National Institute for Plant Biotechnology, Pusa Campus, New Delhi, 110012, India.
Forest Molecular Entomology Laboratory, Excellent Team for Mitigation (ETM), Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Prague, Czech Republic.
Transgenic Res. 2024 Jun;33(3):131-147. doi: 10.1007/s11248-024-00385-x. Epub 2024 May 13.
Δ-pyrroline-5-carboxylate synthetase (P5CS) is one of the key regulatory enzymes involved in the proline biosynthetic pathway. Proline acts as an osmoprotectant, molecular chaperone, antioxidant, and regulator of redox homeostasis. The accumulation of proline during stress is believed to confer tolerance in plants. In this study, we cloned the complete CDS of the P5CS from pearl millet (Pennisetum glaucum (L.) R.Br. and transformed into tobacco. Three transgenic tobacco plants with single-copy insertion were analyzed for drought and heat stress tolerance. No difference was observed between transgenic and wild-type (WT) plants when both were grown in normal conditions. However, under heat and drought, transgenic plants have been found to have higher chlorophyll, relative water, and proline content, and lower malondialdehyde (MDA) levels than WT plants. The photosynthetic parameters (stomatal conductance, intracellular CO concentration, and transpiration rate) were also observed to be high in transgenic plants under abiotic stress conditions. qRT-PCR analysis revealed that the expression of the transgene in drought and heat conditions was 2-10 and 2-7.5 fold higher than in normal conditions, respectively. Surprisingly, only P5CS was increased under heat stress conditions, indicating the possibility of feedback inhibition. Our results demonstrate the positive role of PgP5CS in enhancing abiotic stress tolerance in tobacco, suggesting its possible use to increase abiotic stress-tolerance in crops for sustained yield under adverse climatic conditions.
Δ-吡咯啉-5-羧酸合成酶(P5CS)是参与脯氨酸生物合成途径的关键调节酶之一。脯氨酸作为渗透保护剂、分子伴侣、抗氧化剂和氧化还原平衡调节剂。在应激条件下脯氨酸的积累被认为赋予了植物的耐受性。在这项研究中,我们从珍珠粟(Pennisetum glaucum(L.)R.Br.)中克隆了 P5CS 的完整 CDS,并将其转化为烟草。对具有单拷贝插入的三个转基因烟草植株进行了干旱和热应激耐受性分析。在正常条件下,转基因植株和野生型(WT)植株之间没有观察到差异。然而,在热和干旱条件下,与 WT 植株相比,转基因植株的叶绿素、相对水分和脯氨酸含量更高,丙二醛(MDA)水平更低。在非生物胁迫条件下,还观察到转基因植株的光合参数(气孔导度、胞内 CO 浓度和蒸腾速率)较高。qRT-PCR 分析表明,在干旱和热条件下,转基因的表达分别比正常条件下高 2-10 倍和 2-7.5 倍。令人惊讶的是,只有在热应激条件下 P5CS 增加,表明可能存在反馈抑制。我们的结果表明 PgP5CS 在增强烟草对非生物胁迫的耐受性中具有积极作用,表明其在提高作物对不利气候条件下的非生物胁迫耐受性方面具有潜在的应用价值,以实现持续的产量。