分子标记用于赤眼蜂分子监测的研究

通过对松毛虫赤眼蜂（Trichogramma dedrolimi Matsumura）以及玉米螟赤眼蜂（Trichogramma ostriniae Pang et Chen）rDNA-ITS2基因的克隆测序，并联机检索GenBank核酸序列库中其它赤眼蜂的相关序列，利用核酸分析软件找到松毛虫赤眼蜂和玉米螟赤眼蜂ITS2序列中一段有鉴别意义的标志，并据此设计出蜂种的特异PCR引物，实现了松毛虫赤眼蜂和玉米螟赤眼蜂rDNA特异区带的PCR扩增。研究表明该特异区带具有种的特异性。我们认为该分子标记技术可用于目前我国生防中常用的松毛虫赤眼蜂和玉米螟蒌眼蜂的分子监测，如田间种群动态监控和寄生效果评估。     

阿魏蘑、白灵菇及杏鲍菇亲缘关系的研究

利用PCR产物克隆测序测定了阿魏蘑、白灵菇与杏鲍菇rDNA内转录间隔区(ITS)的序列,并对所测序列进行分析.结果表明,阿魏蘑、白灵菇和杏鲍菇三者之间具有较近的亲缘关系,白灵菇与杏鲍菇具有非常近的亲缘关系,但阿魏蘑、白灵菇和杏鲍菇为3个不同的种.     

白花柽柳nrDNA的ITS序列片段研究

运用PCR直接测序法 ,对白花柽柳 (TamarixalbiflonumM .T .Liu .)的nrDNA的ITS区 (包括ITS -l,5 .8SrDNA和ITS - 2 )进行序列测定。结果表明 ,该植物的ITS序列和长穗柽柳 (Tamarixelongata)的序列差别不大 :ITS - 1片段的长度都是 2 5 9bp ;ITS - 2片段的长度同为 2 44bp ;G C百分含量仅相差 1% ;碱基差异共发生 18处。由ITS及 5 .8SrDNA序列分子证据看 ,白花柽柳更可能是长穗柽柳的变异。     

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。     

昆虫病原线虫rDNA多态性分析

本文对国内外昆虫病原线虫斯氏属和异小杆属的47个品系进行rDNA—ITS PCR—RFLP分析，研究其DNA多态性，并构建了分子系统发育树状图。各品系的ITS区无明显的长度差异，PCR—RFLP分析将47个品系分为斯氏属和异小杆属两大类，两属线虫又分别分为11组和4组。所得结果丰富了ITS PCR—RFLP图谱库，为弄清我国的昆虫病原线虫与国外种类的分子系统发育关系及未定名线虫的鉴定提供重要依据，同时为筛选适合的线虫种类防治害虫奠定基础。     

Identification of Rhizoctonia solani AG 1-IB in Lettuce, AG 4 HG-I in Tomato and Melon, and AG 4 HG-III in Broccoli and Spinach, in Brazil

Fungi isolated in Brazil, from lettuce, broccoli, spinach, melon and tomato, were identified as Rhizoctonia solani. All lettuce isolates anastomosed with both AG 1-IA and IB subgroups and all isolates from broccoli, spinach, melon and tomato anastomosed with AG 4 subgroup HG-I, as well as with subgroups HG-II and HG-III. DNA sequence analyses of ribosomal internal transcribed spacers showed that isolates from lettuce were AG 1-IB, isolates from tomato and melon were AG 4 HG-I, and isolates from broccoli and spinach were AG 4 HG-III. The tomato isolates caused stem rot symptoms, the spinach, broccoli and melon isolates caused hypocotyl and root rot symptoms on the respective host plants and the lettuce isolates caused bottom rot. This is the first report on the occurrence in Brazil of R. solani AG 4 HG-I in tomato and melon, of AG 4 HG-III in broccoli and spinach and of AG 1-IB in lettuce.     

小麦苗枯病菌的ITS分析及PCR检测

 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。     

A coat protein transgene from a Scottish isolate of potato mop-top virus mediates strong resistance against Scandinavian isolates which have similar coat protein genes

Resistance tests were made on seedlings of transformed lines of Nicotiana benthamiana which contain a transgene encoding the coat protein (CP) gene of a Scottish isolate of potato mop-top virus (PMTV). This transgene has been reported to confer strong resistance to the PMTV isolate from which the transgene sequence was derived and also to a second Scottish isolate. Plants of lines of the transgenic N. benthamiana were as resistant to two Swedish and two Danish PMTV isolates as to a Scottish isolate, and of five lines tested, greater than 93.5% of transgenic plants were immune. The coat protein gene sequences of these four Scandinavian isolates were very similar to those of the two Scottish isolates. The greatest divergence between the isolates was three amino acid changes and there was less than 2% change in CP gene nucleotide sequence. It is concluded that the PMTV CP transgene used in these experiments could confer resistance against isolates from different geographical areas because it is becoming apparent that the CP genes of PMTV isolates are highly conserved.     

Isolation of two strains of a new Tombusvirus (Havel river virus, HaRV) from surface waters in Germany

Two virus isolates from water samples — one from a small stream in South Western Germany and another one from the Havel river in North Eastern Germany c. 500 km away, proved to be strains, named S and H, respectively, of a new Tombusvirus for which the name Havel river virus (HaRV) had been suggested previously in a brief account. Immunoelectron microscopical decoration tests and sequence comparisons of the coat proteins indicated that the two HaRV strains are only distantly related to known Tombusviruses. The closest relationships were found to Cucumber necrosis virus. Nothing is known about their natural hosts. Because the S strain of HaRV was isolated in a woody area from a small stream close to its origin, they may be pathogens of trees or wild plants in such habitats.