Research Article

Cloning and Expression Analysis of Tomato SlSPS1 Gene  

Dewen Zeng1,2 , Yahui Liu2 , Hui Zhang2 , Xuedong Yang2 , Longying Zhu2 , Weimin Zhu1,2 , Yingying Zhang2
1 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China
2 Horticultural Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai Key laboratory of Protected Horticulture Technology, Shanghai, 201403, China
Author    Correspondence author
Molecular Plant Breeding, 2022, Vol. 13, No. 2   doi: 10.5376/mpb.2022.13.0002
Received: 05 Jan., 2022    Accepted: 17 Jan., 2022    Published: 26 Jan., 2022
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Zeng D.W., Liu Y.H., Zhang H., Yang X.D., Zhu L.Y., Zhu W.M., and Zhang Y.Y., 2022, Cloning and expression analysis of tomato SlSPS1 gene, Molecular Plant Breeding, 13(2): 1-11 (doi:10.5376/mpb.2022.13.0002)

Abstract

Sucrose is the main product of plant photosynthesis and participates in many physiological and biochemical reactions of plants. Sucrose phosphate synthase is one of the key rate-limiting enzymes necessary for sucrose to enter various metabolic pathways, and its activity directly reflects the ability of sucrose synthesis in plants. In order to study the mechanism of sucrose synthesis in tomato, this study cloned a sucrose phosphate synthase gene from tomato pulp and named it SlSPS1. Through bioinformatics analysis, it was found that its cDNA is 3488bp in length, encoding 1054 amino acids, and there are 3 functional domain. There are 4 SPS genes in tomato, and their homology is relatively high. In Salicaceae crops, SlSPS1 protein is closely related to potatoes, peppers and eggplants. The results of tissue-specific analysis found that SlSPS1 was expressed in all tomato tissues, among which the expression was relatively high in the seedling stage and different development stages of the fruit. In order to further study the mechanism of SlSPS1 in tomato growth and sucrose accumulation, an overexpression and knockout vector of SlSPS1 was constructed and transformed into Micro-Tom to obtain transgenic positive lines. Analysis of the expression level of SlSPS1 on the obtained transgenic lines showed that the expression levels of the overexpression lines at the seedling stage, fruit green ripe stage and ripening stage were significantly up-regulated compared with the wild type, and the enzyme activity and sucrose content increased. Compared with the wild type, the expression of SlSPS1 at the seedling stage, green ripe stage and ripening stage of the knockout line was significantly down-regulated, and the enzyme activity and sucrose content were reduced. As for the mechanism of SlSPS1 in tomato growth and fruit sucrose accumulation, functional verification is needed in the future.

Keywords
Tomato; Clone; SlSPS1; Expression; Analysis
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