割手密醛脱氢酶ScALDH基因的克隆与表达分析

发布时间：2018-06-01 09:50

本文选题：割手密 + 基因克隆　；参考：《广西植物》2017年03期

【摘要】：该研究从甘蔗细茎野生种(割手密Saccharum spontaneum)中克隆抗旱相关的基因Sc ALDH,并分析其在干旱处理条件下的表达情况和序列特征。利用RT-PCR技术克隆甘蔗的ALDH基因片段,并对其核苷酸和氨基酸序列进行分析。使用NCBI Blastx、ORF finder、Mega、NCBI Conserved Domain Search等程序对其分别进行不同物种氨基酸比较、开放阅读框(ORF)寻找、进化树及保守序列分析,并用Real Time-PCR分析所克隆基因在干旱胁迫前后的表达差异。结果表明:克隆出甘蔗的ALDH基因片段,总长度为1996bp,其中蛋白质编码区(CDS)全长1524bp,编码508个氨基酸;与其它物种ALDH类蛋白氨基酸序列有很高的同源性,有ALDH家族的保守序列,并含有完整的开放阅读框,系统进化树分析显示与玉米的蛋白质亲缘关系最近;Real timePCR数据表明,在干旱胁迫下,随着干旱时间的延长,该基因的表达量呈持续积累的表达模式。总体上来说,该基因对干旱胁迫显著表达。利用RT-PCR技术克隆的甘蔗ALDH基因,属于ALDH蛋白家族的一员,具有其典型的功能域,该基因在干旱胁迫过程中参与抗旱作用。该研究结果为野生种资源开发、优良抗旱亲本选择和培育抗旱性强甘蔗品种提供了参考依据。
[Abstract]:In this study, the drought-related gene SC ALDH was cloned from wild sugarcane fine stem species (Saccharum spontaneum), and its expression and sequence characteristics under drought treatment were analyzed. The ALDH gene fragment of sugarcane was cloned by RT-PCR and its nucleotide and amino acid sequences were analyzed. The amino acids of different species were compared by using NCBI Blastx ORF finder Conserved Domain Search and other programs. Open reading frame (ORF) search, phylogenetic tree and conserved sequence analysis were used to analyze the difference of expression of cloned genes before and after drought stress by Real Time-PCR. The results showed that the total length of ALDH gene of sugarcane was 1996bp.The length of protein coding region was 1524bp, encoding 508 amino acids, which had high homology with other species ALDH protein amino acid sequence and conserved sequence of ALDH family. The phylogenetic tree analysis showed that the protein phylogenetic relationship with maize was closely related to Real timePCR data. The results showed that under drought stress, the expression of the gene showed a continuous accumulation pattern with the prolongation of drought time. In general, the gene was significantly expressed under drought stress. The sugarcane ALDH gene cloned by RT-PCR technique is a member of the ALDH protein family and has its typical functional domain. The gene is involved in drought resistance during drought stress. The results provided a reference basis for the exploitation of wild species, the selection of fine drought-resistant parents and the cultivation of sugarcane varieties with strong drought resistance.
【作者单位】：云南农业大学农学与生物技术学院;中国农业科学院作物科学研究所;
【基金】：国家自然科学基金(31560417) 云南省应用基础研究重点项目(2015FA024) 云南省现代农业甘蔗产业技术体系建设项目(2009-2015) 云南省重点新产品开发计划项目(2012BB014) 云南省高原山地作物可持续生产系统研究创新团队项目(云科人发[2012]18号)~~
【分类号】：Q943.2;S566.1

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