Functional Analysis of Sucrose-phosphate Synthase Genes（SPS）in Sweet Cherry

doi:10.16420/j.issn.0513-353x.2020-0630

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (8): 1446-1456.doi: 10.16420/j.issn.0513-353x.2020-0630

• Research Papers • Previous Articles Next Articles

Functional Analysis of Sucrose-phosphate Synthase Genes（SPS）in Sweet Cherry

QI Xiliang, LIU Congli, SONG Lulu, LI Ming()

Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou 450009,China

Received:2020-12-10 Revised:2021-04-26 Online:2021-08-25 Published:2021-09-06
Contact: LI Ming E-mail:liming06@caas.cn

Abstract

Abstract:

In order to ascertain the function of sucrose-phosphate synthase genes（SPS）in the regulation of sweet cherry ripening,four full-length cDNA of sweet cherry SPS genes（PavSPSA1,PavSPSA2,PavSPSB,and PavSPSC）were cloned in sweet cherry genome. Gene expression analysis showed that the expression levels of PavSPSA1 was the highest,followed by PavSPSC,whereas PavSPSA2 and PavSPSB were relatively weakly expressed during the fruit growth and development,and ripening using quantitative real-time PCR. The expression levels of PavSPSA1 and PavSPSA2 coincided with fruit ripening and softening. Furthermore,we used TRV-mediated virus-induced gene silencing（VIGS）to evaluate the function of PavSPSA1,PavSPSA2,PavSPSB,and PavSPSC in the regulation of ripening. The results showed that silencing PavSPSA1 gene decreased sucrose content,delayed fruit coloring,ripening and softening. However,The fruit phenotypes for the PavSPSA2,PavSPSB,and PavSPSC-silenced fruits showed no visible defects when compared with the control fruits. Taken together,our findings determined that PavSPSA1 may be a key gene regulating sucrose synthesis and accumulation in sweet cherry fruit and play an indispensable role in the regulation of sweet cherry fruit ripening and softening.

Key words: sweet cherry, sucrose-phosphate synthase, PavSPSA1, gene expression, fruit ripening, VIGS

CLC Number:

S662.5

QI Xiliang, LIU Congli, SONG Lulu, LI Ming. Functional Analysis of Sucrose-phosphate Synthase Genes（SPS）in Sweet Cherry[J]. Acta Horticulturae Sinica, 2021, 48(8): 1446-1456.

