羊草Δ1-吡咯琳-5-羧酸合成酶(LcP5CS1)基因的克隆与分析

doi:10.11686/cyxb20130424

草业学报 ›› 2013, Vol. 22 ›› Issue (4): 197-204.DOI: 10.11686/cyxb20130424

羊草Δ1-吡咯琳-5-羧酸合成酶(LcP5CS1)基因的克隆与分析

张乐新^1,2,苏蔓¹,马甜^1,3,马兴勇¹,闫学青^1,2,
彭献军¹,陈双燕¹,程丽琴^1*,刘公社^1*

1.中国科学院植物研究所资源植物研发重点实验室,北京100093;
2.中国科学院研究生院,北京 100049;
3.宁夏大学农学院,宁夏银川 750021

出版日期:2013-08-20 发布日期:2013-08-20
通讯作者: E-mail:liugs@ibcas.ac.cn; lqcheng@ibcas.ac.cn
作者简介:张乐新(1986-),男,山东高密人,硕士。E-mail:zhanglexin123@163.com
基金资助:
国家自然科学基金项目(31170316和30970291)资助。

Cloning and analysis of the Δ1-pyrroline-5-carboxylate synthetase (LcP5CS1)from Leymus chinensis

ZHANG Le-xin^1,2, SU Man¹, MA Tian^1,3, MA Xing-yong¹, YAN Xue-qing^1,2,
PENG Xian-jun¹, CHEN Shuang-yan¹, CHENG Li-qin¹, LIU Gong-she¹

1.Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China;
2.Graduate University of Chinese Academy of Sciences, Beijing 100049, China;
3.Agricultural College, Ningxia University, Yinchuan 750021, China

Online:2013-08-20 Published:2013-08-20

摘要/Abstract

摘要： 羊草具有耐寒、耐盐和耐旱等特点,是一种兼具经济价值和生态价值的多年生重要牧草。脯氨酸是植物在渗透胁迫下积累的主要渗透调节物质之一。Δ1-吡咯啉-5-羧酸合成酶(P5CS)是脯氨酸的谷氨酸合成途径中的关键酶。根据前期Solexa测序结果的数据,设计同源引物,从羊草中克隆了一个P5CS基因,该基因cDNA序列全长2 197 bp,推断其编码730个氨基酸,根据序列比对将其蛋白命名为LcP5CS1。同源性分析表明:LcP5CS1与单子叶植物短柄草P5CS的同源性为95％,而与双子叶植物的P5CS同源性较低。表达模式分析表明,在叶鞘、叶和茎中,LcP5CS1均有较高表达;在冷、ABA、盐和干旱胁迫下,该基因明显受胁迫诱导,并随胁迫时间的延长,其表达量总体呈上调趋势。对羊草人工创伤处理后的Solexa测序差异表达数据分析表明:LcP5CS1在处理2和6 h后表达量明显提高,推测羊草创伤处理后体内脯氨酸含量升高可能是其耐牧的一种适应性机制。

Abstract: Leymus chinensis is tolerant to high salinity, drought and cold stresses. It is an important perennial plant with economic and ecological value. Proline is one of the main regulators in many plants under osmotic stresses. Δ1-pyrroline-5-carboxylate synthetase (P5CS) is a key regulatory enzyme that plays a crucial role in proline biosynthesis by the glutamate pathway. Based on analysis of solexa sequencing data, a P5CS gene from L. chinensis was cloned by RT-PCR using homologous primers. The cDNA sequence was 2 197 bp in length and putatively translated to 730 amino acids. Based on the sequence alignment results, the protein was named LcP5CS1. The homology between LcP5CS1 and BdP5CS1 was 95％, but LcP5CS1 had low homology with P5CS from dicotyledons. Analysis expression profiles of LcP5CS1 revealed that LcP5CS1 was highly expressed in leaf sheaths, stems and leaves. In cold conditions, ABA, salt and drought stress, it was significantly induced with time, and the expression levels of LcP5CS1 were up-regulated. The results of Solexa sequencing for wounded L. chinensis showed that LcP5CS1 was up-regulated, and the increased proline in L. chinensis may contribute to its stress tolerance.

中图分类号:

张乐新,苏蔓,马甜,马兴勇,闫学青,彭献军,陈双燕,程丽琴,刘公社. 羊草Δ1-吡咯琳-5-羧酸合成酶(LcP5CS1)基因的克隆与分析[J]. 草业学报, 2013, 22(4): 197-204.

ZHANG Le-xin, SU Man, MA Tian, MA Xing-yong, YAN Xue-qing, PENG Xian-jun, CHEN Shuang-yan, CHENG Li-qin, LIU Gong-she. Cloning and analysis of the Δ1-pyrroline-5-carboxylate synthetase (LcP5CS1)from Leymus chinensis[J]. Acta Prataculturae Sinica, 2013, 22(4): 197-204.

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羊草Δ1-吡咯琳-5-羧酸合成酶(LcP5CS1)基因的克隆与分析

Cloning and analysis of the Δ1-pyrroline-5-carboxylate synthetase (LcP5CS1)from Leymus chinensis

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