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.     

三种微生物杀虫油烟剂成烟后菌体活性的测定

本研究通过电子显微镜扫描和菌体活性测定三种微生物杀虫油烟剂成烟后菌体的活性,结果表明:使用烟雾机分别施放白僵菌、苏云金杆菌和松毛虫质型多角体病毒三种微生物杀虫油烟剂,其成烟后,三种菌体能保持原来的形态和活性,白僵菌孢子能正常萌发,苏云金杆菌芽孢在营养琼脂上能形成晶体,松毛虫质型多角体病毒能有效地感染松毛虫,为进一步研究常规和应急防治松毛虫的简单易行、高效的使用方法提供技术依据。     

稻瘟菌激发子CSB I专化性及相关性质研究

 以一套水稻抗稻瘟病近等基因系为材料,接种稻瘟菌(Magnaporthe grisea)细胞壁来源的糖蛋白激发子CSB I,其诱导植保素的积累在高度非亲和性互作水稻远高于亲和性互作水稻;研究同时表明,CSB I可专化性诱导完全非亲和性互作和高度非亲和性互作水稻的过敏性坏死反应;表明该激发子具有小种-品种专化性。经热、胰蛋白酶和过碘酸钠处理后的生物活性检测结果表明,CSB I的活性位点为糖基部分。经pH稳定性检测,CSB I在酸性及相对弱碱性条件下较稳定;而在强碱性条件下,激发子活性下降较多,甚至完全丧失。对CSB I诱导活性的有效浓度测定表明,激发子诱导水稻叶片酶活性升高的最低有效浓度为0.07~0.70 nmol/L。     

来源于新疆的3 个苹果褪绿叶斑病毒分离物的分子变异研究

Apple chlorotic leaf spot virus (ACLSV) isolates from Korla pear (KI-2), New pear no. 7 (XI-1) and Red Fuji apple (API-4) were collected from XinJiang and characterized by analyzing sequences of their near genomic 3忆-terminal. The RT-PCR products were cloned, and analyzed by single-strand conforma-tion polymorphism (SSCP). Eight out of 39 collected positive clones showing different SSCP patterns were sequenced. The results showed that the amplified products had sizes ranging 676 - 703 bp, including partial coat protein (CP) gene (506 bp, accounts for 87% of the complete cp gene) and 3忆-terminal non-coding re-gion (3忆NCR) sequences. The cp gene sequences from isolate KI-2 showed a high intra-isolate divergence,with 84. 8% - 85. 4% identities at the nucleotide (nt) level, and the intra-isolate identities were 99. 8 % and 92.5% - 99. 8 % for isolate XI-1 and API-4, respectively. Phylogenetic analysis on the nt sequences of cpgene showed that the analyzed ACLSV variants from three isolates fell into two different clusters. A variant KI-2-6 from KI-2 was clustered into a group with an apple isolate aclsv-c from China and a plum isolated from France, and all other variants fell into a large cluster. The 3忆NCR sequences of these variants were identical ranging 80. 6% - 100 % .     

花期低温对水稻叶鞘生理及结实率的影响

盆栽条件下,以耐冷性不同的2个水稻（Oryza sativa L.）品种龙稻5（耐冷型）和龙粳11（冷敏型）为材料,于开花期在人工气候室进行低温（15℃,分别持续1,2,3,4,5 d）处理,研究开花期低温对不同耐冷性水稻结实率及叶鞘膜透性、抗氧化酶等生理指标的影响。结果表明：低温处理2 d时,龙稻5和龙粳11的花粉活力分别下降了8.21%和16.10%,差异达极显著水平;低温处理3 d后,龙稻5的结实率下降到99.90%,与CK相比差异达到显著水平;低温处理1 d后,龙粳11的结实率下降到60.04%,与对照相比差异达到极显著水平。低温处理提高了水稻叶鞘相对电导率和脯氨酸、可溶性蛋白含量,处理5 d时各指标达最大值,与CK相比,龙稻15和龙粳11分别提高了38.01%和20.77%、25.06%和84.85%、25.82%和23.63%。同时低温处理也提高了叶鞘超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性,处理3 d时酶活性达最大值,龙稻15和龙粳11分别较CK提高了10.57%和14.90%、86.26%和68.71%、17.73%和28.25%。综上所述,开花期低温引起水稻叶鞘膜脂过氧化程度升高,但也诱导水稻叶鞘抗氧化系统开启、保护酶活性上升以抵御低温伤害;开花期低温导致水稻花粉活力降低并引起结实率降低,耐冷品种较冷敏品种抵御能力强。     

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。     

山西苹果茎痘病毒遗传多样性分析

苹果茎痘病毒是苹果安全生产的巨大威胁,严重影响苹果产量和质量。探讨苹果茎痘病毒(Apple stem pitting virus, ASPV)在山西省的分布、变异和多样性,可为制定可持续的病毒管理策略提供理论依据。从山西省11个地区采集苹果叶片样品进行RT-PCR检测。结合在线数据库,利用相关软件对文中研究获得的序列进行系统发育、遗传多样性、重组以及种群遗传分化等分析。RT-PCR检测结果显示从山西省11个地区采集的409份苹果叶片样品中,ASPV的检出率为36.33%(临漪)至55.37%(平遥)。选择15个ASPV检测为阳性的样品,分别克隆其CP基因并与22个不同的ASPV分离物进行核苷酸和氨基酸一致性分析,发现37个ASPV分离物的核酸一致率为70.27～96.31%,氨基酸一致率为37.04～96.64%,其中,Shanxi-2与JX673826.1的核酸一致率最高,为90.86%, Shanxi-2与MN617853.1的核酸一致率最低,为70.40%。基于CP的进化树分析结果显示37个ASPV分离物被划分为2个进化群体。遗传多样性分析结果表明ASPV种群发生了收缩,且在两个...     

胶体金免疫层析法快速检测烟草环斑病毒

采用柠檬酸三钠还原法制备胶体金颗粒 ,标记烟草环斑病毒的抗体 ,制成免疫层析检测试纸条。检测粗提纯病毒的灵敏度为 1 0 0 0ng/ml,病汁液稀释 1 0 0 0倍后仍可快速检出。对大豆病种子、烟草冻干病叶等不同材料进行检测也有良好的效果 ,1～ 2min即可出现结果。用 9种不同的病毒进行测试 ,未出现非特异性反应。