Mechanism of PpMADS2 and PpMADS3 Synergistically Regulating Carotenoids Accumulation in Peach Fruit

doi:10.16420/j.issn.0513-353x.2022-0221

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (6): 1173-1186.doi: 10.16420/j.issn.0513-353x.2022-0221

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Next Articles

Mechanism of PpMADS2 and PpMADS3 Synergistically Regulating Carotenoids Accumulation in Peach Fruit

XIAO Xiang¹^,², ZHOU Chujiang¹^,², JIN Shuwan², SHI Liyu², YANG Zhenfeng², CAO Shifeng², CHEN Wei²^,^*()

¹College of Food Science and Technology，Shanghai Ocean University，Shanghai 201306，China
²College of Biological and Environmental Sciences，Zhejiang Wanli University，Ningbo，Zhejiang 315100，China

Received:2023-01-10 Revised:2023-03-15 Online:2023-06-25 Published:2023-06-26
Contact: * （E-mail：vivianchanyee@zwu.edu.cn）

Abstract

Abstract:

To explore the regulation of MADS-box family transcription factors on carotenoid metabolism in yellow fleshed peach, two MADS genes named as PpMADS2（PRUPE_5G208500）and PpMADS3（PRUPE_5G208400）were cloned. The open reading frames of PpMADS2 and PpMADS3 were 768 bp and 756 bp，encoding 255 and 251 amino acids, respectively. Bioinformatics analysis revealed that the N-terminus of both proteins contained typical characteristic domains of MADS-box family. Subcellular localization showed that both PpMADS2 and PpMADS3 were located at the nucleus. The results showed the gene expression levels of PpMADS2，PpMADS3 as well as carotenoid biosynthetic genes such as PpPSY and PpCHYB increased during fruit ripening，which was consistent with the carotenoid accumulation in yellow fleshed peach. Furthermore, the results demonstrated that PpMADS2 could interact with PpMADS3 then synergistically activate the promoters of PpPSY and PpCHYB genes. Therefore，it is speculated that PpMADS2 and PpMADS3 promoted the accumulation of carotenoids in yellow fleshed peach fruit cooperatively through transcriptional activation of PpPSY and PpCHYB. These research provided a theoretical basis for further exploring the regulatory mechanism of MADS-box transcription factors on carotenoid accumulation in peach fruit.

Key words: peach, yellow flesh, carotenoids, MADS-box, protein interaction, transcriptional activation

CLC Number:

S662.1

XIAO Xiang, ZHOU Chujiang, JIN Shuwan, SHI Liyu, YANG Zhenfeng, CAO Shifeng, CHEN Wei. Mechanism of PpMADS2 and PpMADS3 Synergistically Regulating Carotenoids Accumulation in Peach Fruit[J]. Acta Horticulturae Sinica, 2023, 50(6): 1173-1186.

