State Key Laboratory for Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan 430072, China.
J Integr Plant Biol. 2017 Sep;59(9):642-656. doi: 10.1111/jipb.12536. Epub 2017 May 2.
The specific functions of the genes encoding arginine biosynthesis enzymes in plants are not well characterized. We report the isolation and characterization of Arabidopsis thaliana N-acetylglutamate kinase (NAGK), which catalyzes the second step of arginine biosynthesis. NAGK is a plastid-localized protein and is expressed during most developmental processes in Arabidopsis. Heterologous expression of the Arabidopsis NAGK gene in a NAGK-deficient Escherichia coli strain fully restores bacterial growth on arginine-deficient medium. nagk mutant pollen tubes grow more slowly than wild type pollen tubes and the phenotype is restored by either specifically through complementation by NAGK in pollen, or exogenous supplementation of arginine. nagk female gametophytes are defective in micropylar pollen tube guidance due to the fact that female gametophyte cell fate specification was specifically affected. Expression of NAGK in synergid cells rescues the defect of nagk female gametophytes. Loss-of-function of NAGK results in Arabidopsis embryos not developing beyond the four-celled embryo stage. The embryo-defective phenotype in nagk/NAGK plants cannot be rescued by watering nagk/NAGK plants with arginine or ornithine supplementation. In conclusion, our results reveal a novel role of NAGK and arginine in regulating gametophyte function and embryo development, and provide valuable insights into arginine transport during embryo development.
植物中编码精氨酸生物合成酶的基因的具体功能尚未得到很好的描述。我们报告了拟南芥 N-乙酰谷氨酸激酶 (NAGK) 的分离和表征,它催化精氨酸生物合成的第二步。NAGK 是一种定位于质体的蛋白质,在拟南芥的大多数发育过程中表达。在 NAGK 缺陷型大肠杆菌菌株中异源表达拟南芥 NAGK 基因可完全恢复细菌在缺乏精氨酸的培养基中的生长。nagk 突变花粉管比野生型花粉管生长得更慢,通过花粉中 NAGK 的特异性互补或外源补充精氨酸可恢复表型。由于雌性配子体细胞命运特定位点受到影响,nagk 雌性配子体在极孔花粉管导向方面存在缺陷。NAGK 在协同细胞中的表达可挽救 nagk 雌性配子体的缺陷。NAGK 的功能丧失导致拟南芥胚胎不能发育到四细胞胚胎阶段。在 nagk/NAGK 植物中,通过用精氨酸或鸟氨酸补充水分来挽救胚胎缺陷表型是无效的。总之,我们的结果揭示了 NAGK 和精氨酸在调节配子体功能和胚胎发育中的新作用,并为胚胎发育过程中精氨酸转运提供了有价值的见解。