Majumdar Rajtilak, Barchi Boubker, Turlapati Swathi A, Gagne Maegan, Minocha Rakesh, Long Stephanie, Minocha Subhash C
Department of Biological Sciences, University of New Hampshire Durham, NH, USA.
Department of Biological Sciences, University of New HampshireDurham, NH, USA; United States Department of Agriculture Forest Service, Northern Research StationDurham, NH, USA.
Front Plant Sci. 2016 Feb 16;7:78. doi: 10.3389/fpls.2016.00078. eCollection 2016.
The metabolism of glutamate into ornithine, arginine, proline, and polyamines is a major network of nitrogen-metabolizing pathways in plants, which also produces intermediates like nitric oxide, and γ-aminobutyric acid (GABA) that play critical roles in plant development and stress. While the accumulations of intermediates and the products of this network depend primarily on nitrogen assimilation, the overall regulation of the interacting sub-pathways is not well understood. We tested the hypothesis that diversion of ornithine into polyamine biosynthesis (by transgenic approach) not only plays a role in regulating its own biosynthesis from glutamate but also affects arginine and proline biosynthesis. Using two high putrescine producing lines of Arabidopsis thaliana (containing a transgenic mouse ornithine decarboxylase gene), we studied the: (1) effects of exogenous supply of carbon and nitrogen on polyamines and pools of soluble amino acids; and, (2) expression of genes encoding key enzymes in the interactive pathways of arginine, proline and GABA biosynthesis as well as the catabolism of polyamines. Our findings suggest that: (1) the overall conversion of glutamate to arginine and polyamines is enhanced by increased utilization of ornithine for polyamine biosynthesis by the transgene product; (2) proline and arginine biosynthesis are regulated independently of polyamines and GABA biosynthesis; (3) the expression of most genes (28 that were studied) that encode enzymes of the interacting sub-pathways of arginine and GABA biosynthesis does not change even though overall biosynthesis of Orn from glutamate is increased several fold; and (4) increased polyamine biosynthesis results in increased assimilation of both nitrogen and carbon by the cells.
谷氨酸代谢生成鸟氨酸、精氨酸、脯氨酸和多胺是植物氮代谢途径的一个主要网络,该网络还会产生一氧化氮和γ-氨基丁酸(GABA)等在植物发育和胁迫中起关键作用的中间产物。虽然该网络中间产物和产物的积累主要依赖于氮同化,但相互作用的子途径的整体调控尚不清楚。我们检验了以下假设:通过转基因方法将鸟氨酸转向多胺生物合成不仅在调节其自身从谷氨酸的生物合成中发挥作用,还会影响精氨酸和脯氨酸的生物合成。利用两个高腐胺产生的拟南芥品系(含有转基因小鼠鸟氨酸脱羧酶基因),我们研究了:(1)碳和氮的外源供应对多胺和可溶性氨基酸库的影响;以及(2)精氨酸、脯氨酸和GABA生物合成的相互作用途径中关键酶编码基因的表达以及多胺的分解代谢。我们的研究结果表明:(1)转基因产物将鸟氨酸用于多胺生物合成的利用率提高,从而增强了谷氨酸向精氨酸和多胺的整体转化;(2)脯氨酸和精氨酸的生物合成独立于多胺和GABA的生物合成进行调节;(3)尽管从谷氨酸整体合成鸟氨酸增加了几倍,但大多数研究的(28个)编码精氨酸和GABA生物合成相互作用子途径中酶的基因表达并未改变;(4)多胺生物合成增加导致细胞对氮和碳的同化增加。