Lim Benson, Smirnoff Nicholas, Cobbett Christopher S, Golz John F
School of BioSciences, University of Melbourne, Parkville VIC, Australia.
Biosciences, College of Life and Environmental Sciences, University of Exeter Exeter, UK.
Front Plant Sci. 2016 Jul 13;7:1025. doi: 10.3389/fpls.2016.01025. eCollection 2016.
In higher plants the L-galactose pathway represents the major route for ascorbate biosynthesis. The first committed step of this pathway is catalyzed by the enzyme GDP-L-galactose phosphorylase and is encoded by two paralogs in Arabidopsis - VITAMIN C2 (VTC2) and VTC5. The first mutant of this enzyme, vtc2-1, isolated via an EMS mutagenesis screen, has approximately 20-30% of wildtype ascorbate levels and has been reported to have decreased growth under standard laboratory conditions. Here, we show that a T-DNA insertion into the VTC2 causes a similar reduction in ascorbate levels, but does not greatly affect plant growth. Subsequent segregation analysis revealed the growth defects of vtc2-1 mutants segregate independently of the vtc2-1 mutation. These observations suggest that it is the presence of an independent cryptic mutation that affects growth of vtc2-1 mutants, and not the 70-80% decrease in ascorbate levels that has been assumed in past studies.
在高等植物中,L-半乳糖途径是抗坏血酸生物合成的主要途径。该途径的第一个关键步骤由GDP-L-半乳糖磷酸化酶催化,在拟南芥中由两个旁系同源基因——维生素C2(VTC2)和VTC5编码。通过EMS诱变筛选分离出的该酶的第一个突变体vtc2-1,其抗坏血酸水平约为野生型的20%-30%,据报道在标准实验室条件下生长减缓。在此,我们表明,插入VTC2的T-DNA会导致抗坏血酸水平出现类似降低,但对植物生长影响不大。随后的分离分析表明,vtc2-1突变体的生长缺陷与vtc2-1突变独立分离。这些观察结果表明,是一个独立的隐性突变的存在影响了vtc2-1突变体的生长,而不是过去研究中所假设的抗坏血酸水平降低70%-80%。