甘蓝型油菜BnMAPK1超量表达及中油821的转录差异表达分析  

Transcriptional differential expression analysis between BnMAPK1-overexpression and Zhongyou 821 rapeseed(Brassica napus L.)

作　　者：王珍 张晓莉[1] 刘淼 姚梦楠[4] 孟晓静 曲存民 卢坤[1,2] 李加纳[1,2] 梁颖[1,2] WANG Zhen;ZHANG Xiao-Li;LIU Miao;YAO Meng-Nan;MENG Xiao-Jing;QU Cun-Min;LU Kun;LI Jia-Na;LIANG Ying(College of Agronomy and Biotechnology,Southwest University/Chongqing Engineering Research Center for Rapeseed,Chongqing 400715,China;Academy of Agricultural Sciences,Southwest University,Chongqing 400715,China;Key Laboratory of Plant Resource Conservation and Germ-plasm Innovation in Mountainous Region(Ministry of Education)/Institute of Agro-bioengineering College,Guizhou University,Guiyang 550025,Guizhou,China;Jiangsu Yanjiang Institute of Agricultural Sciences/Nantong Academy of Agricultural Sciences,Nantong 210014,Jiangsu,China)

机构地区：[1]西南大学农学与生物科技学院/油菜工程研究中心,重庆400715 [2]西南大学现代农业科学研究院,重庆400715 [3]贵州大学农业生物工程研究院/山地植物资源保护与种质创新教育部重点实验室,贵州贵阳550025 [4]江苏沿江地区农业科学研究所/南通市农业科学研究院,江苏南通210014

出　　处：《作物学报》2023年第3期856-868,共13页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(32101663,31872876);中国博士后科学基金项目(2021M692683);重庆市自然科学基金项目(cstc2021jcyj-bshX0234);重庆市博士后科研特别资助项目(2010010006157688);高等学校学科创新引智基地项目111(B12006)资助。

摘　　要：甘蓝型油菜(Brassica napus L.)是我国重要的油料作物之一,具有较高的产量及营养价值。挖掘油菜广谱抗逆基因并深入探究其分子机制,对提高油菜抗逆性且保证产量及品质具有重要意义。MAPK级联(Mitogen-activated protein kinase cascades)是多种信号跨膜传递的交汇点,参与生长发育、生物及非生物胁迫应答等生物学过程。为了解甘蓝型油菜BnMAPK1基因的生物学功能,本研究以BnMAPK1超量表达(OE)和中油821(CK)油菜为材料进行数字基因表达谱分析。结果显示,与CK相比,650个基因在OE油菜中差异表达;其中上调表达基因243个,下调表达基因407个。GO注释共富集到生物学过程118条途径,其中18条与光合作用相关(包含77个差异表达基因);分子功能7条途径,包括草酸氧化酶活性、锰离子结合、氧化还原酶活性等;细胞组分38条途径,包括叶绿体、质体、类囊体等。KEGG分析结果表明,BnMAPK1参与调控油菜光合作用、碳代谢、次生代谢物生物合成等14条过程。此外,利用实时荧光定量PCR验证8个候选基因在OE和CK油菜中的表达,其表达变化与测序结果基本一致。本研究结果为甘蓝型油菜BnMAPK1在生长发育及光响应等生物学过程中的功能分析奠定了基础,也为农作物广谱抗逆遗传育种提供了理论依据。Rapeseed(Brassica napus L.)is an important oilseed crop in China,with high yield and nutritional value.Exploring the mechanism of the broad-spectrum stress regulatory genes plays key roles for improving stress resistance,yield,and quality in rapeseed.Mitogen-activated protein kinase(MAPK)cascades are the junctions of several transmembrane signaling,and are involved in a wide variety of biological processes,including growth,development,biotic,and abiotic stress responses.In order to survey the biological function of BnMAPK1 gene in rapeseed,BnMAPK1-overexpression(OE)and Zhongyou 821(CK)plants were used as the experimental materials to perform the digital gene expression profiling sequencing at one-month-old seedling stage.Compared with CK,a total of 650 differentially expressed genes were obtained in OE plants,including 243 up-regulated genes and 407 down-regulated genes.GO annotation showed that differentially expressed genes were mainly enriched in 118 pathways in biological process,among which 18 pathways related to photosynthesis(with 77 differentially expressed genes).There were seven pathways in molecular function such as oxalate oxidase activity,manganese ion binding,and oxidoreductase activity;and 38 pathways in cellular component such as chloroplast,plastid,and thylakoid.KEGG indicated that BnMAPK1 was involved in regulating 14 pathways in rapeseed,including photosynthesis,carbon metabolism,and biosynthesis of secondary metabolites.In addition,the relative expression levels of eight candidate genes were validated by qRT-PCR in OE and CK in rapeseed,demonstrating that the expression changes of these eight genes were generally consistent with the sequencing data.Our findings lay a foundation for the function of BnMAPK1 in growth and development and light response in rapeseed and provide a theoretical basis for genetic breeding of crop resistance.

关 键 词：甘蓝型油菜 BnMAPK1 差异表达基因(DEGs) 光合作用 数字基因表达谱测序(DGE-seq) 

分 类 号：S565.4[农业科学—作物学]