利用大白菜DH群体构建AFLP遗传连锁图谱

 以大白菜‘汴早- 26’和‘光90E16’的F₁ 代进行游离小孢子培养所获得的含有59个株系的DH群体为试材, 利用AFLP技术通过63对引物筛选共获得346个AFLP多态性标记, 运用JoinMap 3.0软件构建大白菜遗传连锁图谱。该图谱主要包括10个连锁群, 总图距为708 cM, 平均图距为2.0 cM。     

番木瓜遗传改良研究进展

就番木瓜的株性和抗病性等性状遗传、杂交育种、人工诱变育种、离体培养、分子标记及基因工程育种等现代生物新技术育种方面的研究进展及相关问题进行了评述,重点综述了番木瓜株性遗传、远缘杂交与胚胎学、基因工程在抗环斑型花叶病毒病(PRSV)品种培育及分子标记辅助选择育种应用的研究进展,并分析了今后番木瓜育种的发展趋势。     

柑橘抗CTV转基因与分子标记研究进展

综述了柑橘抗衰退病基因工程中两方面的研究进展。介绍多种来源于柑橘衰退病毒(Citrustriztezavirus,CTV)核酸序列的转基因柑橘和抗性种质资源中抗性基因的分子标记,以及所涉及的方法和遇到的问题。目前研究表明,虽然已成功实现对病毒衣壳蛋白(CP)等基因的转化和Ctv等抗性基因的标记,但尚未获得对CTV有高度抗性的转基因柑橘,而抗性基因亦不能实现定点克隆和转化。因此上述两方面研究还有待深入。     

番茄基因转化中抗生素筛选压力的确定方法

在以抗生素为筛选标记的基因转化中。确定适宜的筛选压力是提高基因转化率的前提。本研究以卡那霉素为例，通过在10mg／L，20mg／L，30mg／L，50mg／L，100mg／L的筛选压力下，转化和非转化番茄子叶外植体的生长表现、愈伤组织和芽的分化率．来确定适宜的筛选浓度。本实验以该方法找到的适宜的卡那霉素筛选浓度为50mg／L。以此浓度为筛选压力进行遗传转化所获得的部分再生苗经southern blot鉴定．均为转化株。     

苜蓿育种几个问题的探讨

在对主要苜蓿遗传资源与遗传特性进行陈述的同时, 重点对苜蓿遗传特性与育种进程的关系、目前常见的育种方法以及苜蓿育种的发展现状进行了阐述,并就近年来我国在苜蓿育种领域内存在的主要问题进行了分析,旨在为今后的苜蓿育种工作提供参考依据。     

Using antisense nucleic acid technology to study the influence of POLκ on genetic stability

AIM:To establish cell line FL-POLκ^- and to study the role ofPOLκ(polymerase kappa) on genetic stability.METHODS:A mammalian expression vector expressing antisense POLκ gene fragment pMAMneo -amp^--POLκ was constructed by cloning the 1 690-1 918 fragment of POLκ gene into the mammalian expression vector pMAMneo-amp^- in antisense orientation. FL cells were fransfected with this antisense RNA expressing vector and selected by G418. Based on the shuttle-plasmid pZ189, the mutation assay was made.RESULTS:The spontaneous mutation frequency of supF tRNA gene in the plasmid replicated in the FL- POLκ^- was 11.2×10^-4, while it was 4.9×10^-4 and 3.7×10^-4 in the control cells FL and FL-M, respectively.CONCLUSION: POLκ playes an important role in maintenance of genetic stability.     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

玫瑰、月季、蔷薇等蔷薇属植物RAPD分析

 对蔷薇属的玫瑰、月季、蔷薇3 个种的15 个品种进行RAPD 分析, 16 个引物对受试品种PCR扩增共获得202 条谱带, 其中123 条(60. 9 %) 表现多态性。利用UPGAMA 法构建分子系统进化树, 以相似性系数0. 5 为阈值, 玫瑰与月季为一个聚类组, 蔷薇为另一个聚类组; 以相似性系数0. 6 为阈值, 4个蔷薇品种分为4 个不同的聚类组, 5 个月季品种为一个聚类组, 5 个玫瑰品种为一个聚类组, 平阴紫枝玫瑰单独为一个聚类组, 表明玫瑰与月季的亲缘关系较近, 两者与蔷薇的亲缘关系较远。     

根癌农杆菌介导绿色荧光蛋白基因转化印度酸桔的研究

 通过根癌农杆菌介导将绿色荧光蛋白基因转入印度酸桔的胚性愈伤组织中, 经潮霉素筛选,获得抗性愈伤组织, 并再生植株。对这些植株进行GUS 染色、PCR 分析、绿色荧光检测和Sourthern 杂交验证, 结果表明绿色荧光蛋白已经在转基因植株中表达。     

十五种柑桔种质资源的RAPD分析

对柑桔15个材料（其中包含14个种，1个为品种）进行了RAPD分析，从结果看，各材料之间具有不同的遗传距离，供试材料聚为8个类群，分别为柠檬，柠檬群；香橼，小香橼群；柚，红心柚群；宜昌橙，香圆群；香橙，大翼橙群；红桔，温州蜜柑群；枳登，富民枳群；枳群。采用RAPD技术可作为品种鉴别，亲缘关系和分类的手段。对所得结果进行了讨论。