部分地区猪传染性胃肠炎病毒株ORF7基因的序列分析

猪传染性胃肠炎（Porcine Transmissible Gastraenteritis,TGE）是猪的一种高度接触性肠道疾病。以不同地区猪传染性胃肠炎病毒株的ORF7基因为靶序列,序列分析不同地区TGEV病毒株ORF7基因的同源性和进化性关系,序列分析包括同源性比较、进化树分析和碱基替换分析等。结果表明：ORF7基...     

小麦穗发芽抗性的4个分子标记有效性检验

为筛选出适宜黄淮麦区和长江中下游麦区种植的抗穗发芽白粒小麦品种或种质资源,以36份黄淮麦区和长江中下游麦区的主要品种（系）及地方品种为研究对象,对已报道的4个与穗发芽抗性相关的分子标记：Vp1B3、Xgwm155、Xgwm269和Xbarc170进行有效性验证。测定参试材料种子萌发指数（GI）,并用上述4种标记进行PCR扩增,对扩增条带进行统计分析。结果表明,GI值显示,红粒品种（GI均值为5.1%）明显较白粒品种（GI均值为28.0%）低;4种标记扩增出的带型中仅Vp1B3的845 bp片段能有效地区分36份小麦品种(系);GI值筛选出6份抗穗发芽品种（系）中,其中3份为Vp1B3标记鉴定,可作为黄淮麦区和长江中下游麦区小麦穗发芽抗性育种中首选基因资源。     

拮抗细菌A2菌株的鉴定及其对小麦白粉病的防治效果

为适应现代无公害农业的发展,针对小麦白粉病的防治,从保护地土壤中分离得到一株生防细菌A2。通过形态学特征、生理生化特性的观察和测定,结合16S rDNA序列分析并构建系统发育树,对菌株A2进行了初步鉴定。同时,以清水为对照,测定了菌株A2对3432品种小麦白粉病的防治效果。结果显示该菌株主要特征是菌体杆状,内生芽孢、芽孢卵圆形,革兰氏染色阳性,好氧,接触酶反应、乙酰甲基甲醇实验为阳性,能利用柠檬酸盐;该菌株与亲缘关系较近菌株B.subtilis isolate G8的同源性达99%。结合以上两点,将菌株A2初步鉴定为枯草芽孢杆菌（B. subtilis）。防效实验显示菌株A2发酵液处理最好防治效果为72.22%;无菌滤液处理最好防治效果为29.24%,与对照相比均具有显著性差异（α=0.05）。表明,拮抗菌株A2菌体与代谢产物对小麦白粉病都具有较强的防治作用,为研制型、无毒、无公害生物农药提供了依据。     

灰葡萄孢霉高效拮抗木霉菌株的筛选及其翻译延伸因子序列分析

为进一步开发利用木霉菌资源,对从菜田土壤中初步分离纯化获得的12株木霉菌株,采用对峙培养和生长速度测定法进行灰葡萄孢霉(Botrytis cinerea)高效拮抗木霉菌株的筛选,并通过翻译延伸因子序列同源性比较对其进行分子鉴定。结果表明菌株Tr-9701和Tr-1108的生长速度快,对病原菌的抑制率高,且协同应用有一定的增效作用。经对Tr9701和Tr1108翻译延伸因子同源序列分析,并结合其形态特征结果表明,Tr9701为绿色木霉(Trichoderma viride)和Tr1108为深绿木霉(Trichoderma atroviride)。绿色木霉和深绿木霉为菜田生境习居菌,2种菌株协同利用对蔬菜灰霉病有较好的协同增效作用,应进一步设计利用多靶位木霉菌来提高防病效果。     

