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。     

潍坊萝卜中芜菁花叶病毒分离物的分子特性及CP基因的表达

 从表现红心病的潍坊萝卜块根上得到一芜菁花叶病毒( Turnip m osaic virus, TuMV) 分离物(TuMV-Ra) , 通过RT-PCR获得了该分离物的外壳蛋白(CP) 基因, 对其进行了序列测定, 并将其核苷酸序列及推导出的氨基酸序列与GenBank中登录的其它20个TuMV萝卜分离物的相应序列进行比较和分析。结果表明, TuMV-Ra与这20个分离物CP基因核苷酸序列的同源性为89.9% ～99.0% , CP氨基酸序列同源性为94.8%～99.7%; 其中与中国浙江杭州萝卜的两个分离物HZLB1、HZLB2同源性最高, 核苷酸序列的同源性为98.7%和99.0% , 氨基酸序列同源性为99.0%和99.7%。TuMV-Ra与1999年从表现相同症状的潍坊萝卜得到的分离物CHINA-WF CP基因核苷酸同源性为93.66% , 氨基酸同源性为96.53% , 说明病毒发生了比较大的变异, 而且变异的位点主要在CP的N末端。将TuMV-Ra CP基因克隆到原核表达载体pET-22b ( + ) , 在大肠杆菌BL21 (DE3) 中表达出分子量为38 kD的融合蛋白。Western blotting分析证明TuMV-Ra CP在大肠杆菌中得到了正确表达。     

桃果肉颜色、果皮茸毛和花粉育性性状的分子标记

 以91-42-51 ×瑞光2号的杂交后代共48个单株为试材, 采用集群分离分析法(BSA) , 对桃果实的有毛/无毛、白肉/黄肉以及无花粉/有花粉3 个性状进行了分子标记研究。研究获得SSR 标记UDP96-018 (300 bp) 与有毛性状连锁, 遗传距离4.5 cM, SSR标记UDP98-407 (680 bp) 与白肉性状连锁, 遗传距离为2.2 cM; 根据拟南芥雄性不育序列标记设计的引物扩增出的两个片段NNJ-I (600 bp ) 和NNJ-I (900 bp) 与桃无花粉性状之间的遗传距离为0 cM。这些标记的获得为桃分子标记辅助育种提供了有效的筛选手段。     

梨果实贮藏中可溶性果胶和半纤维素分子结构的变化

 为了了解分子结构变化与耐藏性的关系，以耐贮性不同的苹果梨和新高梨为试材，利用Ultrogel AcA 22凝胶柱渗漏分离，比较了梨贮藏中可溶性果胶和半纤维素分子大小的变化。结果表明，苹果梨的半纤维素分子大于新高梨。在贮藏过程中，耐贮性强的苹果梨果实可溶性果胶分子大小变化不明显，但半纤维素分子明显变小。相反，新高梨的可溶性果胶分子明显变小，而未观察到半纤维素分子大小的变化。说明梨的耐贮性与可溶性果胶分子结构变化及半纤维素分子大小有关，而与半纤维素分子结构变化没有直接关系。     

桑天牛卵啮小蜂孤雌生殖及体内沃尔巴克氏体(Wolbachia)的检测

桑天牛卵啮小蜂(Aprostocetus prolixus)有孤雌生殖现象,其后代几乎为雄性。沃尔巴克氏体(Wolbachia)是广泛分布于节肢动物体内的一类共生菌,参与多种调控寄主的生殖活动机制。通过对外膜蛋白基因(wsp)、细菌细胞分裂蛋白基因(ftsZ)和核糖体16S rDNA基因的特异性扩增,在米蛾(Corcyra cephalonica Stainton)的DNA中分别扩增出约600、1000和900 bp的片断,验证了米蛾体内已感染了Wolbachia;而在桑天牛卵啮小蜂的DNA中未扩增出任何片断,表明Wolbachia在桑天牛卵啮小蜂体内未被感染或感染率极低。     

Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers

RAPDs and SSRs were used to study the genetic diversity of Iranian almond cultivars and their relationship to important foreign cultivars and three related species. Eight unidentified almond Shahrodi cultivars and three wild almonds (Prunus communis, Prunus orientalis and Prunus scoparia) were also included. Of the primers tested, 42 (out of 80) RAPD and 18 (out of 26) SSR primers were selected for their reproducibility and high polymorphism. A total of 664 polymorphic RAPD bands were detected out of 729 bands. The number of presumed alleles revealed by the SSR analysis ranged from 3 to 10 alleles per locus with a mean value of 6.64 alleles per locus. Both techniques discriminated the genotypes very effectively, but only RAPDs were able to discriminate the cultivars Monagha and Sefied. Results demonstrated an extensive genetic variability within the tested cultivars as well as the value of SSR markers developed in peach for characterization of almond and related species of Prunus. Dice similarity coefficient was calculated for all pair wise comparisons and was used to construct a UPGMA dendrogram. For both markers a high similarity in dendrogram topologies was obtained although some differences were observed. All dendrograms, including that obtained by the combined use of both the marker data, depicted the phenetic relationships among the cultivars and species, depending upon their geographic region and/or pedigree information. Almond cultivars clustered with accession of P. communis showing their close relationship. P. orientalis and P. scoparia were clustered out of the rest of P. dulcis.     

动物性别决定的分子机理及性别鉴定与控制新技术

哺乳动物中位于Ｙ染色体短臂临界区域的ＳＲＹ基因启动雄性性状的发育。该基因在生殖腺脊中的表达可激发下游基因ＭＬＳ的转录,引起缪氏体抑制物和睾酮分泌,促使睾丸组织器官的发育。针对该基因制备特异探针或产该区域片段得到引物对就可用ＦＩＳＨ或ＰＣＲ准确,快速鉴定出植入母体前的胚胎性别,以及对精子筛选分离的结果作出准确评价。与ＰＣＲ相比,ＦＩＳＨ技术更具检测优势。     

基于AT89S51的汽车智能安全报警系统

利用AT89S51设计了一种新型汽车防盗报警系统.当满足报警条件,单片机会向GSM模块发出指令,系统向机主终端发出报警求助短信,同时还发出声光报警、切断发动机电源.本系统具有误报率小,可靠性高,成本低,使用方便等优点.     

Analysis of 18S rRNA gene of Octostigma sinensis (Projapygoidea: Octostigmatidae) supports the monophyly of Diplura

The phylogenetic position of Diplura within Hexapoda has been controversial. There are three major lineages in Diplura: Campodeoidea, Projapygoidea, and Japygoidea. However, most of the previous studies were restricted to Campodeoidea and Japygoidea. Until now, only preliminary morphological study on Projapygoidea was reported, and no sequence data from Projapygoidea was available. The main aim of the present study was to investigate the phylogenetic position of Octostigmatidae, one of the three families of Projapygoidea, in Diplura and to test if Diplura are monophyletic. The complete 18S rRNA gene sequences of Octostigma sinensis (Projapygoidea: Octostigmatidae) from subtropical China, together with representative species of Campodeoidea and Japygoidea, and several species of Protura and Collembola were analyzed. The phylogenetic trees were obtained by different methods (neighbor-joining, maximum parsimony, and maximum likelihood) with a chelicerate species as outgroup. Our results suggested that Octostigma was closer to the genus Parajapyx (Japygoidea: Parajapygidae) than to the representative genus of Campodeidae (Campodeoidea). All phylogenetic trees supported the monophyly of Diplura.