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作 者:王莲 张小娟 王华[1] 贾素琦 李怡珊 徐婉婷 任爱华[2] WANG Lian;ZHANG Xiaojuan;WANG Hua;JIA Suqi;LI Yishan;XU Wanting;REN Aihua(School of Horticulture,Anhui Agricultural University,Hefei 230061;Horticultural Branch,Heilongjiang Academy of Agricultural Sciences,Harbin 150069)
机构地区:[1]安徽农业大学园艺学院,合肥230061 [2]黑龙江省农业科学院园艺分院,哈尔滨150069
出 处:《安徽农业大学学报》2024年第6期967-975,共9页Journal of Anhui Agricultural University
基 金:2021年安徽省重点研究与开发计划项目(202104a06020017);安徽农业大学2023年大学生创新创业训练计划(X202310364593X,X202310364594X,X202310364484)共同资助。
摘 要:为探究杜鹃花(Rhododendron simsii)蔗糖非酵解Ⅰ型相关蛋白激酶(sucrose non-fermenting^(-1)-related protein kinase,SnRK)对非生物胁迫的响应,基于杜鹃花基因组数据,利用生物信息学方法对RsSnRKs进行筛选鉴定和分析,并运用实时荧光定量PCR技术(RT-qPCR)检测该家族基因对干旱胁迫的响应情况。结果鉴定出杜鹃花RsSnRK基因家族共有3个亚族19个成员,它们编码的蛋白均为亲水蛋白,长度为362~555氨基酸,等电点为4.80~9.27,相对分子质量为41025.65~62951.44 Da,二级结构以α-螺旋和不规则卷曲为主,预测其主要位于细胞核或细胞质中。Motif和顺式作用元件分析发现,RsSnRKs同一亚家族基因具有相似的结构和基序,且RsSnRKs上游2000 bp含大量的光响应、激素响应和非生物胁迫响应等元件。共线性分析发现,杜鹃花RsSnRK2/RsSnRK14在内的4组共线性对基因进化选择压力值均小于1,受纯化选择;此外,10个RsSnRKs与15个拟南芥AtSnRKs存在19条共线性关系。RT-qPCR分析发现,杜鹃花RsSnRK第Ⅱ亚家族成员RsSnRK7、RsSnRK9和RsSnRK13的相对表达量随着干旱程度加重而明显上调,复水后又显著下调表达,说明杜鹃花RsSnRKs能够积极响应干旱胁迫。结果为进一步探索杜鹃花RsSnRKs基因功能奠定基础,并为杜鹃花抗旱基因的筛选提供理论依据。In order to explore the response of sucrose non-fermenting^(-1)-related protein kinase(SnRK)to abiotic stress in Rhododendron simsii,based on R.simsii genome data and bioinformatics technology,RsSnRKs were screened,identified and analyzed,and real-time quantitative PCR(RT-qPCR)was used to detect the response of the family genes to drought stress.The results showed that a total of 19 members from 3 subfamilies of the R.simsii RsSnRK gene family were identified.The encoded proteins were all hydrophilic proteins with a length of 362 to 555 amino acids,with an isoelectric point of 4.80 to 9.27 and a relative molecular mass being 41025.65-62951.44 Da.The secondary structures were dominated byα-helices and irregular coil.It was predicted that they were mainly located in the cell nucleus or cytoplasm.Motif and cis-acting element analysis found that genes of the same subfamily of RsSnRKs had similar structures and motifs,and 2000 bp upstream bases in RsSnRKs containing a large number of light response,hormone response,and abiotic stress response elements.Collinearity analysis found that the evolution-ary selection pressure values of four groups of collinear pairs of genes,including Rhododendron RsSnRK2/RsSnRK14,were all less than 1 and were subject to purifying selection;10 RsSnRKs formed 19 collinearity relationships with 15 AtSnRKs.RT-qPCR analysis found that the relative genes expression of RsSnRK7,RsSnRK9 and RsSnRK13 were sig-nificantly upregulated with drought,and significantly downregulated after rehydration,which indicated that RsSnRKs in R.simsii responded actively to drought stress.The research result lay a foundation for further exploring the function of RsSnRKs genes and provides a basis for screening drought resistant genes in Rhododendron.
关 键 词:杜鹃花 RsSnRK基因家族 生物信息学 干旱胁迫 表达分析
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