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[甘薯中蔗糖转运蛋白的功能分析]

[Functional analysis on sucrose transporters in sweet potato].

作者信息

Liu Yiran, Wu Zhengdan, Wu Weitai, Yang Chaobin, Chen Cairui, Zhang Kai

机构信息

College of Agronomy and Biotechnology, Southwest University, Chongqing 400715, China.

Chongqing Key Laboratory of Biology and Genetic Breeding for Tuber and Root Crops, Chongqing 400715, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2023 Jul 25;39(7):2772-2793. doi: 10.13345/j.cjb.220979.

DOI:10.13345/j.cjb.220979
PMID:37584131
Abstract

Sweet potato is an important food crop that can also be used as an industrial raw material. Sucrose is the main form of long-distance carbohydrate transport in plants, and sucrose transporter (SUT) regulates the transmembrane transport and distribution of sucrose during plant growth and metabolism. Moreover, SUT plays a key role in phloem mediated source-to-sink sucrose transport and physiological activities, supplying sucrose for the sink tissues. In this study, the full-length cDNA sequences of and were obtained by rapid amplification of cDNA ends (RACE) cloning according to the transcripts of the two SUT coding genes which were differentially expressed in sweet potato storage roots with different starch properties. Phylogenetic analysis was performed to clarify the classification of IbSUT and IbSUT. The subcellular localization of IbSUT and IbSUT was determined by transient expression in . The function of IbSUT and IbSUT in sucrose and hexose absorption and transport was identified using yeast functional complementarity system. The expression pattern of and in sweet potato organs were analyzed by real-time fluorescence quantitative PCR (RT-qPCR). plants heterologous expressing and genes were obtained using floral dip method. The differences in starch and sugar contents between transgenic and wild-type were compared. The results showed and encoded SUT proteins with a length of 505 and 521 amino acids, respectively, and both proteins belonged to the SUT1 subfamily. IbSUT and IbSUT were located in the cell membrane and were able to transport sucrose, glucose and fructose in the yeast system. In addition, IbSUT was also able to transport mannose. The expression of was higher in leaves, lateral branches and main stems, and the expression of was higher in lateral branches, stems and storage roots. After and were heterologously expressed in , the plants grew normally, but the biomass increased. The heterologous expression of increased the soluble sugar content, leaf size and 1 000-seed weight of plants. Heterologous expression of increased starch accumulation in leaves and root tips and 1 000-seed weight of seeds, but decreased soluble sugar content. The results obtained in this study showed that and might be important genes regulating sucrose and sugar content traits in sweet potato. They might carry out physiological functions on cell membrane, such as transmembrane transport of sucrose, sucrose into and out of sink tissue, as well as transport and unloading of sucrose into phloem. The changes in traits result from their heterologous expression in indicates their potential in improving the yield of other plants or crops. The results obtained in this study provide important information for revealing the functions of and in starch and glucose metabolism and formation mechanism of important quality traits in sweet potato.

摘要

甘薯是一种重要的粮食作物,也可作为工业原料。蔗糖是植物中碳水化合物长距离运输的主要形式,蔗糖转运蛋白(SUT)在植物生长和代谢过程中调节蔗糖的跨膜运输和分配。此外,SUT在韧皮部介导的源库蔗糖运输和生理活动中起关键作用,为库组织提供蔗糖。在本研究中,根据在具有不同淀粉特性的甘薯贮藏根中差异表达的两个SUT编码基因的转录本,通过cDNA末端快速扩增(RACE)克隆获得了 和 的全长cDNA序列。进行系统发育分析以阐明IbSUT和IbSUT的分类。通过在 中的瞬时表达确定IbSUT和IbSUT的亚细胞定位。使用酵母功能互补系统鉴定IbSUT和IbSUT在蔗糖和己糖吸收及运输中的功能。通过实时荧光定量PCR(RT-qPCR)分析 和 在甘薯器官中的表达模式。使用花浸染法获得了异源表达 和 基因的 植株。比较了转基因植株和野生型 植株在淀粉和糖含量上的差异。结果表明, 和 分别编码长度为505和521个氨基酸的SUT蛋白,这两种蛋白均属于SUT1亚家族。IbSUT和IbSUT位于细胞膜上,并且能够在酵母系统中运输蔗糖、葡萄糖和果糖。此外,IbSUT还能够运输甘露糖。 在叶片、侧枝和主茎中的表达较高,而 在侧枝、茎和贮藏根中的表达较高。 和 在 中异源表达后植株生长正常,但生物量增加。 的异源表达增加了 植株的可溶性糖含量、叶片大小和千粒重。 的异源表达增加了叶片和根尖中的淀粉积累以及种子的千粒重,但降低了可溶性糖含量。本研究获得的结果表明, 和 可能是调控甘薯蔗糖和糖含量性状的重要基因。它们可能在细胞膜上执行生理功能,如蔗糖的跨膜运输、蔗糖进出库组织以及蔗糖向韧皮部的运输和卸载。它们在 中的异源表达导致的性状变化表明它们在提高其他植物或作物产量方面的潜力。本研究获得的结果为揭示 和 在甘薯淀粉和葡萄糖代谢以及重要品质性状形成机制中的功能提供了重要信息。

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