曾小玲 赵瑞丽 钟开勤 朱朝辉 谢鑫鑫 温庆放

摘  要：為探讨软腐病菌侵染下菜心转录组功能基因信息，采用BGISEQ-500高通量测序技术对软腐病菌侵染下菜心叶片进行转录组测序，平均每个样获得47.27 M clean reads，Q20值均大于95%，共检测到表达的Unigene为36 760个，其中已知的有35 327个，预测新Unigene有1 433个;共检测出19 549个新转录本，其长度主要集中在300～2000 nt。功能注释结果显示，有32 047个Unigene在Nr数据库获得注释，其中注释到芸薹属白菜上的Unigene最多;GO功能共注释到12 588个Unigene;KEGG数据库注释到18 583个Unigene，涉及到137个代谢通路。共获得21 776个组间差异表达基因（differential expression genes， DEGs），其中对照与发病前期组间的DEGs有15 007个，6 137个上调表达，8 870个下调表达;发病前期与发病中期组间DEGs有13 118个，10 278个上调表达，2 840个下调表达;发病中期与发病后期组间DEGs有11 293个，1 790个上调表达，9 503个下调表达。DEGs的Ven图分析显示5 110个基因在每组间均存在差异，DEGs功能分析显示DEGs参与泛素蛋白降解途径、过氧化物酶体途径、光合碳固定途径、糖酵解途径等多种生命活动。本研究结果为深入开展菜心抗软腐病基因组学和分子生物学研究奠定基础。

关键词：菜心;软腐病;转录组;差异表达基因

中图分类号：S436.34      文献标识码：A

Abstract： To explore the functional gene information in flowering Chinese cabbage （Brassica campestris L. ssp. Chi-nensis （L.） var. utilis Tsen et Lee） inoculated by soft rot disease， the transcriptome of flowering Chinese cabbage was sequenced by BGISEQ-500 high-throughput sequencing technology. The results showed that average obtained 47.27 M clean reads and the Q20 base was more than 95%. A total of 36 760 Unigene transcripts were detected， of which 35 327 were known and 1 433 were predicted. A total of 19 549 new transcripts were detected， the length of which was mainly between 300 and 2000 nt. Functional annotation results showed that 32 047 Unigene were annotated in NR database， among which Unigenes were annotated most on Brassica rapa ssp. Pekinensis. GO function was annotated with 12 588 Unigene. Kegg database was annotated with 18 583 Unigene， involving 137 metabolic pathways. A total of 21 776 differential expression genes （DEGs） were obtained， including 15 007 DEGs， 6 137 up-regulated genes and 8 870 down- regulated genes between the control and premorbid groups. There were 13 118 DEGs， 10 278 up-regulated genes and 2840 down-regulated genes between pre-onset and mid-onset. There were 11 293 DEGs， 1 790 up-regulated genes and 9503 down-regulated genes. Analysis of the DEGs Ven map showed that 5 110 genes were different in each group. Functional analysis of DEGs showed that the differentially expressed genes were involved in various life activities， such as ubiquitin degradation pathway， peroxisome pathway， photosynthetic carbon fixation pathway， glycolytic process pathway and so on. The results of this study would lay a foundation for further studies on the ge-nome and molecular biology of resistance to soft rot of flowering Chinese cabbage.

Keywords： flowering Chinese cabbage; soft rot; transcriptome; differentially expressed genes

DOI： 10.3969/j.issn.1000-2561.2021.05.032

菜心（Brassica campestris L. ssp. Chinensis （L.） var. utilis Tsen et Lee）为十字花科芸薹属白菜亚种的一个变种，又名菜薹，是中国南方的特产蔬菜之一。近年来，随着菜心栽培面积的扩大和重茬现象的出现，菜心病害逐年加重，其中细菌性软腐病是危害菜心的主要病害之一。蔬菜细菌性软腐病是由软腐欧文氏菌胡萝卜亚种（Erwinia carotovora ssp. carotovora， Ecc）引起的一种流行性死体营养型细菌病害，其病原……