Havaux Michel, Ksas Brigitte, Szewczyk Agnieszka, Rumeau Dominique, Franck Fabrice, Caffarri Stefano, Triantaphylidès Christian
Commissariat à l'Energie Atomique, Institut de Biologie Environnementale et de Biotechnologie, Laboratoire d'Ecophysiologie Moléculaire des Plantes, 13108 Saint-Paul-lez-Durance, France.
BMC Plant Biol. 2009 Nov 10;9:130. doi: 10.1186/1471-2229-9-130.
Vitamin B6 is a collective term for a group of six interconvertible compounds: pyridoxine, pyridoxal, pyridoxamine and their phosphorylated derivatives. Vitamin B6 plays essential roles as a cofactor in a range of biochemical reactions. In addition, vitamin B6 is able to quench reactive oxygen species in vitro, and exogenously applied vitamin B6 protects plant cells against cell death induced by singlet oxygen (1O2). These results raise the important question as to whether plants employ vitamin B6 as an antioxidant to protect themselves against reactive oxygen species.
The pdx1.3 mutation affects the vitamin B6 biosynthesis enzyme, pyridoxal synthase (PDX1), and leads to a reduction of the vitamin B6 concentration in Arabidopsis thaliana leaves. Although leaves of the pdx1.3 Arabidopsis mutant contained less chlorophyll than wild-type leaves, we found that vitamin B6 deficiency did not significantly impact photosynthetic performance or shoot and root growth. Chlorophyll loss was associated with an increase in the chlorophyll a/b ratio and a selective decrease in the abundance of several PSII antenna proteins (Lhcb1/2, Lhcb6). These changes were strongly dependent on light intensity, with high light amplifying the difference between pdx1.3 and the wild type. When leaf discs were exposed to exogenous 1O2, lipid peroxidation in pdx1.3 was increased relative to the wild type; this effect was not observed with superoxide or hydrogen peroxide. When leaf discs or whole plants were exposed to excess light energy, 1O2-mediated lipid peroxidation was enhanced in leaves of the pdx1.3 mutant relative to the wild type. High light also caused an increased level of 1O2 in vitamin B6-deficient leaves. Combining the pdx1.3 mutation with mutations affecting the level of 'classical' quenchers of 1O2 (zeaxanthin, tocopherols) resulted in a highly photosensitive phenotype.
This study demonstrates that vitamin B6 has a function in the in vivo antioxidant defense of plants. Thus, the antioxidant activity of vitamin B6 inferred from in vitro studies is confirmed in planta. Together with the finding that chloroplasts contain vitamin B6 compounds, the data show that vitamin B6 functions as a photoprotector that limits 1O2 accumulation in high light and prevents 1O2-mediated oxidative damage.
维生素B6是六种可相互转化的化合物的统称:吡哆醇、吡哆醛、吡哆胺及其磷酸化衍生物。维生素B6作为一系列生化反应中的辅因子发挥着重要作用。此外,维生素B6在体外能够淬灭活性氧,外源施用的维生素B6可保护植物细胞免受单线态氧(1O2)诱导的细胞死亡。这些结果引发了一个重要问题,即植物是否利用维生素B6作为抗氧化剂来保护自身免受活性氧的侵害。
pdx1.3突变影响维生素B6生物合成酶吡哆醛合酶(PDX1),导致拟南芥叶片中维生素B6浓度降低。尽管pdx1.3拟南芥突变体的叶片比野生型叶片含有更少的叶绿素,但我们发现维生素B6缺乏并未显著影响光合性能或地上部和根部的生长。叶绿素损失与叶绿素a/b比值增加以及几种PSII天线蛋白(Lhcb1/2、Lhcb6)丰度的选择性降低有关。这些变化强烈依赖于光照强度,高光会放大pdx1.3与野生型之间的差异。当叶圆片暴露于外源1O2时,pdx1.3中的脂质过氧化相对于野生型增加;超氧化物或过氧化氢未观察到这种效应。当叶圆片或整株植物暴露于过量光能时,相对于野生型,pdx1.3突变体叶片中1O2介导的脂质过氧化增强。高光还导致维生素B6缺乏叶片中1O2水平升高。将pdx1.3突变与影响1O2“经典”淬灭剂(玉米黄质、生育酚)水平的突变相结合,导致高度光敏表型。
本研究表明维生素B6在植物的体内抗氧化防御中具有功能。因此,体外研究推断的维生素B6的抗氧化活性在植物中得到了证实。连同叶绿体含有维生素B6化合物这一发现,数据表明维生素B6作为一种光保护剂,可限制高光下1O2的积累并防止1O2介导的氧化损伤。