普通烟草PYL基因家族成员鉴定及干旱胁迫下基因表达分析  

Identification and Expression Analysis of PYL Gene Family in Common Tobacco(Nicotiana tabacum L.)under Drought Stress

作　　者：扈佳慧 徐腾航 刘杰 王雨果 刘红峰 王世泽 余世洲 王丰 HU Jiahui;XU Tenghang;LIU Jie;WANG Yuguo;LIU Hongfeng;WANG Shize;YU Shizhou;WANG Feng(College of Agriculture,Guizhou University,Guiyang,Guizhou 550025;Institute of Tobacco Science,Guiyang,Guizhou 550081;College of Agriculture and Biotechnology,Hebei Normal University of Science and Technology,Qinhuangdao,Hebei 066004;Guizhou Tobacco Company Guiyang Company,Guiyang,Guizhou 550001)

机构地区：[1]贵州大学农学院,贵州贵阳550025 [2]贵州省烟草科学研究院,贵州贵阳550081 [3]河北科技师范学院农学与生物科技学院,河北秦皇岛066004 [4]贵州省烟草公司贵阳市公司,贵州贵阳550001

出　　处：《核农学报》2025年第1期47-58,共12页Journal of Nuclear Agricultural Sciences

基　　金：中国烟草总公司烟草基因组计划与生物育种重大科技项目(110202101032);中国烟草总公司贵州省公司重点研发项目(2023XM02);贵州省科学技术基金项目(黔科合基础-ZK[2024]一般648)。

摘　　要：PYR/PYL/RCAR(PYL)蛋白作为脱落酸(ABA)的直接受体,在ABA信号转导途径中发挥着关键调控作用。为探究PYL基因在烟草中的系统发育和表达模式,本研究利用生物信息学手段在普通烟草品种K326全基因组范围内进行了鉴定分析,并利用实时荧光定量PCR(qRT-PCR)检测了各基因在遭受干旱胁迫0、1、3、6、12、24、36和48 h的基因表达量。结果共鉴定到26个烟草PYL基因,依据系统发育和结构特点可划分为3个亚家族(Ⅰ~Ⅲ),成员数目分别为9、11和6。蛋白理化性质分析结果表明,所有烟草PYL蛋白都呈亲水性,氨基酸长度在173~586 aa之间,相对分子量在16 763.26~65 709.39 Da之间,等电点(pI)在4.73~9.05之间。顺式作用元件分析结果发现,烟草PYL基因启动子区含有MYB、NAC、WARKY、TCP、ZF-HD等逆境胁迫响应相关元件。基因表达结果显示,烟草PYL基因在遭受干旱胁迫后,除NtPYL8、NtPYL9、NtPYL17和NtPYL21基因外、其余基因表达量整体表现为上调趋势,但各亚家族成员间基因表达具有明显差异。其中亚家族Ⅱ中NtPYL1、NtPYL2、NtPYL4基因和亚家族Ⅲ中NtPYL25基因在遭受干旱胁迫6~12 h后基因表达量达到最高,且相对表达量较0 h上升显著,可作为烟草应答干旱胁迫的重要候选基因。本研究为进一步开展烟草候选抗旱PYL基因功能研究及育种利用奠定了基础。PYR/PYL/RCAR(PYL)proteins serve as direct receptors for abscisic acid(ABA)and play a crucial regulatory role in the ABA signaling pathway.To explore the phylogenetic relationships and expression patterns of PYL genes in tobacco,the complete genome of a common tobacco variety K326 was identified and analyzed with bioinformatics tools.The expression levels of each gene under drought stress conditions at 0,1,3,6,12,24 and 48 h were determined by quantitative real-time PCR(qRT-PCR).The results showed that 26 NtPYL genes were identified and categorized into three subfamilies(Ⅰ-Ⅲ)based on phylogenetic and structural features,with member counts of 9,11 and 6,respectively.Physicochemical analysis of the proteins revealed that all tobacco PYL proteins are hydrophilic,with amino acid lengths ranging form 173 to 586 aa,relative molecular weights between 16763.26 and 65709.39 Da,and isoelectric points(pI)between 4.73 and 9.05.Functional motif analysis identified MYB,NAC,WARKY,TCP,ZF-HD,and other stressresponsive elements in the promoter regions of tobacco PYL genes.Gene expression analysis demonstrated that the overall expression trend of tobacco PYL genes,with the exception of NtPYL8,NtPYL9,NtPYL17,and NtPYL21,was upregulated under drought stress.However,significant differences in gene expression were observed among members of different subfamilies.Notably,NtPYL1,NtPYL2,and NtPYL4 in subfamilyⅡ,as well as NtPYL25 in subfamilyⅢ,exhibited their highest expression levels at 6 to 12 hours after drought stress,showing a significant increase in relative expression compared to that at 0 h.These genes can be considered crucial candidates for drought stress response.This study lays a foundation for further investigation into the functional roles of candidate drought-resistant PYL genes in tobacco and their potential applications in breeding.

关 键 词：烟草 PYL家族 干旱胁迫 基因表达分析 脱落酸 

分 类 号：S572[农业科学—烟草工业]