Figures/Tables 8

References 29

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引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
PavSPSA1-F	ATGGCGAGCAACGATTGGATAAAC	PavSPSA1-J-F	CAGGTTCATGCCCCGCATGGTGA
PavSPSA1-R	CTACGTCTTGACAACTCCGAGTT	PavSPSA1-J-R	ATTGTGGATGCCTTGGTTTGC
PavSPSA2-F	ATGGCGGGAAACGACTGGGTGAAC	PavSPSA2-J-F	GAGGCAGCAGCTTATGGTCTACCT
PavSPSA2-R	CTACCGCTTGAGAATCCCTAGT	PavSPSA2-J-R	CTTGTCCTTCCCTGCTGCCTCA
PavSPSB-F	ATTTGGCATCTTACCCGTAAGAAG	PavSPSB-J-F	ATGAAGGAGGTGGAAATAAGCTGCT
PavSPSB-R	TCACATTCGAGCAGCAGATTTAGAG	PavSPSB-J-R	AGTACATTGGATGGCAACGCAG
PavSPSC-F	ATGGCGGGAAACGACTGGTTAAACAG	PavSPSC-J-F	GAGATTCTGATACAGGTGGTCAGGT
PavSPSA1-q-F	GCAAACCAAGGCATCCACAAT	PavSPSC-J-R	AGCTCCTCAGCCTCGATCCTCTTC
PavSPSA1-q-R	GAACCCTACAGAGCCTTCAGT	PaNCED1-F	CATGTCGGAGGACGACTTGCCGT
PavSPSA2-q-F	TGAGGCAGCAGGGAAGGACAAG	PaNCED1-R	GCGCCGTCTGGAGAGACGTGGA
PavSPSA2-q-R	AGCCCCAACGGACATACAGATACC	PaPG1-F	ATCACCTTCCGCATTGCTG
PavSPSB-q-F	GGCTTGCTTGTGGACCCTCAT	PaPG1-R	TCACCTTAATGTTGTTGGAG
PavSPSB-q-R	AGCAGCTTATTTCCACCTCCTTC	PaXYL1-F	ACAACTGGAACGGTGTCGAT
PavSPSC-q-F	ACCCCACCACCCGCTTTCA	PaXYL1-R	TCCTGGTGTAATGTTGCTCG
PavSPSC-q-R	GACCCTCGCCATTGCCTACG	PaPL-1-F	GATGGTCGCTTCTATGTTGTCA
PavSPSA2-RNAi-F	AGTAAGGTTACCGAATTCTGAGGCAGCAGGGAAGGACAAG	PaPL-1-R	TGAGATGGAGAGCTCCTCACC
PavSPSA2-RNAi-R	GAGCTCGGTACCGGTACCAGCCCCAACGGACATACAGATACC	PaPAL-F	GCCTCACCAGGCAACAAGAGCA
PavSPSB-RNAi-F	AGTAAGGTTACCGAATTCGGCTTGCTTGTGGACCCTCAT	PaPAL-R	TCTGGCCATCTGGTCCAACAGC
PavSPSB-RNAi-R	GAGCTCGGTACCGGTACCAGCAGCTTATTTCCACCTCCTTCAT	PaCHS-F	GTATGTGCGAGTACATGGCA
PavSPSC-RNAi-F	AGTAAGGTTACCGAATTCACCCCACCACCCGCTTTCA	PaCHS-R	GCTTAGTGAGCTGATAGTC
PavSPSC-RNAi-R	GAGCTCGGTACCGGTACCGACCCTCGCCATTGCCTACG	PaDFR-F	CTGCACCGGAGTGTTCCATGT
Histone2-F	GGTGTGCTTCCGCAGATAA	PaDFR-R	CTGGTGCTCTTCGACGTTCAC
Histone2-R	TCCTCCTTGGGTGGTGAAT	PaANS-F	ATCTCCGATGAGCTCATGG
		PaANS-R	CTCAATGTAATCAGCAGGTG

引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
PavSPSA1-F	ATGGCGAGCAACGATTGGATAAAC	PavSPSA1-J-F	CAGGTTCATGCCCCGCATGGTGA
PavSPSA1-R	CTACGTCTTGACAACTCCGAGTT	PavSPSA1-J-R	ATTGTGGATGCCTTGGTTTGC
PavSPSA2-F	ATGGCGGGAAACGACTGGGTGAAC	PavSPSA2-J-F	GAGGCAGCAGCTTATGGTCTACCT
PavSPSA2-R	CTACCGCTTGAGAATCCCTAGT	PavSPSA2-J-R	CTTGTCCTTCCCTGCTGCCTCA
PavSPSB-F	ATTTGGCATCTTACCCGTAAGAAG	PavSPSB-J-F	ATGAAGGAGGTGGAAATAAGCTGCT
PavSPSB-R	TCACATTCGAGCAGCAGATTTAGAG	PavSPSB-J-R	AGTACATTGGATGGCAACGCAG
PavSPSC-F	ATGGCGGGAAACGACTGGTTAAACAG	PavSPSC-J-F	GAGATTCTGATACAGGTGGTCAGGT
PavSPSA1-q-F	GCAAACCAAGGCATCCACAAT	PavSPSC-J-R	AGCTCCTCAGCCTCGATCCTCTTC
PavSPSA1-q-R	GAACCCTACAGAGCCTTCAGT	PaNCED1-F	CATGTCGGAGGACGACTTGCCGT
PavSPSA2-q-F	TGAGGCAGCAGGGAAGGACAAG	PaNCED1-R	GCGCCGTCTGGAGAGACGTGGA
PavSPSA2-q-R	AGCCCCAACGGACATACAGATACC	PaPG1-F	ATCACCTTCCGCATTGCTG
PavSPSB-q-F	GGCTTGCTTGTGGACCCTCAT	PaPG1-R	TCACCTTAATGTTGTTGGAG
PavSPSB-q-R	AGCAGCTTATTTCCACCTCCTTC	PaXYL1-F	ACAACTGGAACGGTGTCGAT
PavSPSC-q-F	ACCCCACCACCCGCTTTCA	PaXYL1-R	TCCTGGTGTAATGTTGCTCG
PavSPSC-q-R	GACCCTCGCCATTGCCTACG	PaPL-1-F	GATGGTCGCTTCTATGTTGTCA
PavSPSA2-RNAi-F	AGTAAGGTTACCGAATTCTGAGGCAGCAGGGAAGGACAAG	PaPL-1-R	TGAGATGGAGAGCTCCTCACC
PavSPSA2-RNAi-R	GAGCTCGGTACCGGTACCAGCCCCAACGGACATACAGATACC	PaPAL-F	GCCTCACCAGGCAACAAGAGCA
PavSPSB-RNAi-F	AGTAAGGTTACCGAATTCGGCTTGCTTGTGGACCCTCAT	PaPAL-R	TCTGGCCATCTGGTCCAACAGC
PavSPSB-RNAi-R	GAGCTCGGTACCGGTACCAGCAGCTTATTTCCACCTCCTTCAT	PaCHS-F	GTATGTGCGAGTACATGGCA
PavSPSC-RNAi-F	AGTAAGGTTACCGAATTCACCCCACCACCCGCTTTCA	PaCHS-R	GCTTAGTGAGCTGATAGTC
PavSPSC-RNAi-R	GAGCTCGGTACCGGTACCGACCCTCGCCATTGCCTACG	PaDFR-F	CTGCACCGGAGTGTTCCATGT
Histone2-F	GGTGTGCTTCCGCAGATAA	PaDFR-R	CTGGTGCTCTTCGACGTTCAC
Histone2-R	TCCTCCTTGGGTGGTGAAT	PaANS-F	ATCTCCGATGAGCTCATGG
		PaANS-R	CTCAATGTAATCAGCAGGTG

