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小麦Em启动子驱动报告基因在转基因大麦和水稻的胚及糊粉层组织中表达。

The wheat Em promoter drives reporter gene expression in embryo and aleurone tissue of transgenic barley and rice.

作者信息

Furtado Agnelo, Henry Robert James

机构信息

Molecular Plant Breeding CRC, Centre for Plant Conservation Genetics, Southern Cross University, PO Box 157, Lismore, NSW 2480, Australia.

出版信息

Plant Biotechnol J. 2005 Jul;3(4):421-34. doi: 10.1111/j.1467-7652.2005.00135.x.

Abstract

The early methionine (Em) proteins are members of the late embryogenesis abundant (LEA) group of proteins that have been considered to be embryo specific. The ability of a 646-bp wheat Em promoter to control green fluorescent protein (gfp) expression was investigated in transgenic barley and rice. Seeds of transgenic plants expressed gfp in the developing embryo but also in the aleurone layer. The 646-bp Em promoter also directed strong gfp expression in cells comprising the junction between the endosperm transfer cells and cells of the aleurone layer. Em-gfp expression in transgenic barley showed differences in spatial and temporal control when compared with that observed in transgenic rice. Em-gfp expression was also detected in mature aleurone cells of transgenic barley and rice with and without abscisic acid (ABA) treatment. Reverse transcriptase-polymerase chain reaction (RT-PCR) results indicated the presence of Em and Em-homologous transcript in embryo, aleurone and endosperm tissues of wheat and of barley and rice, respectively. These results suggest that Em proteins may be expressed in both the embryo and aleurone during seed development, possibly providing protection against desiccation in these two tissues that survive seed drying. They may also have a similar role in these tissues during germination. The Em promoter from wheat may be useful in the expression of novel genes in cereal grains, as an embryo- and aleurone-specific promoter complementing other available endosperm- and pericarp-specific promoters to collectively increase the expression of transgenes in seeds.

摘要

早期甲硫氨酸(Em)蛋白是胚胎后期丰富(LEA)蛋白家族的成员,曾被认为具有胚胎特异性。研究了一个646bp的小麦Em启动子在转基因大麦和水稻中控制绿色荧光蛋白(gfp)表达的能力。转基因植物种子在发育中的胚中以及糊粉层中都表达gfp。646bp的Em启动子还在胚乳转移细胞和糊粉层细胞之间的连接处的细胞中引导强烈的gfp表达。与转基因水稻中观察到的情况相比,转基因大麦中的Em-gfp表达在空间和时间控制上存在差异。在有或没有脱落酸(ABA)处理的转基因大麦和水稻的成熟糊粉细胞中也检测到了Em-gfp表达。逆转录聚合酶链反应(RT-PCR)结果表明,在小麦的胚、糊粉层和胚乳组织以及大麦和水稻中分别存在Em和Em同源转录本。这些结果表明,Em蛋白可能在种子发育过程中在胚和糊粉层中都有表达,可能为这两个在种子干燥后存活的组织提供抗干燥保护。它们在萌发过程中在这些组织中可能也有类似作用。小麦的Em启动子可能有助于在谷物中表达新基因,作为一种胚和糊粉层特异性启动子,补充其他可用的胚乳和果皮特异性启动子,共同增加种子中转基因的表达。

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