马铃薯晚疫病菌分子遗传及与寄主互作研究          下载免费PDF全文

郭军  屈冬玉  王晓武  金黎平  谢开云 《植物保护》2005,31(2):9-13

近年来马铃薯晚疫病的重新暴发再次引起世界各国极大关注,特别对马铃薯晚疫病菌的分子遗传学的研究,包括病菌基因组遗传、转录和物理图谱的构建,病菌致病的分子机制以及马铃薯-马铃薯晚疫病菌互作分子机制等。本文就近几十年来对马铃薯晚疫病菌在生物学和病理学,分子遗传学等研究方面作简要综述,并对其研究趋势进行了展望。  相似文献   

禾谷镰刀菌和稻瘟病菌基因组中的微卫星序列比较   总被引：1，自引：1，他引：1       下载免费PDF全文

李成云  李进斌  刘林  杨静  周晓罡 《植物保护学报》2005,32(3):251-255

利用禾谷镰刀菌Fusarium graminearum和稻瘟病菌Magnaporthe grisea基因组测序结果,对这两种植物病原真菌基因组中的微卫星(SSR)序列进行了系统地分析和比较.结果表明,在禾谷镰刀菌基因组中,共发现4679个SSR序列,总长度为96.2kb,占基因组全长的0.27%.平均7.7kb碱基中有一个大于15 bp的SSR序列.在稻瘟病菌基因组中共发现16398个SSR系列,其总长度达到330kb,约占整个基因全长的0.85%,平均2.36kb碱基中就分布有1个SSR序列.在禾谷镰刀菌基因组中,数量最多的是五碱基重复序列,其次是六碱基重复序列;稻瘟病菌基因组中数量最多的是单碱基重复序列,其次为三碱基重复序列和五碱基重复序列.两基因组中数量最少的都是二碱基重复序列.尽管这两种植物病原真菌都属子囊菌,基因组大小也十分接近,但无论是在SSR的总体数量上,还是在各类SSR的分布上,两种植物病原真菌都存在十分显著的差别.  相似文献   

A Universal `One-tube' RT-PCR Protocol for Amplifying Isolates of Bovine Viral Diarrhoea Virus     

Pfeffer M  Freyburg MV  Kaaden OR  Beer M 《Veterinary research communications》2000,24(7):491-503

The increase in the knowledge of the genetic variability of BVDV and the identification of some of the genetic determinants of its pathogenicity require robust and practical tools for rapid molecular characterization of the various genotypes of this virus. This study was undertaken to develop a standard protocol for RT-PCR that allows the amplification of various parts of the genome of BVDV without the need for optimizing each individual reaction. The reaction set-up is very flexible because it consists of two pre-mixes. These are a master mix, with all the required reagents except the desired primers, which are the components of the second pre-mix and are therefore easily interchangeable between the different reactions. After adding any primer-containing pre-mix to the fixed master mix, a non-interrupted cycling protocol led to the generation of amplicons of up to 4 kbp in size in amounts sufficient for subsequent sequencing reactions. The method was applied to five different regions of the BVDV genome: (i) the well-known 5-UTR to differentiate genotypes I and II; (ii) the entire E2 gene, or an approximately 550 bp region within the E2 gene, in order to find the molecular equivalent of antigenic varieties; (iii) the entire structural protein coding region covering the Npro, capsid, E ^RNS, E1 and E2 genes; (iv) a 2.1 kbp region embracing the NS2/3 junction which is known to be cleaved in cytopathic biotypes of BVDV; and (v) the region covering the entire NS4B and NS5A/B genes. All six RT-PCRs were successfully applied using (i) primers with lengths of between 20 and 52 nucleotides, (ii) an aliquot of RNA extracted from either 10⁶ infected bovine embryonal lung cells or the same number of leukocytes from viraemic cattle, and (iii) all the genotype I and II strains of BVDV tested. The technique described was used to generate various Sindbis virus/BVDV recombinants. The correct processing of the amplicon-derived E2 glycoprotein of BVDV strain PT810 was demonstrated by its reaction with a monoclonal antibody in an immunofluorescence assay. Given the variety of RT-PCRs tested, we conclude that this universal protocol may be useful with other RNA viruses.  相似文献   

Sequence analysis of the genome of OP2, a lytic bacteriophage of Xanthomonas oryzae pv. oryzae     

Yasuhiro Inoue  Takayuki Matsuura  Tatsuji Ohara  Koji Azegami 《Journal of General Plant Pathology》2006,72(2):104-110

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Studies of clomazone mode of action     

Yurdagul Ferhatoglu  Michael Barrett 《Pesticide biochemistry and physiology》2006,85(1):7-14

Intact spinach chloroplasts were used to determine if clomazone, 5-OH clomazone, and/or 5-keto clomazone inhibited the chloroplastic isoprenoid pathway. When isopentenyl pyrophosphate was used as a precursor, neither clomazone nor the clomazone metabolites (5-OH clomazone and 5-keto clomazone) inhibited the formation of products separated by HPLC in the organic phase. However, when pyruvate, a substrate for the first committed step of the pathway, was used as a precursor, both 5-keto clomazone and fosmidomycin reduced the formation of a non-polar product and increased the formation of a polar product in the organic phase. Only 5-keto clomazone, not 5-OH clomazone or clomazone, inhibited the formation of an additional product other than fosmidomycin in the aqueous phase from pyruvate incorporation. In an in vitro assay, 5-keto clomazone inhibited DXP synthase, the enzyme catalyzing the first committed step of the chloroplastic isoprenoid pathway. Therefore, our studies show that neither clomazone nor 5-OH clomazone inhibits the chloroplastic isoprenoid pathway, only 5-keto clomazone does.  相似文献   

