Comparison of genetic variation among accessions of <Emphasis Type="Italic">Aegilops tauschii</Emphasis> using AFLP and SSR markers

Genetic diversity of 54 accessions of Aegilops tauschii from five countries was assessed using sequence-tagged microsatellites (or simple sequence repeats, SSRs) and amplified fragment length polymorphisms (AFLPs). In the case of AFLP analysis, a total of 256 amplification products obtained, 234 of them were polymorphic across all the 54 accessions. A total of 224 fragments were obtained from the 24 SSR primers and 219 of fragments were polymorphic across all the genotypes screened. Based on both AFLP and SSR markers, the highest percentage of polymorphisms were obtained in Iranian and accessions of unknown origin. The highest polymorphic information content (PIC) value was observed for SSRs (0.82) while the highest marker index (MI) value was for AFLPs (8.5) reflecting the hyper-variability of the first and the distinctive nature of the second system. Principal co-ordinate analysis (PCO) revealed congruent patterns of genetic relationships for both data sets, but did not group accessions strictly according to their geographical origins. Poor correlation was found between AFLP and SSR marker loci. This low association may be due to low number of AFLP and SSR markers. These results show that molecular markers can help to organize the genetic variability and expose useful diversity for breeding purposes.     

Genetic diversity of Syrian pistachio (<Emphasis Type="Italic">Pistacia vera</Emphasis> L.) varieties evaluated by AFLP markers

Pistachio (Pistacia vera L.) is a strategic nut tree species in the Middle East which holds comparative advantage over other fruit trees in view of its hardiness, income generation opportunities and benefits for the ecosystem. Yet pistachio cultivation depends on a very narrow genetic base, in spite of the existence of many varieties still marginally exploited. Syria is an important center of diversity for pistachio. A country wide ecogeographic survey in this country was carried out to determine the extent of pistachio genetic diversity and its use. As a whole, 114 accessions were collected from 37 farms to assess diversity at morphological and molecular level. Molecular evaluation was carried out using Amplified Fragment Length Polymorphism (AFLP) technique and performed using seven primer pair combinations. Results from the studies allowed the identification of 25 pistachio female varieties in Syria, some of which unique and described for the first time. Three groups of pistachio diversity were identified by cluster analysis which provides useful information about the distribution of genetic diversity in Syria for enhanced use and sustainable conservation.     

野生绿豆种质资源TC1966抗豆象基因的AFLP标记研究

野生种抗豆象绿豆种质资源TC1966对多个豆象小种均具有很好的抗虫性。通过对抗虫亲本TC1966、感虫亲本绿1号及以中绿1号为轮回亲本经多代回交后获得的3个近等基因系材料进行抗虫鉴定．结果轮回亲本中绿1号的种子被害率〉90％．表现高感（HS）；抗性亲本TC1966及三个高代材料种子被害率〈10％，表现高抗（HR）．说明抗虫亲本TC1966中的抗虫基因呈显性遗传，且能够在后代材料中稳定遗传和表达。本文利用AFLP分子标记技术对该5份试验材料进行了多态性指纹图谱分析．结果从830对AFLP引物组合中筛选出在抗、感虫材料间表现多态性的引物组合100对。将部分多态性AFLP特异扩增条带从聚丙烯酰胺凝胶上回收，然后将二次扩增产物进行测序，并通过同源性序列比较的手段进行了相关生物信息学分析。本研究为今后进一步开展抗豆象基因的AFLP分子标记及其转化研究乃至抗豆象基因的克隆工作提供参考依据。     

人工三倍体桑树新品种嘉陵16号遗传背景的AFLP分析

利用AFLP分子标记技术,从DNA水平对人工三倍体新桑品种嘉陵16号(2n=3x=42)及其亲本进行了遗传背景分析。获得了它们的指纹图谱、遗传距离及遗传相似系数,并绘制出了它们的UP-GMA聚类图。研究结果发现:嘉陵16号的母本西庆一号(4X;♀)与父本育二号(2X;♂)之间的遗传相似系数为0.6607。这为今后更具强大杂交优势和多倍体优势的新型人工三倍体新桑品种选育时杂交亲本的选择与组配提供了较有用的参考依据。     

