Identification and Expression Analysis of Spinach PSY Gene Family

doi:10.13560/j.cnki.biotech.bull.1985.2023-0392

Abstract

Abstract:

PSY（phytoene synthase, PSY）is the first rate-limiting enzyme in the plant carotenoid synthesis pathway and plays an important role in plant growth and development. In order to understand the function of PSY gene family in spinach, the number, structure, promoter elements, amino acid characteristics and gene evolution of PSY gene family members in spinach were identified by bioinformatics method, and the expression pattern of PSY genes in spinach tissue and in response to the changes in the ratio of red to blue light was analyzed by real-time fluorescence quantitative PCR（RT-qPCR）. Meanwhile, the expressions of R1B3（red∶blue light = 1∶3）and R3B1（red light∶blue light = 3∶1）and the correlation between SoPSYs gene expression and total carotenoid content were analyzed. The results showed that there were 4 PSY genes in spinach, containing SQS_PSY conserved domain, and the SoPSY protein was a hydrophilic protein. PSY1 and PSY2 of spinach were closely related to beet in Amaranthaceae, and PSY3 and PSY4 were closely related to PSY3 in tomato. There were various light-responsive and phytohormone-related elements in the promoter sequence of spinach PSY genes. SoPSY genes had tissue expression specificity, SoPSY1, SoPSY2, and SoPSY3 were only expressed in the leaves, and SoPSY4 was mainly expressed in the roots. Spinach carotene content decreased after treatment with R1B3 and R3B1. Among spinach genotypes, the expression patterns of each SoPSY gene in response to the ratio of red to blue light were different. The expression of SoPSY1 was not significantly changed in cultivated spinach, the expression patterns of SoPSY2 were reversed in both spinach genotypes, and the relative expression patterns of SoPSY3 were consistent in the wild and cultivated spinach, and the relative expressions of these three genes was closely related to the change of total carotenoid content. This study lays the foundation for an in-depth understanding of the biological function of the PSY genes in spinach and their regulatory mechanism in response to changes in the ratio of red to blue light.

Key words: PSY, Spinacia oleracea, identification of gene family, gene expression pattern, red and blue light, carotenoids, correlation analysis

REN Li, QIAO Shu-ting, GE Chen-hui, WEI Zi-tong, XU Chen-xi. Identification and Expression Analysis of Spinach PSY Gene Family[J]. Biotechnology Bulletin, 2023, 39(12): 169-178.

Figures/Tables 10

References 32

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引物名称 Primer name	上游引物 Upstream primer（5'-3'）	下游引物 Downstream primer（5'-3'）
18S	CTGAGAAACGGCTACCACA	CCCAAGGTCCAACTACGAG
SoPSY1	CGGTGTAGTGAAGTTTGTGCT	TGCCCAAATGGCTTTTCTCTTT
SoPSY2	CGGTGTAGTGAAGTTTGTGCTGA	TATAGCCCAAATGGCTTTTCGCTTC
SoPSY3	GCTCTTTCCTTGGGTACTGCT	GAGCTCGTCTTGTGGAAGGT
SoPSY4	ATGGCCGAACTTCGTTGATG	CCCTTATTGGATAGAGCAAGAAAGC

引物名称 Primer name	上游引物 Upstream primer（5'-3'）	下游引物 Downstream primer（5'-3'）
18S	CTGAGAAACGGCTACCACA	CCCAAGGTCCAACTACGAG
SoPSY1	CGGTGTAGTGAAGTTTGTGCT	TGCCCAAATGGCTTTTCTCTTT
SoPSY2	CGGTGTAGTGAAGTTTGTGCTGA	TATAGCCCAAATGGCTTTTCGCTTC
SoPSY3	GCTCTTTCCTTGGGTACTGCT	GAGCTCGTCTTGTGGAAGGT
SoPSY4	ATGGCCGAACTTCGTTGATG	CCCTTATTGGATAGAGCAAGAAAGC

基因名称 Gene name	基因ID Gene ID	染色体编号 Chrom No.	氨基酸长 Length/aa	分子质量 Molecular weight/kD	等电点 pI	重力值 GRAVY	亚细胞定位 Location
SoPSY1	Spo03700	SpoScf_03339	418	47.15	8.88	-0.262	Chloroplast
SoPSY2	Spo13982	SpoScf_02365	418	47.14	8.89	-0.262	Chloroplast
SoPSY3	Spo00660	chr4	359	41.48	9.17	-0.494	Chloroplast
SoPSY4	Spo15086	chr4	351	40.24	9.11	-0.363	Chloroplast

基因名称 Gene name	基因ID Gene ID	染色体编号 Chrom No.	氨基酸长 Length/aa	分子质量 Molecular weight/kD	等电点 pI	重力值 GRAVY	亚细胞定位 Location
SoPSY1	Spo03700	SpoScf_03339	418	47.15	8.88	-0.262	Chloroplast
SoPSY2	Spo13982	SpoScf_02365	418	47.14	8.89	-0.262	Chloroplast
SoPSY3	Spo00660	chr4	359	41.48	9.17	-0.494	Chloroplast
SoPSY4	Spo15086	chr4	351	40.24	9.11	-0.363	Chloroplast

处理 Treatment	含量变化量 Content variation	24 h基因相对表达量24 h relative expression				12 d基因相对表达量12 d relative expression
处理 Treatment	含量变化量 Content variation	SoPSY1	SoPSY2	SoPSY3	SoPSY4	SoPSY1	SoPSY2	SoPSY3	SoPSY4
CK	US362	-0.849	0.615	-0.102	-1.000**	0.999*	-0.593	0.745	-0.999*
CK	KS4	-0.362	-0.475	-0.104	-0.920	-0.754	-0.686	-0.348	-0.146
R1B3	US362	-0.058	0.749	0.673	0.570	-0.042	0.999*	0.953	0.765
R1B3	KS4	-0.476	-0.490	-0.479	-0.726	-0.978	-0.955	-0.011	0.011
R3B1	US362	-0.999*	-0.485	0.954	-0.485	-0.970	0.492	-0.970	-0.338
R3B1	KS4	0.128	-0.380	0.534	0.765	-0.697	0.260	-0.061	-0.156