Expression and cellular localization of rab28 mRNA and Rab28 protein during maize embryogenesis
Expression and cellular localization of rab28 mRNA and Rab28 protein during maize embryogenesis
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DOI：
10.1046/j.1365-313x.1996.09040549.x
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发表时间：
1996-04-01
期刊：
影响因子：
7.2
通讯作者：
Pages, M
中科院分区：
文献类型：
--
作者：
Niogret, MF;CulianezMacia, FA;Pages, M
玉米脱落酸（阿坝）应答基因rab28在胚和营养组织中是ABA诱导的。制备了Rab28蛋白的多克隆抗血清。利用免疫印迹和免疫沉淀，抗血清特异性地识别约30 kDa和pl 6的蛋白质，这与推导的氨基酸序列预测的分子量和pl密切一致。rab28基因产物在胚胎发育后期积累。在营养组织中，脱水胁迫诱导rab28基因表达在光和在黑暗中。利用地高辛标记的Rab28探针进行原位杂交，并利用抗Rab28抗体对Rab28蛋白进行免疫化学定位，研究胚胎发生过程中Rab28 mRNA表达的时空格局。rab28 mRNA的表达仅限于provascular组织在年轻的胚胎，并在发展的后期阶段，最普遍的积累发生在分生组织和在幼芽，根和盾片的维管元件。使用免疫电镜，Rab28蛋白已定位于不同细胞类型的核仁中。根据这些结果，rab28的应激调节和这种蛋白在胚胎发生后期可能发挥的作用进行了讨论。
The maize abscisic acid (ABA) responsive gene rab28, has been shown to be ABA-inducible in embryos and vegetative tissues. A polyclonal antiserum was raised against Rab28 protein. Using immunoblotting and immunoprecipitation, the antiserum specifically recognized a protein of about 30 kDa and pl 6 which is in close agreement with the molecular weight and pl predicted by the deduced amino acid sequence. The rab28 gene product accumulated during late embryogenesis. In vegetative tissues, dehydration stress induced rab28 gene expression both in the light and in the dark. The spatial and temporal pattern of rab28 mRNA expression during embryogenesis was investigated by in situ hybridization using digoxigenin-labelled rab28 probes, and the immunochemical localization of Rab28 protein using anti-Rab28 antibodies. Expression of rab28 mRNA is restricted to provascular tissues in young embryos, and at later stages of development the most prevalent accumulation occurred in meristem and in the vascular elements of the plumule, root and scutellum. Using immunoelectron microscopy the Rab28 protein has been located in the nucleolus of different cell types. In light of these results the stress regulation of rab28 and a likely role for this protein during late embryogenesis are discussed.