甜菜DAMD引物的筛选及不同凝胶系统对扩增产物多态性的影响

为了筛选适宜于甜菜品种纯度鉴定的DAMD引物,笔者利用12 个不同的甜菜品种分别对26 条DAMD引物进行扩增,并分别采用6%聚丙烯酰胺凝胶电泳和2%的琼脂糖凝胶电泳对扩增产物进行检测。结果表明,从26 条DAMD引物中筛选出16 条扩增清晰的引物,这16 条引物共扩增出139 条带,其中多态性条带为122 条,多态性百分比为87.7%,这16 条引物可以作为甜菜品种纯度和真实性鉴定的核心引物,同时发现聚丙烯酰胺凝胶电泳与琼脂糖凝胶电泳的检测结果差异不大,而琼脂糖凝胶具有制作方便、检测简单以及不使用任何有毒药剂等优点,推荐甜菜DAMD扩增产物的检测使用琼脂糖凝胶电泳。     

Analysis of genetic diversity among wild pomegranates in Western Himalayas,using PCR methods

The genus Punica (Punicaceae) is distributed in Iran, Afghanistan, India and Mediterranean countries. Iran is considered to be its primary center of origin. In India Punica granatum is found in wild only in Western Himalayan regions comprising, Jammu and Kashmir, Himachal Pradesh and Uttarakhand states. However, there is little information available about the genetic variation present in pomegranates in the regions. In this paper we describe the use of DAMD and RAPD methods that generate the profiles, to study genetic diversity in wild genotypes of the P. granatum in India. Forty-nine accessions representing two regions of Western Himalaya were analyzed. Similarity coefficient value varied from 0.08 to 0.79 across different accessions. The results indicate that DAMD (97.08%) revealed more polymorphism in comparison to RAPD (93.72%). The results show that these methods are sufficiently informative to unravel the genetic variations in wild pomegranates in Western Himalayas.     

Molecular analyses of genetic variability in soap nut (Sapindus mukorossi Gaertn.)

Sapindus mukorossi Gaertn. (Sapindaceae) is commercially an important plant much valued for its fruits commonly known as soap nut as well as for the seed oil used as a feed-stock for biodiesel production. Molecular polymorphism, determined, using three single primer amplification reaction (SPAR) methods (RAPD, DAMD and ISSR) is used to assess the genetic variability in 69 accessions of S. mukorossi Gaertn., encompassing different geographical regions in India. Cumulative band data generated using the three SPAR methods resulted in 82.49% polymorphism across all genotypes of S. mukorossi Gaertn. UPGMA tree showed two major clusters that were in accordance with the geographical diversity. The intra-population genetic diversity (0.16), Shannon information index (0.24) and polymorphic loci (45.62%) were highest in AS population, while the corresponding lowest values were recorded in HP population. The inter-population average distance ranged from 0.05 (GA and HP) to 0.26 (AP and HP). Based on these distances, the UPGMA tree for the populations was computed, which showed the presence of two major clusters. Cluster I contained all the populations from Himachal Pradesh (HP) and Uttarakhand (GA and KU) and cluster II grouped the populations from North Eastern region (AS, AP, MG). The present study showed that SPAR methods are informative and useful to unravel the diversity among different populations of the soap nut plants and is a prelude for further utilization of promising and genetically divergent materials in the breeding programmes.     

利用DAMD分子标记构建甜菜品种的分子身份证

为了根据遗传背景快速区分和鉴定甜菜品种的真实性和特异性,补充和打破形态学鉴定法的短板和局限,以96份甜菜登记品种为试材,采用DAMD分子标记技术构建了甜菜品种的分子身份证。结果显示,有7条扩增条带清晰、多态性较好的DAMD引物可用于鉴别甜菜品种,这7条引物共检测出103条带,其中多态性条带101条,多态性98.1%;96个品种间的最小遗传距离为0.039,最大为0.485,平均遗传距离0.237。通过聚类分析,96个甜菜品种被分为两类群,类群I仅有54号品种,类群II包含其余95个品种,类群II进一步又分两个亚群。仅利用URP1F、URP17R和URP2R这3条引物组合将96个供试甜菜品种完全区分。利用DAMD引物构建甜菜品种的分子身份证,为快速、高效地鉴定甜菜品种提供了一种新的途径,也为保护育种家和农民的利益提供了保障。