玉米ZmCBL2-2基因的克隆及其表达分析

doi:10.3969/j.issn.1000-7091.2012.02.006

摘要/Abstract

摘要： 为通过转基因植物等方法分析玉米ZmCBL2-2基因在耐逆性中的功能,运用RT-PCR(Reverse transcriptionPCR)方法克隆了该基因的编码区cDNA片段,并将其构建到植物表达载体p3301(pCAMBIA3301)上。同时,运用荧光实时定量PCR(Real-time quantitative PCR,RT-qPCR)技术对玉米幼苗期该基因在高盐、低温和养分缺乏等6种非生物胁迫下的表达动态进行了分析,发现其对于缺N、低K、低温、外源ABA(Abscisic acid)和水分胁迫均有显著的应答反应,而对于盐胁迫无明显的响应。

关键词: 玉米, CBL, 基因克隆, 表达分析

Abstract: To analyze the role of ZmCBL2-2 gene in the tolerance of maize to abiotic stresses by methods such?as transgenic plants，the complete coding region cDNA fragment of the gene was cloned by reverse transcription polymerase chain reaction (RT-PCR) and constructed into the plant expression vector p3301 (pCAMBIA3301)． Moreover，the expression profiles of ZmCBL2-2 under six abiotic stresses such as high salinity，low temperature and nutritional deficiency were analyzed by real time quantitative PCR (RT-qPCR) method，and it was found that the gene?responded significantly to nitrogen deprivation，low potassium，low temperature，exogenous abscisic acid (ABA) and?water stress，but did not respond obviously to salt stress．

Key words: Maize, CBL, Gene cloning, Expression analysis

中图分类号:

S513
Q78

李记园, 王存芳, 赵欢欢, 王茅雁. 玉米ZmCBL2-2基因的克隆及其表达分析[J]. 华北农学报, 2012, 27(2): 30-34. doi: 10.3969/j.issn.1000-7091.2012.02.006.

LI Ji-yuan, WANG Cun-fang, ZHAO Huan-huan, WANG Mao-yan. Cloning and Expression Analysis of ZmCBL2-2 Gene from Maize (Zea mays． L)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(2): 30-34. doi: 10.3969/j.issn.1000-7091.2012.02.006.

http://www.hbnxb.net/CN/Y2012/V27/I2/30

参考文献

[1] Reddy V S，Reddy A S N． Proteomics of calcium-signaling?components in plants[J]． Phytochemistry，2004，65:1750?－1776．
[2] Batistic O，Kudla J． Integration and channeling of calcium signaling through the CBL calcium sensor/CIPK protein?kinase network[J]． Planta，2004，219:915－924．
[3] Kolukisaoglu U，Wein S，Blazevic D，et al． Calcium sensors and their interacting protein kinases:genomics of the?Arabidopsis and rice CBL-CIPK signaling networks[J]．?Plant Physiol，2004，134:43－58．
[4] Luan S，Kudla J，Rodriguez-Concepcion M，et al． Calmodulins and calcineurin B-like proteins:calcium sensors for?specific signal response coupling in plants[J]． Plant?Cell，2002，14(Suppl):389－400．
[5] ?Liu J，Zhu J K． A calcium sensor homolog required for?plant salt tolerance[J]． Science，1998，280:1943－1945．
[6] Guo Y，Halfter U，Ishitani M，et al． Molecular characterization of functional domains in the protein kinase SOS2?that is required for plant salt tolerance[J]． Plant Cell, 2001，13:1383－1399．
[7] Guo Y，Qiu Q S，Quintero F J，et al． Transgenic evaluation?of activated mutant alleles of SOS2 reveals a critical requirement for its kinase activity and C-terminal regulatory?domain for salt tolerance in Arabidopsis thaliana [J]．?Plant Cell，2004，16:435－449．
[8] Zhu J K． Regulation of ion homeostasis under salt stress
[J]． Curr Opin Plant Bio，2003，16:441－445．
[9] Mart nez-Atienza J，Jiang X，Garciadeblas B，et al． Conservation of the Salt Overly Sensitive pathway in rice[J]．?Plant Physiol，2007，143:1001－1012．
[10] Cheong Y H，Kim K N，Pandey G K，et al． CBL1，a calcium sensor that differentially regulates salt，drought，and?cold responses in Arabidopsis[J]． Plant Cell，2003，15:?1833－1845．
[11] Cheong Y H，Pandey G K，Grant J J，et al． Two calcineurin B-like calcium sensors，interacting with protein?kinase CIPK23，regulate leaf transpiration and root potassium uptake in Arabidopsis[J]． Plant Journal，2007，52:?223－239．
[12] Xu J，Li H D，Chen L Q，et al． A protein kinase，interacting with two calcineurin B-like proteins，regulates K+?transporter AKT1 in Arabidopsis[J]． Cell，2006，125:?1347－1360．
[13] 李利斌，刘开昌，王殿峰，等．玉米 CBL 基因的生物信?息学分析[J]．玉米科学，2010，18 (1):6－11．
[14] ?Wang Maoyan，Gu Dan，Liu Tingsong，et al． Overexpression of a putative maize calcineurin B-like protein in?Arabidopsis confers salt tolerance[J]． Plant Mol Bio, 2007，65:733－746．
[15] Zhao Jinfeng，Sun Zhenfei，Zheng Jun，et al． Cloning and?characterization of a novel CBL-interacting protein kinase from maize[J]． Plant Mol Biol，2009，69:661－?674．
[16] 高大中，林清芳，王存芳，等．玉米 ZmCBL6 基因的克?隆及其植物表达载体构建[J]．内蒙古农业大学学?报，2009，30 (4):153－162．
[17] 王存芳，李记园，王茅雁．玉米 ZmCBL5-2 基因的克隆?及其转化拟南芥[J]．内蒙古农业大学学报，2011，32?(1):1－5．
[18] Livak K J，Schmittgen T D． Analyzing real-time PCR data by the 2－ ΔΔCTmethod[J]． Nature Protocols，2001，3:?1101－1108．
[19] Girdhar K，Pandey． Emergence of a novel calcium signaling pathway in plants:CBL-CIPK signaling network[J]．?Physiol Mol Biol Plants，2008，14:1－2．
[20] Kudla J，Xu Q，Harter K，et al． Genes for calcineurin Blike proteins in Arabidopsis are differentially regulated by?stress signals[J]． Proc Natl Acad Sci USA，1999，96:?4718－4723．
[21] ?Zhang Hechen，Yin Weilun，Xia Xinli． Calcineurin BLike family in Populus: comparative genome analysis?and expression pattern under cold，drought and salt?stress treatment[J]． Plant Growth Regul，2008，56:129?－140．
[22] Pandey G K，Cheong Y H，Kim K N，et al． The calcium?sensor calcineurin B-like 9 modulates Abscisic Acid sensitivity and biosynthesis in Arabidopsis [J]． Plant Cell, 2004，16:1912－1924．
[23] Hwang Y S，Bethke P C，Cheong Y H，et al． A gibberellin-regulated calcineurin B in rice localizes to the tonoplast and is implicated in vacuole function[J]． Plant?Physiol，2005，138:1347－1358．

选择文件类型/文献管理软件名称

选择包含的内容