Faculty of Life Science and Biotechnology, Kunming University of Science and Technology, Chenggong, Kunming, 650500, China.
Planta. 2018 Feb;247(2):339-354. doi: 10.1007/s00425-017-2790-9. Epub 2017 Oct 7.
Over-expression of AtFDH controlled by the promoter of Rubisco small subunit in chloroplasts increases formaldehyde uptake and metabolism in tobacco leaves. Our previous study showed that formaldehyde (HCHO) uptake and resistance in tobacco are weaker than in Arabidopsis. Formate dehydrogenase in Arabidopsis (AtFDH) is a key enzyme in HCHO metabolism by oxidation of HCOOH to CO, which enters the Calvin cycle to be assimilated into glucose. HCHO metabolic mechanism in tobacco differs from that in Arabidopsis. In this study, AtFDH was over-expressed in the chloroplasts of transgenic tobacco using a light inducible promoter. C-NMR analysis showed that the carbon flux from HCHO metabolism was not introduced into the Calvin cycle to produce glucose in transgenic tobacco leaves. However, the over-expression of AtFDH significantly enhanced the HCHO metabolism in transgenic leaves. Consequently, the productions of [4-C]Asn, [3-C]Gln, [U-C]oxalate, and HCOOH were notably greater in transgenic leaves than in non-transformed leaves after treatment with HCHO. The increased stomatal conductance and aperture in transgenic leaves might be ascribed to the increased yield of oxalate in the guard cells with over-expressed AtFDH in chloroplasts. Accordingly, the transgenic plants exhibited a stronger capacity to absorb gaseous HCHO. Furthermore, the higher proline content in transgenic leaves compared with non-transformed leaves under HCHO stress might be attributable to the excess formate accumulation and Gln production. Consequently, the HCHO-induced oxidative stress was reduced in transgenic leaves.
叶绿体中 Rubisco 小亚基启动子控制的 AtFDH 过表达增强了烟草叶片对甲醛的摄取和代谢。我们之前的研究表明,与拟南芥相比,烟草对甲醛的摄取和抗性较弱。拟南芥中的甲酸脱氢酶(AtFDH)是通过氧化 HCOOH 为 CO 将 HCHO 代谢为甲醛进入卡尔文循环并被同化到葡萄糖中的关键酶。烟草中的 HCHO 代谢机制与拟南芥不同。在这项研究中,使用光诱导启动子在转基因烟草的叶绿体中过表达 AtFDH。13C-NMR 分析表明,来自 HCHO 代谢的碳通量没有被引入 Calvin 循环以在转基因烟草叶片中产生葡萄糖。然而,AtFDH 的过表达显著增强了转基因叶片中的 HCHO 代谢。因此,在 HCHO 处理后,转基因叶片中[4-C]Asn、[3-C]Gln、[U-C]草酸盐和 HCOOH 的产量明显高于非转化叶片。转基因叶片中增加的气孔导度和孔径可能归因于叶绿体中过表达的 AtFDH 在保卫细胞中产生的草酸盐增加。因此,转基因植物对气态 HCHO 的吸收能力更强。此外,在 HCHO 胁迫下,与非转化叶片相比,转基因叶片中的脯氨酸含量较高可能归因于过量的甲酸积累和 Gln 产生。因此,在转基因叶片中减少了 HCHO 诱导的氧化应激。