mRNA差异显示技术分离水稻铵态氮营养诱导表达基因

Identification of Differentially Expressed Genes Induced by Ammonium Nitrogen in Rice Using mRNA Differential Display

应用mRNA差异显示技术对不同氮素营养下水稻叶片基因表达差异进行了分析,分离了一些差异表达片段。Reverse Northern和Northern杂交确认了部分阳性克隆,结果显示片段A-02(Oryza sativadrought stress related gene)和A-03(Zea mayspartial mRNA for TFⅡB-related protein)受到铵态氮营养诱导表达。生理学测定结果表明,与硝态氮营养相比,水稻在铵态氮营养下具有更高的过氧化氢酶、过氧化物酶活性和还原型谷胱甘肽含量,因此,更利于自身自由基的清除,从而提高抗逆性。

RNAs isolated from ammonium- and nitrate-treated rice leaves were used to screen for differentially expressed genes through mRNA differential display. A total of 72 bands appeared significant difference and some of them were further confirmed by reverse Northern and Northern blot. Two genes, A-02 （Oryza sativa drought stress related gene） and A-03 （Zea rnays partial mRNA for TF Ⅱ B-related protein） were highly up-regulated in the ammonium-treated rice leaves. The enzyme assays showed that the activities of the two anti-oxidative enzymes, catalase and peroxidase, and the content of glutathi-one, were significantly higher in the ammonium-treated rice leaves than those in the nitrate-treated rice leaves, indicating that the ammonium nutrition might improve stress resistance in rice during growth and development.