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采用半定量RT-PCR技术对甜菜胞液型谷氨酰胺合成酶(GS1)和质体型谷氨酰胺合成酶(GS2)基因进行mRNA的表达检测,同时进行谷氨酰胺合成酶活性的测定,研究不同氮素处理对谷氨酰胺合成酶(GS)基因表达的调控,并以三磷酸甘油醛脱氢酶(GAPDH)为内参照进行基因表达水平的半定量分析。结果显示,NO3-:NH4+>1和NO3-:NH4+=50:50较NO3-:NH4+<1更能促进GS活性;NO3-:NH4+=80:20和NO3-:NH4+=50:50较NO3-:NH4+<1更能促进GS1基因的表达;NO3-:NH4+>1和NO3-:NH4+=50:50较NO3-:NH4+<1更能促进GS2基因的表达。说明硝态氮比例较高的混合态氮较铵态氮比例较高的混合态氮及单一铵态氮更能促进GS活性和GS基因的表达,氮素能有效地在转录水平上调控甜菜幼苗叶片GS基因的表达。
Semi-quantitative RT-PCR was used to detect mRNA expression of cytosolic glutamine synthetase (GS1) and plastid glutamine synthetase (GS2) in beet, and the activity of glutamine synthetase The effects of different nitrogen treatments on the gene expression of glutamine synthetase (GS) were studied. Semi-quantitative analysis of gene expression levels was performed with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal reference. The results showed that NO3-: NH4 +> 1 and NO3-: NH4 + = 50:50 promoted GS activity more than NO3-: NH4 + <1; NO3-: NH4 + = 80:20 and NO3-: NH4 + -: NH4 + <1 can promote the expression of GS1 gene; NO3-: NH4 +> 1 and NO3-: NH4 + = 50:50 can promote the expression of GS2 gene more than NO3-: NH4 + <1. The results showed that the mixed nitrogen with higher proportion of nitrate nitrogen promoted the GS activity and the expression of GS gene better than the mixed type nitrogen and single ammonium nitrogen with a higher proportion of ammonium nitrogen. Nitrogen could effectively regulate the growth of sugar beet seedlings Leaf GS gene expression.