摘要
【目的】研究芒果基因组SSR的总体特征，开发芒果SSR标记，为SSR标记在芒果种质资源鉴定、品种选育及遗传连锁图谱构建奠定基础。【方法】利用新一代454GSFLX高通量测序技术结合SSR文库构建法，得到19611条无冗余的序列，对其进行SSR搜索，共获得8831条SSR。利用Primer5．0软件设计SSR引物，并以筛选的引物对24份芒果种质进行多样性分析。【结果】在芒果SSR中，二核苷酸（5980个SSR）和三核苷酸（2451个SSR）占主导地位，所占比例分别为67，85％和27，81％；出现频率最高的二核苷酸重复基元是AC／GT，占总数的17．94％，出现最高的三核苷酸重复基元为TTC／GAA，占5．07％。随机选择50个SSR位点合成引物，经24份芒果种质筛选鉴定，21对引物可扩增获得产物，有效扩增率为42．00％；其中17对引物表现出良好多态性，占有效引物的80．95％，占总引物的34．00％。在17对多态性引物中，每对引物扩增等位基因数2—12个，产生的多态性片段为2-7个，多态性比例为50％-100％。【结论】454GSFLX高通量测序开发SSR引物有较好的实用性，开发获得的17个具有多态性的芒果SSR标记可用于研究芒果及其相关近缘物种的遗传变异。
[Objective]The present study was conducted to characterize and develop SSR markers in mango genome in order to lay a foundation for germplasm resource evaluation, linkage mapping and genetic improvement of mango. [Method ]The new-generation 454 GSFLX Titanium pyrosequencing technology was used for isolating a microsatellite-en- riched library mango genome. In total, 19611 non-redundant sequences were isolated and 8831 SSRs were identified. SSR primers were designed by using software Primer 5. Fifty randomly selected SSR markers were used for analyzing the genetic diversity among 24 mango accessions. [ Result ]The analysis results of SSR motifs revealed that, the dinucleotides （5980 SSRs） and trinucleotides（2451 SSRs） were predominante in all motifs, which accounted for 67.85% and 27.81%, respec- tively. Among the dinucleotide motifs, AC/GT presented the highest frequency, which accounted for 17.94%. TTC/GAA （5.07%） was the most frequent trinucleotide motif. Among the 50 microsateUite loci, 21 primer pairs were successfully amplified for genome DNA of 24 mango resources, accounting for 42.00%. Of which, 17 primers presented good polymor- phism, accounting for 80.95% of effective primers and 34.00% of all primers; 2-12 gene loci were found for each pair of primer with 2-7 polymorphic bands, and the polymorphic rate ranged from 50% to 100%. [Conclusion]It was concluded that the SSR primers developed by 454 GSFLX Titanium pyrosequencing technology is of better practicability, ,and the 17 polymorphic primers of mango SSRs could be used for studying genetic variation of mango and its sibling species.
出处
《南方农业学报》
CAS
CSCD
北大核心
2015年第10期1745-1749,共5页
Journal of Southern Agriculture
基金
海南省自然科学基金项目(34128)
中国热带农业科学院基本科研业务费专项项目(1630032013031)
作者简介
王明（1981-），博士，主要从事热带作物种子种苗分子检测研究工作，E-mail：mingwang23@gmail．com.
通讯作者，张如莲（1968-），研究员，主要从事热带作物种子种苗质量监督与检测研究工作，E-mail：zhang23300406@126．com。
