干旱胁迫下枇杷叶片的转录组分析

Transcriptome Profiling of Loquat Leaves under Drought Stress

分析干旱胁迫下枇杷叶片的转录组,挖掘功能基因并对其差异表达基因进行筛选和分析,为枇杷抗旱提供理论依据。利用新一代高通量测序技术测序,对测序结果进行de novo拼接、功能注释和ORF预测,将差异表达基因在COG、GO和KEGG数据库中进行比对注释。测序结果表明,获得转录本共88 530个,平均长度为740.64 bp,ORF41 748条。COG、GO和KEGG数据库将转录本分别划分为24,54个功能类别及291条代谢通路中。25 197个差异表达的基因在30条代谢通路中显著富集,与酶活性、激素合成代谢和信号转导等相关的差异基因积极响应枇杷干旱,其中双萜类、油菜素类固醇和类胡萝卜素3条生物合成途径中的差异基因呈现出较为一致的表达。采用实时荧光定量(qRT-RCR)对选取的差异基因进行验证,其中过氧化物酶、蛋白激酶byr2、丝氨酸苏氨酸蛋白激酶、脱落酸8'-羟化酶、吲哚-3-乙酰乙酸合酶相关基因上调表达,细胞色素P450 734A1基因下调表达。为枇杷提供了较为全面的基因信息和代谢途径数据,为干旱胁迫下枇杷分子调控机制的深入研究奠定了基础。

The transcriptome of loquat leaves under drought stress were sequenced by Illumina Hi Seqhighthroughput platform which is a new generation of sequencing technology. This study was conducted to elucidate functional gene,analyzed its differential expressed gene and provided a theoretical basis for its genetic improvement. The sequencing data was assembled by de novo assembly and the differential expressed gene screened was annotated in COG,GO and KEGG database. The sequencing results showed that 88 530 transcripts with the mean length of 740. 64 bp were conducted. 41 748 ORFs were predicted by opening reading fragment. Clusters of orthologous groups performed the transcripts into 24 categories. All assembled transcripts and ORFs were divided into 54 branches by Gene ontology and 291 pathways by the Kyoto Encyclopedia of Genes and Genomes annotations analyses. 25 197 DEGs were enriched in 30 metabolic pathways and numerous genes related to enzyme activity,hormone metabolism and signal transduction inferred that might be connected with drought stress of loquat. Moreover,DEGs in carotenoid biosynthesis,brassinosteroid biosynthesis and diterpenoid biosynthesis showed a similar expression.Furthermore,DEGs were confirmed by qRT-PCR and peroxidase,protein kinase byr2,Serine/threonine protein kinase,abscisic acid 8＇-hydroxylase and indole-3-acetic acid-amido synthetase were down regulated,Cytochrome P450 734A1 was on the contrary. This study not only provided valuable genetic resources and a comprehensive map of drought-responsive genes in this species but also a data basis for further explorer molecular response mechanism to drought stress in loquat.