利用IBV重组N蛋白建立ELISA抗体检测试剂盒的研究   总被引：3，自引：0，他引：3  

王宏俊  陈福勇 《中国兽医杂志》2003,39(9):3-6

利用表达鸡传染性支气管炎病毒Gray株N蛋白的基因工程重组菌，表达IBV—N蛋白，并利用表达载体上带有组氨酸标记的特点，应用金属螯合填料将其纯化。用纯化的IBV—N蛋白作为包被抗原，成功地建立了一种检测血清中IBV抗体的间接ELISA方法。确定抗原最佳包被浓度为0．58μg／ml；被检血清的最佳稀释度为1：100；酶标二抗的工作浓度为1：1000。确定了血清样品阴、阳性临界0D值为0．14。与IDEXX公司IBV抗体检测试剂盒比较，结果表明自建ELISA试剂盒具有较高的敏感性和特异性。  相似文献   

Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers   总被引：6，自引：1，他引：6  

F. M. Abu-El Samen  G. A. Secor  N. C. Gudmestad† 《Plant pathology》2003,52(3):314-325

Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed.  相似文献   

Genes for resistance to powdery mildew in European barley cultivars registered in the Czech Republic from 2011 to 2015          下载免费PDF全文

Antonín Dreiseitl 《Plant Breeding》2017,136(3):351-356

Barley (Hordeum vulgare) is cultivated on 49.1 million hectares worldwide with 50.2% of the area located in Europe. Powdery mildew, caused by Blumeria graminis f. sp. hordei (Bgh), occurs wherever barley is grown. Cultivar resistance plays an important role in global barley production, especially in parts of Europe where high concentrations of both spring and winter types are grown. The aim of this report was to postulate specific resistance genes in barleys from nine European countries registered in the Czech Republic from 2011 to 2015. Thirty‐five spring cultivars and 27 winter barleys were tested with 56 diverse Bgh isolates. Twenty‐five known resistance genes were postulated, and unknown genes were detected in Sandra, Saturn and Zeppelin. Unidentified specific resistance genes were also present in winter hybrids Hobbit and Wootan. Spring cultivars Arthur and Francin consisted of three and two genotypes, respectively. Resistance gene mlo was present in 26 spring cultivars, and the proportion of cultivars with this gene increased from 62.9% in 2006–2010 to 75.7% in 2011–2015. The gene Mlp1 was identified for the first time in German winter cultivar Saturn. Five spring cultivars registered in Slovakia were included in the tests. All the cultivars that were tested contained one or more specific resistance genes to powdery mildew. Adaptability of the pathogen and possibilities for breeding winter barleys are discussed.  相似文献   

青贮玉米促生菌的鉴定及生物学特性的研究   总被引：2，自引：0，他引：2  

李梅  刁治民  赵子倩 《青海畜牧兽医杂志》2001,31(1):3-5

根据布坎南.R.E等著《伯杰细菌鉴定手册》（第八版）,对4株青贮玉米促进菌进行鉴定。结果表明,4株促生菌分属于短芽胞杆菌（Bacillus brevis),蜡质芽孢杆菌（Bacillus cereus）,放射形土壤杆菌（Agrobacteria radiobacter）和假单胞杆菌（Pseudomonas sp）。此餐,还对4株促生菌的生物学特性进行了比较观察。  相似文献   

Assessment of biological and molecular variability between and within field isolates of Plasmodiophora brassicae     

M. J. Manzanares-Dauleux†  I. Divaret  F. Baron  G. Thomas 《Plant pathology》2001,50(2):165-173

Plasmodiophora brassicae is an obligate biotroph that causes clubroot, one of the most damaging diseases of crucifers. Differential cultivars and random amplified polymorphic DNA markers were used to assess the extent of genetic diversity among nine single-gall populations of P. brassicae and 37 single-spore isolates (SSI) derived from four of those field samples. Isolates were classified into eight pathotypes, and each isolate was associated with a unique molecular genotype. Virulence and DNA polymorphisms were detected within and between field isolates, and among SSIs from different pathotypes, hosts and geographical origins. The relatively high level of genetic diversity among field isolates was similar to that among SSIs derived from a single-club field isolate. Molecular and pathogenicity-based classifications were not clearly correlated, but isolates belonging to pathotype P1 were clustered. Two RAPD markers were specific to pathotype P1. The finding that genetic differences can occur in P. brassicae field isolates will be an important consideration in resistance genetic studies and in choosing breeding strategies to develop durable clubroot resistance.  相似文献   

