Analysis of the expression of sucrose phosphate synthase genes during the development of blackberry fruit

YAN Zhixiang, YANG Haiyan, FAN Sufan, WU Wenlong, LYU Lianfei, LI Weilin

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (1) : 179-186.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (1) : 179-186. DOI: 10.12302/j.issn.1000-2006.202007067

Analysis of the expression of sucrose phosphate synthase genes during the development of blackberry fruit

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Abstract

【Objective】Sucrose phosphate synthase (SPS) is a key enzyme that regulates the synthesis pathway of sucrose metabolism in plants. Although it plays an important role in the accumulation and distribution of photosynthetic products, systematic research on SPS genes in blackberry has rarely been carried out. This study aimed to investigate the phylogenetic relationship between SPS genes in blackberry, the protein properties SPS expression pattern in different tissues at various developmental stages, and its relationship with blackberry development.【Method】 The blackberry cultivar, ‘Boysenberry’, was used in the tests, where three SPS gene family members were cloned and identified. The amino acid sequences, conservative elements, encoded protein characteristics, protein structure, evolutionary relationships, and spatio-temporal expression of RuSPS1, RuSPS2 and RuSPS1 genes were analyzed using bioinformatics and quantitative real-time polymerase chain reaction (qRT-PCR). Correlation analysis was conducted between gene expressions and phosphate synthase activity.【Result】The multiple amino acid sequence comparisons showed that the blackberry SPS protein has two conserved domains and two relatively conserved unique protein phosphate sites to the plant SPS family. Evolutionary analysis showed that the three RuSPS genes were divided into two subfamilies, RuSPS1 and RuSPS3 belong to subfamily A RuSPS2 becong to subfamily B. The conserved elements analysis showed that the RuSPS1 and RuSPS3 genes exhibited fragment deletions to different degrees, with the exception of the basic conserved elements of RuSPS2. The different characteristics of blackberry SPS genes were revealed with other families using sequence analysis and comparison of blackberry SPS genes. The qRT-PCR analysis showed that RuSPS were expressed in all organs. RuSPS1 expression in leaves and fruits was higher than in flowers, RuSPS2 expression was high in mature fruits although much lower in other tissues, while RuSPS3 was highly expressed in all organs. These results indicate that SPS genes had tissue-specific expression in blackberry. Additionally, the expression of three RuSPS genes had increased during fruit and leaf development. Correlation analysis showed that the expression of RuSPS2 showed a significant positive correlation with leaf SPS activity (P < 0.05), and the expression of RuSPS1 was significantly negatively correlated (P < 0.05) with fruit SPS activity.【Conclusion】 There is a close relationship among RuSPS genes, sucrose synthesis and metabolism during blackberry fruit development. This is indicative of the involvement of blackberry SPS genes in the regulation of blackberry growth and development. It is inferred that the change of SPS enzyme activity in leaves was regulated by RuSPS2, while changes to SPS activity in fruits was regulated by RuSPS1. These results lay a foundation for in-depth analysis of the function of the blackberry SPS family genes in future.

Key words

blackberry / fruit development / sucrose phosphate synthase gene (SPS) / expression analysis

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YAN Zhixiang , YANG Haiyan , FAN Sufan , et al . Analysis of the expression of sucrose phosphate synthase genes during the development of blackberry fruit[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2022, 46(1): 179-186 https://doi.org/10.12302/j.issn.1000-2006.202007067

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