DNA分子标记检测家蚕苏菊、明虎品种和纯度的研究

利用单粒卵微量DNA抽提法,从江苏省家蚕主要品种苏菊、明虎及其一代杂交种蚕卵快速提取出基因组DNA,用筛选出的 RAPD随机引物S77稳定扩增出苏菊、明虎的DNA特异性片段RS和RM,克隆和测序后大小分别为1694bp和1303bp,使用Blastn程序在NCBI 数据库检索发现,Rs的nt30-429与一个家蚕WGS(whole genome shotgun sequence)(dbj|BAAB01119085．1,contis504766)的nt472-874有92％的同源性,其中存在11个空缺位点(Gaps);RM的nt1070-1301与另一个家蚕WGS(dbj|BAAB01141578．1,contig554782)的nt262-31有91％的碱基相同,其中存在2个Gaps。进一步根据RS和RM序列合成SCAR标记引物PS-1(FP:5’ttccccccagtacgcaataac3’,RP:5’ ttccccccagacgtcacgtact3’)和PM-1(FP:5’ttcccccagtgatggaatttacg3’,RP:5’ttccccccagtccaatgttaca3’),能够准确、快速地鉴别所标记的苏菊、明虎及其F1蚕品种。     

Regular Production of Transgenic Wheat Mediated by Agrobacterium tumefaciens

Wheat is the number one crop both in acreage and total yield in the world. Therefore, it is very important to improve wheat by gene engineering techniques even though it belongs to the plants insensitive to gene transformation, especially to Agrobacterium-mediated transformation. Wheat immature embryos of 1.0-1.5mm in size, C58c1 of Agrobacterium strain harboring pPTN249, pPTN270, pPTN254, and pSIS-GFP respectively (all the vectors contain the aphA selectable gene driven by enhanced 35S promoter and a target gene controlled by ubi promoter or E35S promoter), AB medium for Agrobacterium activate culture, WCC medium for co-culture, were used in this study. The embryos with 4 days of pre-culture were transferred onto selection medium with 10mg/L geneticin, 50mg/L ticarcillin, 50mg/L vancomycin, and 50mg/L cefotaxine after 30 minutes of infection and 2 days of co-cultivation with Agrobacterium. Followed callus production,shoot regeneration on selection medium, 114 resistant plantlets were obtained from 10 transformation experiments of four genotypes. By nptⅡ ELISA (nptⅡ enzyme assay), PCR, Southern blot and leaf bleach,29 positive plants were identified from two genotypes of Bobwhite and Yanglmai 10, with an average transformation efficiency of 0.82 %. The result tested by Southern blot also showed that the transgenic plants with single- copy integration of target gene took 65.52% among total positive plants. The ELISA value of transgenic plant was also related to the copies of alien DNA integrating into wheat chromosomes, the transgenic plants with single copy integration giving higher ELISA value than the ones with 2 or 3 copies integration.     

黄瓜全雌性基因连锁的AFLP和SCAR分子标记

 本研究以全雌品种‘戴多星’自交系和弱雌品种‘北京截头’自交系为双亲杂交获得F₁ ,然后得到F₂ 性型分离群体, 利用分离群体分组分析法(Bulked Segregant Analysis, BSA) 构建全雌和弱雌两个基因池, 筛选了64对AFLP选择性引物EcoR I-NN +Mse I-NNN组合, 发现EcoR I-TG +Mse I-CAC引物组合在全雌基因池中扩增出一条分子量为234 bp的特异带。经F₂ 代单株验证, 该特异条带能在全雌单株中稳定出现。以MAP MAKER (Version 310) 软件分析, 该标记与全雌性位点的连锁距离在617 cM。命名该连锁标记为TG/CAC₂₃₄。将该特异条带回收、克隆、测序, 设计特异SCAR引物, 再对F₂ 代单株基因组DNA进行扩增, 仅在全雌单株中扩增出1条分子量为166 bp 的特异带, 表明已成功地将与黄瓜全雌性连锁的AFLP标记转化为操作简便、表现稳定的SCAR标记, 该标记命名为SA₁₆₆。     