Figures/Tables 14

References 26

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用途 Function	名称 Name	基因ID Gene ID	引物序列（5′-3′） Primer sequence
高保真PCR High fidelity PCR	gPpMADS2	PRUPE_5G208500	F：ATGGGGAGGGGAAGGGTGCAGCTGAAGAG R：CTATTCATTAAGGTGGCGGAGCATCCATGGG
	gPpMADS3	PRUPE_5G208400	F：ATGGGAAGAGGTAGAGTTGAGCTGAAGAGGAT R：TCAAAGCATCCACCCAGGAATGAATCCAT
荧光定量PCR qRT-PCR	PpMADS2	PRUPE_5G208500	F：TGGCAAAGAAGGTAAAGGAGAAG R：GAAGGGTGGAGGAGCAGTC
	PpMADS3	PRUPE_5G208400	F：CCAGAACAAGGAACAAATGC R：AGTGAGGATGATGAGGAAGG
	PpPSY	PRUPE_3G013200	F：TATTATGTTGCTGGGACTG R：GTGTTTGTGAGCTGATTCG
	PpCHYB	PRUPE_2G300800	F：AGGTTGCTGCTGCTCATC R：CGCTTTGATTCTCCTTTC
	PpPDS	PRUPE_1G174100	F：CCGTTGAAGGTCGTGATTG R：CTTTCCGCCCAGAACATC
	PpLCYB	PRUPE_7G046100	F：CGTGGCACAGCAAGTCTCAG R：CAGGTAGTGTCCAGGCAATCAAG
	PpZEP	PRUPE_7G133100	F：AGAAGCAGAACAAGAAGTG R：AACCAGTCCTCCAATTCC
	PpZDS	PRUPE_6G340000	F：AGAAGCAGAACAAGAAGTG R：AACCAGTCCTCCAATTCC
载体构建 Carrier construction	PpMADS2-GFP	PRUPE_5G208500	F：ACGGGGGACTCTTGACCATGGCCATGGGGAGGGGAAGGGTG R：GCCCTTGCTCACCATACTAGTTTCATTAAGGTGGCGGAGCA
	PpMADS3-GFP	PRUPE_5G208400	F：ACGGGGGACTCTTGACCATGGCCATGGGAAGAGGTAGAGTTGAGCTG R：GCCCTTGCTCACCATACTAGTAAGCATCCACCCAGGAATGA
	PpMADS2-AD	PRUPE_5G208500	F：GCCATGGAGGCCAGTGAATTCATGGGGAGGGGAAGGGTG R：CAGCTCGAGCTCGATGGATCCCTATTCATTAAGGTGGCGGAGC
	PpMADS3-AD	PRUPE_5G208400	F：GCCATGGAGGCCAGTGAATTCATGGGAAGAGGTAGAGTTGAGCTG R：CAGCTCGAGCTCGATGGATCCTCAAAGCATCCACCCAGGAA
	PpMADS2-BD	PRUPE_5G208500	F：ATGGCCATGGAGGCCGAATTCATGGGGAGGGGAAGGGTG R：CTAGTTATGCGGCCGCTGCAGCTATTCATTAAGGTGGCGGAGC
	PpMADS3-BD	PRUPE_5G208400	F：ATGGCCATGGAGGCCGAATTCATGGGAAGAGGTAGAGTTGAGCTG R：CTAGTTATGCGGCCGCTGCAGTCAAAGCATCCACCCAGGAA
	PpMADS2-62-sk	PRUPE_5G208500	F：CGCTCTAGAACTAGTGGATCCATGGGGAGGGGAAGGGTG R：GTCGACGGTATCGATAAGCTTCTATTCATTAAGGTGGCGGAGC
	PpMADS3-62-sk	PRUPE_5G208400	F：CGCTCTAGAACTAGTGGATCCATGGGAAGAGGTAGAGTTGAGCTG R：GTCGACGGTATCGATAAGCTTTCAAAGCATCCACCCAGGAA
载体构建（启动子克隆） Carrier construction	PpPSYpro-LUC	PRUPE_3G013200	F：ATCCCGGGTTGCTCAGGATCTGCCACGTGTA R：ATCCATGGGTTGTTTGAACTTGAAACCTCAGCC
（Promoter cloning）	PpCHYBpro-LUC	PRUPE_2G300800	F：GCGGATCCTCTGGAAGTGAAGGAATTGTCGTAC R：GCCCATGGAAAAGATGCCTGGTTTGTGGAC

