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通过转录组测序技术鉴别普通油茶(Camellia oleifera)EST-SSR位点,开发筛选适用于普通油茶种质资源评价与利用的多态性SSR标记,对普通油茶的遗传变异研究和油茶良种的分子标记辅助育种具有重要意义。利用MISA工具从普通油茶转录组67 434条Unigenes(63.39 MB)中筛选得到26 751个包含1~6个核苷酸重复类型的SSR位点,SSR的分布频率为39.67%,发生频率为1/2.33 kb;SSR位点中的主导类型是二核苷酸重复,占总SSR的44.28%,其次是单核苷酸(35.28%)和三核苷酸重复(18.27%)。通过SSR标记的琼脂糖凝胶电泳和荧光标记毛细管电泳,从143对SSR引物中遴选出了20对扩增效率高、稳定性好的多态性SSR引物,并对来自浙江产区的57个普通油茶品种进行遗传关系分析。结果显示:20对SSR引物共检测出129个等位基因,每对引物2~17个,平均6.45个;多态性信息量(PIC)为0.0499~0.8635,平均0.4355。57个普通油茶品种在Dice遗传相似系数大约0.47时聚为一类,在大约0.60时可分为七个大类。本研究印证了利用普通油茶转录组数据开发SSR标记的可行性,也证明了茶属树种SSR标记的可转移性;不仅为普通油茶遗传多样性分析和遗传图谱构建提供了有价值的候选标记,也为普通油茶良种的鉴别与分子选育提供了分子技术手段。
The EST-SSR loci of common Camellia oleifera were identified by transcriptome sequencing, and SSR markers for evaluation and utilization of common Camellia oleifera germplasm resources were developed. The genetic variation of common Camellia oleifera and the molecular markers of Camellia oleifera Assisted breeding is of great importance. A total of 26 751 SSR loci with 1 ~ 6 nucleotide repeat types were screened from 67 434 Unigenes (63.39 MB) of common Camellia oleifera transcriptome using MISA tool. The frequency of SSR was 39.67% 2.33 kb. The dominant type in SSR loci was dinucleotide repeats, accounting for 44.28% of the total SSRs, followed by mononucleotides (35.28%) and trinucleotide repeats (18.27%). Twenty SSR primers with high efficiency and good stability were selected from 143 pairs of SSR primers by SSR-labeled agarose gel electrophoresis and fluorescence-labeled capillary electrophoresis. 57 SSR primers from Zhejiang province Genetic analysis of common oil tea varieties. The results showed that a total of 129 alleles were detected with 20 pairs of SSR primers, with 2 to 17 primers per primer (average 6.45). The polymorphism information content (PIC) ranged from 0.0499 to 0.8635 with an average of 0.4355.57 Dice clustered together at about 0.47 and were classified into seven broad categories at about 0.60. This study confirms the feasibility of developing SSR markers using common oil-tea transcriptome data and also demonstrates the transferability of SSR markers in Camellia species. It not only provides valuable candidate markers for the genetic diversity analysis and genetic map construction of Camellia oleifera, It also provides a molecular technique for the identification and molecular breeding of common Camellia.