基于转录组数据的杜仲红叶性状SSR分子标记分析

Analysis of Red Leaf Color SSR Molecular Markers by Transcriptome Sequencing of Eucommia ulmoides

对‘华仲12号’杜仲的幼嫩叶片(绿色)和成熟叶片(红色)及‘华仲11号’杜仲成熟叶片(绿色)进行转录组测序,进行测序数据的拼接和组装,且对转录组获得的基因(Unigenes)进行SSR分析。研究得到54 517条平均长度为806.90 bp的Unigenes,其中25 993条Unigenes在Nr、Swiss-Prot、KEGG和COG蛋白数据库获得功能注释,占所有Unigenes的47.68%。参照KEGG数据库,可将注释到的6 910条Unigenes划分到122个代谢途径分支,其中花色苷代谢途径相关酶基因39个,类黄酮代谢途径38个,类胡萝卜素合成途径34个。54 517条Unigenes中共包含17 010个完整型SSR位点,占总SSR位点的96.28%。完整型SSR位点共包含67种重复基元,其中出现频率最高的重复基元类型为单核苷酸重复中的A/T(7 747个),其次是AG/CT(5 039个)和AT/AT(850个)从花色苷代谢途径、类黄酮代谢途径及类胡萝卜素代谢途径中共找到13个SSR位点。为今后杜仲遗传多样性分析、遗传图谱构建及杜仲红叶性状分子标记开发等方面奠定了分子基础。

The transcriptomes of Eucommia ulmoides ＇ Huazhong 12＇ young and mature leaves and ＇ Huazhong 11＇ mature leaves were sequenced. The transcriptome data was assembled and classified by function, and microsatellites characteristics from obtained Unigenes and analyzed. The Unigenes were assembly generated a total of 54 517 Unigenes with an average length of 806.90 bp. Among them, 25 993 Unigenes accounted for 47.68% were annotated by comparing the assembled Unigenes with the Nr, Swiss-Prot, KEGG, and COG protein databases. KEGG pathway analysis presented that 6 910 annotated Unigenes divided into 122 classes according to its function. Among them, the Unigenes of ascorbate and aldarate metabolism was 39, flavonoid biosynthesis was 38, carotenoid biosynthesis was 34. There were 17 010 complete SSR located in 54 517 Unigenes accounted for 96.28% of the total SSR. The complete SSR included 67 frequent motifs, and the highest repeat of complete SSR type was A/T （7 747 ）, following by AG/CT （ 5 039 ）, AT/AT （850）. There were 13 SSR located in ascorbate and aldarate metabolism, flavonoid biosynthesis and carotenoid biosynthesis. Our study would provide the molecular basis for genetic diversity analysis, genetic map construction and red leaf molecular marker development of E. ulmoides.