Identification of divergent variants of Grapevine virus A     

D.E. Goszczynski  A.E.C. Jooste 《European journal of plant pathology / European Foundation for Plant Pathology》2003,109(4):397-403

Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3 terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3 UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.  相似文献   

Biological activity and characterization of nucleopolyhedrovirus isolates ofSpodoptera litura     

C. M. Senthil Kumar  R. J. Rabindra  N. Sathiah 《Phytoparasitica》2006,34(1):92-101

Geographical isolates ofSpodoptera litura (Fabricius) (Noctuidae: Lepidoptera) nucleopolyhedrovirus (SpltNPV), collected from different parts of India and maintained at Tamil Nadu Agricultural University, were compared for their biological activity and subjected to Restriction Endonuclease (REN) analysis. Neonate and second instar bioassay studies revealed similarity in biological activity as shown by the overlapping fiducial limits of LC₅₀ values. However, there were differences in yield among isolates: significantly higher yields were obtained from isolates UAS and CBE than from the BARC isolate. REN analysis of the four isolates withPst I,Hind III,Bam HI andEco RI enzymes indicated genotypic variation among the isolates. Based on the commonality of the bands, the isolates could be broadly divided into two groups: isolates AU and CBE formed one group, and the other group comprised UAS and BARC based on genetic relatedness. http://www.phytoparasitica.org posting Nov. 21, 2005.  相似文献   

四环素耐药基因在鸡源沙门氏菌中的分布和传播     

王晓泉  王彦红  吴双  焦新安  潘志明  刘秀梵 《中国家禽》2007,29(9):10-12,18

试验分析了84株鸡源沙门氏菌分离株的四环素耐药性,用PCR方法检测四环素耐药基因在分离株中的分布情况。结果显示,四环素耐药率为49%(41/84),鸡白痢和鸡伤寒沙门氏菌仅携带tet(A)基因(23/23),肠炎沙门氏菌和德尔卑沙门氏菌携带tet(A)(8/18)、tet(B)(17/18)或tet(G)(10/18)三种基因,tetC基因在这些沙门氏菌中都没有检测到。该类基因多数位于结合性质粒上,但是不在整合子范围内。  相似文献   

鸡传染性支气管炎病毒N蛋白研究进展     

邓小敏  叶柱德  王晶钰  罗艳 《动物医学进展》2007,28(9):78-82

鸡传染性支气管炎病毒(IBV)是严重危害养鸡业的重要病原之一.近年来国内外学者在分子生物学领域方面的研究不断深入,在病毒基因组的结构与功能及其N蛋白的研究方面取得了重要进展.对N蛋白如何特异性地识别病毒基因组RNA、参与病毒RNA的复制和包装、诱导IBV体液和细胞免疫应答的机制、IBV新型基因工程疫苗研制以及IBV的抗体水平监测等方面的研究,为IBV基因工程疫苗设计及其免疫防控提供了科学依据.  相似文献   

应用RT-PCR技术扩增禽传染性支气管炎病毒S_1基因的研究     

黄勇  王红宁  王林川  廖明 《中国预防兽医学报》1998,(5)

选择Ｓ１基因做为目的基因,运用ＲＴ－ＰＣＲ技术分别扩增１２株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的ＩＢＶ,所用引物为ＩＢＶＢｅａｕｄｅｔｔｅ株Ｓ１基因两侧的对应序列,跨幅为１．７２ｋｂ。结果６株ＩＢＶ毒株（ＳＡＩＢ３、Ｆ、Ｄ４１、Ｍ４１、Ｈ５２、Ｈｏｌｔｅ）扩增出了长约１．７ｋｂ的Ｓ１基因片段,而另６株ＩＢＶ毒株虽经４次ＲＴ－ＰＣＲ重复后也显阴性。用ＨａｅⅢ内切酶对６个毒株的ＲＴ－ＰＣＲ产物进行酶切,结果显示Ｍ４１、Ｈ５２、Ｄ４１３个ＩＢＶ毒株的Ｓ１基因包含两个ＨａｅⅢ位点,Ｆ株与Ｈｏｌｔｅ株Ｓ１基因包含１个ＨａｅⅢ位点,而ＳＡＩＢ３株Ｓ１基因则已丢失ＨａｅⅢ位点。实验结果表明：运用ＲＴ－ＰＣＲ技术检测ＩＢＶ时,由于Ｓ１基因的核酸序列具有较高的变异率,因此Ｓ１基因不宜做为目的基因。  相似文献