草莓角斑病菌传入中国的风险分析

草莓角斑病菌是一种严重为害草莓生产的细菌。本文从其分布状况、寄主、经济重要性、传入与定殖可能性和风险管理难度等方面进行了综合分析评估,结果表明该病菌属于特别危险的有害生物。并根据风险分析的结果提出了防止其传入我国的管理措施。     

应用PCR方法快速检测黄瓜细菌性角斑病菌

黄瓜细菌性角斑病是黄瓜上的一种重要细菌病害,其病原为丁香假单胞菌黄瓜致病变种(Pseudomonas syringae pv.lachrymans),目前未见到该病害特异性PCR检测方法的报道。通过分析丁香假单胞菌(P.syringae)不同致病变种glyceraldehyde-3-phosphate dehydrogenase 1(gap1)基因序列设计得到一对Psl特异性PCR引物。利用该引物对丁香假单胞菌不同致病变种、假单胞菌属其他种及其他属的共46株菌株进行了PCR扩增,结果表明,所有不同来源的12株黄瓜细菌性角斑病菌均得到179bp的目标片段,而所有其他参试菌株均无扩增条带,PCR检测的灵敏度为7.5×103cfu/mL。利用该方法可从接种后发病的黄瓜叶片总DNA中检测到特异条带,而健康叶片无条带。该引物的PCR检测方法可直接用于植株总DNA的检测,无需进行病原菌的分离培养,快速简便,适用于进出境检验检疫及种苗健康检测等。     

黄瓜细菌性角斑病菌交叉引物恒温扩增检测技术

黄瓜细菌性角斑病是我国黄瓜生产上的重要病害之一,其病原菌为Pseudomonas syringae pv.lachrymans。根据该病原菌甘油醛-3-磷酸脱氢基因保守序列设计引物和探针,建立了交叉引物恒温扩增和核酸试纸条检测技术。菌体DNA检测灵敏度可达0.55 ng,纯菌直接检测灵敏度基本可达到单个细菌。所测试的5株黄瓜细菌性角斑病菌和染病黄瓜叶片均为阳性,其他13株对照菌株均为阴性。该方法灵敏度高,且操作简单,对设备要求低等,适合基层实验室应用。     

瓜类果斑病菌胶体金试纸条的检测应用

瓜类果斑病是一种严重为害西甜瓜作物的细菌病害,胶体金试纸条是对其进行现场检测的主要手段.将中国检验检疫科学研究院研制的瓜类果斑病菌胶体金检测试纸条与市场上商品化的美国Agdia公司同类试纸条进行比较,发现两种试纸条的灵敏度和特异性相当,适用于检测出现疑似果斑病症状的叶片和果实;也可用于种子带菌检测,但易出现假阳性结果....     

基于环介导等温扩增技术检测瓜类细菌性果斑病菌

本文利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP),针对西瓜嗜酸菌(Acidovorax citrulli)基因组中Aave_4063和Aave_4064序列设计了2对特异引物Ac-F3/Ac-B3和Ac-FIP/Ac-BIP,建立了西瓜嗜酸菌的LAMP检测体系。利用该体系在65℃保温1h,通过荧光显色即可完成检测。设计的引物特异性强,其检测灵敏度为2.0×101 cfu/mL。该方法为西瓜嗜酸菌的检疫及其所致病害的快速诊断提供了新的技术。     

利用GICA-PCR快速检测瓜类果斑病菌

瓜类果斑病菌(Acidovorax avenae subsp.citrulli,Aac)是瓜类作物上重要的病原细菌,为我国进境植物检疫性有害生物。胶体金免疫层析试纸条方便快捷,应用广泛。该方法使用不当会出现假阳性问题,仅适用于病原菌的初筛检测。本研究将Aac胶体金免疫层析方法(GICA)与PCR技术相结合,建立了GICA-PCR检测方法。检测结果表明,该方法在蛋白与核酸2个层面上从发病西瓜叶片上检测到瓜类果斑病菌,有效解决了试纸条检测的假阳性问题,提高了瓜类果斑病菌检测的准确性,值得推广应用。     

玉米大斑病菌菌丝蛋白电泳分析及抗体检测

 研究表明,玉米大斑病菌1号小种(原命名法为2号小种)产生的毒素对带有Ht1基因的玉米具有高度的致毒活性,且与0号小种(原命名法为1号小种)毒素的成分不同,1号小种毒素含有一种克服Ht1基因玉米的特异性毒性组分。     

免疫检测试纸条法检测西瓜子中的瓜类果斑病菌

将西瓜子进行表面消毒后破碎,用无菌水于4℃浸泡4h,取300μL浸提液加入250mL细菌液体培养基中,28℃150r/min培养3d,取培养液0.7mL与Agdia公司提供的瓜类果斑病菌检测试纸条样品浸提网袋中的缓冲液混匀,取混合液1mL用于免疫检测试纸条检测.该方法可有效检测出西瓜子中是否带有瓜类果斑病菌(Acidovorax avenae subsp.citrulli(Schaad et al.)Willems et al.).     

