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园艺学报 ›› 2018, Vol. 45 ›› Issue (7): 1382-1392.doi: 10.16420/j.issn.0513-353x.2017-0661

• 研究报告 • 上一篇    下一篇

人心果转录组De novo组装及奇可胶途径相关基因的分析

刘慧敏1,庆 军1,胡晶晶2,杜红岩1,乌云塔娜1,*   

  1. (1中国林业科学研究院经济林研究开发中心,郑州 450003;2懿奈(上海)生物科技有限公司,上海 200335)
  • 出版日期:2018-07-25 发布日期:2018-07-25

De novo Transcriptomes and Genes Involved in Chicle Biosynthesis of Manilkara zapota

LIU Huimin1,QING Jun1,HU Jingjing2,DU Hongyan1,and WUYUN Tana1,*   

  1. (1Non-timber Forestry Research and Development Center,Chinese Academy of Forestry,Zhengzhou 450003,China;2Inertia Shanghai Biotechnology Co.,Ltd.,Shanghai 200335,China)
  • Online:2018-07-25 Published:2018-07-25

摘要: 为了探索人心果(Manilkara zapota L.)的转录组及奇可胶合成相关的基因,使用Illumina测序平台,对人心果果实、树皮和叶片分别进行转录组测序,使用Trinity等软件进行De novo组装和注释以及计算表达量,采用实时荧光定量PCR对转录组数据进行验证。经过组装得到162 455条unigene,总长度达139 792 553 nt,平均长度达861 nt,N50达1 544 nt。通过与NR、NT、Swiss-Prot、KEGG、COG和GO等数据库比对,共计89 628条unigene得到注释。以组装出来的unigene作为参考序列,在人心果转录组中共鉴别出57 362个SSR位点,果实、叶片和树皮表达的基因中分别检测到99 925、65 989和129 109个SNP位点。在人心果转录组中共鉴别出105个与奇可胶合成相关的unigene,参与甲羟戊酸(MVA)途径的基因在果实和树皮中表达量较高,磷酸盐(MEP)途径中的基因则在叶片中的表达量较高,实时荧光定量PCR结果与转录组计算得到的表达量一致。初步推测MVA途径可能是奇可胶合成的主要途径。

关键词: 人心果, 转录组, 奇可胶

Abstract: To study the transcriptomes and genes related to chicle biosynthesis of Manilkara zapota L.,we conducted the RNA sequencing and analysis of M. zapota. The transcriptomes of fruit,bark,and leaf from M. zapota were sequenced on the Illumina platform. The sequencing data were then analyzed by Trinity and other software. qRT-PCR was performed to validate the transcriptome data. The raw data were assembled into 162 455 unigenes,with the total length,mean length,and N50 being 139 792 553 nt,861 nt,and 1 544 nt,respectively. Through mapping all the unigenes to databases,like NR,NT,Swiss-Prot,KEGG,COG,and GO,89 628 unigenes were annotated. In total,57 362 SSR sites were identified taking all the unigenes as references. A total of 99 925,65 989,and 129 109 SNPs were identified in fruit,leaf,and bark expressed genes. In all,105 unigenes were indentified to be chicle-related. The unigenes involved in the MVA pathway were highly expressed in fruit and bark,whereas,the unigenes involved in the MEP pathway were highly expressed in leaf. The gene expression profiles calculated by qRT-PCR were in accordance with those by transcriptome data. The transcriptome was assembled well and the gene expression level calculated by RNA-sequencing was reliable. Plenty of SSRs were indentified,which provides an important basis for the development of molecular markers. According to the expressional profiles of genes involved into chicle biosynthesis,we proposed that the MVA pathway might be the mainly produced initiators for chicle.

Key words: Manilkara zapota, transcriptome, chicle

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