用途 Function	名称 Name	基因ID Gene ID	引物序列（5′-3′） Primer sequence
高保真PCR High fidelity PCR	gPpMADS2	PRUPE_5G208500	F：ATGGGGAGGGGAAGGGTGCAGCTGAAGAG R：CTATTCATTAAGGTGGCGGAGCATCCATGGG
	gPpMADS3	PRUPE_5G208400	F：ATGGGAAGAGGTAGAGTTGAGCTGAAGAGGAT R：TCAAAGCATCCACCCAGGAATGAATCCAT
荧光定量PCR qRT-PCR	PpMADS2	PRUPE_5G208500	F：TGGCAAAGAAGGTAAAGGAGAAG R：GAAGGGTGGAGGAGCAGTC
	PpMADS3	PRUPE_5G208400	F：CCAGAACAAGGAACAAATGC R：AGTGAGGATGATGAGGAAGG
	PpPSY	PRUPE_3G013200	F：TATTATGTTGCTGGGACTG R：GTGTTTGTGAGCTGATTCG
	PpCHYB	PRUPE_2G300800	F：AGGTTGCTGCTGCTCATC R：CGCTTTGATTCTCCTTTC
	PpPDS	PRUPE_1G174100	F：CCGTTGAAGGTCGTGATTG R：CTTTCCGCCCAGAACATC
	PpLCYB	PRUPE_7G046100	F：CGTGGCACAGCAAGTCTCAG R：CAGGTAGTGTCCAGGCAATCAAG
	PpZEP	PRUPE_7G133100	F：AGAAGCAGAACAAGAAGTG R：AACCAGTCCTCCAATTCC
	PpZDS	PRUPE_6G340000	F：AGAAGCAGAACAAGAAGTG R：AACCAGTCCTCCAATTCC
载体构建 Carrier construction	PpMADS2-GFP	PRUPE_5G208500	F：ACGGGGGACTCTTGACCATGGCCATGGGGAGGGGAAGGGTG R：GCCCTTGCTCACCATACTAGTTTCATTAAGGTGGCGGAGCA
	PpMADS3-GFP	PRUPE_5G208400	F：ACGGGGGACTCTTGACCATGGCCATGGGAAGAGGTAGAGTTGAGCTG R：GCCCTTGCTCACCATACTAGTAAGCATCCACCCAGGAATGA
	PpMADS2-AD	PRUPE_5G208500	F：GCCATGGAGGCCAGTGAATTCATGGGGAGGGGAAGGGTG R：CAGCTCGAGCTCGATGGATCCCTATTCATTAAGGTGGCGGAGC
	PpMADS3-AD	PRUPE_5G208400	F：GCCATGGAGGCCAGTGAATTCATGGGAAGAGGTAGAGTTGAGCTG R：CAGCTCGAGCTCGATGGATCCTCAAAGCATCCACCCAGGAA
	PpMADS2-BD	PRUPE_5G208500	F：ATGGCCATGGAGGCCGAATTCATGGGGAGGGGAAGGGTG R：CTAGTTATGCGGCCGCTGCAGCTATTCATTAAGGTGGCGGAGC
	PpMADS3-BD	PRUPE_5G208400	F：ATGGCCATGGAGGCCGAATTCATGGGAAGAGGTAGAGTTGAGCTG R：CTAGTTATGCGGCCGCTGCAGTCAAAGCATCCACCCAGGAA
	PpMADS2-62-sk	PRUPE_5G208500	F：CGCTCTAGAACTAGTGGATCCATGGGGAGGGGAAGGGTG R：GTCGACGGTATCGATAAGCTTCTATTCATTAAGGTGGCGGAGC
	PpMADS3-62-sk	PRUPE_5G208400	F：CGCTCTAGAACTAGTGGATCCATGGGAAGAGGTAGAGTTGAGCTG R：GTCGACGGTATCGATAAGCTTTCAAAGCATCCACCCAGGAA
载体构建（启动子克隆） Carrier construction	PpPSYpro-LUC	PRUPE_3G013200	F：ATCCCGGGTTGCTCAGGATCTGCCACGTGTA R：ATCCATGGGTTGTTTGAACTTGAAACCTCAGCC
（Promoter cloning）	PpCHYBpro-LUC	PRUPE_2G300800	F：GCGGATCCTCTGGAAGTGAAGGAATTGTCGTAC R：GCCCATGGAAAAGATGCCTGGTTTGTGGAC

名称 Name	基因ID Gene ID	FPKM
名称 Name	基因ID Gene ID	第2次膨大期 Second stage of enlargement（S3）	完熟期 Ripening stage（S5）
PpMADS2	PRUPE_5G208500	38.45 ± 1.14 b	63.50 ± 2.05 a
PpMADS3	PRUPE_5G208400	51.14 ± 1.33 b	127.24 ± 2.17 a

名称 Name	基因ID Gene ID	FPKM
名称 Name	基因ID Gene ID	第2次膨大期 Second stage of enlargement（S3）	完熟期 Ripening stage（S5）
PpMADS2	PRUPE_5G208500	38.45 ± 1.14 b	63.50 ± 2.05 a
PpMADS3	PRUPE_5G208400	51.14 ± 1.33 b	127.24 ± 2.17 a

Mechanism of PpMADS2 and PpMADS3 Synergistically Regulating Carotenoids Accumulation in Peach Fruit

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