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

doi:10.12302/j.issn.1000-2006.202007067

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (1): 179-186.doi: 10.12302/j.issn.1000-2006.202007067

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

YAN Zhixiang¹(), YANG Haiyan²^,^*(), FAN Sufan², WU Wenlong², LYU Lianfei², LI Weilin¹^,^*()

1. Co-Innovation Center for the Sustainable Forstry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
2. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China

Received:2020-07-30 Accepted:2021-06-04 Online:2022-01-30 Published:2022-02-09
Contact: YANG Haiyan,LI Weilin E-mail:1096019134@qq.com;haiyanyang_025@126.com;wlli@njfu.edu.cn

Abstract

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

CLC Number:

S66
Q786

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

Figures/Tables 8

Table 1

Table 2

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References 23

[1]	甘四明. 林木分子育种研究的基因组学信息资源述评[J]. 南京林业大学学报(自然科学版), 2020, 44(4):1-11.
	GAN S M. A review on genomics information resources available for molecular breeding studies in forest trees[J]. J Nanjing For Univ (Nat Sci Ed), 2020, 44(4):1-11.DOI: 10.3969/j.issn.1000-2006.202005036. doi: 10.3969/j.issn.1000-2006.202005036
[2]	柴松琳, 钟洋敏, 程远, 等. 辣椒蔗糖磷酸合成酶和磷酸蔗糖磷酸酶基因家族鉴定及表达[J]. 分子植物育种, 2021, 19(13):4259-4267.
	CHAI S L, ZHONG Y M, CHENG Y, et al. Genome wide identification and expression of SPS and SPP genes in pepper[J]. Mol Plant Breed, 2021, 19(13):4259-4267.DOI: 10.13271/j.mpb.019.004259. doi: 10.13271/j.mpb.019.004259
[3]	HUBER S C, HUBER J L. Role and regulation of sucrose-phosphate synthase in higher plants[J]. Annu Rev Plant Physiol Plant Mol Biol, 1996, 47:431-444.DOI: 10.1146/annurev.arplant.47.1.431. doi: 10.1146/annurev.arplant.47.1.431
[4]	黄东亮, 李双喜, 廖青, 等. 植物蔗糖磷酸合成酶研究进展[J]. 中国生物工程杂志, 2012, 32(6):109-119.
	HUANG D L, LI S X, LIAO Q, et al. Advances on sucrose phosphate synthase in plants[J]. China Biotechnol, 2012, 32(6):109-119.DOI: 10.13523/j.cb.20120618. doi: 10.13523/j.cb.20120618
[5]	LANGENKÄMPER G, FUNG R W, NEWCOMB R D, et al. Sucrose phosphate synthase genes in plants belong to three different families[J]. J Mol Evol, 2002, 54(3):322-332.DOI: 10.1007/s00239-001-0047-4. doi: 10.1007/s00239-001-0047-4
[6]	GROF C P L, SO C T E, PERROUX J M, et al. Research Note:The five families of sucrose-phosphate synthase genes in Saccharum spp.are differentially expressed in leaves and stem[J]. Funct Plant Biol, 2006, 33(6):605.DOI: 10.1071/fp05283. doi: 10.1071/fp05283
[7]	SUN J, ZHANG J, LARUE C T, et al. Decrease in leaf sucrose synjournal leads to increased leaf starch turnover and decreased RuBP regeneration-limited photosynjournal but not Rubisco-limited photosynjournal in Arabidopsis null mutants of SPSA1[J]. Plant Cell Environ, 2011, 34(4):592-604.DOI: 10.1111/j.1365-3040.2010.02265.x. doi: 10.1111/j.1365-3040.2010.02265.x
[8]	JIANG J, ZHANG Z, CAO J. Pollen wall development: the associated enzymes and metabolic pathways[J]. Plant Biol (Stuttg), 2013, 15(2):249-263.DOI: 10.1111/j.1438-8677.2012.00706.x. doi: 10.1111/j.1438-8677.2012.00706.x
[9]	LIN I W, SOSSO D, CHEN L Q, et al. Nectar secretion requires sucrose phosphate synthases and the sugar transporter SWEET9[J]. Nature, 2014, 508(7497):546-549.DOI: 10.1038/nature13082. doi: 10.1038/nature13082
[10]	CHEN S, HAJIREZAEI M, BÖRNKE F. Differential expression of sucrose-phosphate synthase isoenzymes in tobacco reflects their functional specialization during dark-governed starch mobilization in source leaves[J]. Plant Physiol, 2005, 139(3):1163-1174.DOI: 10.1104/pp.105.069468. doi: 10.1104/pp.105.069468
[11]	WANG D, ZHAO J, HU B, et al. Identification and expression profile analysis of the sucrose phosphate synthase gene family in Litchi chinensis Sonn[J]. PeerJ, 2018, 6:e4379.DOI: 10.7717/peerj.4379. doi: 10.7717/peerj.4379
[12]	YU XY, WANG X F, FAN J D, et al. Cloning and characterization of a sucrose phosphate synthase-encoding gene from muskmelon[J]. J Amer Soc Hort Sci, 2007, 132(4):557-562.DOI: 10.21273/jashs.132.4.557. doi: 10.21273/jashs.132.4.557
[13]	VERMA A K, UPADHYAY S K, VERMA P C, et al. Functional analysis of sucrose phosphate synthase (SPS) and sucrose synthase (SS) in sugarcane (Saccharum) cultivars[J]. Plant Biol, 2011, 13(2):325-332.DOI: 10.1111/j.1438-8677.2010.00379.x. doi: 10.1111/j.1438-8677.2010.00379.x
[14]	吴文龙, 顾姻. 新经济植物黑莓的引种[J]. 植物资源与环境, 1994, 3(3):45-48.
	WU W L, GU Y. Blackberry introduction and cultivation[J]. J Plant Resour Environ, 1994, 3(3):45-48.
[15]	张春红, 熊振豪, 吴文龙, 等. 黑莓果实发育成熟期木质素合成CAD酶及其基因表达[J]. 南京林业大学学报(自然科学版), 2019, 43(1):141-148.
	ZHANG C H, XIONG Z H, WU W L, et al. CAD enzyme activity and gene expression in connection with lignin synjournal during fruit development and ripening process of blackberry[J]. J Nanjing For Univ (Nat Sci Ed), 2019, 43(1):141-148.DOI: 10.3969/j.issn.1000-2006.201805004. doi: 10.3969/j.issn.1000-2006.201805004
[16]	CASTLEDEN C K, AOKI N, GILLESPIE V J, et al. Evolution and function of the sucrose-phosphate synthase gene families in wheat and other grasses[J]. Plant Physiol, 2004, 135(3):1753-1764.DOI: 10.1104/pp.104.042457. doi: 10.1104/pp.104.042457
[17]	CHUA T K, BUJNICKI J M, TAN T C, et al. The structure of sucrose phosphate synthase from Halothermothrix orenii reveals its mechanism of action and binding mode[J]. Plant Cell, 2008, 20(4):1059-1072.DOI: 10.1105/tpc.107.051193. doi: 10.1105/tpc.107.051193
[18]	GROF C L, SO C T E, PERROUX J M, et al. Research note:the five families of sucrose-phosphate synthase genes in Saccharum spp.are differentially expressed in leaves and stem[J]. Funct Plant Biol, 2006, 33(6):605.DOI: 10.1071/fp05283. doi: 10.1071/fp05283
[19]	WINTER H, HUBER S C. Regulation of sucrose metabolism in higher plants:localization and regulation of activity of key enzymes[J]. Crit Rev Biochem Mol Biol, 2000, 35(4):253-289.DOI: 10.1080/10409230008984165. doi: 10.1080/10409230008984165
[20]	SZOPA J. Transgenic 14-3-3 isoforms in plants:the metabolite profiling of repressed 14-3-3 protein synjournal in transgenic potato plants[J]. Biochem Soc Trans, 2002, 30(4):405-410.DOI: 10.1042/bst0300405. doi: 10.1042/bst0300405
[21]	OKAMURA M, AOKI N, HIROSE T, et al. Tissue specificity and diurnal change in gene expression of the sucrose phosphate synthase gene family in rice[J]. Plant Sci, 2011, 181(2):159-166.DOI: 10.1016/j.plantsci.2011.04.019. doi: 10.1016/j.plantsci.2011.04.019
[22]	HUANG D L, QIN C X, GUI Y Y, et al. Role of the SPS gene families in the regulation of sucrose accumulation in sugarcane[J]. Sugar Tech, 2017, 19(2):117-124.DOI: 10.1007/s12355-016-0454-x. doi: 10.1007/s12355-016-0454-x
[23]	CHEN S, HAJIREZAEI M, BÖRNKE F. Differential expression of sucrose-phosphate synthase isoenzymes in tobacco reflects their functional specialization during dark-governed starch mobilization in source leaves[J]. Plant Physiol, 2005, 139(3):1163-1174.DOI: 10.1104/pp.105.069468. doi: 10.1104/pp.105.069468

