首页|期刊导航|华北农学报|杜梨PbKRP2序列特征与表达特性分析

杜梨PbKRP2序列特征与表达特性分析OA

Analysis of Sequence Characteristics and Expression Patterns of PbKRP2 in Pyrus betulaefolia

中文摘要英文摘要

KRP基因家族作为细胞周期调控的关键因子,通过抑制周期蛋白依赖性激酶(CDK)活性调节细胞分裂与核内复制进程,并参与植物非生物胁迫响应.为探索杜梨PbKRP2的结构特征和对环境胁迫的响应特性,采用逆转录PCR技术进行PbKRP2的基因克隆,并结合生物信息学方法对其序列特征、蛋白互作及亲缘关系等进行分析,通过荧光定量PCR技术检测杜梨PbKRP2的组织表达特性和在不同逆境胁迫下的表达模式.结果表明,该基因全长585 bp,包含4个外显子和3个内含子,编码194个氨基酸,含有25个潜在磷酸化位点和1个O-糖基化修饰位点,且在该蛋白C端具有周期蛋白依赖性激酶抑制因子家族特有的CDI结构域;进化树分析显示,杜梨PbKRP2与裸花东京樱花PyKRP2、桃PpKRP2和苹果MdKRP2的亲缘关系较为接近;蛋白质互作网络筛选发现,PbKRP2与10个蛋白存在潜在互作关系,互作蛋白静态结构分析发现,PbKRP2与其靶蛋白PbCDKB1;1和PbCYCD7;1形成空间互作构型;该基因启动子区存在多个响应环境因子与激素信号的顺式调控元件;实时荧光定量PCR结果表明,PbKRP2基因在茎和叶中表达量显著高于根、花瓣和果实,且该基因的转录水平在低温胁迫和ABA处理时持续上调,脱水和盐胁迫下显著下调.综上所述,通过解析PbKRP2的序列特性及其对非生物胁迫的差异化响应模式,为梨抗逆育种及果树遗传改良提供了候选基因.

The Kip-related protein(KRP)gene family acts as a key regulator of the cell cycle,modulating cell division and endoreduplication by inhibiting cyclin-dependent kinase(CDK)activity,and participates in plant abiot-ic stress responses.To investigate the structural characteristics and environmental stress response profile of PbKRP2 in Pyrus betulifolia,the gene was cloned using reverse transcription PCR(RT-PCR).Bioinformatic analysis was con-ducted to elucidate its sequence features,predicted protein interactions,and phylogenetic relationships.Quantitative Real-time PCR(qRT-PCR)was employed to assess the tissue-specific expression characteristics of PbKRP2 and its expression patterns under various stress conditions.Results showed that PbKRP2 was 585 bp in length,contained 4 exons and 3 introns,and encoded a 194-amino acid protein.The predicted protein had 25 potential phosphorylation sites,a O-glycosylation site,and a conserved C-terminal cyclin-dependent kinase inhibitor(CDI)domain.Phyloge-netic analysis indicated that PbKRP2 was closely related to PyKRP2 in Prunus yedoensis var.nudiflora,PpKRP2 in Prunus persica,and MdKRP2 in Malus domestica.Protein interaction network screening identified potential interac-tions between PbKRP2 and 10 proteins.Analysis of the static structure of potential interacting proteins suggested that PbKRP2 formed spatial interaction conformations with its target proteins PbCDKB1;1 and PbCYCD7;1,respec-tively.The promoter region of PbKRP2 contained multiple cis-acting regulatory elements responsive to environmental factors and hormonal signals.qRT-PCR analysis demonstrated that PbKRP2 transcript levels were significantly high-er in stems and leaves compared to roots,petals,and fruits.Furthermore,PbKRP2 transcription levels were consist-ently upregulated under low-temperature stress and abscisic acid(ABA)treatment,but significantly downregulated under dehydration and salt stress.In summary,these findings provide a candidate gene for stress-resistant breeding of pear and genetic improvement of fruit trees by elucidating the sequence characteristics of PbKRP2 and its differ-ential response patterns to abiotic stresses.

靳丛;徐连理;刘琦;张晓小;张宇双;徐源;陈钰祺;任雅楠;王纪忠

淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003淮阴工学院生命科学与食品工程学院,江苏淮安 223003

农业科技

杜梨PbKRP2基因基因克隆非生物胁迫表达分析

Pyrus betulaefoliaPbKRP2 geneGene cloningAbiotic stressExpression analysis

《华北农学报》 2026 (2)

36-46,11

国家自然科学基金青年基金项目(31801829)江苏省自然科学基金青年基金项目(BK20170463)江苏省大学生创新训练项目(S202511049090)淮阴工学院博士科研启动基金项目(Z301B16532)

10.7668/hbnxb.20196258

评论