Plant Molecular Biology Laboratory, Department of Biotechnology, Dr Harisingh Gour Central University, Sagar, 470003, Madhya Pradesh, India.
Metagenomics and Secretomics Research laboratory, Department of Botany, Dr Harisingh Gour Central University, Sagar, 470003, Madhya Pradesh, India.
Plant Sci. 2018 Jul;272:267-275. doi: 10.1016/j.plantsci.2018.04.024. Epub 2018 May 1.
Vitamin B6 is a vital metabolite required for living organisms as a cofactor in several metabolic biochemical reactions and recognized as a potent antioxidant molecule which modulates the expression of the proteins responsible for the scavenging of cellular reactive oxygen species. It is well established that the microorganisms and plants can synthesize the B6 de novo, therefore, all the animals including humans must acquire it from the plant dietary resources. However, the bioavailability of the vitamin in the edible portions of the commonly consumed plants is insufficient to meet the daily recommended doses. Genetic engineering techniques have proven successful in increasing the vitamin B6 content in the model plants. Present study describe the development of transgenic potato (Solanum tuberosum L. cv. Kufri chipsona) overexpressing key vitamin B6 pathway gene, the PDXII (NCBI database Ref. ID- NM_125447.2) isolated from Arabidopsis thaliana under the control of CaMV 35S constitutive promoter. The stable integration and expression of transgene in the transgenic lines were confirmed by PCR, Southern blot and RT-PCR analysis. Transgenic tubers exhibited considerably improved vitamin B6 accumulation (up to 107-150%) in comparison to the untransformed controls potato. This increase in vitamin B6 was also correlated with the increased mRNA expression of PDXII gene. The prominent increase in the B6 content of transgenic potato was also associated with the capability to survive under abiotic stresses, therefore, the transgenic lines were able to withstand various abiotic stresses imposed by salinity (NaCl) or methyl viologen (MV). We thus demonstrated that overexpression of PDXII gene under the control of a constitutive promoter enhanced the accumulation of the vitamin B6 which also augmented the tolerance under various abiotic stresses in potato (Solanum tuberosum L.).
维生素 B6 是一种重要的代谢物,作为几种代谢生化反应的辅助因子,是生物体所必需的,并且被认为是一种有效的抗氧化分子,可以调节负责清除细胞内活性氧的蛋白质的表达。微生物和植物可以从头合成 B6,因此,包括人类在内的所有动物都必须从植物饮食资源中获取它。然而,在常见食用植物的可食用部分中,维生素的生物利用度不足以满足每日推荐剂量。基因工程技术已成功地用于增加模式植物中的维生素 B6 含量。本研究描述了在拟南芥中分离的关键维生素 B6 途径基因 PDXII(NCBI 数据库参考 ID-NM_125447.2)在 CaMV 35S 组成型启动子的控制下,在转基因马铃薯(Solanum tuberosum L. cv. Kufri chipsona)中过表达。通过 PCR、Southern blot 和 RT-PCR 分析证实了转基因株系中转基因的稳定整合和表达。与未转化的对照马铃薯相比,转基因块茎表现出相当大的维生素 B6 积累增加(高达 107-150%)。这种 B6 增加与 PDXII 基因的 mRNA 表达增加相关。转基因马铃薯中 B6 含量的显著增加也与适应非生物胁迫的能力有关,因此,转基因株系能够耐受盐度(NaCl)或甲基紫精(MV)施加的各种非生物胁迫。因此,我们证明了在组成型启动子的控制下过表达 PDXII 基因可以增强维生素 B6 的积累,从而提高马铃薯(Solanum tuberosum L.)在各种非生物胁迫下的耐受性。