家蚕杂交分离系的AFLP分析初探

以日系家蚕品种"湘晖"和"872"为亲本进行杂交,从 F2代分别向茧丝量高(A)、中(B)、低(C)3个方向进行定向选择.经过F2～F6共5代的同蛾区交配和选择,获得了茧丝量性状有显著差异的A,B,C共3个家系.以中系蚕品种芙蓉、菁松及其杂交后代为参照群体,对3个杂交分离系进行AFLP分析,获得了617个AFLP分子标记.结果显示:选择到F4代时,A,B,C各杂交分离系的AFLP分子标记表现出一定的差异,到F6代时,A,B,C各杂交分离系间的分子标记差异十分显著且稳定.利用UPGMA法进行聚类分析的结果也显示有很强的规律性,所有个体均按家系有规律地聚在一起.从分子水平证明家蚕同蛾区杂交后代系统选择到F6代即可获得遗传性稳定的不同杂交分离系.     

分子标记在花生中的应用现状及前景

分子标记反映了DNA分子的多态性,可以作为研究生物遗传变异和进化关系的重要手段。介绍了常用分子标记技术的原理、方法、特点及其在花生上的应用现状,同时展望了分子标记技术在花生上的应用前景。     

AFLP技术及其常见问题与解决方案

结合工作实践及当前最新研究进展对AFLP分析过程中模板DNA的制备、基因组DNA的酶切与连接、酶切连接产物的预扩增与选择性扩增、扩增产物的变性聚丙烯酰胺凝胶电泳等过程中的常见问题及其解决办法作了简要综述。     

空间诱变辣椒AFLP反应体系的优化

为了建立适合空间诱变辣椒的AFLP反应体系,以3～4叶期的辣椒叶片为试材,对体系中的DNA用量、酶切与连接、预扩增及产物的稀释倍数等关键因素进行了优化,确立了适合空间诱变辣椒的AFLP反应体系.结果表明:采用该体系对空间诱变辣椒进行AFLP分析,能够得到清晰稳定的分子标记图谱.     

Analysis of Genetic Diversity and Relationships in Waxy Rice (Oryza sativa L.) using AFLP and ISSR Markers

Opaque endosperm is the main phenotypic indicator for waxy rice, but other phenotypic and genotypic variation among waxy rice accessions has largely been ignored. Previous studies showed that wide diversity in starch physiochemical properties exists in both indica and japonica waxy rices, especially for starch gelatinization temperature (GT) which could be divided into a high- and a low-GT group. In the present study, amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) molecular markers were employed to examine genetic diversity and relationships of 56 waxy rice accessions. A total of 358 AFLP fragments were amplified with five primer combinations, showing a high level of polymorphism (78.3%). A total of 190 ISSR bands were generated with a single primer and a primer pair, showing a very high level of polymorphism (92.2%). The genetic distance matrices obtained from the two sets of markers were significantly correlated (r = 0.731, P = 0.004). The dendrogram generated with combined AFLP and ISSR markers could clearly differentiate the indica and japonica groups. Newly released varieties and breeding lines within each subspecies tended to be clustered together, whereas landraces were more distantly placed in the dendrogram. Only one AFLP band was found specific to the indica type, while no specific bands were found for starch GT. The implications for the conservation and breeding of waxy rice are discussed.     

Diversity of Nordic Landrace Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Revealed by AFLPs and Morphological Characters

The management of genebank collections of cultivated potato is costly due to the need for in vitro maintenance and virus eradication. Therefore, it is important to set up conservation strategies, which prevent duplicates entering the collections. In this study, 32 Nordic potato landraces were studied for 57 morphological traits and analysed for amplified fragment length polymorphism (AFLP). Most landraces could be distinguished based on the morphological characters, except five accessions. Using five primer combinations to generate 114 reproducible AFLPs, of which 63 (55%) were polymorphic, the five morphologically indistinguishable accessions were placed into two groups with identical AFLP patterns, suggesting that some of the accessions were redundant for long-term preservation. The AFLP data showed that the Nordic collection of potato landraces is composed of genetically different clones, and morphological analysis revealed a wide range of variability. This variability seems to be distributed randomly over the Nordic region since the cluster analysis based on AFLPs and morphological traits revealed no grouping based on the country of origin. Principal component analysis suggests that fewer morphological traits than used in this study will be sufficient to discriminate between different genotypes of cultivated potato (Solanum tuberosum L.). Future possibilities for rationalising potato collections are discussed.