基因名称 Gene name	cDNA/ bp	氨基酸 Amino acid	GenBank登录号 ID in GenBank	染色体位置 Chromosome location	拟南芥同源基因 Homologous gene in Arabidopsis
基因名称 Gene name	cDNA/ bp	氨基酸 Amino acid	GenBank登录号 ID in GenBank	染色体位置 Chromosome location	TAIR位置 Locus in TAIR	氨基酸相似度/% Similarity of amino acid
PavSPSA1	3 174	1 057	XM_021953379.1	Chr7：18 650 177 ~ 18 655 575	AtSPS1F（At5g20280）	77.69
PavSPSA2	3 180	1 059	XM_021966658.1	Chr1：35 009 999 ~ 35 016 230	AtSPS2F（At5g11110）	71.36
PavSPSB	2 844	947	XM_021946233.1	chrUn：91 085 245 ~ 91 090 791	AtSPS3F（At1g04920）	67.35
PavSPSC	3 072	1 023	XM_021957268.1	Chr8：324 800 ~ 330 670	AtSPS4F（At4g10120）	72.10

基因名称 Gene name	cDNA/ bp	氨基酸 Amino acid	GenBank登录号 ID in GenBank	染色体位置 Chromosome location	拟南芥同源基因 Homologous gene in Arabidopsis
基因名称 Gene name	cDNA/ bp	氨基酸 Amino acid	GenBank登录号 ID in GenBank	染色体位置 Chromosome location	TAIR位置 Locus in TAIR	氨基酸相似度/% Similarity of amino acid
PavSPSA1	3 174	1 057	XM_021953379.1	Chr7：18 650 177 ~ 18 655 575	AtSPS1F（At5g20280）	77.69
PavSPSA2	3 180	1 059	XM_021966658.1	Chr1：35 009 999 ~ 35 016 230	AtSPS2F（At5g11110）	71.36
PavSPSB	2 844	947	XM_021946233.1	chrUn：91 085 245 ~ 91 090 791	AtSPS3F（At1g04920）	67.35
PavSPSC	3 072	1 023	XM_021957268.1	Chr8：324 800 ~ 330 670	AtSPS4F（At4g10120）	72.10

Functional Analysis of Sucrose-phosphate Synthase Genes（SPS）in Sweet Cherry

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