The Complete Nucleotide Sequence of a Japanese Isolate of White clover mosaic virus Strain RC

The complete nucleotide sequence was determined for genomic RNA of White clover mosaic virus (WClMV-RC) isolated from red clover (Trifolium pratense) in Japan, It is 5843 nucleotides in length, excluding the poly(A) tail at the 3' terminus. Similar to other potexviruses, it contains five open reading frames (ORFs 1 through 5), which putatively encode an RNA-dependent RNA polymerase (RdRp) (147 kDa), a triple gene block (TGB) (26 kDa/13 kDa/7 kDa), and a coat protein (CP) (22 kDa), respectively. The deduced amino acid sequence of the WClMV-RC CP was identical to that of WClMV-O, one of two New Zealand isolates, but only 85% identical to that of WClMV-M, the other New Zealand isolate, because of heterogeneity in the C-termini of CP amino acid sequences. The implication of this CP heterogeneity is discussed. Received 30 August 2001/ Accepted in revised form 11 January 2002     

SSR技术及其在果树上的应用

SSR(Simple sequence repeat)技术以其丰富的多态性、共显性遗传、重复性好和操作简便等优点日益受到重视,已成为植物遗传和育种研究中不可缺少的分子标记。对SSR技术的原理和特点作了简要的介绍,较详细地分析了如何获得SSR引物,特别是综述了果树上SSR引物的研究现状,同时将其与其它几种主要的分子标记进行了比较分析,认为SSR标记检测的位点多态性水平明显高于RFLP,而且重复性优于RAPD;着重介绍了SSR技术在果树种质资源和构建果树遗传图谱及基因定位等研究中的应用现状;指出SSR技术将在果树科研上起到重要的作用。     

SSR-based genetic linkage analysis of resistance to crown rust (Puccinia coronata f. sp. lolii) in perennial ryegrass (Lolium perenne)

Crown rust (caused by Puccinia coronata f. sp. lolii) is a serious foliar disease of the pasture and turfgrass perennial ryegrass (Lolium perenne). Previous genetic studies have detected both qualitative and quantitative resistance mechanisms, and interpretation of the genetic system is complicated by variation within the sexually reproducing pathogen. Resistant and susceptible parental genotypes of ryegrass were identified using a composite urediniospore population collected from three geographically distinct locations. A two-way pseudo-testcross mapping population was obtained as the F₁ progeny of the pair-cross between ryegrass parental genotypes Vedette₆ and Victorian₉. Both parents showed intermediate resistance against a pathogen population collected in a single geographical zone (Hamilton, Victoria), but in the F₁ population, significant variation for a range of resistance-associated characters was detected. Statistical analysis of phenotypic data suggested a major gene effect, hence bulked segregant analysis with map-assigned simple sequence repeat (SSR) markers was used to scan the genome. A marker showing strong association with resistance was assigned to linkage group (LG) 2 of perennial ryegrass. Analysis of 11 LG2 SSR markers defined an interval between loci xlpssrh03f03 and xlpssrk02e02 as containing the gene or genes (LpPc1) conferring crown rust resistance. Resistance gene determinants were inherited from both parents, with up to 80% of the total phenotypic variation explained by markers segregating from Vedette₆ and up to 26% of the variation explained by markers segregating from Victorian₉. The two contributions together resulted in an additive increase in effect, with fully resistant individuals requiring determinants from both parents. A conserved syntenic relationship was observed with linkage group B of Avena strigosa, which is the location of a cluster of resistance genes to the oat form of crown rust. The implications of this study for marker-assisted selection of disease resistance in perennial ryegrass are discussed.     

小菜蛾对溴氢菊酯抗药性的随机扩增多态性DNA分析

利用室内选育119代的抗溴氰菊酯小菜蛾Plutella xylostella品系和敏感品系，通过反复回交和自交建立近等基因系。用Operon公司合成的20个引物对近等基因系基因组DNA进行PCR扩增。3个引物在抗性品系中分别产生1条特异扩增带，4个引物在敏感品系中分别产生了1～5条特异扩增带。由于在近等基因系中，除了与抗性相关的基因区外，其它的遗传背景是相同的，因此可以认为这些特异带与小菜蛾对溴氰菊酯的抗性有关。     

Biological and molecular characterization of Tomato spotted wilt Virus in Israel

Received April 24, 1997; received in final form June 29, 1997. Symptoms resembling tomato spotted wilt virus (TSWV) infections were documented among ornamental and vegetable crops in commercial greenhouses and open fields in Israel. Plants exhibiting these symptoms were collected from January 1992 to December 1996. Among cultivated plants analyzed for TSWV by enzyme-linked immunosorbent assay (ELISA), 19 species representing five families were found to be infected; natural infection was also recorded in six plant species of weeds. Virus identity was characterized by host range, serology and electron microscopy. Serological reaction with the isolates, found in Israel, using antisera from different sources as well as the sequence analysis of the nucleocapsid gene, demonstrated that the Israeli isolates of TSWV are a member of tospovirus serogroup I, type I (BR-01 strain). No virus transmission was found in seeds collected from virus-infected vegetable and ornamental crops. A non-radioactive molecular probe derived from the cloned nucleocapsid isolate enables specific detection of the virus in crude sap from infected plants. The detection of TSWV in Israel constitutes a severe potential threat to the ornamental and vegetable industry.     

