摘要
为解决白皮松分子标记资源相对匮乏的问题,对其针叶转录组测序结果进行检测分析及引物开发,结果在针叶转录组数据中共检测到精确型SSR位点3 268个以及复合型SSR位点104个,序列长度为10~27 bp的短序列(2 954,93.36%)占主导地位;SSR重复单元的重复次数在4~30次之间,重复次数为9次的重复单元数量最多,所占比例最大(684,20.93%);在所有142个重复单元中,A/T所占比例最大(1 508, 46.14%),其次为AT/CA(280,8.57%)和TA/TC(245,7.50%);从所设计的1 151对引物中,随机选择60对EST-SSR引物,对来自不同地区的50株白皮松优树进行PCR扩增,有28对引物能扩增出清晰的条带,有效扩增率为46.67%,其中多态性引物2对,引物多态率为7.14%,多态性信息含量 PIC分别为0.375、 0.305 。为了检验引物的多态性,又选择了7对已发表的对白皮松基因型具有多态性的SSR引物进行验证分析,结果发现,这7对引物均能扩增出清晰的条带,但对50株白皮松优树均无多态性,也侧面说明了白皮松基因序列的相对保守性。该研究结果为白皮松种质资源评价、分子标记辅助育种等方面的研究提供了参考。
In order to solve the problem of relative scarcity of molecular marker resources in Pinus bungeana , the results of coniferous transcriptome sequencing were analyzed and primers were developed. A total of 3 268 exact SSR loci and 104 complex SSR loci were detected in coniferous transcriptome data. Short sequences (2 954, 93.36%) with 10-27bp length were dominant. Repetition number of repetition units in Pinus bungeana transcription group ranged from 4 to 30, and the number of repetitive units with 9 repetitions was the largest, accounting for the largest proportion (684, 20.93%). Among all 142 repetitive units, A/T accounted for the largest proportion (1 508, 46.14%), after which were the AT/CA (280, 8.57%)and TA/TC (245, 7.50%). Sixty EST-SSR primers were randomly selected from 1151 pairs of primers designed for PCR amplification. Among them, 28 pairs of primers could amplify clear bands,and the effective amplification rate was 46.67%. Among them, there were 2 pairs of polymorphic primers, and the polymorphism rate was 7.14%. The polymorphic information content was 0.375 and 0.305, respectively. In order to test the polymorphism of primers, seven pairs of published polymorphic primers of Pinus bungeana were selected for validation and analysis. The results showed that these seven pairs of primers could amplify clear bands, but none of them had polymorphism in 50 superior Pinus bungeana , which also reflected the relative conservativeness of Pinus bungeana gene sequence. The results provide a reference for the evaluation of Pinus bungeana germplasm resources and molecular marker-assisted breeding.
作者
李昕蔓
金卓颖
苏安然
杨林夕
苏晟
王磊
张军
LI Xinman;JIN Zhuoying;SU Anran;YANG Linxi;SU Sheng;WANG Lei;ZHANG Jun(College of Forestry,Hebei Agricultural University,Baoding 071000,China)
出处
《林业与生态科学》
2019年第3期266-272,共7页
Forestry and Ecological Sciences
基金
河北农业大学科研发展基金计划(JY2018006)
关键词
白皮松
转录组
EST-SSR
多态性
引物开发
Pinus bungeana Zucc.ex Endl.
transcriptome
EST-SSR
polymorphism
primer development
