铃薯纺锤块茎类病毒是影响中国马铃薯生产重要的检疫性病害。该病害具有广泛的自然寄主,可侵染马铃薯、番茄和辣椒等重要作物。该病害可通过植物学种子、花粉或卵细胞、无性繁殖以及机械等方式传播。马铃薯纺锤块茎类病毒可造成马铃薯产量降低,块茎畸形、变小,商品性和商品薯率下降。该病害主要可通过培育抗病品种、使用脱毒种薯、加强种质资源和种薯卫生管理、加强检验检疫以及实行马铃薯种薯质量认证制度等措施进行综合防治。     

用聚丙烯酰胺凝胶电泳鉴定马铃薯纺锤块茎类毒病(综述)

五十多年前,就已经发现了马铃薯纺锤块茎病。半个世纪以来,由于缺少适宜的指示植物和鉴定方法,与同一时期发现的其它马铃薯病毒病相比,研究进展得相当缓慢。1964年,Raymer等人发现番茄(Lycopersicum esculentum Mill.)可以作为马铃薯纺锤块茎类病毒(PSTV)的指示植物,但后来Fernow发现,由于存在一个在番茄上不表症状的PSTV     

准确、简易的检测马铃薯纺锤块茎类病毒的双向聚丙烯酰胺凝胶电泳技术

准确、简易的检测马铃薯纺锤块茎类病毒的双向聚丙烯酰胺凝胶电泳技术姜成模，金顺福，崔哲官，崔永一（延边州农业科学研究所龙井市１３３４００）马铃薯纺锤块茎类病毒（ＰｏｔａｔｏＳｐｉｎ－ｄｌｅＴｕｂｅｒＶｉｒｏｉｄ．ＰＳＴＶ－ｄ）是危害我国马铃薯生产的重要...     

马铃薯纺锤块茎类病毒不同株系及不同接种方法对产量的影响

马铃薯纺锤块径类病毒(Potato spindle tuber viroid)是危害黑龙江省马铃薯生产的一种重要病害。为了查明不同类病毒变株对主栽品种克新1号和4号所引起的产量损失和症状反应,应用往复聚丙烯胺凝胶电泳法(Peturn-Polyacmdegel electophoresis)鉴定类病毒。试验结果表明,接种当年,接种类病毒的处理平均减产37%,按商品产量计算减产达47%。当年表现症状的植株平均为59.6%,块茎产生症状的植株平均为57.4%。用于接种的3个马铃薯纺锤块茎类病毒变株的致病力,以当地的类病毒弱系为最强,其次为北美的类病毒强系,北美的类病毒弱系的致病力较弱。类病毒对产量的影响与马铃薯品种和类病毒变株的组合有极为密切的关系。克新1号接种当地的类病毒弱系时薯块严重畸变,商品产量只为不接种对照的21%。用往复电泳法检测各个试验处理被类病毒侵柴的情况,芽接种处理,于出苗后1周即可检测到类病毒,叶接种处理,于接种后2周即能检测到类病毒。出苗后7周,植株中类病毒浓度达到高峰,随即急剧下降,到出苗后10周,大部分接种植株已检测不到类病毒。块茎直径达1厘米时,即能检测到类病毒,随着块茎膨大,类病毒浓度也相应增高,到收获前,块茎中的类病毒含量只略低于植株含类病毒高峰期的浓度。用于本试验的克新1号、4号无类病毒和主要马铃薯病毒的核心材料,在1988年已交付给省内外的良种场繁殖使用。于当年秋季抽样检测克山第二良种场连续种植二年的种薯,当年种植试管苗生产的种薯,以及试管苗等,均未检测到类病毒。     

类病毒在花卉类种子组织内分布的检测

<正>花卉研究所松下阳介研究员以番茄花芽形成组织中两种类病毒的分布差异为主题,报告了其研究结果。1类病毒是病原由马铃薯纺锤块茎类病毒(PSTVd)侵染马铃薯种子内部的胚珠等生殖器官可推测,PSTVd病毒不仅污染番茄种子的表面,甚至还可侵入到种子内部组织。有关PSTVd导致园艺花卉种子病害已有报道,期待本文对花卉种子类病毒的侵染分布研究及其检测技术的开发有所帮助。2确认新病害     

应用NASH方法检测马铃薯类病毒(PSTVd)

1　前　言马铃薯纺锤块茎类病毒 (ＰｏｔａｔｏＳｐｉｎｄｌｅＴｕｂｅｒＶｉｒｏｉｄ ,简称ＰＳＴＶｄ)是引起马铃薯纺锤块茎病的原因 ,该病害发现于 192 2年 ,但直到 196 7年及以后的研究中才发现该病由一种叫类病毒的病原引起的。分布十分广泛 ,是危害我国北方马铃薯生产的重要病害之一 ,严重影响马铃薯的质量和产量 ,一般弱系可造成减产 2 0 %～ 35 % ,强系可达 6 0 %。对其检测有指标植物方法 (利用指示植物鲁格斯番茄Ｒｕｔｇｅｒｓ ,新莨蓿等 ) ,在我国 ,目前较常用的检测方法为聚丙烯酰胺凝胶电泳法 ,而随着分子生物学的发展 ,杂交…     

