Characterisation of a maize-infecting potyvirus from Spain

In order to characterise and classify an unknown maize-infecting potyvirus isolated from fields in northeast Spain, the entire coat protein gene and the C-terminal twothirds of the large nuclear inclusion protein (NIb) gene were cloned and sequenced. Protein sequencing enabled the cleavage site between the two proteins to be deduced and also revealed that on storage the viral coat protein undergoes a specific degradation in which the N-terminal 39 amino acids are removed. Comparison of the nucleotide sequence of the 3 non-coding region of the viral RNA and the predicted amino acid sequence of the coat protein with the equivalent regions of other members of the potyvirus group revealed that the Spanish virus is closely related to maize dwarf mosaic virus strain A.     

晋南冬麦区大麦黄矮病毒流行株系监测及防治策略探讨

连续5年(1996～2000年)采集晋南冬麦区小麦黄矮病标样,采用生物学和血清学(酶联免疫吸附法)相结合的诊断方法对该地区的大麦黄矮病毒流行株系进行了鉴别。结果表明,该小麦黄矮病流行区近五年以GAV株系为主流株系,兼有少量GPV、PAV和混合株系存在。同时对小麦抗黄矮病新品种“临抗1号”进行了GPV和GAV两种株系的抗性测定,明确了该品种兼抗GPV和GAV两种株系。根据小麦黄矮病发生现状,提出了一套以选育推广抗耐病品种为主,以药剂防治为辅的综合防治措施。以期为当地小麦生产服务。     

Phylogenetic relationships between rice dwarf phytoreovirus isolates from five countries

Rice dwarf virus isolates were collected from several locations in Japan, the Philippines, China, Nepal and Korea. Genomic dsRNA segment profiles in polyacrylamide gel electrophoresis differed among the isolates. There were less differences in the profiles between isolates from Japan and Korea than in those between these two Countries and others. Nucleic acid hybridization was used to examine the extent of genomic variation. Full-length cDNAs to all genomic segments encoding non-structural proteins (S4, S6, S9, S10, S11 and S12) were synthesized from two Japanese isolates, and were used for dot-blot hybridization. Hybridizations using probes generated from the full-length cDNA clones failed to differentiate isolates from different geographical areas. However, cDNA probes covering a variable region of S12 were able to distinguish Japanese and Korean isolates from those of other countries. Phylogenetic tree analysis based on the amino acid sequence of P12 encoded by S12 grouped Japanese and Korean isolates together. The Chinese isolates from two different locations (Yunnan and Fujian) were closely related to each other, and were the most distantly related to Japanese and Korean isolates.     

Mitochondrial DNA analysis of Nepalese domestic dwarf cattle Lulu*

Dwarf Lulu cattle, the only Bos Taurus type of cattle in Nepal, are raised under severe environments in the mountainous zone of that country. In the present study, the body measurement traits, cytogenetic and molecular genetic characteristics of the Lulu cattle are investigated. Blood samples were collected from 31 animals in four villages (altitudes 2590–3550 m) in the southern part of Mustang. The Lulu cattle had a normal karyotype with 2n = 60, XY or XX. Only one male examined had a large submetacentric X‐chromosome and a small submetacentric taurine type Y‐chromosome. The mitochodrial DNA (mtDNA) genotypes were analyzed by PCR mediated restriction fragment length polymorphisms, displacement (D)‐loop region PCR mediated single strand conformation polymorphisms, and D‐loop region sequences. Many base substitutions were found in the D‐loop region, suggesting that the Lulu cattle originated from at least 10 maternal lines. Three types of mtDNA from these cattle were found, the Bos taurus type (n = 23), the Bos indicus type (n = 6), and the Bos grunniens type (n = 2). In the village at the lowest altitude, four of the five cows were of the Bos indicus type. These results indicated that mtDNA types of the Lulu cattle mostly belong to Bos taurus, but have been hybridized with Bos indicus cattle in lower‐elevation regions in their maternal lineage.     

浙江中部地区水稻黑条矮缩病的发生流行

报道了水稻黑条矮缩病再次回升流行的发生规律、分布格局、介体昆虫灰飞虱的种群动态及其相互关系、病害与产量的关系。感病秧苗、大田前期介体灰飞虱密度以及本田期再侵染是本田病害流行的３个重要部分,但早、晚稻有所不同;秧田期介体密度与移栽时、移栽后病害程度,本田初前期介体密度与抽穗期病害程度,株发病率与丛病率,病害程度与最终产量均有显著的关系。     

