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转基因烟草植株中硝酸盐还原酶宿主基因和转基因的共抑制

Co-suppression of nitrate reductase host genes and transgenes in transgenic tobacco plants.

作者信息

Dorlhac de Borne F, Vincentz M, Chupeau Y, Vaucheret H

机构信息

Laboratoire de Biologie Cellulaire, INRA, Versailles, France.

出版信息

Mol Gen Genet. 1994 Jun 15;243(6):613-21. doi: 10.1007/BF00279570.

Abstract

Constructs carrying the entire or part of the tobacco nitrate reductase cDNA (NIA) cloned between the promoter and terminator sequences of the 35S RNA of the cauliflower mosaic virus were introduced into tobacco, in an attempt to improve nitrate assimilation. Several transgenic plants that had elevated NIA mRNA and nitrate reductase (NR) activity were obtained. In addition, a few plants that exhibited a chlorotic phenotype characteristic of NR-deficient mutants were also obtained. One of these plants contained no NIA mRNA, no NR activity and accumulated nitrate. This phenotype was therefore assumed to result from co-suppression of 35S-NIA transgenes and host NIA genes. NR-deficient plants were also found among the progeny of transformants overexpressing NIA mRNA. Genetic analyses indicated that these NR-deficient plants were homozygous for the 35S-NIA transgene, although not all homozygous plants were deficient for NR. The ratio of normal to NR-deficient plants in the progeny of homozygous plants remained constant at each generation, irrespective of the state of expression of the NIA genes (active or inactive) in the previous generation. This ratio also remained unchanged when field trials were performed in two areas of France: Versailles and Bergerac. The analysis of homozygous plants revealed that co-suppression was reversible at some stage of sexual reproduction. Indeed, host genes and transgenes reactivated at each generation, and co-suppression always appeared after a lag period of normal growth, suggesting that the phenomenon is developmentally regulated.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为了提高硝酸盐同化能力,将携带克隆于花椰菜花叶病毒35S RNA启动子和终止子序列之间的全部或部分烟草硝酸还原酶cDNA(NIA)的构建体导入烟草。获得了几种NIA mRNA和硝酸还原酶(NR)活性升高的转基因植物。此外,还获得了一些表现出NR缺陷型突变体典型黄化表型的植物。其中一株植物不含NIA mRNA,没有NR活性且积累了硝酸盐。因此,这种表型被认为是由35S-NIA转基因和宿主NIA基因的共抑制导致的。在过表达NIA mRNA的转化体后代中也发现了NR缺陷型植物。遗传分析表明,这些NR缺陷型植物对于35S-NIA转基因是纯合的,尽管并非所有纯合植物都缺乏NR。纯合植物后代中正常植物与NR缺陷型植物的比例在每一代都保持恒定,与上一代NIA基因的表达状态(活跃或不活跃)无关。当在法国的凡尔赛和贝热拉克两个地区进行田间试验时,这个比例也保持不变。对纯合植物的分析表明,共抑制在有性生殖的某个阶段是可逆的。实际上,宿主基因和转基因在每一代都会重新激活,并且共抑制总是在正常生长的滞后期后出现,这表明该现象受发育调控。(摘要截短至250字)

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