遗传连锁方法的历史,现状与发展

本文简要介绍了遗传学当中的连锁分析方法的历史，现状及其近几十年的发展，并对这些方法的优缺点进行了评价。     

QTL Analysis for Plant Height with Molecular Markers in Maize

Plant height has become one of important agronomic traits with the increase of planting densityrecently and the rapid developments of molecular markers have provided powerful tools to localize importantagronomic QTL at the genomic level. The purposes of this investigation are to map plant height QTL with mo-lecular markers and to analyze their genetic effects in maize. An F2:3 population from an elite combination(Zong3 × 87-1) was utilized for evaluating plant height in two locations, Wuhan and Xiangfan, with a ran-domized complete block design. The mapping population included 266 F2:3 family lines. A genetic linkagemap, containing 150 SSR and 24 RFLP markers, was constructed, spanning a total of 2 531.6 cm with an av-erage interval of 14.5 cm. Totally 10 QTL affecting plant height were mapped on six different chromosomeswith the composite interval mapping. Seven of 10 QTL were detected in two locations. The contributions tophenotypic variations for the single QTL varied between 5.3 and 17.1%. Additive, partial dominance, domi-nance, and overdominance actions existed among all detected QTL affecting plant heights. A large number ofdigenic interactions for plant height were detected by two-way analyses of variance. 107 and 98 two-locus com-binations were found to be significant at a 0.01 probability level in two locations respectively. 23 of them weresimultaneously detected in both locations. They accounted for phenotypic variations of 4.5 -11%. It was no-ticed that a locus, umc1122, had digenic interactive effects with other four different loci for plant height,which distributed on three chromosomes. A few of plant height QTL was involved in significant digenic inter-actions, but most significant interactions occurred between markers that are not adjacent to mapped QTL.These results demonstrated that epistatic interactions might play an equal importance role as the single-locuseffects in determining plant height of maize.     

利用分子标记和接种鉴定分析美国水稻育种亲本的稻瘟病抗性

稻瘟病是由真菌Magnaporthe oryzae引起的,水稻与稻瘟病菌的互作符合经典的“基因对基因”学说,抗稻瘟病基因Pi-ta可有效防治携带A VR-Pita1稻瘟菌的侵染.本研究利用位于Pi-ta基因前端的显性 .分子标记YLI53/YL153和位于中间区域的显性分子标记YL155/YL87对39份来自美国的水稻...     

分子印迹技术在药学中的发展应用及前景

介绍了分子印迹技术在药学中药物分离和分析、中药活性成分的分离纯化、药物残留检测(环境检测、食品药品残留分析检测)、药物传递系统中的应用,并对分子印迹技术在药学领域中的发展前景进行了展望。     

细菌IV型分泌系统结构的研究进展

革兰氏阴性菌的IV型分泌系统（T4SS）是一个由ATP供能,多种蛋白质复合构成的体系,该体系能够跨过整个细胞外膜、分泌大量底物。详细阐述了T4SS组成元件的结构,如底物转运通道、核心复合物、外膜复合物、内膜复合物、菌毛等。T4SS更大组成元件、其他结构、组成元件间相互作用的机理等有待进一步研究。     

牦牛肺炎病原多杀性巴氏杆菌和殊异支原体的分子鉴定与分析

青海省刚察县某牦牛产业基地牦牛发生咳嗽,鼻孔有分泌物流出,呼吸困难等症状的疾病。为快速确诊发病牛的致病原因,及时防控和治疗,从发病和病死的牦牛中,采集鼻腔拭子和肺脏、胸腔积液,利用PCR方法进行分子鉴定与分析。结果显示:此牛场中引发肺炎的病原是A型多杀性巴氏杆菌和殊异支原体两种病原体混合感染,测序结果显示5个有效样品中鉴定的为完全相同的病原,P.multocida-GC1的Kmt基因与参考序列中国株CP031554/3/2/1同源性在99%以上,M.dispar-GC1与参考序列的16S rRNA基因同源性在97%以上。同时在遗传进化角度再次分析确认了鉴定的P.multocida与中国和俄罗斯的A型多杀性巴氏杆菌聚成一大支;鉴定的M.dispar与殊异支原体聚为一支。结果表明,青海刚察地区牦牛存在感染多杀性巴氏杆菌和殊异支原体的潜在风险,该研究为牦牛呼吸道疾病与肺炎的病原学调查和病原的流行病学调查提供了参考数据。     

芸薹属蔬菜霜霉病研究概况及进展

霜霉病是世界范围内芸薹属蔬菜生产中常见的重要病害,病原菌为卵菌门无色霜霉属寄生无色霜霉(Hyaloperonos poraparasitica),叶片是病菌最主要的为害部位,植株发病后叶片出现枯黄症状,导致芸薹属蔬菜品质和产量下降,严重时可减产20%~60%,造成大量的经济损失,制约我国芸薹属蔬菜产业的发展。虽然霜霉病为害严重,但目前对于芸薹属霜霉病的发生规律和防治方法的认知还存在不足。本文从芸薹属蔬菜霜霉病的发生为害、防治措施、抗性遗传规律、分子标记等方面对其研究现状进行总结概括,并对未来芸薹属霜霉病的研究方向进行展望,旨在为相关研究提供参考。