以20份高油野生花生为材料,从252对SSR引物中筛选出46对扩增条带清晰且多态性丰富的引物对其基因组DNA进行扩增,46对引物共扩增出425条带,全部为多态性条带,多态条带比例为100%。每对引物扩增获得的条带数为2~21条,平均9条。其中引物2E6的扩增效率最高,能将20份材料中的14份区分开。双引物组合2E6/PM403的鉴别能力最强,能将20份材料中的18份区分开。5组三引物组合能将20份材料完全区分,包括2E6/PM403/1B9、2E6/PM403/9A7、2E6/PM403/10H1A、2E6/PM403/PM201、2E6/PM403/PM458,其中三引物组合2E6/PM403/10H1A为涉及引物中的最佳引物组合。综合使用野生花生材料的国家统一编号、引物名称、分子数据建立了20份野生花生高油种质DNA指纹身份数据库,为野生花生的安全保存和保护以及有效利用奠定了基础。
Establishment of DNA fingerprint identity(ID) of peanut wild relatives is useful for maintaining and utilizing the resource.In the present study,20 accessions of peanut wild relatives with high oil content were analyzed by SSR technique to establish their DNA fingerprint identity.Forty-six SSR primer pairs with polymorphism were selected from 252 primers to amplify the genomic DNA.Totally 425 polymorphic bands were produced through the 46 primers.These SSR primers amplified 2 to 21 polymorphic bands with average of 9 bands.Neither single-primer nor double-primer combinations were able to distinguish all the involved accessions.However primer 2E6 of 46 single-primers and dowble-primer combination 2E6/PM403 could distinguish the 14 and 18 genetypes of the all 20 accessions respectively.Based on the results of three-primer combination detection,five three-primer combinations were able to identify the all 20 accessions involved,of which 2E6/PM403/10H1A combination was the most efficient.Based on SSR data of the primer combination 2E6/PM403/10H1A,and integrated the accession number and primer name,the DNA fingerprint ID of the 20 Arachis three-wild accessions with high oil content was established.The DNA fingerprint information is useful for peanut breeding and resource storing.