邱爽　张军　何佳琦　李铭杨　周雨明　邬长乐　袁洪淼　刘嘉仪　翟莹

摘要： 肌醇半乳糖苷合成酶（GolS）是棉籽糖系列寡糖（RFO）生物合成途径中的关键酶，在植物应对非生物胁迫过程中发挥重要作用。实时荧光定量RT-PCR结果显示，高温胁迫可以诱导GmGolS2-1在大豆幼苗中的表达。将GmGolS2-1基因构建到植物表达载体pRI101上并通过叶盘法转化烟草，经卡那霉素抗性筛选，PCR及qRT-PCR检测共获得6株阳性转基因烟草植株（OE1～OE6）。对野生型烟草植株和GmGolS2-1转基因烟草植株进行高温胁迫处理，结果显示野生型烟草的电解质渗透率和丙二醛含量均高于转基因烟草。由此推测GmGolS2-1可以提高转基因烟草的耐热性。

关键词： 大豆;肌醇半乳糖苷;GolS基因;高温胁迫;转基因烟草

中图分类号： S565.1 文献标识码： A 文章编号： 1000-4440（2021）01-0038-06

Expression of soybean GmGolS2-1 induced by heat stress and identification of GmGolS2-1 transgenic tobacco

QIU Shuang1， ZHANG Jun2， HE Jia-qi1， LI Ming-yang1， ZHOU Yu-ming3， WU Chang-le1， YUAN Hong-miao1， LIU Jia-yi1， ZHAI Ying1

（1.College of Life Science and Agro-Forestry， Qiqihar University， Qiqihar 161006， China;2.Branch of Animal Husbandry and Veterinary of Heilongjiang Academy of Agricultural Sciences， Qiqihar 161005， China;3.Jilin Zhongzhi Jiufang Consulting Co.， Ltd.， Changchun 130000， China）

Abstract： Galactinol synthase （GolS） is the key enzyme in the biosynthetic pathway of raffinose family oligosaccharides （RFOs）， which plays an important role in the response to abiotic stresses of plants. The results of real-time fluorescence quantitative RT-PCR showed that the expression of GmGolS2-1 could be induced by high temperature stress in soybean seedlings. The GmGolS2-1 gene was constructed into expression vector pRI101 in plants and was transformed into tobacco using leaf disc method. Six positive transgenic tobacco plants （OE1-OE6） were obtained by kanamycin resistance screening， PCR and qRT-PCR. The wild-type and GmGolS2-1 transgenic tobacco plants were treated with heat stress. The results showed that the electrolyte leakage and malondialdehyde content of wild-type tobacco were both higher than that of transgenic tobacco. These data indicate that GmGolS2-1 can increase the tolerance to heat stress of transgenic tobacco.

Key words： soybean;galactinol;GolS gene;heat stress;transgenic tobacco

植物在遭受不良環境条件后，可以诱导合成大量渗透调节物质来增加植物细胞的渗透压，提高植物抵抗胁迫的能力，从而维持植物自身的代谢和生长发育。其中棉子糖系列寡糖（Raffinose family oligosaccharides， RFO）就是这些渗透调节物质的典型代表。它们是高等植物中含量仅次于蔗糖的一类可溶性糖，在逆境胁迫条件下可以维持细胞膨压，保持细胞稳定性，降低植物氧化性损伤，维持光合作用正常进行[1-3]。肌醇半乳糖苷合成酶（Galactinol synthase， GolS）是RFO生物合成途径中的关键酶，它所催化的UDP-半乳糖和肌醇合成肌醇半乳糖苷的反应既是RFO代谢通路中的第一步，也是限速步骤[4]。

GolS基因在植物中以多基因家族形式存在，不同GolS基因在抵御非生物胁迫过程中的作用也各不相同。……