Dorbe M F, Caboche M, Daniel-Vedele F
Laboratoire de Biologie Cellulaire, INRA, Versailles, France.
Plant Mol Biol. 1992 Jan;18(2):363-75. doi: 10.1007/BF00034963.
A nitrate reductase (NR) deficient mutant of Nicotiana plumbaginifolia totally impaired in the production of functional nia transcript and protein was restored for NR activity by transformation with a cloned tomato nia gene. The transgenic plants expressed from undetectable to 17% of the control NR activity in their leaves. Restoration of growth rates comparable to the wild type was obtained for transgenic plants expressing as little as 10% of the wild-type activity showing that nitrate reduction is not a growth-limiting factor in the wild-type plant. The analysis of the transgene expression showed that the tomato nia gene transcription was regulated by light, nitrate and a circadian rhythm as in tomato plants. These results suggest that all the cis-acting sequences involved in these regulations are contained in the 3 kb upstream region of the tomato nia gene and are still functional in transgenic N. plumbaginifolia plants. The amount of NR transcript synthesized from the tomato nia gene was reduced when a functional N. plumbaginifolia nia locus was introduced by sexual crosses. These data support the hypothesis that nitrate reduction is regulated by nitrate-derived metabolites as demonstrated in fungi.
一个在功能性nia转录本和蛋白质产生方面完全受损的烟草硝酸还原酶(NR)缺陷型突变体,通过用克隆的番茄nia基因进行转化而恢复了NR活性。转基因植物叶片中的NR活性从不可检测到对照活性的17%不等。对于表达量低至野生型活性10%的转基因植物,其生长速率恢复到了与野生型相当的水平,这表明硝酸盐还原不是野生型植物生长的限制因素。对转基因表达的分析表明,番茄nia基因的转录受光、硝酸盐和昼夜节律的调控,就像在番茄植株中一样。这些结果表明,参与这些调控的所有顺式作用序列都包含在番茄nia基因上游3 kb的区域内,并且在转基因烟草植株中仍然具有功能。当通过有性杂交引入功能性烟草nia基因座时,从番茄nia基因合成的NR转录本数量减少。这些数据支持了这样一种假说,即硝酸盐还原受硝酸盐衍生的代谢产物调控,这在真菌中已得到证实。