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玉米自交系干旱应答基因的鉴定

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为开发耐旱分子选择标记提供有用信息,用mRNA差异显示技术分离耐旱玉米自交系'81565'在干旱胁迫与灌溉对照之间差异表达的基因,发现MD1、MD2和MD3三个在干旱胁迫下差异表达的片段.MD1和MD2为下调表达,MD3为上调表达.序列分析和同源性比对表明,MD1与编码成熟酶的玉米叶绿体基因matK有97%的相似性,MD2与极端耐旱植物Sporobolus stapfianus编码丝氨酸/苏氨酸蛋白磷酸酶的PP2C基因有99%的相似性,MD3与属精氨酸/赖氨酸特异性半胱氨酸蛋白酶类的水稻metacaspase酶基因有99%的相似性.根据MD2片段序列,结合电子克隆和RT-PCR方法,克隆出一条1731 bp的全长cDNA序列,它编码388个氨基酸.此氨基酸序列包含丝氨酸/苏氨酸蛋白磷酸酶2C的催化中心结构域,被认定为玉米PP2C基因族的新成员,命名为ZmPP2Ca.实时荧光定量PCR结果表明,在干旱胁迫下,ZmPP2Ca基因在3个耐早自交系中呈下调表达,在2个不耐旱自交系中呈上调表达.
Identification of Drought-responsive Genes from Maize Inbred Lines
To provide a useful piece of information for the choice of molecular markers to be used in selection of drought tolerance,mRNA differential display was used to isolate genes from a drought-tolerant maize inbred line '81565'.After drought stress,two down-regulated expression gene fragments(MD 1 and MD2)and one upregulated expression fragment(MD3)were obtained.Results of sequence and homology analysis show that MD 1 has 97% similarity with mark in maize chloroplast genome,a gene encoding RNA maturase involved in group Ⅱ intron splicing of RNA transcript;MD2 has 99% similarity with the gene serine/threonine phosphorylase 2C in Sporobolus stapfianus;and MD3 has 99% similarity with rice the gene encoding metacaspase,an arginine/lysine-specific cysteine protease.Based on the sequence of fragment MD2,a new member of maize PP2C gene family,ZmPP2Ca,was cloned by in silicon cloning and reverse transcription polymerase chain reaction(RT-PCR).Real-time fluorescence quantitative polymerase chain reaction(FQ-PCR) showed that expression of the gene was down-regulated in the three drought

mRNA differential displaymaizedrought stressserine/threonine protein phosphatase 2CZmPP2Careal-time fluorescence quantitative polymerase chain reaction(FQ-PCR)

李富华、付风玲、沙莉娜、李晚忱

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四川农业大学玉米研究所/西南作物基因资源与遗传改良教育部重点实验室,四川雅安,625014

mRNA差异表达 玉米 干旱胁迫 丝氨酸/苏氨酸蛋白磷酸酶2C ZmPP2Ca 实时荧光定量PCR

国家自然科学基金美国洛克菲勒兄弟基金会资助项目教育部长江学者和创新团队发展计划

305711722004FS047IRT0453

2007

植物生理与分子生物学学报
中国植物生理学会

植物生理与分子生物学学报

CSTPCD
ISSN：1671-3877
年,卷(期)：2007.33(6)
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