L-galactono-1, 4-lactone dehydrogenase(GLDH) is an important enzyme that catalyzes the last step of the ascorbate biosynthetic pathways in plants. A full-length c DNA clone encoding GLDH was isolated from potato(Solanum tuberosum L. ‘Favorita') leaf and subcloned into a binary vector, p BI121, to construct sense and antisense recombinant plant expression vectors. The recombinants were introduced into potato via Agrobacterium-mediated transformation, and plants were confirmed as transgenic using PCR and quantitative real-time PCR. Two anti-sense potato lines(G1 and G2) and three sense lines(G3, G4 and G5) were obtained. The GLDH activity of G4 and G5 were increased in vivo. Moreover, the ascorbic acid(As A) and dehydroascorbate(DHA) contents were up-regulated in both leaves and tubers. However, the shoots of G1 were suppressed and its leaves were deformed. Additionally, the As A contents in G1 leaves and tubers decreased by 28.8% and 10.3%, respectively. The GLDH activity in leaves treated with L-galactono-1,4- lactone(L-Gal L) increased in all lines, as did the As A and DHA contents. These results indicate that GLDH activity plays an important role in regulating the As A level as well as the growth and development of potato plants.
GDP-D-mannose pyrophosphorylase (GMPase) catalyses the synthesis of GDP-D-mannose and represents the first committed step in the synthesis of ascorbate. In the present study, the GMPase gene of tomato was introduced into potato by Agrobacterium-mediated transformation. Two transgenic lines with higher GMPase expression were selected using qPCR and protein blot analyses. The results showed that the content of L-ascorbic acid (AsA) and the ratio of AsA/ DHA (dehydroascorbate) significantly increased in both leaves and tubers of transgenic potato plants. Both pigment content and photosynthetic rate were much higher in transgenic plants than in wild-type plants. Transgenic plants showed a distinguishable change in phenotype from the wild-type plants. Furthermore, transgenic plants showed delayed senescence.
LIN Ling-lingSHI Qing-huaWANG Hua-senQIN Ai-guoYU Xian-chang
