Construction of a genetic map with SRAP markers and localization of the gene responsible for the first-flower-node trait in cucumber (Cucumis sativus L.)

Cucumber (Cucumis sativus L.) is an annualclimber originated from the tropic rain forest area inthe south of Himalayas, which belongs to the Cucur bitaceae family. Cucumber is one of the importantvegetables in the world, and its planting area is sec ond only to that of tomato[1]. However genetics re search on cucumber obviously drops behind that of thelatter. The classic genetic map of cucumber, with sixlinkage groups, is composed of more than 100 genesfor color, morphology, and dise…     

黄瓜杂交种子纯度的RAPD鉴定

鉴定和分析了黄瓜品种真实性，并建立了应用于鉴定亲本及其杂交种子的RAPD指纹图谱。从102条引物中筛选出了应用于黄瓜种子纯度鉴定的引物32条：13条偏母型引物，9条偏父型引物，5条互补型引物，2条特异型引物及3条缺失型引物。利用4条引物S293，SS87，S297，S300鉴定3份黄瓜种子的纯度，在94粒种子中有父本混杂2粒，母本混杂1粒及6粒其它种子混杂，从而表明该份黄瓜种子的纯度为90．04％。建立了黄瓜早青二号父本T03雄性系、早青二号母本B35雌性系以及其杂交种子的RAPD的指纹图谱。     

黄瓜全雌性状相关的RAPD分子标记筛选

用CTAB法提取黄瓜总基因组DNA,采用RAPD技术探寻与黄瓜全雌性相关的分子标记.共筛选39条随机引物,其中有12条引物显示多态性.进一步筛选表明：S10引物能在全雌系黄瓜中扩增出一条约2000 bp的稳定、清晰的特异条带,而在雌雄同株的单株中不存在.经回收、克隆并测序,获得全长2003 bp的序列.在NCBI上进行Blast分析,表明为新发现序列,含有编码62个氨基酸的开放阅读框.其编码短肽的功能尚不确定.     

不同性别类型黄瓜品种基因组DNA的提取及RAPD标记的初步研究

采用改良的异丙醇沉淀法成功地提取了11个不同性别类型黄瓜品种的基因组DNA.以此作为PCR扩增反应模板进行RAPD分析,结果从22个10bp随机寡核苷酸引物中筛选出2个引物S23和S91在全雌性品种MT700和其他性别类型的品种之间稳定地扩增出多态性.至于该多态性标记是否与黄瓜雌性性别有关,还有待进一步分析.     

乌头基因组DNA提取与RAPD反应体系优化

采用改进方法提取乌头基因组DNA,建立乌头RAPD分析的PCR反应体系,成功地进行了RAPD扩增,筛选出乌头RAPD的最佳方案为:25μl反应体系中包括dNTPs 0.06 mmol·L-1,引物0.2μmol.μL-1,模板DNA40 ng,Taq酶1U,Mg2+ 4 mmol·L-1,10×buffer缓冲液2.5μl,其余部分为无菌超纯水。PCR扩增循环为95℃3 min,38 ℃1 min,72 ℃2min 1次循环;94 ℃=1 min,38℃1 min,72 ℃2 min,45次循环,最后72℃延伸10 min。     

不同生态型麦冬特异分子标记的选择与鉴定

采用RAPD技术对四川、湖北、浙江及上海等地 9个不同生态型的麦冬进行遗传多样性研究 ,筛选出 8个可以扩增出清晰、稳定、重复性好和多态性高的随机引物 ,通过对 8个有效引物的指纹图谱分析 ,发现 5个特异的RAPD分子标记 ,可以对 5个不同生态型的麦冬进行区别鉴定     

Isolation and genetic characterization of a fragile plant mutant in rice (itOryza sauva) L.

The fragile rice mutant was isolated from an M₂ population of indica variety Shuang Ke Zao (SKZ) treated with g-rays, and designated as fp1 (fragile plant 1) because of its fragile leaves and culms. To map FP1 locus, an F² mapping population was derived from a cross between the fp1 and C-bao, a polymorphic japonic variety. The primary mapping result places the FP1 locus in an interval between two molecular markers, microsatellite marker RM16 (3.1 cM proximal to FP1) and STS marker G144a (9.1 cM distal to FP1) in the centromere region of chromosome 3. A CAPS marker C524a was further developed between RM16 and G144a, with 0.4 cM genetic distances to the FP1locus, providing a practical starting point for constructing a BAC contig spanning the FP1 locus and cloning the fp1 gene. Allelism test demonstrated that fp1 is allelic to bc1, a fragile rice mutant reported previously.     