禽腺联病毒全基因组的克隆及感染性病毒的拯救     

王建业  孙怀昌  朱国强 《中国预防兽医学报》2007,29(11):825-829

为了克隆禽腺联病毒(Avian adeno-associated virus,AAAV)全基因组用于构建基因转移载体研究,以鸡胚致死孤儿病毒(CELO)作为辅助病毒与AAAV共接种SPF鸡胚进行AAAV的增殖,将AAAV约4.7kb双链基因组DNA与pCR2.1载体连接,构建了含AAAV全基因组的重组质粒pAAAV并进行了测序。序列分析表明,AAAV YZ-1株的基因组为4684bp,两端具有141bp的末端倒置重复序列和Rep蛋白结合位点特征序列,与GenBank中收录的AAAV DA-1株和VR-865株的核苷酸序列同源性分别为95.0%和92.2%。将pAAAV质粒转染CELO病毒感染的鸡胚肝细胞系,获得了感染性AAAV病毒粒子,结果证明克隆的AAAV基因组中存在与病毒复制和包装相关的正确关键序列,可用于重组AAAV载体的构建。  相似文献   

口蹄疫病毒WFL株基因组全长cDNA的克隆及序列分析   总被引：1，自引：0，他引：1  

任林柱  王兴龙  李莉  李晓艳  段小宇  梅英武  刘锴  王学理 《中国预防兽医学报》2007,29(1):22-26

根据口蹄疫病毒基因组的结构特点以及GenBank上公布的全序列，用DNAMAN分别设计了涵盖整个基因组序列的3对引物，从接种口蹄疫病毒WFL株的细胞培养液中提取了病毒基因组RNA，采用RT-PCR方法和RACE法分别扩增了3条基因片段，并将扩增片段分别与T载体连接，在体外分别进行了5-半分子和3’半分子的构建。最后将5’半分子和3’半分子连接成基因组全长cDNA分子。经PCR鉴定、酶切鉴定及全长cDNA测序，证实成功构建了口蹄疫病毒wFL株基因组全长eDNA分子。序列分析结果表明，供试口蹄疫病毒基因组全长为8155nt，5＇UTR长1059nt；具有一个大的读码框，其核苷酸长度为6969nt。包括201aa的前导蛋白基因和2122aa的聚合蛋白基因；3＇UTR长127nt，包括34nt的polv（A）。  相似文献   

一株山羊支原体山羊肺炎亚种的分离鉴定与分子特征   总被引：8，自引：0，他引：8  

辛九庆  李媛  张建华  胡守萍  王亮 《中国预防兽医学报》2007,29(4):243-248

从送检的山羊肺炎肺脏中成功分离到一株支原体，经过3次克隆纯化后进行生化试验、电镜观察、PCR及酶切、基因特征鉴定，结果显示分离物SD3属于山羊支原体山羊肺炎亚种成员。将培养物经气管接种2只山羊可引起1只山羊典型发病，体温升高至41．5℃，IgG和IgM抗体效价明显升高，其中IgG抗体变化与临床表现基本同步。剖检发现肺脏发生严重病变，并从中再次分离到该病原体。  相似文献   

香蕉A基因组品种间遗传关系的SSR检测   总被引：3，自引：2，他引：3  

冀小蕊  陈业渊  王静毅  郑丽珊  魏守兴  谢子四  武耀廷 《果树学报》2007,24(6):783-787

应用SSR技术,对32个香蕉A基因组类型品种(系)的遗传关系进行了检测。40对SSR引物在32个品种(系)中分别扩增带数在3～15个,平均每个SSR座位可检测2.99个多态性带;引物的多态信息量(PIC)在0.00～0.88,平均0.62。依据SSR数据计算的品种间遗传距离在0.00%～34.27%,平均12.45%,大多数品种间的遗传变异非常有限,但也存在着遗传差异突出的品种:FHIA25、Yangambi KM5、Pisang Jari Buaya、Rose和皇帝蕉。依据26%的遗传距离,除了FHIA25和Pisang Jari Buaya单独化成1组外,其它30个品种可以分为2组:品种间遗传差异相对较高的组I和品种间遗传差异相对较低的组Ⅱ。Williams与引进的洪都拉斯3号、M931之间,洪都拉斯1号和洪都拉斯2号之间,高脚青芽蕉和高脚顿地雷分别没有区分开来,这可能是同物异名,也可能是同一品种未能分辨的突变体。  相似文献   

Identification and evaluation of chloroplast uni- and trinucleotide sequence repeats in citrus     

Ziniu Deng  Stefano La Malfa  Yuming Xie  Xingyao Xiong  Alessandra Gentile 《Scientia Horticulturae》2007

A cpDNA fragment of 34 genotypes belonging to Citrus and four related genera was amplified and sequenced. Chloroplast microsatellites were revealed with the length of repeats ranging from 25 to 44 bases. Other than the normal uninucleotide poly(A) repeats, a trinucleotide poly(TAA) motif was also found, the first report of such repeats in a plant chloroplast genome. According to SSR structure variations, 18 Chloroplast SSR Types (CST) were identified. The CST sequences were informative for better understanding the genetic relationships of chloroplast genomes among the analyzed genotypes and confirmed some previous hypotheses about the female parent of several hybrid accessions.  相似文献