水分胁迫下斑茅3个逆境代谢相关基因表达的实时PCR分析

摘要/Abstract

摘要： 【研究目的】初步探讨斑茅3个关键逆境适应相关基因在水分胁迫下的表达机制。【方法】已克隆S-腺苷蛋氨酸脱羧酶、鸟氨酸氨基转移酶、甜菜碱脱氢酶3个基因和内参基因的基础上，应用定量PCR技术，对这3个基因在不同胁迫时间下的表达水平进行分析。【结果】斑茅的S-腺苷蛋氨酸脱羧酶、鸟氨酸氨基转移酶基因、甜菜碱脱氢酶基因表达在初期均受抑制，后期表达持续上调，但甜菜碱脱氢酶基因受水分诱导表达较前两个基因明显。【结论】这可能暗示着在斑茅中，脯胺酸和甜菜碱的合成受水分胁迫诱导，而甜菜碱的合成相对脯胺酸和多胺受水分胁迫诱导影响更大。这些研究为斑茅抗旱的分子机制提供初步的理论基础。

关键词: 丘陵山地, 丘陵山地, 果园, 节水技术

Abstract: 【OBJECTIVE】 In order to study the molecular mechanism of Saccharum arundinaceum Retz under water-stress.【METHOD】 In this paper, three gene S-adenosylmethionine decarboxylase (SAMDC), ornithine aminotransferase (OAT) and betaine aldehyde dehydrogenase (BADH) plus one house-keeping gene had been cloned and their expression levels under different stresses were evaluated by real-time PCR technique. 【RESULTS】 The results showed that the expression of SAMDC was inhibited at the beginning and unregulated at later stage by water stress, as the same to that of OAT was inhibited at the beginning and unregulated at later stage, and the expression of BADH was prohibited at the earlier stage along with the subsequent continuous more obvious unregulation. 【CONCLUSION】 From above, it can be speculated spm, proline and betaine could be induced by water stress, of which the influence on betaine was the biggest. These studies aim to provide the theory base of molecular mechanism for drought resistance breeding in Saccharum arundinaceum Retz.

中图分类号:

Q786

张积森,阙友雄,李伟,阮妙鸿,张木清,陈如凯. 水分胁迫下斑茅3个逆境代谢相关基因表达的实时PCR分析[J]. 中国农学通报, 2007, 23(5): 49-49.

Zhang Jisen, Que Youxiong, Li Wei, Ruan Miaohong, Zhang Muqing, Chen Rukai. Analysis of the Expression of Three Adversity Genes under Water-stress in Saccharum arundinaceum through Realtime PCR[J]. Chinese Agricultural Science Bulletin, 2007, 23(5): 49-49.

参考文献

[1] 戴艺民,卢川北,林江波,等.福建斑茅核型分析初报.福建农业学报,2002,17(3):148-150.
[2] George P, Mandy J, Christopher, et al. Molecular contribution to selection of intergeneric hybrids between sugarcane and the wild species Erianthus arundinaceus. Genome, 2000,43(6):1033-1037.
[3] 郑雪芳,张木清,李奇伟,等.甘蔗斑茅的杂交利用及其杂种后代鉴定系列研究(二)甘蔗斑茅远缘真实杂种的分子鉴定[J].分子植物育种,2004,(1):35-42.
[4] 肖凤迥.斑茅种与甘蔗、蔗茅二属几个种的植物形态学和同工酶比较研究[J].甘蔗,1994,1(1):22-27.
[5] 张木清,洪艺珣,李奇伟,等.中国斑茅种质资源分子多态性分析.植物资源与环境学报,2004,13(1):1-6.
[6] 文建成,蔡青,范源洪,等.甘蔗属割手密(Saccharum Spontaneum),近缘属斑茅(Sclerostachya)及河八王(Narenga)的染色体数目研究.甘蔗糖业,2001,3:12-15.
[7] 刘文荣,邓祖湖,张木清,等.甘蔗斑茅的杂交利用及其杂种后代鉴定系列研究Ⅲ.甘蔗斑茅远缘杂交后代细胞遗传分析.作物学报,2004,30(11):1093-1096.
[8] 郭莺,余爱丽,张木清,等.甘蔗斑茅的杂交利用及其杂种后代鉴定系列研究(三)拔地拉与斑茅耐盐性差异分析.热带作物学报,2005,26(2):88-93.
[9] 张木清,潘婕翁,笑艳,等,甘蔗和斑茅的杂交利用及其杂种后代鉴定系列研究(四)甘蔗与斑茅抗旱生理生化差异性分析.热带作物学报,2004,25(2):50-54.
[10] Walden R, Cordeiro A, Tiburcio AF. Polyamines:small molecules triggering pathways in plant growth and development. Plant Physiol, 1997,113:1009-1013.
[11] Bhavna W, Manchikatla VR. Effect of increased polyamine bioxynthesis on stress responses in transgenic tobacco by introduction of human S-adenosylmethionine gene. Plant Science, 2003,164(5):727-734.
[12] Delauney A J, HU C AA, KISHOR P B K, et a1. Cloning of ornithine-aminotransferase cDNA from Vigna aconitifolia by trans-complementafion in Escherichia coli and regulation of prollne biosynthesis. J Biol Chem, 1993,268:18673-18678.
[13] Weretilnyk E A, Hanson A D. Betaine dldehyde dehygrogenase polymorphism in spinach: genetic and biochemical characterization.Biochem Genet, 1988,26(1-2):143-151.
[14] Weretilnyk E A, Hanson A D. Betaine dldehyde dehygrogenase from spinach leaves:purification, in vitro translation of the mRNA, and regulation by salinity. Arch Biochem Biophys, 1989,271(1):56-63.
[15] Livak K J, Flood S J, Marmaro J, et al. Oligonucleotides with fluorescent dyes at opposite ends provide aquenched probe system useful for detecting PCR product and nucleic acid hybridization[J]. PCR Methods Appl, 1995,4(6):357-362.
[16] Gibson U E, Heid C A, Williams P M. A novel method for real time quantitative RT-PCR[J]. Genome Res, 1996,6(10):995-1001.
[17] Heid C A, Stevens J, Livak K J, et al. Real time quantitative PCR[J]. Genome Res, 1996,6(10):986-994.
[18] Ishitani M, Nakamura T, Han S Y, et al. Expression of the betaine aldehyde dehydrogenase gene in barley in response to osmotic stress and abscisic acid. Plant Molecular biology, 1995,27:307-315.
[19] 许雯,孙梅好,朱亚芳,等.甘氨酸甜菜碱增强青菜抗盐性的作用.植物学报,2001,43(8):809-814.
[20] 张士功,高吉寅,宋景芝,等.甜菜碱对NaCl胁迫下小麦细胞保护酶活性的影响.植物学报,1999,16(4):429-432.