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引物 primers	序列(5'-3') sequences (5'-3')	预期片段/bp predicted length	退火温度/℃ annealing temperature
SPS1-F	CCACAGGAGAGGAACACCAGATTG	214	59.4
SPS1-R	CACCATGAGCCGATACATCAGTCA	214	59.4
SPS2-F	CGACGGCGATTGAGGAACAGA	233	58.7
SPS2-R	CCACTCCAACTGCTTCTTCTTACG	233	58.7
SPS3-F	GGTGACTCTGCCGCTCTTCTATC	166	59.0
SPS3-R	GCTCCTCTGCTTCTATCCTTCTCA	166	59.0
Actin-F	GGTGTCAGCCATACAGTCCC	783	64.0
Actin-R	CAGTGAGGTCACGACCAGCA	783	64.0

基因 gene	基因长度/bp gene length	开放阅读框长度/bp length in ORF	蛋白长度/aa protein length	分子质量/ ku molecular weight	等电点 IP
RuSPS1	3 030	3 027	1 009	113.53	6.16
RuSPS2	3 207	3 204	1 068	119.97	6.00
RuSPS3	2 622	2 553	851	93.98	5.96

基因 gene	果实蔗糖磷酸合成酶活 sucrose phosphate synthase activity in fruits				叶片蔗糖磷酸合成酶活 sucrose phosphate synthase activity in leaves
基因 gene	21 d	27 d	33 d	39 d	21 d	27 d	33 d	39 d
RuSPS1	-0.016^*	-0.530^*	-0.228^**	-0.381^*	-0.406^**	0.230^*	0.250^*	-0.135^*
RuSPS2	0.066	-0.268^*	-0.696^**	-0.085	0.186^**	0.023^*	0.021^**	0.159^*
RuSPS3	-0.197	-0.322^*	0.137	-0.189	0.097	0.131	0.329^*	-0.370^*

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

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