杜梨PbCBL10基因表达与启动子功能分析Expression of the PbCBL10 gene and functional analysis of its promoter in pear plants(Pyrus betulaefolia)
许园园;蔺经;李晓刚;李慧;常有宏;
摘要(Abstract):
【目的】类钙调磷酸酶B亚基蛋白(Calcineurin B-like proteins,CBLs)参与调控植物生长发育及在响应逆境胁迫过程发挥重要作用。深入探明杜梨CBLs家族成员PbCBL10在逆境下的表达模式并在转录水平上揭示表达差异,为揭示梨抗逆机制提供理论依据。【方法】以杜梨(Pyrus betulaefolia Bunge)幼苗为试材,运用实时荧光定量RT-PCR技术分析了PbCBL10在Na Cl、Ca Cl2、甘露醇(Mannitol)、聚乙二醇(PEG6000)、脱落酸(ABA)、6-苄氨基腺嘌呤(6-BA)、萘乙酸(NAA)和温度诱导下的表达特性;通过基因组步移技术分离PbCBL10启动子序列,并利用生物信息学方法对其功能进行预测;构建启动子植物表达载体并通过农杆菌介导法转化烟草,研究多种胁迫及激素诱导下GUS蛋白瞬时表达情况。【结果】RT-qPCR分析表明:PbCBL10表达受Na Cl、Ca Cl2、甘露醇、PEG6000、ABA、6-BA、NAA和温度诱导;获得了长度为919bp的PbCBL10启动子序列,该序列含有典型调控元件与多个胁迫顺式元件;瞬时表达结果表明,多种胁迫与激素作用因子均可诱导PbCBL10启动子调控GUS基因的表达。【结论】PbCBL10参与多种激素和逆境胁迫过程的信号转导,在调控梨树生长发育及响应环境胁迫方面发挥重要作用。
关键词(KeyWords): 杜梨;胁迫响应;PbCBL10;RT-qPCR;启动子;GUS
基金项目(Foundation): 国家自然科学基金(31372051);; 江苏省博士后基金(1302006B)
作者(Author): 许园园;蔺经;李晓刚;李慧;常有宏;
Email:
DOI: 10.13925/j.cnki.gsxb.20140199
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