轮枝镰孢SSR标记开发及在玉米分离群体遗传多样性分析中的应用

【目的】开发轮枝镰孢（Fusarium verticillioides）的SSR标记,为轮枝镰孢遗传多样性研究提供技术支持。【方法】利用Fastpcr在轮枝镰孢基因组中查找符合条件的SSR位点,采用Primer5.0软件设计引物,利用NTSYS及Popgene分析来自玉米的轮枝镰孢单孢分离物群体的PCR扩增结果。【结果】共设计有效扩增SSR引物158对,经筛选,109对（69.0%）可扩增出2条及以上的条带,其中55对引物（34.8%）多态性较好,可扩增出3条及以上的条带。从11条已组装的轮枝镰孢染色体上各选出2对多态性引物,计22对引物,对66株轮枝镰孢玉米分离物进行扩增,共获得125个等位变异,变异范围为2-11个,平均为5.68个。轮枝镰孢玉米分离物之间Nei’s基因多样性指数为0.1139-0.8687,平均为0.6199,表现出较高的遗传多样性。【结论】基于轮枝镰孢基因组序列开发的SSR标记具有很好的多态性,可用于轮枝镰孢的遗传多样性分析。用22对SSR引物扩增,以遗传相似系数0.3进行划分,可将66株轮枝镰孢玉米分离物分为3群;中国轮枝镰孢玉米分离物的遗传多样性与地理分布无相关性。     

Vegetative compatibility of cotton-defoliating <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in Israel and its pathogenicity to various crop plants

Verticillium dahliae isolates recovered from a new focus of severe Verticillium wilt of cotton in the northeast of Israel were tested for vegetative compatibility using nitrate non-utilizing (nit) mutants and identified as VCG1, which is a new record in Israel. Other cotton isolates of V. dahliae from the northern and southern parts of the country were assigned to VCG2B and VCG4B, respectively. VCG1 isolates induced severe leaf symptoms, stunting and defoliation of cotton cv. Acala SJ-2, and thus were characterized as the cotton-defoliating (D) pathotype, whereas isolates of VCG2B and VCG4B were confirmed as the earlier described defoliating-like (DL) and non-defoliating (ND) pathotypes, respectively. This is the first record of the D-pathotype in Israel. The host range of representative isolates of each VCG-associated pathotype was investigated using a number of cultivated plants. Overall, the D isolates were more virulent than DL isolates on all tested host plants, but the order of hosts (from highly susceptible to resistant) was the same: okra (Hibiscus esculentus local cultivar), cotton (Gossypium hirsutum cv. Acala SJ2), watermelon (Citrullus lanatus cv. Crimson Sweet), safflower (Carthamus tinctorius cv. PI 251264), sunflower (Helianthus annuum cv. 2053), eggplant (Solanum melongena cv. Black Beauty), and tomato (Lycopersicon esculentum cv. Rehovot 13). The pattern of virulence of ND isolates differed from that of D and DL isolates, so that the former were highly virulent on eggplant but mildly virulent on cotton. Tomato was resistant to all cotton V. dahliae isolates tested. RAPD and specific PCR assays confirmed that the D isolates from Israel were similar to those originating from other countries.     

应用抑制性差减杂交技术分离椪柑枯水相关基因

【目的】探索椪柑枯水分子机理,寻找柑橘枯水应答基因,为柑橘枯水防治奠定基础。【方法】利用抑制性差减杂交技术以椪柑正常果肉cDNA作为Driver,以枯水果肉cDNA作为Tester构建正向差减文库。【结果】挑选了300个有效克隆进行测序分析,260个测序成功。经BLASTx比对分析,197条表达序列标签（ESTs）找到了分属于52个基因的同源序列;63条ESTs同源性较低或没有同源性。对文库中编码β-微管蛋白、衰老相关蛋白、细胞色素c、半胱氨酸蛋白酶、磷酸烯醇式丙酮酸羧激酶、转运蛋白、天冬氨酸蛋白酶前体、WRKY转录因子21的基因进行实时定量RT-PCR验证,结果显示这8个基因均明显上调表达。这些差异性表达基因主要涉及衰老、抗逆防御、几丁质和细胞壁代谢、蛋白降解等过程。【结论】通过构建椪柑果肉枯水正向文库,得到一些与枯水相关的基因,这些基因反映了果实对枯水的调控情况,可作为今后防治枯水病害的候选基因。     

