Cloning and Expression Analysis of Tomato &lt;i&gt;SlSPS1&lt;/i&gt; Gene

Dewen Zeng,Hui Zhang,Longying Zhu,Weimin Zhu,Yahui Liu,Yingying Zhang,Xuedong Yang

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.

Highlights

Solanum lycopersicum belongs to Lycopersicon of Solanaceae, commonly known as tomato, and is a fruit and vegetable that is very popular among consumers
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
1.1 Cloning and bioinformatics analysis of SlSPS1 gene Using tomato fruit genomic DNA and the whole gene cDNA obtained by reverse transcription as templates, primers were designed to clone the full length of SlSPS1 gene and cDNA

Summary

Introduction

Solanum lycopersicum belongs to Lycopersicon of Solanaceae, commonly known as tomato, and is a fruit and vegetable that is very popular among consumers. Carbohydrates affect the growth and development of fruit, and as signal molecules participate in regulating the expression of genes related to fruit ripening and the process of stress tolerance (Liao et al, 2020). The current research results have found that SPS participates in plant growth, fruit ripening and other processes, and has the effect of responding to abiotic stress. At present, the transcriptional regulation process of SPS in plants, the molecular mechanisms and pathways involved in various biological functions are still unclear. It is of great significance to analyze the function and regulation mechanism of SPS in sucrose metabolism and plant growth, which can provide a theoretical basis for increasing crop yield. SlSPS1 gene was cloned from tomato fruit and analyzed by bioinformatics. SlSPS1 expression analysis, SPS enzyme activity determination and sucrose determination were performed on the obtained transgenic lines

Methods

Results

Conclusion