木本油料植物光皮树SSR特征分析及其SSR-PCR反应体系优化  

Analysis of SSR Characteristics and the Optimization of the SSR-PCR Reaction System of the Woody Oil Plant Swida wilsoniana

作　　者：周宵 彭映辉[1] 蒋丽娟[1] 陈景震[2] 王明怀[3] 何祯祥[4] 陈勇 康向阳[6] 张良波[2,6] Zhou Xiao;Peng Yinghui;Jiang Lijuan;Chen Jingzhen;Wang Minghuai;He Zhenxiang;Chen Yong;Kang Xiangyang;Zhang Liangbo(Central South University of Forestory and Technology,College of Life Sciences and Technology,Changsha,410002;Hunan Academy of Forestry Sciences,Bioenergy Research Institute,Changsha,410002;Guangdong Academy of Forestry,Guangzhou,510000;School of Life Sciences,Nanjin University,Nanjin,210000;Xishan Forestry Centre,Dazu District,Chongqing,Chongqing,402360;Beijing Forestry University,Institute ofBiologi・cal Sciences and Technology,Beijing,100083)

机构地区：[1]中南林业科技大学生命科学与技术学院,长沙410002 [2]湖南省林业科学院生物能源研究所,长沙410004 [3]广东省林业科学研究院,广州510000 [4]南京大学生命科学学院,南京210000 [5]重庆市大足区西山林场,重庆402360 [6]北京林业大学生物科学与技术学院,北京100083

出　　处：《分子植物育种》2020年第24期8155-8162,共8页Molecular Plant Breeding

基　　金：湖南省重点研发计划项目(2018NK2044,2017WK2021);国家外国专家项目计划(GS20190129002);长株潭国家自主创新示范区专项(2018XK2005);国家光皮树山苍子良种基地良种繁育补助项目(湘财资环指(2019)15号)共同资助。

摘　　要：本研究基于木本油料植物光皮树叶片转录组数据,利用软件MISA (1.0)对所有Unigene进行SSR搜索、检测和特征分析。结果显示,光皮树叶片转录组36 839条Unigene,有8 945条Unigene含有SSR。一共有SSR位点12 538个,出现频率约为2 514 bp/个。其中完整型和混合型SSR位点个数分别为11 050和1 488,一条Unigene中含有5、4、3、2、1个SSR位点的分别有11、37、199、1 329、8 540条。SSR丰富度依次为二、一、三、四、六、五核苷酸,分别有7 282、3 388、1 675、96、59、38个。SSR共有210种重复基元,从一核苷酸到六核苷酸分别有4、12、60、52、27、55种。其中SSR重复基元的重复次数均在5~44次,重复次数主要集中在4~15次,占94.47%。不同类型SSR重复基元频率最高的4类碱基序列依次为AG/CT、A/T、AT/AT和AC/CT,以二核苷酸为主。通过正交实验,得出SSR-PCR反应体系最佳组合为:退火温度55℃+循环次数34+模板浓度50 ng/μL+引物量0.5μL。SSR-PCR反应体系优化保证了后续分子标记实验数据的可靠性,为光皮树SSR遗传多样性、表型性状关联等分析提供了依据。This study is based on the data of woody oil plant Swida wilsoniana transcriptome. The SSR search,detection and characteristic analysis of all unigene are carried out by using the software Misa. The results showed that there were 8 945 Unigene containing SSR in 36 839 Unigenes. The total SSR loci was 12 538. The frequency of occurrence is about 2 514 bp per loci. The number of complete and mixed SSR loci is 11 050 and 1 488,respectively. The Unigenes contains 5, 4, 3, 2, 1 SSR loci in one Unigene is 11, 37, 1 999, 1 329, 8 540, respectively. The richness of SSR is Dinucleotide, Mononucleotide, Trinucleotide, Tetranucleotide, Hexanucleotide, Pentanucleotide, with 7 282, 3 338, 1 675, 96, 59, 38 SSRs, respectively. SSR contains a total of 210 repeating units,from Mononucleotide to Hexanucleotide is 4, 12, 60, 52, 27, 55, respectively. The number of repeats of SSR repetition unit is from 5 to 44 times. And the repetition times of units were mainly 4 to 15, accounting for 94.47%.Four kinds of base sequence with the highest repetition units frequency in different types of SSR is AG/CT, A/T,AT/AT and AC/CT. The optimum combination of SSR-PCR reaction system was obtained by orthogonal experiment: 55℃(Annealing temperature) +34(Cycle index) +50 ng/μL(Template concentration) +0.5 μL(Primer quantity). The optimization of the SSR-PCR reaction system ensures the reliability of the following molecular marker experimental datas. It lays the foundation for the analysis of the genetic diversity and correlation analysis of phenotypic traits of Swida wilsoniana.

关 键 词：木本油料植物 转录组 特征分析 SSR PCR反应体系 

分 类 号：S565.9[农业科学—作物学]