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利用RAPD和ISSR标记分析揭示的桑树(桑属)遗传多样性及亲缘关系

Genetic diversity and relationships in mulberry (genus Morus) as revealed by RAPD and ISSR marker assays.

作者信息

Awasthi Arvind K, Nagaraja G M, Naik G V, Kanginakudru Sriramana, Thangavelu K, Nagaraju Javaregowda

机构信息

1Seribiotech Research Laboratory, Central Silk Board, CSB Complex, Kodathi, Carmelram post, Bangalore- 560035, India.

出版信息

BMC Genet. 2004 Jan 10;5:1. doi: 10.1186/1471-2156-5-1.

Abstract

BACKGROUND

The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species.

RESULTS

RAPD analysis using 19 random primers generated 128 discrete markers ranging from 500-3000 bp in size. One-hundred-nineteen of these were polymorphic (92%), with an average of 6.26 markers per primer. Among these were a few putative species-specific amplification products which could be useful for germplasm classification and introgression studies. The ISSR analysis employed six anchored primers, 4 of which generated 93 polymorphic markers with an average of 23.25 markers per primer. Cluster analysis of RAPD and ISSR data using the WINBOOT package to calculate the Dice coefficient resulted into two clusters, one comprising polyploid wild species and the other with domesticated (mostly diploid) species.

CONCLUSION

These results suggest that RAPD and ISSR markers are useful for mulberry genetic diversity analysis and germplasm characterization, and that putative species-specific markers may be obtained which can be converted to SCARs after further studies.

摘要

背景

桑属植物,即桑树,是一种雌雄异株且异花授粉的植物,是家养桑蚕(Bombyx mori)的唯一食物来源。由于环境对目标性状的影响,利用形态特征进行分类的传统方法在确定不同桑树物种间的多样性和关系方面大多并不成功。作为一种更可靠的替代方法,采用了基于PCR的标记分析,包括随机扩增多态性DNA(RAPD)和简单序列重复区间(ISSR),来研究12个家养桑树品种和3个野生桑树品种的遗传多样性及相互关系。

结果

使用19条随机引物进行RAPD分析,产生了128个离散标记,大小在500 - 3000 bp之间。其中119个具有多态性(92%),每个引物平均产生6.26个标记。其中有一些假定的物种特异性扩增产物,可用于种质分类和渐渗研究。ISSR分析使用了6条锚定引物,其中4条产生了93个多态性标记,每个引物平均产生23.25个标记。使用WINBOOT软件包计算Dice系数,对RAPD和ISSR数据进行聚类分析,得到两个聚类,一个包含多倍体野生种,另一个包含家养(大多为二倍体)种。

结论

这些结果表明,RAPD和ISSR标记可用于桑树遗传多样性分析和种质鉴定,并且可能获得假定的物种特异性标记,经过进一步研究后可转化为序列特征扩增区域(SCAR)标记。

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