Wang R, Guegler K, LaBrie S T, Crawford N M
Division of Biology, University of California at San Diego, La Jolla, California 92093-0016, USA.
Plant Cell. 2000 Aug;12(8):1491-509. doi: 10.1105/tpc.12.8.1491.
Microarray and RNA gel blot analyses were performed to identify Arabidopsis genes that responded to nitrate at both low (250 microM) and high (5 to 10 mM) nitrate concentrations. Genes involved directly or indirectly with nitrite reduction were the most highly induced by nitrate. Most of the known nitrate-regulated genes (including those encoding nitrate reductase, the nitrate transporter NRT1, and glutamate synthase) appeared in the 40 most strongly nitrate-induced genes/clones on at least one of the microarrays of the 5524 genes/clones investigated. Novel nitrate-induced genes were also found, including those encoding (1) possible regulatory proteins, including an MYB transcription factor, a calcium antiporter, and putative protein kinases; (2) metabolic enzymes, including transaldolase and transketolase of the nonoxidative pentose pathway, malate dehydrogenase, asparagine synthetase, and histidine decarboxylase; and (3) proteins with unknown functions, including nonsymbiotic hemoglobin, a senescence-associated protein, and two methyltransferases. The primary pattern of induction observed for many of these genes was a transient increase in mRNA at low nitrate concentrations and a sustained increase when treated with high nitrate concentrations. Other patterns of induction observed included transient inductions after both low and high nitrate treatments and sustained or increasing amounts of mRNA after either treatment. Two genes, AMT1;1 encoding an ammonium transporter and ANR1 encoding a MADS-box factor, were repressed by nitrate. These findings indicate that nitrate induces not just one but many diverse responses at the mRNA level in Arabidopsis.
进行了微阵列和RNA凝胶印迹分析,以鉴定在低(250微摩尔)和高(5至10毫摩尔)硝酸盐浓度下对硝酸盐有反应的拟南芥基因。直接或间接参与亚硝酸盐还原的基因受硝酸盐诱导程度最高。大多数已知的硝酸盐调节基因(包括编码硝酸还原酶、硝酸盐转运蛋白NRT1和谷氨酸合酶的基因)出现在所研究的5524个基因/克隆的至少一个微阵列上硝酸盐诱导最强的40个基因/克隆中。还发现了新的硝酸盐诱导基因,包括编码(1)可能的调节蛋白的基因,如一个MYB转录因子、一个钙反向转运蛋白和假定的蛋白激酶;(2)代谢酶的基因,如非氧化戊糖途径的转醛醇酶和转酮醇酶、苹果酸脱氢酶、天冬酰胺合成酶和组氨酸脱羧酶;以及(3)功能未知的蛋白的基因,如非共生血红蛋白、一种衰老相关蛋白和两种甲基转移酶。观察到许多这些基因的主要诱导模式是在低硝酸盐浓度下mRNA短暂增加,而在高硝酸盐浓度处理时持续增加。观察到的其他诱导模式包括低硝酸盐和高硝酸盐处理后均出现短暂诱导,以及任一处理后mRNA持续增加或量增加。两个基因,编码铵转运蛋白的AMT1;1和编码MADS盒因子的ANR1,被硝酸盐抑制。这些发现表明,硝酸盐在拟南芥的mRNA水平上不仅诱导一种,而且诱导多种不同的反应。