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作 者:王飞飞[1] 李勇[1] 王学东[1] 朱延明[1] 才华[1] 纪巍[1] 柏锡[1]
机构地区:[1]东北农业大学植物生物工程研究室,黑龙江哈尔滨1500302
出 处:《作物学报》2011年第11期1984-1990,共7页Acta Agronomica Sinica
基 金:国家自然科学基金项目(30940005);黑龙江省教育厅科技项目(11521024);东北农业大学博士启动基金项目(190106)资助
摘 要:拟南芥AT2G14260基因编码脯氨酸亚氨基肽酶,基因芯片表达谱数据显示该基因响应高盐、低温等非生物胁迫。为研究其功能和表达特性,本研究采用野生型和突变体植株pip-1进行低温、干旱和盐胁迫处理,结果显示在正常培养基中野生型与pip-1植株表型没有区别。而在逆境处理下pip-1植株的根比野生型明显短,且在干旱胁迫下,突变体的叶子比野生型植株明显变黄萎蔫。在正常环境、冷、干旱、盐胁迫下,野生型植株脯氨酸含量的平均值分别是突变体的1.11、1.23、1.10和1.34倍,这说明AT2G14260基因对提高非生物胁迫的耐性具有一定的作用。同时分离了该基因的启动子,构建了表达GUS基因的载体并转化拟南芥,该启动子在正常环境中不表达。胁迫处理时从两叶期开始在根、叶、茎中表达,到达花期后,干旱处理下的花瓣、花枝和冷处理下的花柱头中也有表达。GUS相对活性试验也证明对胁迫的响应能力。可见AT2G14260基因启动子是逆境胁迫诱导表达启动子,同时具有组织表达特异性,因此,AT2G14260基因及其启动子在转基因工程育种中具有潜在的应用价值。The AT2G14260 gene encodes proline iminopeptidase in Arabidopsis thaliana. Genechip expression profile indicate AT2G14260 responses to abiotic stress, such as high salt or cold. To analyze the function and expression specificity, we compared the wild type with mutants pip-1 under cold, drought and salt stresses. Under normal conditions, mutant plants showed similar growth performance with wild type plants. However, after stress treatments, mutant plants exhibited much shorter roots compared with wild type plants. In addition, the leaves of mutant plants were notably more yellow than wild-type leaves under drought stress. The average relative proline contents in wild type plants were 1.11, 1.23, 1.10, and 1.34 times of those in mutants. All these results showed that AT2G14260 gene plays an important role in plant tolerance to abiotic stress. Meanwhile, we isolated the gene promoter and constructed plant vector in which the GUS gene was expressed by AT2G14260 promoter, and then transformated into Arabidopsis thaliana. The results of the GUS staining revealed that the GUS gene did not express under normal conditions. After treatments, GUS gene expressed in the roots, stems and leaves of the transgenic seedlings at the two-leaf stage and the blooming stage, especially in the petals and flowers under drought stress or in the stigma under cold stress. Results of GUS relative activity measurement suggested this gene had the response ability to stress. Taken together, the promoter of AT2G14260 is stress inducible and tissue specific. Thus, AT2G14260 gene and its promoter have application potentials in genetic engineering.
关 键 词:AT2G14260 逆境胁迫诱导表达启动子 组织特异性表达
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