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为了探讨番茄GDP-L-半乳糖磷酸酶对烟草抗坏血酸(AsA)含量及抗氧化能力的影响,从番茄叶片中分离了GDP-L-半乳糖磷酸酶基因(LeGGP),并转入到烟草中。以野生型(WT)和转正义LeGGP烟草株系T1-3和T1-15为试材,测定了甲基紫精(MV)处理下AsA、脱氢抗坏血酸(DHA)、H2O2、O2.和叶绿素含量、抗坏血酸过氧化物酶(APX)活性、光合速率和叶绿素荧光参数等。Northern杂交分析表明LeGGP的表达受MV的诱导,在MV处理下,野生型烟草的离体叶圆片发生比转基因烟草更严重的光漂白,转基因烟草的AsA含量及清除H2O2和O2.的能力明显强于野生型,过表达LeGGP提高了烟草的生长量。并且转基因烟草比野生型具有更高的净光合效率(Pn)和光系统Ⅱ(PSⅡ)最大光化学效率(Fv/Fm)。结果表明,LeGGP的过表达有助于提高烟草AsA含量及抗氧化胁迫能力。
In order to investigate the effect of tomato GDP-L-galactose phosphatase on the content of AsA and the antioxidant capacity of tobacco, the GDP-L-galactose phosphatase gene (LeGGP) was isolated from tomato leaves and transferred into tobacco . AsA, dehydroascorbic acid (DHA), H2O2, O2 (subscript x), were measured under wild-type (WT) and transgenic LeGGP tobacco lines T1-3 and T1-15 Chlorophyll content, ascorbate peroxidase (APX) activity, photosynthetic rate and chlorophyll fluorescence parameters. Northern hybridization analysis showed that the expression of LeGGP was induced by MV. Under MV treatment, the leaves of wild-type tobacco produced more severe photobleaching than transgenic tobacco, the AsA content of transgenic tobacco and the ability to clear H2O2 and O2. Significantly stronger than the wild-type, overexpression of LeGGP increased tobacco growth. And transgenic tobacco had higher net photosynthetic efficiency (Pn) and maximum photochemical efficiency (Fv / Fm) than that of wild type (PSⅡ). The results showed that overexpression of LeGGP could improve the content of AsA and the ability of anti-oxidation in tobacco.