甜菜抗逆基因P5CS的克隆及盐胁迫下的表达分析
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摘要
脯氨酸在植物适应逆境胁迫中起重要作用,脯氨酸合成酶(Proline synthetase)是脯氨酸合成代谢的关键酶,深入研究编码该酶的基因,对了解逆境下脯氨酸积累的分子机理、提高其抗逆性意义重大。本研究采用RT-PCR技术从甜菜中克隆了△1-吡咯啉-5-羧酸合成酶(P5CS)基因,利用生物信息学方法对序列进行了比对及其理化特性、结构域分析,该基因ORF为2 151 bp,编码由716个氨基酸组成的蛋白。序列已提交到GENBANK(ID:KY019201)。采用实时荧光定量PCR技术测定不同时间高盐(300 mmol/L NaCl)胁迫下对甜菜幼苗叶片P5CS基因相对表达量的影响,结果表明:随着处理时间的延长,幼苗受胁迫程度加深,P5CS基因的表达上调明显,进一步说明该基因响应逆境胁迫,本研究为今后深入研究提供了依据。
Proline plays an important role in plant adaptation to stress. Proline synthetase is the key enzyme in proline synthesis and metabolism. It is of great significance to study the gene encoding proline synthesis in order to understand the molecular mechanism of proline accumulation under stress and improve its stress resistance. In this study, P5CS gene was cloned from beet by RT-PCR technology, moreover, sequence distribution, physical and chemical characteristics and structural domain of the gene were analyzed via bioinformatics method. The ORF length of the gene was 2151 bp, encoding a protein composed of 716 amino acids. The sequence has been submitted to GENBANK(gene ID: KY019201). The expression pattern of P5CS gene in seedlings leaf during salt-stress(300 mmol/L NaCl) in Beta vulgaris was studied by qRT-PCR, the results showed that the expression of P5CS was upregulated obviously with the extension of treatment times and the intensification of stress, which further demonstrated that this gene could responded to stress. This study provides a basis for further study.
Proline plays an important role in plant adaptation to stress. Proline synthetase is the key enzyme in proline synthesis and metabolism. It is of great significance to study the gene encoding proline synthesis in order to understand the molecular mechanism of proline accumulation under stress and improve its stress resistance. In this study, P5CS gene was cloned from beet by RT-PCR technology, moreover, sequence distribution, physical and chemical characteristics and structural domain of the gene were analyzed via bioinformatics method. The ORF length of the gene was 2151 bp, encoding a protein composed of 716 amino acids. The sequence has been submitted to GENBANK(gene ID: KY019201). The expression pattern of P5CS gene in seedlings leaf during salt-stress(300 mmol/L NaCl) in Beta vulgaris was studied by qRT-PCR, the results showed that the expression of P5CS was upregulated obviously with the extension of treatment times and the intensification of stress, which further demonstrated that this gene could responded to stress. This study provides a basis for further study.
引文
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[2]吕春华,王宇光,於丽华,等.甜菜耐盐性分子及育种研究进展[J].中国糖料,2019,41(2):65-70.
[3]冯瑞军,伍国强.甜菜耐盐性生理及其分子水平研究进展[J].中国糖料,2015,37(6):60-65.
[4]史淑芝,崔杰,鲁兆新,等.甜菜种质资源耐盐性筛选[J].中国甜菜糖业,2008(4):7-9.
[5]李鹤男.DDRT-PCR技术分析甜菜盐胁迫相关基因差[D].哈尔滨工业大学,2013:46-48.
[6]田庆斌,崔杰,李俊良,等.甜菜盐胁迫相关蛋白的初步筛选[J].中国甜菜糖业,2016(2):1-3.
[7]Jie Cui,Zongyan Sun,Junliang Li,et,al.Characterization of miRNA160/164 and Their Targets Expression of Beet(Beta vulgaris)Seedlings Under the Salt Tolerance[J].Plant Molecular Biology Reporter,2018,36:790-799.
[8]Kubala S,?ukasz Wojtyla,Quinet M,et al.Enhanced expression of the proline synthesis gene P5CSA,in relation to seed osmopriming improvement of Brassica napus,germination under salinity stress[J].Journal of Plant Physiology,2015,183(4):1-12.