摘要
甘薯[Ipomoea batatas (L.) Lam.]是重要的粮食、保健、工业原料和能源作物,而一些甘薯品种因叶型或叶色丰富被作为观赏植物或绿化植物。CHLI1基因编码叶绿素合成途径中镁离子螯合酶(MgCh)的Chl I亚基,对叶绿素的合成以及叶绿体的发育至关重要,并影响植物叶片着色。本实验以‘徐薯29号’为材料,采用RT-PCR方法克隆CHLI1基因对其进行生物信息学分析,并进行了不同部位的表达模式分析。结果表明,甘薯IbCHLI1基因最大阅读框(ORF)长度为1 281 bp,编码426个氨基酸,蛋白质相对分子质量为46 184.90 Da,其分子式为C_(2032)H_(3332)N_(568)O_(632)S_(11),原子总数为6 575,理论等电点为5.68;该蛋白属于亲水性非分泌蛋白,定位在叶绿体中,稳定性较好,不存在跨膜结构域以及信号肽,有多个磷酸化位点;该蛋白二级结构主要包括α-螺旋(42.25%)、无规卷曲(35.21%)、延伸链(15.96%)和β-转角(6.57%),三级结构预测结果显示该蛋白含有核苷三磷酸水解酶的P-loop环,保守结构域预测结果显示该蛋白是含有AAA结构域的ATP结合蛋白,属于Chl I超家族;系统进化分析发现该蛋白与三裂叶野牵牛(Ipomoea triloba)的CHLI1蛋白亲缘关系最近,序列相似性达到99.30%。组织特异性表达分析结果显示Ib CHLI1基因在甘薯不同组织中存在明显的组织表达特异性,在叶片中表达量最高,在根中表达量最低。以上对IbCHLI1基因在甘薯中的初步分析将为后续利用基因工程手段创制观赏甘薯新种质提供理论依据。
Sweetpotato [Ipomoea batatas(L.) Lam.] is an important food, health, industrial raw materials and energy crop. Among them, some sweetpotato varieties are used as ornamental or green plants because of their rich leaf shapes or colors and ornamental values. The CHLI1 gene encodes the Chl I subunit of magnesium ion chelatase(MgCh) in the chlorophyll synthesis pathway, which is essential for chlorophyll synthesis and it is essential for chlorophyll synthesis and chloroplast development, and affects plant leaf coloration. In this study, the CHLI1 gene was cloned using RT-PCR method for bioinformatics analysis of 'Xushu29' as the material and the expression pattern of different parts was analyzed. The results showed that the gene contains an open reading frame(ORF) with1 281 bp which encoding 426 amino acids and the relative molecular weight is 46.18 kD. Its molecular formula is C_(2032)H_(3332)N_(568)O_(632)S_(11), the total number of atoms is 6 575 and the theoretical isoelectric point is 5.68. This protein is a hydrophilic non-secreted protein with good stability, localized in chloroplast and no transmembrane domain and signal peptide. It has multiple phosphorylation sites and locates in chloroplasts. The secondary structure of the protein mainly includes alpha helix(42.25%), random coil(35.21%), extended strand(15.96%) and beta turn(6.57%). The prediction of conserved domain has demonstrated that the protein is an ATP-binding protein containing AAA domain and belongs to the Chl I superfamily. The prediction result of the tertiary structure has shown that the protein contained the P-loop of nucleoside triphosphate hydrolase. Phylogenetic tree analysis has shown that the protein is the most closely homologous to Ipomoea triloba, with a sequence similarity of 99.30%. Tissue specific expression analysis showed that IbCHLI1 gene had obvious tissue expression specificity in different tissues of sweetpotato, with the highest expression level in leaves and the lowest expression level in roots. These results would provide a
作者
高兵倩
王珧
刘洋
赵路宽
邓逸桐
曹清河
邹华文
Gao Bingqian;Wang Yao;Liu Yang;Zhao Lukuan;Deng Yitong;Cao Qinghe;Zou Huawen(College of Agriculture,Yangtze University,Jingzhou,434023;Jiangsu Xuhuai Reigion Xuzhou Institute of Agricultural Sciences,Sweetpotato Research Institute,Chinese Academy of Agricultural Sciences,Xuzhou,221132)
出处
《分子植物育种》
CAS
北大核心
2024年第22期7327-7336,共10页
Molecular Plant Breeding
基金
农业农村部物种资源保护项目(19210138)
海南省重点研发项目(ZDYF2020226)
江苏省根茎类种质资源库项目(JSGB2018-03)共同资助。