应用反向聚丙烯酰胺凝胶电泳检测马铃薯纺锤块茎类病毒

1 前言马铃薯纺锤块茎类病毒(PSTV)在马铃薯上广泛存在,目前几乎遍及世界各地,对马铃薯生产威胁极大。     

马铃薯纺锤块茎类病毒检测技术比较与选择

马铃薯纺锤块茎类病毒(PSTVd)是威胁我国马铃薯生产的重要病害之一,目前有多种技术可应用于PSTVd的检测,如生物学方法、电子显微镜技术、往返聚丙烯酰胺凝胶电泳(R-PAGE)、核酸斑点杂交(NASH)、RT-PCR和FQ RT-PCR。这些方法各有利弊,互相补充,构成了PSTVd的检测技术体系。日常工作中,应根据自身条件及目的等科学地选择适宜的检测技术,确保脱毒种薯/种苗的质量。     

Detection,distribution and long-term persistence of potato spindle tuber viroid in true potato seed from heilongjiang,China

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.     

Survey of potato spindle tuber viroid in seed potato growing areas of the United States

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.     

Chemiluminescent detection of potato spindle tuber viroid in true potato seed using a digoxigenin labelled DNA probe

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.     

Detection of potato viruses A,M, S,X, Yand leafroll and potato spindle tuber viroid from tissue culture plantlets using single leaf discs

Using enzyme-linked immunosorbent assay (ELISA) and dotimmunobinding assay (DIBA) for potato viruses A (PVA), M (PVM), S (PVS), X (PVX), Y^N (PVY^N), Y^O (PVY^O) 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.     

Detection on polyacrylamide gel of a diagnostic nucleic acid from tissue infected with potato spindle tuber viroid

A diagnostic procedure for potato spindle tuber disease is described which allows detection of the causal viroid in small amounts of potato and tomato tissue. The method involves extraction of cellular nucleic acids, their separation by polyacrylamide gel electrophoresis and staining with toluidine blue O. The procedure has been used to index greenhouse and field potatoes for mild and severe strains of the pathogen     

Detection of potato spindle tuber viroid by nucleic acid spot hybridization: Evaluation with tuber sprouts and true potato seed

A diagnostic test for the potato spindle tuber viroid (PSTV) that is based on hybridization of highly radioactive, recombinant DNA complementary to PSTV with PSTV bound to a nitrocellulose membrane and autoradiographic detection of the resulting DNA-RNA hybrids has been evaluated with tubers from 20 potato clones maintained at the International Potato Center (CIP) and with true seed obtained from healthy or PSTV-infected potato plants. The nucleic acid spot hybridization test detected PSTV not only in tuber sprouts from 10 clones that had previously tested positive in polyacrylamide gel electrophoretic (PAGE) analysis, but also in tuber sprouts from 9 clones that had previously tested negative in PAGE analysis. Further tests confirmed the presence of PSTV in these clones. The spot hybridization test detected PSTV in mixtures of seed extracts equivalent to as few as one seed from a PSTV-infected plant and 80 seeds from healthy plants. The spot hybridization test was shown to be more sensitive and reliable than PAGE analysis; it is suitable for the testing of large numbers of samples with a minimum expenditure of labor and materials.     

Ineractions between a mild and a severe strain of potato spindle tuber viroid in doubly infected potato plants

Potato (Solarium tuberosum) plants co-infected with a mild and a severe strain of potato spindle tuber viroid (PSTVd) were analyzed by return-polyacrylamide gel electrophoresis for the presence of both strains in vegetative and reproductive plant parts. Both strains were detected in the anthers, flowers, inflorescences, leaves, ovaries, ovules, petals, pistils, roots, sepals, stolons and tubers. Only mild strain was detected from pollen in the cultivars tested. True potato seed (TPS) were not doubly infected when they were obtained from co-infected maternal parent plants pollinated with pollen from healthy plants. Also, when maternal plants infected with severe strain were pollinated with pollen from healthy plants or from those infected with the mild strain, TPS were not doubly infected. A small number of TPS with double infection was obtained when co-infected or mild-strain-infected plants were pollinated with pollen containing the severe strain (singly or doubly infected). The number of TPS containing mild strain predominated in a ratio of 7:1 over TPS containing the severe strain. This study indicates that segregation of strains from doubly-infected plants probably takes place during pollen formation and persists through seed development.