猪大肠杆菌水肿毒素SLT-IIeA基因的克隆和序列分析

研究以本地猪水肿病大肠杆分离物ED1株为材料 ,利用 PCR克隆了含猪水肿病大肠杆菌 (VTEC) slt-IIe A基因 987bp的片段 ,并测定了含该克隆片段的 Bam HI、Hind 酶切片段的核苷酸序列。结果表明 ,slt-IIe A基因的编码区全长 960 bp,编码 3 1 9个氨基酸的蛋白质 ,序列与国外报导的 S1 1 79株进行比较发现其核苷酸同源性为 98.9%。经推导的氨基酸序列的同源性为 99.7%。这为进一步研究 slt-IIe A的生物学特性、水肿病的分子诊断及其防制打下基础     

Cortical gene expression pattern in rat focal cerebral ischemia

AIM: To investigate the genes differential expression in cortex during rat focal cerebral ischemia.METHODS: cDNA microarray chips containing numerous cDNAs were used to investigate the gene expression pattern between samples of focal cerebral ischemia and sham-control operation rats. RESULTS: Two hundred and eleven genes differentially expressed were screened out, among these genes, up-and down-regulated genes were 199 and 12, respectively. CONCLUSIONS: The analysis of gene expression pattern of focal cerebral ischemia based on cDNA microarray can realize high-throughput screening of the genes associated with the focal cerebral ischemia. The differential expression of genes may be related to the pathogenesis of focal cerebral ischemic diseases.     

Seasonal variation in pigmentation and anthocyanidin phenetics in commercial Eustoma flowers

The seasonal change in petal color and pigmentation of 29 commercial Eustoma cultivars was studied. The flowers are basically divided into four groups according to the major anthocyanidin phenotype in association with petal coloration, i.e., delphinidin (Dp)-based (purple flower), cyanidin (Cy)-based (reddish purple flower), pelargonidin (Pg)-based (pink flower), and none (white flower) groups. The constitution of petal anthocyanidins was not changed by forcing treatment in most of the flowers. Lightness (L^*) and chroma (C^*, color saturation) showed a change along with the increase/decrease of hue angle difference (ΔH^*), thus simultaneously the chromatic tonalities tended to move to redder and bluer, respectively. Floral pigment clustering described two flower groups in a dendrogram, based on anthocyanidin constitutions as phenetic markers, which are apparently the Dp- and Pg-based phenotypes of anthocyanidin syntheses. The Cy-based flowers made a subcluster with the Pg-based flowers, indicating a close relationship in the biosynthesis of the two anthocyanidins, and suggesting the Dp- and Pg-syntheses complement one another.     

丝状霉形体丝状亚种SC型脂蛋白LppQ N末端基因克隆与序列分析

脂蛋白LPPQ是丝状霉形体丝状亚种SC型(MmmSC)非洲株、欧洲株和疫苗株所特有的。LPPQ N末端域具有良好的免疫原性，在牛体内可诱导产生强大、特异、早期、持续的免疫反应。本研究根据已发表的LPPQ基因序列设计引物，用Pyrobest^TM高保真DNA聚合酶从MmmSC HVRI X株中扩增出了LPPQ N末端基因序列，并进行了克隆与序列测定。核苷酸序列比较结果显示，HVRI X株的LPPQ N末端基因序列与国外发表的序列同源性为99．7％，由其推导的氨基酸序列同源性为99，1％，为脂蛋白LPPQ N末端基因体外表达奠定了基础。     

禽大肠杆菌外膜蛋白基因C(ompC)的序列分析

根据 Gen Bank中人源大肠杆菌 K- 12外膜蛋白基因 C(omp C)的核苷酸序列设计引物 ,应用 PCR方法从禽大肠杆菌 O2 、O78株及它们的融合双价弱毒菌株 O2 ,78(Norr Chlr)中分别扩增得到 omp C基因 ,序列测定及分析比较发现 ,3个菌株的 om p C基因均由 170 2 nt组成 ,核苷酸序列完全相同 ,只有 1个大的开放性阅读框 (ORF) ,长 10 92 bp,编码由 36 3个氨基酸组成的前 Om p C蛋白 ,前 2 1个氨基酸残基组成信号肽 ,成熟的 Omp C蛋白由 342个氨基酸残基组成 ,Mr为 4 0 0 0 0。其氨基酸序列也完全相同。从基因水平上证明了禽大肠杆菌 O2 、O78株及融合双价弱毒菌株 O2 ,78(NorrChlr)存在相同的外膜蛋白 C抗原 ,从而为进一步研究 Omp C蛋白的免疫原性奠定了基础