Genetic analysis and gene mapping of leafy head （lhd）, a mutant blocking the differen-tiation of rachis branches in rice （Oryza sativa L.）

Flowers, fruits and seeds are products of plant re- productive development and provide the important sources of foods for humans. Therefore, the moleculargenetic mechanisms of floral development have been ahotspot of research of plant developmental biology[1]. Rice is one of the most important staple food crops. Theoutcome of its reproductive development would determine the yield and quality of grains. Rice is also a model plantof cereals. Hence, the study of rice reproductivedevelopment, esp…     

24份普通白菜遗传多样性的形态特征和RAPD分析

利用形态特征与RAPD分析技术,分析了国内不同地区的24份普通白菜(Brassica campestris L.ssp.chinensis(L.)Makino var.communis Tsee et Lee.)种质的遗传多样性.结果表明,13个形态学性状的变异系数为12.4%~69.9%,其中叶柄变异最大,子叶变异最小.24份普通白菜材料在形态学聚类的距离5处聚为4类.利用筛选出的15条RAPD引物从24份普通白菜材料中共扩增出113条带,其中多态性条带83条,多态率73.4%,平均每条引物条带7.53条、多态性条带5.53条.RAPD数据聚类在相似系数0.77处,分为8类,与形态聚类间有重叠和细分.以形态聚类4个类别计算的Nei's基因多样性指标H和Shannon信息指数I,均为第Ⅳ大类第Ⅰ大类第Ⅲ大类第Ⅱ大类,第Ⅳ类的H和I分别为0.2395和0.3523.而不同地区则以江淮地区的Nei's基因多样性H和Shannon信息指数I最高,分别为0.2076和0.3098.这些结果与普通白菜种质资源多样性特征以及地区分布相吻合.     

菠菜叶绿体基因组定点整合表达载体构建及原核表达

分析菠菜叶绿体基因组全序列,选用了rbcL基因和accD基因的间隔区作为外源基因的定点整合位点,并从菠菜叶绿体基因组中克隆了rbcL基因全长和accD基因的5′端部分,长度分别为1956 bp和1320 bp。以这2个DNA片段作为同源重组片段,以烟草叶绿体基因的启动子Prrn和终止子psbA3′控制外源基因的转录,构建了包含筛选标记基因aadA基因(编码氨基糖苷-3′-腺苷酸转移酶,具有壮观霉素和链霉素抗性)和报告基因GFP(编码绿色荧光蛋白)的菠菜叶绿体基因组定点整合表达载体pRAGA。酶切结果显示构建正确。将该载体转化大肠杆菌,在激光扫描共聚焦显微镜下用488 nm蓝光激发,发现大肠杆菌发出强烈的绿色荧光,而对照菌体没有荧光,表明GFP基因在原核大肠杆菌中已经成功表达。实验结果说明构建的菠菜叶绿体定点整合表达载体pRAGA可以用于菠菜叶绿体转化。     

Genetic analysis and gene fine mapping of a rolling leaf mutant (<Emphasis Type="Italic">rl</Emphasis><Subscript><Emphasis Type="Italic">11(t)</Emphasis></Subscript>) in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The exploration of new genes controlling rice leaf shape is an important foundation for rice functional genomics and plant archi-tecture improvement. In the present study, we identified a rolling leaf mutant from indica variety Yuefeng B, named rl_11(t), which exhibited reduced plant height, rolling and narrow leaves. Leaves in rl_11(t) mutant showed abnormal number and morphology of veins compared with those in wild type plants. In addition, rl_11(t) mutant was less sensitive to the inhibitory effect of auxin than the wild type. Genetic analysis suggested that the mutant was controlled by a single recessive gene. Gene Rl_11(t) was initially mapped between SSR markers RM6089 and RM124 on chromosome 4. Thirty-two new STS markers around the Rl_11(t) region were developed for fine mapping. A physical map encompassing the Rl_11(t) locus was constructed and the target gene was finally delimited to a 31.6 kb window between STS4-25 and STS4-26 on BAC AL606645. This provides useful information for cloning of Rl_11(t) gene.