Effect of Heterologous Expression of StSWEET16b Gene on Fructose Content and Resistance to Late Blight in Tobacco (Nicotiana tabacum)
FAN Xi-De1, HE Miao-Miao1,2, YE Guang-Ji1,2, WANG Jian1,2, ZHOU Yun1,2,*
1 Academy of Agriculture and Forestry Sciences,Qinghai University, Xining 810016, China; 2 Qinghai Academy of Agriculture and Forestry Sciences/State key Laboratory of Plateau Ecology and Agriculture, Qinghai University/Key Laboratory of Qinghai-Tibetan Plateau Biotechnology of Ministry of Education, Qinghai University/Key Laboratory of Potato Breeding of Qinghai, Xining 810016, China
摘要糖转运蛋白(sugars will eventually be exported transporters, SWEETs)广泛存在于真核生物中,在植物糖类的长距离转运、激素转运、花粉发育、果实形成、种子灌浆及植物-病原体互作等生理过程中发挥重要作用。为解析StSWEET16b基因在糖的转运及病原菌互作中的功能,本研究以马铃薯(Solanum tuberosum) '青薯9号'为材料,采用qRT-PCR技术分析了该基因在不同糖处理下的表达规律,结果发现,StSWEET16b在3%的蔗糖、果糖和葡萄糖分别处理下表达模式均发生变化,表明该基因在马铃薯中可能同时参与蔗糖、果糖及葡萄糖的转运。通过分析转基因烟草(Nicotiana tabacum)在不同糖培养基上的生长状况,并测定植株体内蔗糖、果糖及葡萄糖的含量发现,在葡萄糖和蔗糖培养基上,转基因烟草的长势弱于野生型植株,并且转基因植株中的蔗糖和葡萄糖含量均低于野生型,而在果糖培养基上,转基因烟草的长势明显优于野生型植株,且转基因植株中的果糖含量高于野生型(P<0.05),表明在烟草中StSWEET16b的异源表达可促进果糖的转运,同时抑制蔗糖和葡萄糖的转运。瞬时表达结果表明,StSWEET16b基因参与了本氏烟-晚疫病菌之间的互作,抑制了晚疫病菌病斑的扩展,提高了植株的抗病性。研究结果表明,StSWEET16b蛋白在果糖转运和植物-病原病菌互作过程中发挥着重要作用。本研究为马铃薯品质的改良和抗病品种的选育提供理论依据。
Abstract:Sugars will eventually be exported transporters (SWEETs), a novel family of membrane-transporters widely distributed in eukaryotes, which performed essential roles in plant growth and development processes, such as phloem loading for long distance sugar transport, hormone transport, pollen development, fruit development, seed filling and plant-pathogen interactions. To analyze the function of StSWEET16b in sugar transport and plant-pathogen interactions, in this study, potato (Solanum tuberosum 'Qingshu 9') was used as material, after treated with sucrose, fructose and glucose individually, the expression of StSWEET16b was analyzed by qRT-PCR in the leaves of 'Qingshu 9'. The expression pattern of StSWEET16b was changed under 3% sucrose, fructose and 3glucose treatments respective, suggesting that may be involved in the translocation of sucrose, fructose and glucose in potato. By analyzing the growth condition of the transgenic tobacco (Nicotiana benthamiana) on different sugar medium and measuring the contents of sucrose, fructose and glucose in the plants, it was found that the transgenic tobacco grew weaker than the wild-type plants on glucose and sucrose medium, and the contents of sucrose and glucose in the transgenic plants were lower than those in the wild-type plants. However, transgenic tobacco grew significantly better than wild-type plants on fructose medium (P<0.05), and the fructose content of transgenic plants was higher than that of wild-type plants, indicated that the heterologous expression of StSWEET16b in tobacco could promote fructose transport, and inhibit the transport of sucrose and glucose. Transient expression results showed that StSWEET16b gene was involved in the interaction between tobacco and Phytophthora infestans. Furthermore, transient expression of StSWEET16b enhanced the resistance to P. infestans in tobacco, resulting in inhibiting the expansion of late blight disease spots. It inhibited the expansion of the disease spot of P. infestans and improved the disease resistance of the plant. The results suggesed that StSWEET16b protein played an important role in sugar transport and plant-pathogen interactions for resistance to late blight. This study provides a theoretical basis for improving potato quality and breeding of disease-resistant varieties.
范希德, 贺苗苗, 叶广继, 王舰, 周云. StSWEET16b基因在烟草中异源表达对果糖含量和晚疫病抗性的影响[J]. 农业生物技术学报, 2021, 29(6): 1031-1039.
FAN Xi-De, HE Miao-Miao, YE Guang-Ji, WANG Jian, ZHOU Yun. Effect of Heterologous Expression of StSWEET16b Gene on Fructose Content and Resistance to Late Blight in Tobacco (Nicotiana tabacum). 农业生物技术学报, 2021, 29(6): 1031-1039.
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