基于SSR分子标记的福建百香果品种(系)鉴定及指纹图谱构建

Research on the identification of passion fruit varieties in Fujian province and construction of fingerprint based on SSR markers

【目的】基于SSR分子标记鉴别福建百香果品种的遗传多样性,构建福建百香果品种的分子指纹图谱。【方法】基于转录组测序,开发西番莲SSR分析标记,并筛选出15个重复性好、多态性高的分子标记用于17个福建百香果品种(系)鉴定及遗传多态性分析,构建百香果品种(系)的SSR分子指纹图谱。【结果】利用MISA软件对1 kb以上的24319条西番莲(Passiflora caerulea L.)Unigene进行搜索,于8742条Unigene上检测出11385个SSR位点,出现频率为46.82%,平均分布距离为7.15 kb。单核苷酸、二核苷酸和三核苷酸分别占总SSR的63.72%、20.40%和14.28%,为数量较大的优势重复基序,其优势重复基元分别为A/T、AG/CT和AAG/CTT。通过Primer 3.0共获得出28257对SSR引物。从26对有效扩增引物中选择重复性好的15对引物,对17份百香果品种(系)进行多态性验证分析。15对SSR引物共产生235条多态性条带,PIC平均值为0.95。利用UPGMA进行聚类分析并作图,在遗传距离0.765处,可将17个品种(系)分为7个类群。根据15对多态性引物扩增电泳图,选出PFSSR20和PFSSR29引物组合用于构建DNA指纹图谱,可鉴别出17个百香果品种(系)。【结论】利用西番莲转录组数据构建的SSR分子标记具有较高的多态性潜能,类型丰富,本研究基于此构建的DNA指纹图谱可分辨出17个百香果品种(系)。

【Objective】Based on SSR markers,the genetic diversity of passion fruit varieties in Fujian province were analyzed,and SSR fingerprints of passion fruit varieties were constructed.【Method】The SSR markers of passion fruit were developed using transcriptome sequencing.15 SSR markers with good repeatability and high polymorphism were screened for the identification and genetic polymorphism analysis of 17 Fujian passion fruit varieties,and the SSR molecular fingerprint was constructed.【Result】Using the MISA software,24319 unigenes above 1 kilo base pairs of passion fruit were analyzed,and a total of 11385 SSRs were identified from 8742 unigenes,with an appearing frequency of 46.82%and an average distribution distance of 7.15 kb.Mononucleotide,dinucleotide and trinucleotide accounted for 63.72%,20.40%and 14.28%of the total SSR,respectively.Furthermore,the A/T、AG/CT and AAG/CTT were the predominant dinucleotide repeat types for mononucleotide,dinucleotide and trinucleotide repeat types,respectively.There were 28257 SSR primers designed using Primer 3.0.15 pairs of primers with good repeatability were selected from 26 pairs of effective primers to verify the polymorphism of 17 passion fruit cultivars.A total of 235 polymorphic bands were generated by 15 pairs of SSR primers,and the average PIC was 0.95.The 17 cultivars were divided into 7 groups at the genetic distance of 0.765 by UPGMA cluster analysis.According to the 15 pairs of polymorphic primer amplification electrophoresis map,PFSSR20 and PFSSR29 primer combinations were selected to construct DNA fingerprinting,and 17 passion fruit varieties could be identified.【Conclusion】The SSR molecular markers constructed using passion fruit transcriptome data have high polymorphism potential and rich types.17 Passiflora varieties(lines)were successfully identified based on the DNA fingerprints constructed in this study.