利用转录组挖掘羊肚菌干旱胁迫相关基因  

Transcriptome Mining of Genes Involved in Drought Stress Response in Morchella sextelata

作　　者：仝乐涛 徐爱国 鲍大鹏[2] 王伟霞 李福后 唐利华[2] TONG Letao;XU Aiguo;BAO Dapeng;WANG Weixia;LI Fuhou;TANG Lihua(College of Marine Food and Bioengineering,Jiangsu Ocean University,Lianyungang 222005,Jiangsu,China;Institute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China;Institute of Plateau Biology,Tibet Autonomous Region,Lasa 850000,Xizang,China)

机构地区：[1]江苏海洋大学海洋食品与生物工程学院,江苏连云港222005 [2]上海市农业科学院食用菌研究所,上海201403 [3]西藏自治区高原生物研究所,西藏拉萨850000

出　　处：《食用菌学报》2024年第2期26-33,共8页Acta Edulis Fungi

基　　金：西藏自治区重点研发计划(XZ202101ZY0005N)。

摘　　要：为挖掘羊肚菌(Morchella sextelata)干旱胁迫相关基因,设置对照组和干旱组,每组3畦,待对照组子实体生长至黑褐色时,每畦随机取1个子实体样品,通过转录组测序和基因功能注释分析对照组与干旱组的差异表达基因,采用GO和KEGG富集分析显著富集的GO条目和KEGG代谢通路,再根据GO和KEGG富集分析结果,通过基因注释信息确定与干旱胁迫相关的关键差异表达基因,随机选择5个与干旱胁迫相关的关键差异表达基因进行荧光定量PCR验证。结果表明:对照组子实体颜色白至淡黄、外观丰满,干旱组子实体颜色偏黑褐色、外观干瘪,且子实体较小;对照组与干旱组相比,共获得2 817个差异表达基因,其中上调表达基因1 492个,下调表达基因1 325个;GO富集结果表明,差异表达基因显著富集于DNA复制、DNA依赖性DNA复制、单糖运输、蛋白质-DNA复合物、核糖体、MCM复合体、核糖体的结构成分、FAD结合能力和溶质:阳离子同向转运蛋白活性等;KEGG富集结果表明,差异表达基因主要参与DNA复制、错配修复、类固醇生物合成、核糖体、剪辑切除修复、核苷酸切除修复、甲烷代谢、淀粉和蔗糖代谢、精氨酸和脯氨酸代谢、硫铵代谢等过程;与干旱胁迫相关的18个关键差异表达基因包括转运蛋白、热激蛋白、转录因子、激酶和自噬相关基因;荧光定量PCR测定结果与转录组测序分析的关键差异表达基因结果一致。Genes associated with drought stress in Morchella sextelata were identified by comparative transcriptome analysis.Two groups,control group and drought-stressed group,were established,and then three fruiting body samples(one per plot)were randomly taken from each group when fruiting bodies of the control group turned dark brown.Differentially expressed genes(DEGs)between the control and drought-stressed groups were analyzed through transcriptome sequencing and gene functional annotation.GO and KEGG enrichment analyses were used to identify significantly enriched GO terms and KEGG metabolic pathways.Based on the above analyses,key DEGs related to drought stress were determined,and then five key DEGs were randomly selected to be validated by qRT-PCR.The results showed that fruiting bodies in the control group were white to light yellow with a plump appearance,while those in the drought-stressed group were predominantly dark brown,shriveled,and smaller than the control group.A total of 2817 DEGs were identified,including 1492 up-regulated genes and 1325 down-regulated genes.GO enrichment analysis showed that the DEGs were significantly enriched in DNA replication,DNA-dependant DNA replication,monosaccharide transport,protein-DNA complex,ribosome,MCM complex,structural constituent of ribosome,FAD binding,and solute:cation symporter activity,etc..KEGG enrichment analysis revealed the involvement of the DEGs in DNA replication,mismatch repair,steroid biosynthesis,ribosome base excision repair,nucleotide excision repair,methane metabolism,starch and sucrose metabolism,arginine and proline metabolism,and sulfur metabolism,etc..Eighteen key DEGs associated with drought stress were identified,including genes related to transporter proteins,heat shock proteins,transcription factors,kinases,and autophagy.The results of qRT-PCR were consistent with those of the transcriptome sequencing analysis for the five randomly selected DEGs.

关 键 词：羊肚菌 干旱 转录组 基因差异表达 

分 类 号：S646.7[农业科学—蔬菜学]