布顿大麦TB1基因的克隆及内生真菌对其表达量的影响  

Cloning of TB1 from Hordeum bogdanii and the effect of endophytic fungi on its expression

作　　者：韩丹 龙凤 陈胜 户萌菲 王栋[2] 陈水红[1] HAN Dan;LONG Feng;CHEN Sheng;HU Meng-fei;WANG Dong;CHEN Shui-hong(School of Life Science and Technology,Tarim University,Key Laboratory of Tarim Basin Biological Resources Conservation and Utilization Corps,Alaer 843300,China;School of Animal Science and Technology,Tarim University,Alaer 843300,China;Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China)

机构地区：[1]塔里木大学生命科学与技术学院,塔里木盆地生物资源保护利用兵团重点实验室,新疆阿拉尔843300 [2]塔里木大学动物科学与技术学院,新疆阿拉尔843300 [3]中国科学院深圳先进技术研究院,广东深圳518055

出　　处：《草业学报》2023年第8期176-185,共10页Acta Prataculturae Sinica

基　　金：国家自然科学基金(31802132,32260356,31660689);塔里木大学校长基金(TDZKZD202201)资助

摘　　要：分蘖生长在茎基部不伸长的节间,有不定根,能独立于主茎存活,禾本科作物布顿大麦主要通过分蘖来提高产量;内生真菌和基因均能调控布顿大麦分蘖,为了探究内生真菌对宿主植物分蘖相关基因TB1的影响,本研究利用RT-PCR法对布顿大麦TB1基因进行克隆并测序,采用生物信息学方法对该基因的序列结果进行分析,用SYBR Green荧光染料法对新疆温宿县和青海柴达木盆地带内生真菌(E+)和不带内生真菌(E-)的布顿大麦TB1基因进行q-PCR相对表达量分析。结果显示,克隆的TB1基因蛋白质编码区(CDS)全长为804 bp,编码267个氨基酸残基;TB1基因无启动子和PolyA位点;经亚细胞定位,TB1蛋白预计在液泡,TB1蛋白无跨膜结构域、无信号肽、不属于跨膜蛋白、存在于TCP家族;推测TB1蛋白为不稳定蛋白质。布顿大麦TB1基因与大麦亚种的TB1同源性最高,为96.72%,且与大麦亚种的TB1处于同一支系。温宿县和柴达木盆地布顿大麦根、茎、叶中TB1基因表达量趋势一致,内生真菌显著降低了植株分蘖发生部位茎基部的TB1基因表达量,表明内生真菌侵染影响了宿主TB1基因表达进而调控宿主植物分蘖。研究结果为后续布顿大麦分蘖机制研究奠定了理论基础。The tillers of the Poaceous crop plant Hordeum bogdanii develop from non-elongating internodes at the base of the stem,form adventitious roots,and can survive independently of the main stem.The yield of H.bogdanii is mainly increased by tillering,which is regulated by both endophytic fungi and genes of the host.To explore the effect of endophytic fungi on the tiller-related gene TB1 in the host plant,the TB1 gene of H.bogdanii was cloned by RT-PCR,sequenced,and then analyzed using bioinformatic methods.The transcript levels of TB1 were determined by PCR using SYBR Green fluorescent dye.These PCR analyses were conducted for H.bogdanii with endophytic fungi(E+)and without endophyte fungi(E-)growing in Wensu County,Xinjiang and Tsaidam Basin,Qinghai.The full-length coding sequence of the TB1 gene clonedfrom H.bogdanii was 804 bp,encoding a polypeptide consisting of 267 amino acid residues.The TB1 gene had no promoter and did not encode a polyA tail.A subcellular localization analysis predicted that TB1 localizes in the vacuole.The TB1 protein,which belongs to the TCP family,has no transmembrane domains and no signal peptide,so it is not a transmembrane protein.It was predicted to be an unstable protein.The TB1 gene of H.bogdanii showed the highest homology(96.72%)with TB1 of Hordeum vulgare subsp.,and was located in the same branch as TB1 from H.vulgare subsp.in the phylogenetic tree.The transcript profiles of TB1 in the roots,stems,and leaves were similar between plants growing in Wensu County and those growing in the Tsaidam Basin.Endophytic fungi significantly reduced the expression of TB1 at the base of stems(the tillering site).These results show that endophytic fungal infection affects the expression of the host’s TB1 gene,thereby regulating tillering.These findings provide a theoretical foundation for further research on the tillering mechanism of H.bogdanii.

关 键 词：内生真菌 布顿大麦 TB1基因 荧光定量PCR 

分 类 号：S543.9[农业科学—作物学]