小麦不孕病菌的检测和鉴定

小麦不孕病菌是国外发生的一种检测危险的性病害,广州局截获该病菌在全国口岸尚属首次。本文介绍了有关检验,鉴定情况,并比较了几个常见近似种之间的差异,最后对不孕病菌的潜在危害性进行了讨论。     

东北春大豆灰斑病菌生理小种鉴定结果初报

 大豆灰斑病病原菌具有高度的变异性,能分化出许多生理小种,美国的Athow 1952年首次报道了大豆灰斑病病原菌的生理分化现象。     

Sensitive real-time PCR detection of Xanthomonas fragariae in strawberry plants

Xanthomonas fragariae , the causal agent of angular leaf spot on strawberry, is a quarantine organism in strawberry propagation material in the European Union. For the reliable screening of planting material for latent infections, a real-time PCR assay based on Taqman® chemistry for the detection of X. fragariae was developed. Primers and probe sequences were based on a DNA fragment amplified by a previously reported X. fragariae -specific technique. The sequence of this genomic fragment had no significant similarity with any published GenBank sequence. Specificity of the designed assay was tested with an extended range of X. fragariae collection strains and isolates, with other Xanthomonas spp. and with unidentified bacterial isolates from strawberry plants. A nested PCR, which until now was the reference method for sensitive detection in planta , cross-reacted with the reference strain of Xanthomonas campestris pv. campestris . In combination with an elaborated DNA extraction procedure, the Taqman® PCR enabled reliable detection down to 300 colony forming units in a 100 mg strawberry leaf sample. The assay offers a new tool for epidemiological research and for sanitary control of plant material with low level or latent infections of X. fragariae .     

Nested Pcr (polymerase chain reaction) for detection of Xanthomonas fragariae in symptomless strawberry plants

Journal of Plant Diseases and Protection - For rapid and sensitive detection of Xanthomonas fragariae, the causal agent of angular leaf spot of strawberry, a nested polymerase chain reaction (Pcr)...     

基于PCR技术的植物病原菌分子定量检测技术研究进展

植物病原菌的菌源量是病害发生和流行的重要因子之一,对其精准的定量测定或检测可大大提高植物病害预测的准确性,本文对实时荧光定量PCR (qPCR)与数字PCR在植物病原菌定量检测、以及基于RNA水平的real-time PCR和基于核酸染料(EMA/PMA)与qPCR相结合的技术在植物病原菌活体定量检测中的应用进行了综述,并展望其在植物病害流行和预测中的应用前景。     

红掌细菌性疫病病原菌的PCR特异性检测

 红掌细菌性疫病(Xanthomonas axonopodis pv. dieffenbachiae,简称Xad)是红掌等天南星科花卉毁灭性病害,该病通过带菌种苗调运不断在我国扩散蔓延,国内尚未有检测方法。通过筛选和重新设计引物,建立了Xad 的PCR 检测方法,结果表明,利用引物Xad-F / Xad-R 进行PCR,能扩增出检测Xad 的特异性DNA 片断,其灵敏度可达1 × 102 CFU / mL,DNA 的最低检出限为0. 44 ng / μL,可用于红掌苗的带菌检测和红掌细菌病害的鉴定。     

Eradication of the first outbreak of Xanthomonas fragariae in the United Kingdom

Xanthomonas fragariae was identified in the UK in strawberry fruiting crops in October 2004. As this pathogen had not been confirmed in the UK before and is listed as a quarantine organism by the EU and EPPO, emergency official action was taken to contain and eventually eradicate this pest. In order to eradicate this disease the affected growers were given the option of either destroying the crop to eradicate the disease immediately or maintaining the infected plants for the life of the crop for fruit production with hygiene measures to prevent the spread of the disease to uninfected crops. The affected growers chose to maintain the crops with hygiene measures to contain the disease. The crops continued to be monitored and no further symptoms were identified in either the infected crop or other crops on the farms.     

Detection of Xanthomonas fragariae in symptomless strawberry plants by nested PCR*

Xanthomonas fragariae spreads in symptomlessly infected strawberry plantlets and a method for detection of latent infections is necessary. It is a very slow‐growing bacterium in culture and is easily overgrown by saprophytic bacteria. Therefore, plating is not a suitable method for detecting low numbers of bacteria in symptomless plants. In addition, selective media are not available. Serological assays like immunofluorescence are useful for testing in‐vitro plants, but they are not suitable for field‐grown plants, as cross reactions are common with the available antisera. For these plants, nested PCR with primers from Pooler ( Pooler et al., 1996 ) and Zimmermann ( Zimmermann et al., 2004 ) has proved to be a valuable method. The method was successfully applied for a survey of strawberry plants from fields in Germany and for testing imported plants (frigo and green plants).     

PCR-based detection of Xanthomonas campestris pathovars in Brassica seed

Xanthomonas campestris is a seedborne bacterium that causes black rot of crucifers. Substantial crop losses may result from the rapid spread of the bacteria under favourable conditions, especially those occurring during seedling production. A PCR-based method has been developed for the rapid and sensitive detection of the pathovars of X. campestris that affect crucifers. Primers were designed to specifically amplify a 619 bp fragment of the hrpF gene from X. campestris . Amplification products were not detected from other Xanthomonas species, or from other pathogenic or epiphytic bacteria occurring on these plants. To avoid false-negative results arising from the presence of amplification inhibitors in plant extracts, primers targeting a 360 bp section of the internal transcribed spacer (ITS) region from Brassica spp. were included in a multiplex PCR. The assay readily detected X. campestris pv. campestris infections in diseased plants and from bacterial colonies isolated on growth media, and was more sensitive and specific than traditional plating methods and a commercially available ELISA. A seed-washing protocol was optimized to allow the detection of a single artificially infected seed among 10 000 healthy seeds using the multiplex PCR.