Characteristics of the Transcription Factor BrWRKY57 and Its Regulation on BrPPH1 and BrNCED3

doi:10.16420/j.issn.0513-353x.2020-0432

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (3): 518-530.doi: 10.16420/j.issn.0513-353x.2020-0432

• Research Papers • Previous Articles Next Articles

Characteristics of the Transcription Factor BrWRKY57 and Its Regulation on BrPPH1 and BrNCED3

ZENG Zexiang¹, XIAO Xianmei¹, TAN Xiaoli¹, FAN Zhongqi²^,^*(), CHEN Jianye¹^,^*()

¹Engineering Research Center of Southern Horticultural Products Preservation,Ministry of Education/Guangdong Provincial Key Laboratory of Postharvest Science of Fruits and Vegetables,College of Horticulture,South China Agricultural University,Guangzhou 510642,China
²Key Laboratory of Postharvest Biology for Subtropical Special Agricultural Products of Fujian Province,College of Food Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China

Received:2020-07-17 Online:2021-03-25 Published:2021-04-02
Contact: FAN Zhongqi,CHEN Jianye E-mail:ffanzqi@163.com;chenjianye@scau.edu.cn

Abstract

Abstract:

In this study,a WRKY transcription factor termed BrWRKY57,was obtained from Chinese flowering cabbage. Amino acid sequence alignment and phylogenetic analysis revealed that BrWRKY57 contained one WRKY conserved domain,exhibited the highest homology with Arabidopsis thalianaAtWRKY57,and belonged to sub-group IIc. Real-time quantitative PCR analysis demonstrated that BrWRKY57 was up-regulated during leaf senescence of Chinese flowering cabbage,and its expression was significantly enhanced by abscisic acid（ABA）treatment. Subcellular localization and transcriptional activity analysis suggested that BrWRKY57 was a nuclear protein and had transcriptional activation activity. Moreover,dual-luciferase transient expression assay revealed that BrWRKY57 could activate the promoter activities of chlorophyll catabolic gene BrPPH1and ABA biosynthetic gene BrNCED3. These results indicate that BrWRKY57 might be associated with Chinese flowering cabbage leaf senescence via effecting the genes expression of chlorophyll degradation and ABA synthesis.

Key words: Chinese flowering cabbage, leaf senescence, abscisic acid, WRKY transcription factor, transcriptional regulation

CLC Number:

S634.5

ZENG Zexiang, XIAO Xianmei, TAN Xiaoli, FAN Zhongqi, CHEN Jianye. Characteristics of the Transcription Factor BrWRKY57 and Its Regulation on BrPPH1 and BrNCED3[J]. Acta Horticulturae Sinica, 2021, 48(3): 518-530.

Figures/Tables 10

References 32

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引物用途 Primer assay	引物名称 Primer name	引物序列（5′-3′） Primer sequence	限制性内切酶 Enzyme
全长扩增Full- length amplification	BrWRKY57	F:ATGAACGATCCTAAAGATCCCGAT; R:TCAAGGATTGCGCATGGTTTGAG
实时荧光定量PCR Real-time quantitative PCR	BrAct1 BrWRKY57 BrPPH BrNCED3	F:CGCTTAACCCGAAAGCTAAC;R:TACGCCCACTAGCGTAAAG F:ACGGAGGAGAAGAAGAAGGC;R:ACTCTCTTCTTCACCGTGCA F:ATCGAGTATGGGCGGTTGAT;R:CCTGCAATGTACACTGGCTC F:GTCCTCTGCGAGATTCGGCT;R:GGAGACTTTAGGCCACGGCT
亚细胞定位 Subcellular localization	BrWRKY57	F:ATCTAGAGCAGTCGACGGTACCATGAACGATCCTAAAGATCCCGAT R:CTCCTCGCCCTTGCTCACCATAGGATTGCGCATGGTTTGAGGTAC	KpnⅠ BamHⅠ
酵母体内转录活性分析Transcriptional activity in yeast cells	BrWRKY57-pGBKT7	F:CATGGAGGCCGAATTCATGAACGATCCTAAAGATCCCGAT R:GCCGCTGCAGGTCGACGTCAAGGATTGCGCATGGTTTGAG	EcoR Ⅰ SalⅠ
双荧光素酶瞬时表达 Dual-luciferase transient expression assay	62SK-BD-BrWRKY57 62SK-BrWRKY57 0800-BrPPH1 pro 0800-BrNCED3 pro	F:CGCCGTCTAGAACTAGTGGATCCATGAACGATCCTAAAGATCCCGAT R:TCGATAAGCTTGATATCGAATTCTCAAGGATTGCGCATGGTTTGAG F:GGCCGCTCTAGAACTAGTGGATCCATGAACGATCCTAAAGATCCCGAT R:ATCGATAAGCTTGATATCGAATTCTCAAGGATTGCGCATGGTTTGAGG F:TATAGGGCGAATTGGGTACCCTTTAGAGCATACTGAGGCTG R:TTGGCGTCTTCCATGGTACACAAATCAATCCTGTAAGGAAC F:TATAGGGCGAATTGGGTACCCTTAGAGACCAAGTCATATGATGTA R:TTGGCGTCTTCCATGGTTTAAATATGATTAGTCAGTATTGGTGC	BamHⅠ EcoRⅠ BamHⅠ EcoRⅠ KpnⅠ NcoⅠ KpnⅠ NcoⅠ

