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水孔蛋白协助玉米副卫细胞中H2O2的跨膜转运
王巧艳1, 李 思1, 张 莉,等2
1.西北农林科技大学 生命科学学院;2.中国科学院水利部 水土保持研究所
摘要:
【目的】探索玉米气孔副卫细胞中H2O2的来源,以阐明禾本科植物气孔运动过程中保卫细胞和副卫细胞协同调节的机理。【方法】采用细胞化学方法对H2O2进行亚细胞荧光定位,以TubulinGAPDH为内参基因,利用qPCR技术,对玉米水孔蛋白基因ZmPIP2;4、ZmPIP2;5和ZmPIP2;6表达量进行分析,从而确定水孔蛋白协助玉米副卫细胞中H2O2的跨膜转运。【结果】暗处理组,加水孔蛋白抑制剂AgNO3与不加AgNO3副卫细胞中的H2O2积累相反;外源H2O2引起副卫细胞中H2O2积累,先加入水孔蛋白抑制剂AgNO3再加外源H2O2理后副卫细胞中H2O2不积累;短细胞发生后,副卫细胞中的H2O2开始积累,同时ZmPIP2;5基因的相对表达量上调;随着水分胁迫程度增加,ZmPIP2;4ZmPIP2;5、ZmPIP2;6基因的相对表达量随短细胞发生上调。【结论】水孔蛋白具有转运H2O2的功能,玉米叶片下表皮副卫细胞中的H2O2是外源的,其积累和清除可能与水孔蛋白ZmPIP2;5的转运有关。
关键词:  副卫细胞  水孔蛋白  H2O2  跨膜转运
DOI:
分类号:
基金项目:国家高技术研究发展计划“863计划”项目(2011AA100504)
Aquaporin facilitates transmembrane diffusion of H2O2 in maize subsidiary cells
WANG Qiaoyan,LI Si,ZHANG Li,et al
Abstract:
【Objective】This study researched the sources of H2O2 in maize subsidiary cells to clarify the co regulation mechanism of guard cells and subsidiary cells in grassy stomatal movement.【Method】This study researched the cell localization of H2O2 by cytochemical fluorescent probe and analyzed the gene expression of ZmPIP2;4,ZmPIP2;5,ZmPIP2;6 by qPCR technology,with Tubulin and GAPDH as reference genes.Then the fact of aquaporin facilitates transmembrane diffusion of H2O2 in maize subsidiary cells was confirmed.【Result】Under light and dark treatments,the accumulation of H2O2 in subsidiary cells was opposite to the control group supplemented with aquaporin inhibitor AgNO3.Exogenous H2O2 caused H2O2 accumulation in subsidiary cells,but aquaporin inhibitor AgNO3 added before the addition of exogenous H2O2 did not cause H2O2 accumulation.After short cells appeared,H2O2 started to accumulate in subsidiary cells,at the same time the gene expression of ZmPIP2;5 was upregulated.After short cells appeared,the expressions of ZmPIP2;4,ZmPIP2;5 and ZmPIP2;6 were upregulated when increased soil water shortages.【Conclusion】Aquaporin can transport H2O2 in leaf epidermis of maize and H2O2 in subsidiary cells was exogenous,whose accumulation and elimination were possibly mediated by aquaporin ZmPIP2;5.
Key words:  subsidiary cells  aquaporin  H2O2  transmembrane diffusion

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