粉蕉、大蕉和龙牙蕉的AFLP分类研究

 应用AFLP 技术, 对32 个粉蕉、大蕉、龙牙蕉品种进行了遗传多样性分析及分类研究。在分子水平上, 可将供试品种聚为6 个独立群体, 其中包括3 个分别由单一品种组成的群体(群体Ⅳ: 酸大蕉;群体Ⅴ: 孟变; 群体Ⅵ: 千旁培) ; 可以将群体Ⅰ (大蕉) 和群体Ⅱ (粉蕉) 分别划分为3 个品种群, 将群体Ⅲ (龙牙蕉) 划分为2个品种群。参照Simmonds 的形态学分类方法, 研究结果显示粉蕉、大蕉以及龙牙蕉各个品种之间存在丰富的遗传多样性, 将其划分为AAB 或ABB 等类型, 不能反映品种间的亲缘关系; 粉蕉与大蕉两个类群之间遗传距离差异明显, 将其简单的归为ABB 类型是不妥当的。     

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.     

Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers

Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed.     

Rapid detection of Phytophthora cinnamomi using PCR with primers derived from the Lpv putative storage protein genes

Phytophthora cinnamomi is an ecologically and economically important pathogen. In this study, PCR assays were developed with primer pair LPV2 or LPV3 for rapid detection and identification of this organism. Both primer pairs were selected from putative storage protein genes. The specificity of these primer pairs was evaluated against 49 isolates of P. cinnamomi , 102 isolates from 30 other Phytophthora spp., 17 isolates from nine Pythium spp. and 43 isolates of other water moulds, bacteria and true fungi. PCR with both primer pairs amplified the DNA from all isolates of P. cinnamomi regardless of origin. The LPV3 primers showed adequate specificity among all other species tested. The LPV2 primers cross-reacted with some species of Pythium and true fungi, but not with any other Phytophthora species. PCR with the LPV3 primers detected the pathogen at levels of a single chlamydospore or 10 zoospores in repeated tests. The PCR assay was at least 10 times more sensitive than the plating method for detection of the pathogen from artificially infested soilless medium, and, to a lesser extent, from naturally infected plants. PCR with LPV3 primers can be a useful tool for detecting P. cinnamomi from soilless media and plant tissues at ornamental nurseries, whereas the LPV2 primers can be an effective alternative for identification of this species from pure culture. Applications of these assays for detection of P. cinnamomi in other environments were also discussed.     

棉花黄萎病菌致病型的AFLP分析

 选用41个棉花黄萎病菌(Verticillium dahliae)代表菌系,在温室条件下,对4个棉花品种鄂荆1号(感)、中棉所12(耐)、文-5(抗)和唐棉2号(抗)进行致病性测定,结果可将供试菌系分为落叶型与非落叶型2类。选取8对AFLP引物PCR扩增的结果中,统计带型稳定、清晰且有多态性的条带,共169条作系统聚类分析,将上述菌系分为2大类,第一类为非落叶型菌系,包括10个非落叶型菌系和1个过渡菌系;第二类为30个落叶型菌系。根据聚类分析建立树状图,发现菌系与地理来源存在一定的相关性,而依据菌系致病力强弱分类则相关关系不大。选用25对EcoRⅠ和MseⅠ引物组合,对供试的41个V.dahliae进行AFLP扩增,筛选到2对引物E64(GACTGCGTACCAATTCGAC)、M53(GATGAGTCCTGAGTAACCG)和E49(GACTGCGTACCAATTCCAG)、M65(GAT-GAGTCCTGAGTAAGAG),能分别扩增出433bp和110bp2条仅为V.dahliae非落叶型菌系独有的特异片段,可将落叶型与非落叶型菌系分开,这2条特异片段被命名为EM₄₃₃和EM₁₁₀。     

荔枝品种亲缘关系的AFLP分析

 应用AFLP技术，对39个荔枝品种进行了遗传多样性分析及分类研究。筛选出适于荔枝AFLP分析的最佳引物组合3对，分别为EcoRI AAC+MseI CTG，EcoPd AGC+MseI CAT，EcoRI ACC+Msel CAT。在分子水平上，荔枝品种的遗传多样性并非形态学性状所体现的那样丰富。AFLP分析将39个荔枝品种分成了6组，与传统的以果皮龟裂片隆起类型为分类标准的分类没有一致性。初步建立起荔枝主要栽培品种的分子标记标准图谱。应用AFLP技术对来自两个不同地方的两个荔枝品种(桂味、妃子笑)进行了鉴别。     

扩增片段长度多态性（AFLP）在动物遗传育种中的应用进展

AFLP是一种新的分子遗传标记技术，它结合了RFLP和PCR技术的优点，具有RFLP的稳定性和PCR的高效性，被称为“最有威力的分子遗传标记”或“下一代分子标记”。相对于国外而言，我国在这方面的研究还比较少。本文综述了扩增片段长度多态性(AFLP)这种遗传标记的原理、技术特点、技术发展及其在动物遗传多样性、种质鉴定、遗传图谱构建、基因定位和标记辅助育种等研究中的应用。     

棉铃虫核型多角体病毒分子生物学和基因工程研究进展

棉铃虫核型多角体病毒(HaSNPV)是棉铃虫专一性病原物,隶属于杆状病毒科核型多角体病毒属.对其分子生物学和基因工程的研究主要包括以下几个方面:测定了HaSNPV G4株和C1株基因组核苷酸全序列,并与其它病毒进行了同源性比较;研究了HaSNPV部分基因的结构、转录、表达及其功能.构建了HaSNPV Bac to Bac杆状病毒表达系统;重组病毒杀虫剂的研究为HaSNPV大面积防治棉铃虫展示了广阔的前景.随着棉铃虫核型多角体病毒分子生物学和基因工程研究的不断深入,重组病毒杀虫剂将在棉铃虫综合防治中发挥更为重要的作用.     

小麦叶锈菌生理小种MFR的分子鉴定研究

 用AFLP方法对来自中国和墨西哥的23个小麦叶锈菌生理小种进行分析,共筛选了64对引物,获得一对引物(M₀₅/E₀₃)可在MFR小种中扩增出一条特异性DNA片段,进行回收、克隆、测序,结果表明该片段具有325个碱基。根据特异性片段序列设计出SCAR标记引物,对60个叶锈菌生理小种分离物进行回检结果表明,研制的SCAR标记能够准确区分MFR生理小种。本实验结果为小麦锈菌生理小种分子检测体系的建立奠定了基础     

苹果基因组分子生物学研究进展

借助分子标记手段苹果基因组研究取得了巨大成就,涉及苹果的抗性、生长发育、果实品质等各方面基因,综合有关文献,将其研究的主要成果,包括已发现的主要苹果遗传标记、已构建的苹果遗传连锁图谱及苹果的转基因研究进行综述,并对目前苹果基因组研究存在的问题和对策进行了分析,以期为苹果遗传育种提供参考。