Genotypic diversity and migration of clonal lineages of Botrytis cinerea from chickpea fields of Bangladesh inferred by microsatellite markers

An analysis of allelic diversity at nine microsatellite loci provided an insight into the population structure of Botrytis cinerea from four fields (sampled in 2003 and 2004) that represented important regional locations for chickpea production in Bangladesh. Although three populations were limited by sample size after clone‐correction, a total of 51 alleles were amplified among 146 B. cinerea isolates from Bangladesh, which revealed a high amount of within‐population and overall genetic diversity (H_S = 0·48 and H_T = 0·54, respectively). The percentage of maximal genotypic diversity (G) ranged between populations (G = 23–40), with a total of 69 haplotypes detected (G = 25). Bayesian cluster analysis depicted two major clusters distributed among the four Bangladesh populations, indicating population admixture from two origins that have spread throughout these regions. Genotype flow between regions was detected and indicated the spread of clonal lineages, consistent with relatively low differentiation among the four populations (mean G_ST = 0·1, P < 0·05). These results highlighted the potential threat of host resistance breakdown as a result of considerable genetic diversity, genotype flow and the evolutionary potential of B. cinerea.     

Biochemical and histological alterations of cellular metabolism from jerboa (Jaculus orientalis) by 2,4-dichlorophenoxyacetic acid: Effects on d-3-hydroxybutyrate dehydrogenase

2,4-Dichlorophenoxyacetic acid (2,4D) is one of the widely used herbicide of the phenoxy family with possible startling number of adverse effects on species other than the weeds which is designed to kill. The effects of 2,4D were investigated in jerboa (Jaculus orientalis), a wild animal of subdesert highlands. The jerboas have been daily treated intraperitonally with 2,4D 3 mg/kg body weight for 4 weeks. Plasmatic markers, and antioxidants defences systems were assessed and histological alterations were evaluated. The in vivo and in vitro effects of 2,4D on the mitochondrial d-3-hydroxybutyrate dehydrogenase (BDH) were also determined. Our results showed a strong decrease of triglycerides level and HDL cholesterol and an increase in GOT level and LDL cholesterol. The microscopic evaluation showed that 2,4D induced necrosis of seminiferous tubules cells in testis, hyperplasia of hepatocytes in liver and presence of multinucleated giant cells in brain. The results show also an inhibitory effect on BDH in terms of activity and kinetic parameters. All of these results show that 2,4D induces toxicity which affects energy metabolism, morphological perturbation and oxidative stress.     

西瓜花叶病毒2号新疆昌吉分离物外壳蛋白基因核苷酸序列分析

 利用RT-PCR从新疆昌吉地区表现花叶、疱斑、扭曲等症状的南瓜病株上检测到西瓜花叶病毒2号新疆昌吉分离物(简称WMV-2-XJ-CJ),并测定了该分离物外壳蛋白(CP)基因序列。序列分析表明,新疆昌吉分离物CP基因全长850个核苷酸,编码197个氨基酸。与国内外报道的12个WMV-2CP基因相比,其核苷酸序列同源性为92.6%~98.3%,由此推导的氨基酸序列同源性为94.7%~99.3%。新疆昌吉分离物在CP N'端可变区明显不同于国内外报道的核苷酸序列。WMV-2新疆昌吉分离物与日本和郑州分离物较其它国家和地区的分离物多出6个核苷酸,但其核苷酸及其推导的氨基酸序列差异较大。新疆昌吉分离物外壳蛋白有2个氨基酸残基明显不同于其它分离物,其中蚜传株系的特征结构域DAG突变为DAE。