摘要
为了获得姜荷花叶绿素降解的关键酶基因PPH信息,本试验在前期通过姜荷花全长转录组测序获得大量转录组信息的基础上,进行筛选分析,获得了2条PPH基因,分别命名为PPH1和PPH2。PPH1基因(GenBank:MT077178),cDNA序列全长1 795 bp,开放阅读框1 437 bp (138~1 574 bp),编码一条478 aa的氨基酸序列。PPH2基因(GenBank:MT077179),c DNA序列全长1 393 bp,开放阅读框1 227 bp (70~1 296 bp),编码一条408 aa的氨基酸序列。采用BLAST、Translate tool (ExPASy)、Clustal Omega、Find Conserved Domains(NCBI)、ProtParam、TMHMM Server、SOPMA、SWISS-MODEL、ClustalX (1.81)、MEGA 4.1等预测和分析了这2个基因编码蛋白氨基酸的一级结构(包括氨基酸序列的组成,保守区,理化特征等)、二级结构、三级结构及分子系统进化关系。PPH1和PPH2的核苷酸与蛋白氨基酸序列与其它物种的PPH基因都具有很高的同源性,两者都包含有水解酶(Hydrolase)特征PLN02578的保守区。分子系统进化分析表明,姜荷花的PPH1和PPH2聚为一小类,然后与小果野芭蕉PPH (XP018677219.1)的亲缘关系最为接近,而与双子叶植物的关系较远。本研究为后期通过遗传转化手段改良姜荷花不育苞片的颜色提供了分子基础。
To obtain key enzyme gene PPH in chlorophyll degradation process of Curcuma alismatifolia, on the basis of obtaining a large amount of transcriptome information by sequencing the full-length transcriptome, the study had screened and analyzed these transcriptome information and obtained 2 PPH genes which named PPH1 and PPH2. The PPH1 gene(GenBank: MT077178) has a full-length cDNA sequence of 1 795 bp in length, an open reading frame of 1 437 bp(from 138 to 1 574 bp), and encode a sequence with 478 aa amino acid. The PPH2 gene(GenBank: MT077179) has a full-length c DNA sequence of 1 393 bp, an open reading frame of 1 227 bp(from70 to 1 296 bp), and encode a sequence with 408 aa amino acid. Using BLAST, Translate tool(ExPASy), Clustal Omega, Find Conserved Domains(NCBI), ProtParam, TMHMM Server, SOPMA, SWISS-MODEL, ClustalX(1.81), MEGA 4.1 and so on. Their amino acid composition, physical and chemical properties, conserved domains,secondary structures, tertiary crystal structures, and molecular phylogeny were predicted and analyzed. The nucleotide and protein amino acid sequences of PPH1 and PPH2 have high homology with these PPH genes of other species, and both of them contain a conserved region PLN02578 with hydrolase characteristic. Molecular phylogenetic analysis showed that a small cluster of PPH1 and PPH2 were closest to Musa acuminate PPH(XP01867-7219.1), but far away from dicotyledons.This study provided a molecular basis for improving color of Curcuma alismatifolia sterile bracts by genetic transformation in the future.
作者
丁华侨
毛俐慧
胡伟
董青
刘建新
Ding Huaqiao;Mao Lihui;Hu Wei;Dong Qing;Liu Jianxin(Xiaoshan Cotton and Bast Fiber Crops Research Institute of Zhejiang Province,Hangzhou,311202;Yuanpei College,Shaoxing University,Shaoxing,312000)
出处
《分子植物育种》
CAS
北大核心
2021年第8期2521-2526,共6页
Molecular Plant Breeding
基金
浙江省省级重点研发计划(2019C02025)资助。