引物用途 Primer assay	引物名称 Primer name	引物序列（5′-3′） Primer sequence	限制性内切酶 Enzyme
全长扩增Full- length amplification	BrWRKY57	F:ATGAACGATCCTAAAGATCCCGAT; R:TCAAGGATTGCGCATGGTTTGAG
实时荧光定量PCR Real-time quantitative PCR	BrAct1 BrWRKY57 BrPPH BrNCED3	F:CGCTTAACCCGAAAGCTAAC;R:TACGCCCACTAGCGTAAAG F:ACGGAGGAGAAGAAGAAGGC;R:ACTCTCTTCTTCACCGTGCA F:ATCGAGTATGGGCGGTTGAT;R:CCTGCAATGTACACTGGCTC F:GTCCTCTGCGAGATTCGGCT;R:GGAGACTTTAGGCCACGGCT
亚细胞定位 Subcellular localization	BrWRKY57	F:ATCTAGAGCAGTCGACGGTACCATGAACGATCCTAAAGATCCCGAT R:CTCCTCGCCCTTGCTCACCATAGGATTGCGCATGGTTTGAGGTAC	KpnⅠ BamHⅠ
酵母体内转录活性分析Transcriptional activity in yeast cells	BrWRKY57-pGBKT7	F:CATGGAGGCCGAATTCATGAACGATCCTAAAGATCCCGAT R:GCCGCTGCAGGTCGACGTCAAGGATTGCGCATGGTTTGAG	EcoR Ⅰ SalⅠ
双荧光素酶瞬时表达 Dual-luciferase transient expression assay	62SK-BD-BrWRKY57 62SK-BrWRKY57 0800-BrPPH1 pro 0800-BrNCED3 pro	F:CGCCGTCTAGAACTAGTGGATCCATGAACGATCCTAAAGATCCCGAT R:TCGATAAGCTTGATATCGAATTCTCAAGGATTGCGCATGGTTTGAG F:GGCCGCTCTAGAACTAGTGGATCCATGAACGATCCTAAAGATCCCGAT R:ATCGATAAGCTTGATATCGAATTCTCAAGGATTGCGCATGGTTTGAGG F:TATAGGGCGAATTGGGTACCCTTTAGAGCATACTGAGGCTG R:TTGGCGTCTTCCATGGTACACAAATCAATCCTGTAAGGAAC F:TATAGGGCGAATTGGGTACCCTTAGAGACCAAGTCATATGATGTA R:TTGGCGTCTTCCATGGTTTAAATATGATTAGTCAGTATTGGTGC	BamHⅠ EcoRⅠ BamHⅠ EcoRⅠ KpnⅠ NcoⅠ KpnⅠ NcoⅠ

Characteristics of the Transcription Factor BrWRKY57 and Its Regulation on BrPPH1 and BrNCED3

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