尾叶紫薇全长转录组测序及微卫星标记开发  

Full-length Transcriptome Sequencing and Microsatellite Marker Development in Lagerstroemia caudata

作　　者：章寒 吴宜静 张一鸣 续言 陆泓锦 赵天祎 蔡明[1] 张曼 蔡军火[2] 潘会堂[1] 张启翔[1] ZHANG Han;WU Yijing;ZHANG Yiming;XU Yan;LU Hongjin;ZHAO Tianyi;CAI Ming;ZHANG Man;CAI Junhuo;PAN Huitang;ZHANG Qixiang(Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular Breeding,National Engineering Research Center for Floriculture,Beijing Laboratory of Urban and Rural Ecological Environment,Engineering Research Center of Landscape Environment Ministry of Education,Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education,College of Landscape Architecture,Beijing Forestry University,Beijing 100083,China;College of Landscape and Art,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]北京林业大学花卉种质创新与分子育种北京市重点实验室/国家花卉工程技术研究中心/城乡生态环境北京实验室/园林环境教育部工程研究中心/林木花卉遗传育种教育部重点实验室/园林学院,北京100083 [2]江西农业大学园林与艺术学院,江西南昌330045

出　　处：《江西农业大学学报》2021年第4期919-930,共12页Acta Agriculturae Universitatis Jiangxiensis

基　　金：“十三五”国家重点研发计划项目(2019YFD1001000);国家自然科学基金青年科学基金项目(31500574);江西省林业局林业科技创新专项([2019]13)。

摘　　要：【目的】尾叶紫薇是培育香花紫薇的重要种质资源,目前花香功能基因的研究及已开发的SSR分子标记较少,无法满足分子育种研究需求。【方法】采用PacBiosequel平台对尾叶紫薇3个开花时期混样进行全长转录组测序,通过公共数据库对Unigene进行功能注释,利用MISA软件检索全长转录组的SSR位点,Primer3批量设计SSR引物并检测。【结果】测序获得39087条非冗余、高质量全长转录本,平均长度为2109 bp,N50长度为2427 bp,长度主要分布在500-4000 bp。有38559条序列(98.64%)获得功能注释,功能主要集中于细胞进程及代谢进程。其中参与花香相关萜类物质合成的有56个,参与脂肪酸衍生物合成的有922个。获得的16425个SSR位点分布在12060条序列中,平均每5.02 kb出现1个SSR序列,3141条序列含有多个SSR位点。SSR位点主要重复类型是二核苷酸和三核苷酸,分别占所有SSR位点的41.95%和31.58%,重复基序类型有96种,AG/CT重复类型的出现频率最高。SSR的重复次数以5-15次为主,长度为10-86 bp,平均为16.84 bp。随机选取52对SSR引物进行PCR检测,有40对引物能产生单一、清晰的条带,有效扩增率为76.92%。【结论】获得与花香物质合成相关基因序列和大量有效SSR标记,将为深入了解尾叶紫薇花香物质合成与代谢、紫薇属植物种质资源评价与保护、分子标记辅助育种等研究奠定基础。[Objective]Lagerstroemia caudata is an important germplasm resource for cultivating fragrant flowering varieties of crape myrtle.There are few researches on floral functional genes and SSR molecular markers that have been developed so far,which cannot meet the needs of molecular breeding research.[Method]In this study,the PacBio sequel platform was used to sequence the full-length transcriptome of the mixed flowers of L.caudata at three stages,and the Unigene was functionally annotated through public databases.The SSR locus was searched in the full-length transcriptome using MISA software and the SSR primers were designed by Primer3 and tested.[Result]Sequencing obtained 39087 non-redundant,high-quality full-length transcripts,with an average length of 2109 bp,the N50 length was 2427 bp,and the lengths were mainly distributed from 500 to 4000 bp.38559 sequences(98.64%)were functionally annotated.Their functions were mainly focused on cell processes and metabolic processes.There were unigenes annotated,in which 56 were involved in the synthesis of terpenoids and 922 were involved in the synthesis of fatty acid derivatives.The 16425 SSR loci obtained from the search were distributed among 12060 sequences.The average per SSR loci appeared every 5.02 kb,and 3141 sequences contained multiple SSR loci.The main repeat types of SSR loci in the full-length transcriptome of L.caudata were dinucleotide and trinucleotide,which accounted for 41.95%and 31.58%of the total SSRs,respectively.There were 96 types of repeat motifs,AG/CT repeat types appeared most frequently.The numbers of repetitions of SSR were mainly 5 to 15 times,the lengths were distributed in the range of 10 to 86 bp,and the average length was 16.84 bp.52 pairs of SSR primers were randomly selected for PCR detection,40 pairs of the primers could produce a single,clear band,and the effective amplification rate was 76.92%.[Con⁃clusion]This study obtained gene sequences related to the synthesis of floral substances and a large number of effective SSR markers,whi

关 键 词：尾叶紫薇 全长转录组 分子标记 SSR 

分 类 号：S685.99[农业科学—观赏园艺]