Graham I A, Smith L M, Leaver C J, Smith S M
Department of Botany, University of Edinburgh, U.K.
Plant Mol Biol. 1990 Oct;15(4):539-49. doi: 10.1007/BF00017829.
The cucumber malate synthase (MS) gene, including 1856 bp of 5' non-transcribed sequence, has been transferred into Petunia (Mitchell) and Nicotiana plumbaginifolia plants using an Agrobacterium binary vector. The transferred gene is found in variable copy number in different transformants, and is stably transmitted in each case as a single Mendelian character. Transgene mRNA accumulates in the seedling during the first three days of germination, then declines in amount as the cotyledons emerge from the seed. The decline is more pronounced in light-grown seedlings than in dark-grown seedlings. Expression of the MS transgene is also detected at a low level in petals of transformed Petunia plants. In these respects the pattern of MS gene expression is similar in cucumber and in transformed plants, showing that the transferred DNA fragment contains a functional MS gene. A 1076 bp fragment of 5' sequence was linked to the beta-glucuronidase reporter gene and transferred into Nicotiana, where it was shown to direct temporal and spatial patterns of expression similar to that of the complete MS gene. However, histochemical localisation of beta-glucuronidase activity demonstrated that the chimaeric gene is expressed not only in cotyledons of transgenic plants, but also in endosperm and some hypocotyl cells during early germination. The relevance of these findings to the control of malate synthase gene expression is discussed.
黄瓜苹果酸合酶(MS)基因,包括1856 bp的5'非转录序列,已通过农杆菌双元载体转入矮牵牛(Mitchell)和垂花烟草植株中。在不同的转化体中,转入的基因以可变拷贝数存在,并且在每种情况下都作为单个孟德尔性状稳定遗传。转基因mRNA在萌发的前三天在幼苗中积累,然后随着子叶从种子中长出,其数量下降。在光照下生长的幼苗中这种下降比在黑暗中生长的幼苗中更明显。在转化的矮牵牛植株的花瓣中也检测到MS转基因的低水平表达。在这些方面,黄瓜和转化植株中MS基因的表达模式相似,表明转入的DNA片段包含一个功能性的MS基因。一个1076 bp的5'序列片段与β-葡萄糖醛酸酶报告基因相连并转入烟草,结果表明它指导的表达时空模式与完整的MS基因相似。然而,β-葡萄糖醛酸酶活性的组织化学定位表明,嵌合基因不仅在转基因植物的子叶中表达,而且在早期萌发期间在内胚乳和一些下胚轴细胞中也表达。讨论了这些发现与苹果酸合酶基因表达调控的相关性。