芥蓝SPL基因家族鉴定及表达分析  

Identification and Expression Analysis of SPL Gene Family in Chinese Kale

作　　者：曾东琳 唐为玲 曾家晶 Gefu WANG-PRUSKI 赖钟雄[1] 郭容芳[1,2] ZENG Donglin;TANG Weiling;ZENG Jiajing;Gefu WANG-PRUSKI;LAI Zhongxiong;GUO Rongfang(Institute of Horticultural Plant Bioengineering, Fujian Agriculture and Forestry University, Fuzhou 350002;2Joint Laboratory of Daierhausi University, Fujian Agriculture and Forestry University, Fuzhou 350002, China)

机构地区：[1]福建农林大学园艺植物生物工程研究所,福州350002 [2]福建农林大学戴尔豪西大学联合实验室,福州350002

出　　处：《西北植物学报》2022年第3期361-370,共10页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31772310,31401859);博士后启动经费(132300155);博士后第10批特别资助(2017T100464)。

摘　　要：SPL存在于所有绿色植物中,在植物生长发育及叶片形成中起着重要的作用。该研究以‘改良香菇’芥蓝为材料,采用生物信息学方法,对芥蓝SPL家族(BoSPL)进行全基因组鉴定及分析;采用PCR技术克隆BoSPL3-1、BoSPL10-2和BoSPL11-2基因,并对芥蓝不同生长时期及不同组织部位进行定量分析,为揭示‘改良香菇’芥蓝叶片发育过程的分子机制以及培育芥蓝新品种奠定基础。结果表明:(1)BoSPL家族共有31个成员,编码区蛋白质氨基酸长度在157~1028 aa之间,BoSPL蛋白质的氨基酸均没有信号肽,属于非分泌蛋白;亚细胞定位预测结果显示,所有的BoSPL家族成员均有定位在细胞核上,少数成员有定位在液泡、叶绿体和细胞质中;BoSPL家族基因包含内含子数量为1~9个,所有成员均包含SBP保守结构域;系统进化分析显示,BoSPL可以分为8个进化群亚组,各群基因数量不等,且BoSPL进化关系与AtSPL相似。(2)成功克隆到BoSPL3-1、BoSPL10-2和BoSPL11-2基因,并成功构建pCAMBIA1302-35S-BoSPL3-1-GFP、pCAMBIA1302-35S-BoSPL10-2-GFP和pCAMBIA1302-35S-BoSPL11-2-GFP重组质粒,经根癌农杆菌GV3101介导瞬时转化烟草下表皮细胞,激光共聚焦显微镜检测表明,BoSPL3-1、BoSPL10-2和BoSPL11-2均定位在细胞核中,与预测结果一致。(3)qRT-PCR分析显示,BoSPL3-1、BoSPL10-2和BoSPL11-2在‘改良香菇’芥蓝叶片中相对表达量均较高,但3个基因在叶肉中的相对表达量无明显差异,而BoSPL10-2在花状变形叶中却显著高表达,推测其可能与花状变形叶形成相关。SPL exists in all green plants and plays an important role in plant growth and leaf formation.In this study,‘Gailiangxianggu’Chinese kale was used as material,and the whole genome of BoSPL family was identified and analyzed by bioinformatics method.The genes BoSPL3-1,BoSPL10-2 and BoSPL11-2 were cloned by PCR,and the quantitative analysis of different growth stages and different tissue parts of Chinese kale was carried out,which laid a foundation for revealing the molecular mechanism of the leaf development process of‘Gailiangxianggu’Chinese kale and cultivating new varieties of Chinese kale.The results show that:(1)there are 31 members of BoSPL family,and the amino acid length of the coding region proteins are 157-1028 aa.The amino acid of BoSPL protein has no signal peptide,which belongs to non-secretory protein;The results of subcellular localization prediction showed that all BoSPL family members were located in the nucleus,and a few members were located in vacuoles,chloroplasts and cytoplasm.BoSPL family genes contain 1-9 introns,and all members contain SBP conserved domains.Phylogenetic analysis shows that BoSPL can be divided into 8 subgroups of evolutionary groups,with different number of genes in each group,and the evolutionary relationship of BoSPL is similar to AtSPL.(2)Successfully cloned the BoSPL3-1,BoSPL10-2 and BoSPL11-2 genes of Chinese kale,and successfully constructed pCAMBIA1302-35S-BoSPL3-1-GFP,pCAMBIA1302-35S-BoSPL10-2-GFP and pCAMBIA1302-35S-BoSPL11-2-GFP.Transient transformation of tobacco lower epidermal cells mediated by agrobacterium tumefaciens GV3101,and laser confocal microscopy showed that BoSPL3-1,BoSPL10-2 and BoSPL11-2 were all located in the nucleus,which was consistent with the predicted results.(3)qRT-PCR analysis showed that the relative expression levels of BoSPL3-1,BoSPL10-2 and BoSPL11-2 were all higher in the leaves of‘Gailiangxianggu’Chinese kale,but there was no significant difference in the relative expression levels of the three genes in mesophyll,while Bo

关 键 词：芥蓝 BoSPL 全基因组鉴定 亚细胞定位 

分 类 号：Q785[生物学—分子生物学] Q786