大豆GmGolS2-2提高转基因烟草耐旱性

Soybean GmGolS2-2 improves drought resistance of transgenic tobacco

摘要:

肌醇半乳糖苷合成酶（galactinol synthase，GolS）基因在植物应对非生物胁迫时发挥重要作用。本研究构建大豆GmGolS2-2基因植物表达载体并转化烟草，对转基因烟草的耐旱性进行鉴定。通过逆转录-聚合酶链式反应（reverse transcription-polymerase chain reaction，RT-PCR）从大豆叶片中克隆了975 bp的GmGolS2-2基因编码序列。通过限制性内切酶位点Nde I和EcoR I将GmGolS2-2基因与植物表达载体pRI101相连，通过叶盘法转化烟草。基因组DNA PCR和实时荧光定量PCR结果显示，共获得3棵GmGolS2-2转基因烟草。干旱胁迫下GmGolS2-2转基因烟草的生长状态好于野生型烟草。干旱胁迫处理后，转基因烟草的电解质渗透率和丙二醛含量低于野生型烟草，脯氨酸含量和可溶性糖含量则高于野生型烟草。实时荧光定量PCR结果显示，GmGolS2-2的异源表达提高了转基因烟草中抗逆相关基因NtERD10C和NtAQP1的表达量。以上结果表明，GmGolS2-2提高了转基因烟草的耐旱性。

Abstract:

Galactinol synthase (GolS) genes play important roles in plant response to abiotic stress. In this research, the plant expression vector of soybean GmGolS2-2 gene was constructed and transformed into tobacco to study the drought tolerance of transgenic tobacco. A GmGolS2-2 gene with 975 bp coding sequence was cloned from soybean leaves by reverse transcription-polymerase chain reaction (RT-PCR). GmGolS2-2 was linked to the plant expression vector pRI101 by restriction enzyme sites Nde I and EcoR I, and transformed into tobacco by leaf disc method. Genomic DNA PCR and real-time PCR showed that three GmGolS2-2 transgenic tobacco plants were obtained. The growth status of GmGolS2-2 transgenic tobacco under drought stress was better than that of wild-type tobacco. After drought stress treatment, the electrolyte leakage and malondialdehyde content of transgenic tobacco were lower than those of wild-type tobacco, but the proline content and soluble sugar content were higher than those of wild-type tobacco. The results of real-time PCR showed that the heterologous expression of GmGolS2-2 increased the expression of stress-related genes NtERD10C and NtAQP1 in transgenic tobacco. The above results indicated that GmGolS2-2 improved drought resistance of transgenic tobacco.