基于转录组测序的红树秋茄叶片发育中差异表达基因分析  

Analysis of Differentially Expressed Genes of Kandelia During Development with High-throughput RNA-seq Technology

作　　者：张存家 刘小青 韩婧 刘冰 聂磊[1] Zhang Cunjia;Liu Xiaoqing;Han Jing;Liu Bing;Nie Lei(Shenzhen Institute of Information Technology,Shenzhen,518172)

机构地区：[1]深圳信息职业技术学院,深圳518172

出　　处：《分子植物育种》2023年第24期8075-8083,共9页Molecular Plant Breeding

基　　金：深圳信息职业技术学院博士课题启动项目(SZIIT2021KJ009)资助。

摘　　要：为探索秋茄(Kandelia obovata)叶片发育过程中物质合成和基因表达情况,本研究对红树叶片三个发育时期的m RNA进行提取和高通量测序,获得高质量序列(Clean reads)64189053个,高质量碱基19.29 Gb,共组装获得转录本108077个,转录本长度大于2000 bp的为37206个,200~300 bp的转录本11797个,Q30均在95.00%左右。通过对红树秋茄叶片三个不同时期(初期,中期,成熟期)的差异基因进行KEGG富集分析,结果发现,差异基因主要富集在以下代谢途径:碳水化合物代谢途径(淀粉和蔗糖代谢),苯丙氨酸代谢,能量代谢途径,植物激素信号调控,生物碱合成,脂质代谢途径,昼夜节律信号转导途径。GO富集分析表明,差异基因主要富集在生物学过程(包括转录调控,DNA复制,蛋白磷酸化等),细胞组分(包括细胞核,细胞膜,叶绿体,线粒体等),分子功能(包括DNA结合,蛋白结合等),另外在秋茄叶片发育的三个时期共统计到22333个SSR位点,占比最高的为单核苷酸,数量为12268条,其次为三核苷酸和二核苷酸,分别6410条和3245条,这些结果为红树秋茄分子标记辅助育种提供了重要依据,由于秋茄是非泌盐性红树品种,通过对成熟红树叶片进行扫描电镜的观察,发现红树叶片上表面无气孔结构,下表面分布有大量气孔,同时对叶片表面进行了元素种类和含量的分析,发现红树叶片下表面的钠离子和硫离子含量比叶片上表面高,同时在靠近叶片下表面的细胞中,发现了可能储存盐分的结构,这些结果为研究非泌盐性红树的叶片的生长发育提供了一定的理论基础。In order to explore the mechanism of material synthesis and gene expression during the development of Kandelia obovata,this study extracted and high-throughput sequenced the mRNA of mangrove leaves at three development stages,and obtained 64 189 053 high-quality sequences(Clean reads) and 19.29 Gb of high-quality bases.A total of 108 077 transcripts were obtained by assembly,37 206 transcripts with a length of more than2 000 bp,11 797 transcripts with a length of 200~300 bp,and Q30 were about 95.00%.Through KEGG enrichment analysis of the differential genes in the leaves of mangrove candel at three different stages,it was found that the differential genes were mainly enriched in the following metabolic pathways:carbohydrate metabolism pathway(starch and sucrose metabolism),phenylalanine metabolism,and energy metabolism pathway,Plant hormone signal regulation,amino acid metabolism pathway,lipid metabolism pathway,signal transduction pathway.GO enrichment analysis showed that the biological processes(including transcriptional regulation,DNA replication,protein phosphorylation,etc.),cell components(including nucleus,cell membrane,chloroplast,mitochondria,etc.),and molecular functions(including DNA binding,protein binding,etc.),in the three stages of the development of the leaves of the eggplant,a total of 22 333 SSR sites were counted,with the highest proportion of single nucleotide,12 268,followed by trinucleotide and dinucleotide,6 410 and 3 245,respectively.These results provide an important basis for molecular marker-assisted breeding of mangrove Kandelia.Because Kandelia is a non-saline-secreting mangrove variety,through scanning electron microscope observation of mature mangrove leaves,it is found that there is no stomatal structure on the upper surface of mangrove leaves,and a large number of stomata are distributed on the lower surface.At the same time,the element type and content of the leaf surface are analyzed,It was found that the content of sodium and sulfur ions on the lower surface of mangrove leaves

关 键 词：红树秋茄 转录组 差异表达基因 盐分转运蛋白 

分 类 号：S567.19[农业科学—中草药栽培]