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草业学报 ›› 2019, Vol. 28 ›› Issue (5): 132-142.DOI: 10.11686/cyxb2018304

• 研究论文 • 上一篇    下一篇

海滨雀稗自交结实突变体及野生型幼穗组织的转录组分析

钱晨1,2, 刘智微1,2, 钟小仙1,2,*, 吴娟子1,2, 张建丽1,2, 潘玉梅1,2   

  1. 1.江苏省农业科学院畜牧研究所, 江苏 南京 210014;
    2.农业部种养结合重点实验室, 江苏 南京 210014
  • 收稿日期:2018-05-09 修回日期:2018-08-24 出版日期:2019-05-20 发布日期:2019-05-20
  • 通讯作者: E-mail: xiaoxian@jaas.ac.cn
  • 作者简介:钱晨(1984-),男,江苏扬州人,助理研究员,博士。E-mail: 53891604@qq.com
  • 基金资助:
    江苏省农业科学院国家牧草育种创新基地项目和江苏省农业科技自主创新资金项目(CX(14)2049)资助

Transcriptomic analysis of the self-incompatibility mechansim in Paspalum vaginatum by comparison with an artificial self-compatible mutant

QIAN Chen1,2, LIU Zhi-wei1,2, ZHONG Xiao-xian1,2,*, WU Juan-zi1,2, ZHANG Jian-li1,2, PAN Yu-mei1,2   

  1. 1.Laboratory of Forage Breeding, Institute of livestock Science, Jiangsu Academy of Agricultural Science, Nanjing 210014, China;
    2. Key Laboratory of Crop and Livestock Integrated Farming, Ministry of Agriculture, Nanjing 210014, China
  • Received:2018-05-09 Revised:2018-08-24 Online:2019-05-20 Published:2019-05-20
  • Contact: E-mail: xiaoxian@jaas.ac.cn

摘要: 二倍体海滨雀稗Adalayd作为自交不亲和的暖季型草坪草, 在环境保护及修复上发挥着重要的作用, 但由于自身自交不亲和性减缓了其大面积推广。至今海滨雀稗自交不亲和的机制尚且未知并且基因数据库资源十分匮乏。为了研究海滨雀稗自交不亲和的机制, 以二倍体海滨雀稗自交亲和体细胞突变体SP2008-3和自交不亲和野生型Adalayd为材料, 采用Illumina Hiseq 2000高通量测序技术进行转录组测序。获得2个材料幼穗组织的表达谱, 共获得68175654个Raw reads片段, 测序结果de novo拼接获得117619个单基因簇(Unigene), 其中50%的Unigene被注释。比较两个基因表达谱, 发现1303个差异表达基因并对这些差异基因进行了基因本体论(gene ontology, GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)分类, 筛选出了22个与植物自交不亲和反应相关的基因, 其中14个钙离子信号通路(钙调素、钙依赖蛋白激酶、类钙调素互作蛋白激酶)、3个F-box和5个硫氧还蛋白(thioredoxin)基因在体细胞突变体SP2008-3和野生型Adalayd之间存在差异表达, 测序结果获得了很多与海滨雀稗自交不亲和相关的遗传资源信息。本研究首次将转录组学研究应用于海滨雀稗自交不亲和研究, 为海滨雀稗自交不亲和进一步研究提供了宝贵且有价值的基因信息基础。

关键词: 二倍体海滨雀稗, 自交不亲和, 高通量测序, 差异表达基因

Abstract: The warm-season turfgrass seashore paspalum (Paspalum vaginatum) is rapidly gaining recognition as having a potentially important contribution in a range of turf and environment conservation applications. However, self-incompatibility (SI) creates issues for seed production in seashore paspalum, consequently limiting the feasibility of large scale planting. The mechanism of SI response in P. vaginatum is still largely unknown. In addition, the lack of genomic and transcriptomic information makes it highly challenging to clarify the mechanism of SI in seashore paspalum. In order to further understand the mechanism of SI in seashore paspalum, we created a self-compatible (SC) mutant (SP2008-3) and compared the first transcriptome of SI seashore paspalum (variety Adalayd) and the SC mutant using Illumina sequencing technology. Transcriptome analysis using RNA-sequencing was performed to profile gene expression patterns in Adalayd and SP2008-3 (the SC mutant). A total of 68.17 million raw reads were obtained, and de novo assembly produced 117619 Unigenes. All assembled Unigenes were annotated by querying against public databases. 58810 Unigenes (50%) were found to be homologous to genes in the NCBI non-redundant protein (Nr) database. In further orthologous analyses, comparison of the transcriptomes found 1303 Unigenes that showed significant differences in transcript abundance between Adalayd and SP2008-3. These Unigenes were functionally annotated within the Gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) pathways. A large number of notable genes potentially involved in SI responses showed differential expression. We conclude these genes may encode critical regulators of SI responses. Examples of these genes include 14 CaM (Calmodulin, calcium-dependent protein kinases and CBL-interacting protein kinases), 3 F-box and 5 THL (Thioredoxin). Our data represent a genetic resource for the discovery of genes related to SI in seashore paspalum. To our knowledge, this is the first study of the transcriptome of seashore paspalum focusing on SI. This resource will be very useful for future studies on the mechanisms of SI in seashore paspalum.

Key words: Seashore paspalum, self-incompatibility, transcriptome, differential expression gene