猕猴桃属植物通用型SSR分子标记引物的筛选及应用

【目的】基于猕猴桃全基因组数据，开发、筛选一批多态性高、通用性强的SSR引物，为猕猴桃属种质资源遗传多样性分析、品种鉴定等奠定基础。【方法】基于‘红阳’猕猴桃全基因组序列，设计并合成435对SSR引物，采用荧光标记毛细管电泳进行等位变异检测。首先，采用遗传差异较大的5份猕猴桃种质资源对引物进行有效性筛选；其次，选择9个物种或杂交组合的16份猕猴桃种质资源开展引物复筛；最后，利用所选引物对国家猕猴桃种质资源圃内猕猴桃属51个类型共225份种质资源进行基因分型及亲缘聚类分析。【结果】从435对引物中初筛到216对有效性引物，经复筛确定分布于29条染色体上的67对引物为最终核心引物。67对引物在16份种质中共得到842个等位变异，每个位点检测到等位变异6—18个，平均等位基因数（Na）为12.57个；有效等位基因数（Ne）为3.27(A-Geo-149)—13.84(A-Geo-407)个，平均为8.18个；观测杂合度（Ho）为0.60(A-Geo-073)—0.93(A-Geo-158)，平均为0.77；期望杂合度（He）为0.72(A-Geo-149)—0.92(A-Geo-101、A-...

Screening and Application of Universal SSR Molecular Marker Primers in Actinidia

【Objective】 Based on the whole genome data of kiwifruit, a batch of SSR primers with high polymorphism and strong universality were developed and screened to lay a foundation for genetic diversity analysis and variety identification of kiwifruit germplasm resources. 【Method】 Based on the whole genome sequence of Hongyang kiwifruit, 435 pairs of SSR primers were designed and synthesized, and the allelic variation was detected by fluorescence labeled capillary electrophoresis. Firstly, five kiwifruit germplasm resources with large genetic differences were used to screen the effectiveness of primers. Secondly, 16 kiwifruit germplasm resources from 9 species or hybrid combinations were selected for primer re-screening. Finally, 225 germplasm resources belonging to 51 types of Actinidia in the National Actinidia Germplasm Repository were analyzed by core primers.【Result】 From 435 pairs of primers, 216 pairs of valid primers were initially screened, and 67 pairs of primers distributed on 29 chromosomes were identified as the final core primers after re-screening. Using 67 SSR markers, a total of 842 observed alleles (Na) were detected with 6-18 alleles (mean = 12.57) per locus; the effective number of allele (Ne) ranged from 3.27 (A-Geo-149) to 13.84 (A-Geo-407) with an average of 8.18; the observed heterozygosity (Ho) ranged from 0.60 (A-Geo-073) to 0.93 (A-Geo-158) with an average of 0.77; the expected heterozygosity (He) ranged from 0.72 (A-Geo-149) to 0.92 (A-Geo-101 and A-Geo-158) with an average of 0.85; the polymorphism information content (PIC) ranged from 0.67 (A-Geo-149) to 0.92 (A-Geo-158) with an average of 0.84; the Shannon diversity index (I) range was 1.47 (A-Geo-149) to 2.73 (A-Geo-101) with an average of 2.22. The above results indicated that the polymorphism of primers was very high, so it is suitable for the analysis of genetic relationship and genetic diversity of kiwifruit germplasm resources. The clustering results of 225 germplasm resources could clearly reveal the genetic relationship of Actinidia.【Conclusion】 The selected SSR primers were stable, reliable, polymorphic and universal, which could be used as core primers for germplasm resources identification, fingerprint construction, core germplasm mining and genetic diversity analysis of Actinidia.