Pajuelo P, Pajuelo E, Forde B G, Márquez A J
Departamento de Bioquímica Vegetal y Biología Molecular, Facultad de Química, Universidad de Sevilla, Spain.
Planta. 1997 Dec;203(4):517-25. doi: 10.1007/s004250050221.
We have investigated the regulation of ferredoxin-glutamate synthase (Fd-GOGAT) in leaves of barley (Hordeum vulgare L. cv. Maris Mink) at the mRNA, protein and enzyme activity levels. Studies of the changes in Fd-GOGAT during plant development showed that the activity in shoots increases rapidly after germination to reach a maximum (on a fresh-weight basis) at day 10 and then declines markedly to less than 50% of the maximal activity by day 30, this decline being correlated with an equivalent loss of Fd-GOGAT protein. Growing the plants in darkness reduced the maximum activity attained in the shoots, but did not affect the overall pattern of the changes or their timing. The activity of Fd-GOGAT increased two- to three-fold within 48 h when etiolated leaves were exposed to light, and Northern blots indicated that the induction occurred at the mRNA level. However, whilst a carbon source could at least partially substitute for light in the induction of nitrate reductase activity, no induction of Fd-GOGAT activity was seen when etiolated leaves were treated with either sucrose or glucose. Interestingly, the levels of Fd-GOGAT mRNA and activity remained high up to a period of 16 h or 72 h darkness, respectively. Compared with plants grown in N-free medium, light-grown plants supplied with nitrate had almost two-fold higher Fd-GOGAT activities and increased Fd-GOGAT mRNA levels, but nitrate had no effect on the abundance of the enzyme or its mRNA in etiolated plants, indicating that light is required for nitrate induction of barley Fd-GOGAT.
我们在mRNA、蛋白质和酶活性水平上研究了大麦(Hordeum vulgare L. cv. Maris Mink)叶片中铁氧还蛋白-谷氨酸合酶(Fd-GOGAT)的调控。对植物发育过程中Fd-GOGAT变化的研究表明,发芽后芽中的活性迅速增加,在第10天达到最大值(以鲜重计),然后显著下降,到第30天时降至最大活性的50%以下,这种下降与Fd-GOGAT蛋白的等量损失相关。在黑暗中种植植物降低了芽中达到的最大活性,但不影响变化的总体模式或其时间。当黄化叶片暴露在光下时,Fd-GOGAT的活性在48小时内增加了两到三倍,Northern印迹表明诱导发生在mRNA水平。然而,虽然碳源在诱导硝酸还原酶活性方面至少可以部分替代光,但用蔗糖或葡萄糖处理黄化叶片时,未观察到Fd-GOGAT活性的诱导。有趣的是,Fd-GOGAT mRNA和活性水平分别在长达16小时或72小时的黑暗期内保持较高。与在无氮培养基中生长的植物相比,供应硝酸盐的光生长植物的Fd-GOGAT活性几乎高出两倍,且Fd-GOGAT mRNA水平增加,但硝酸盐对黄化植物中该酶或其mRNA的丰度没有影响,这表明光对于硝酸盐诱导大麦Fd-GOGAT是必需的。