麦二叉蚜传播玉米矮花叶病毒的机制

用胶体金标记法和荧光抗体标记法研究了麦二叉蚜(Schizaphis graminum)传播玉米矮花叶病毒(Maize dwaft mosaic virus,MDMV)的机制。麦二叉蚜传播玉米矮花叶病毒需要辅助成份-蛋白酶(Helper component-proteinase,HC-Pro)的参与。在电镜下观察到HC-Pro可以与MDMV粒子结合。用FITC标记的HC-Pro抗体和MDMV抗体证明,HC-Pro可以直接结合到蚜虫口针上;而MDMV粒子不能直接结合到蚜虫口针,必须在HC-Pro的辅助下才能结合到蚜虫口针上。这为HC-Pro在蚜虫传毒过程中起桥梁作用提供了新的证据。MDMV粒子主要吸附在蚜虫口针的尖端和中间部分。     

我国水稻黑条矮缩病和玉米粗缩病研究进展

上世纪60年代初我国华东和华北地区分别发现了水稻黑条矮缩病和玉米粗缩病,后来都分别发生了2次大流行,同时都以病害防治为目标分别开展了研究,探明了当地病害发生规律,提出了相应的防治方法,获得了不同程度的防病效果.80年代以来,水稻黑条矮缩病的发生报道仅限于华东地区局部地市,玉米粗缩病的发生涉及华北、东北、西北、西南和华中地区13个省市.根据各地对两病病原形态、寄主及症状、介体昆虫及传病特性等方面相似性的报道,提出了我国玉米粗缩病与水稻黑条矮缩病病原异同性问题.经近10年来对两病用生物学和分子生物学方法进行比较鉴定,证明我国玉米粗缩病和水稻黑条矮缩病病原同属水稻黑条矮缩病毒(RBSDV).同时基本探明了水稻黑条矮缩病在浙江杂交稻区和华北玉米区的再次流行成灾的原因,提出了相应的防治措施,并有效地控制了病害的流行危害.     

玉米抗粗缩病遗传及分子标记研究

Maize rough dwarf disease caused by Rice black-streaked dwarf virus (RBSDV) is transmitted by planthopper in China. Identification and development of resistant hybrids are complicated because of the inconsistencies in viral disease pressure every year. Marker-assisted selection can provide means for main-taining virus resistance alleles even in the absence of disease. In this paper a F₂ segregation population was constructed to identity the molecular markers linked to the resistance gene using a cross between a resistant and a susceptible parents (Qi319×Ye107). Fifteen-day-old seedlings of F₂ population were exposed to small brown planthoppers carrying RBSDV for 3 days in specific inoculation chamber. The inoculated plants were transplanted to screenhouse after removing the insects completely. In plant maturity stage the disease resistance of all the individuals were visually assessed. The results showed that 17, 8, 11, 51 and 122 plants were scaled from 0-4 respectively, in which 0 means no symptoms and 4 represents highly susceptible. Chi-square test demonstrated that the segregation ratio of phenotype was 1∶15 (resistant: susceptible) or 1∶6∶9 (resistant∶moderate∶susceptible) in the F₂ population, indicating RBSDV resistance of maize was controlled by two recessive genes. The F₂ individuals DNA were extracted and 261 SSR (simple sequence repeat) primers derived from maize genome ten chromosomes were selected from maize GDB database to construct genetic linkage map. The linkage map consisted of 71 polymorphic SSR markers, spanning a genetic distance of 996.6 cM with an average interval of 14.0 cM between adjacent markers. The resistant and susceptible gene pools were set up for BSA (bulked segregant analysis) and 6 polymorphism markers were obtained with BSA-SSR method between the two pools. The F₂individuals were further analyzed with 6 polymorphism markers. Chi-square test showed that phi 051, umc1407 and umc1432, mapped on chromosome 7 and 10, exhibited segregation distortion significantly and very significantly in susceptible individuals. These three SSR markers were identified as potential markers linked to the resistant loci.     

水稻瘤矮病毒P12蛋白抗血清制备及其应用

为了进一步研究水稻瘤矮病毒(Rice gall dwarf virus,RGDV)P12的功能,将RGDV-GX P12编码区亚克隆至原核表达载体,并导入大肠杆菌诱导表达;重组的P12融合蛋白经Ni-NTA His·Bind Resins纯化后用于免疫小鼠制备抗血清,并进行Western blotting分析。结果显示,约41 kD大小的RGDV P12融合蛋白可在大肠杆菌中高效表达;利用该重组蛋白所制备的抗血清可特异地与融合表达和非融合的P12蛋白发生强烈的免疫学反应。利用该特异性抗血清在病毒粒子样品中检测不到P12蛋白,而在病株总蛋白样品中可检测到1条与预期大小一致的明显条带,表明RGDV P12是一种非结构蛋白。     

我国粮食作物病毒病发生与防控现状

近年来,我国水稻、小麦、玉米病毒病发生严重,造成重大经济损失。本文就近年来水稻、小麦和玉米等主要粮食作物上水稻条纹叶枯病、水稻黑条矮缩病、南方水稻黑条矮缩病、小麦黄花叶病以及玉米粗缩病的发生现状、防控情况,以及防控过程中存在的问题作简要概述,并展望下一阶段防控任务。