Cloning and Functional Analysis of MdWRKY74 in Apple

doi:10.16420/j.issn.0513-353x.2021-0244

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (3): 482-492.doi: 10.16420/j.issn.0513-353x.2021-0244

• Research Papers • Previous Articles Next Articles

Cloning and Functional Analysis of MdWRKY74 in Apple

XIANG Li¹, ZHAO Lei¹, WANG Mei¹, LÜ Yi², WANG Yanfang³, SHEN Xiang¹, CHEN Xuesen¹, YIN Chengmiao¹^,^**(), MAO Zhiquan¹^,^**()

¹State Key Laboratory of Crop Biology,College of Horticulture Science and Engineering,Shandong Agricultural University,Tai’an,Shandong 271018,China
²Weihai Acedemy of Agricultural Sciences,Weihai,Shandong 264200,China
³College of Chemistry and Material Science,Shandong Agricultural University,Tai’an,Shandong 271018,China

Received:2021-04-26 Revised:2021-05-10 Online:2022-03-25 Published:2022-03-25
Contact: YIN Chengmiao,MAO Zhiquan E-mail:yinchengmiao@163.com;mzhiquan@sdau.edu.cn

Abstract

Abstract:

In this study,a WRKY transcription factor gene,named MdWRKY74（XM_029090147.1）was obtained from apple rootstock M9T337（Malling 9 NAKBT337）. The open reading frame of MdWRKY74 was 939 bp,which encoded 312 amino acids,and contained a typical WRKY domain. Amino acid sequence alignment and phylogenetic tree analysis showed that MdWRKY74 had the highest homology with Pyrus bretschneideri WRKY74. Subcellular localization assays showed that MdWRKY74 was located in the nucleus. Quantitative real-time PCR analysis showed that MdWRKY74 was expressed in different apple tissues. Overexpression of MdWRKY74 in Arabidopsis could improve the salt resistance of plants,and promote the expression of SOS1 and NHX1. The results showed that MdWRKY74 can be induced by salt stress,so it may participate in the regulation of salt resistance in apple.

Key words: apple, MdWRKY74, subcellular localization, salt stress, function analysis

CLC Number:

S661.1

XIANG Li, ZHAO Lei, WANG Mei, LÜ Yi, WANG Yanfang, SHEN Xiang, CHEN Xuesen, YIN Chengmiao, MAO Zhiquan. Cloning and Functional Analysis of MdWRKY74 in Apple[J]. Acta Horticulturae Sinica, 2022, 49(3): 482-492.

Figures/Tables 10

References 47

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用途 Use	引物名称 Primer name	序列（5′-3′） Sequence
CDs	MdWRKY74	F：ATGGAGGAGGTTGAGGCAGC;R：CCAGGCCGATTGTGAATGC
qRT-PCR	MdWRKY74	F：GGAAGAGGAAGAGTGAGA;R：GGAAAGTAGGAGGGAGTAA
qRT-PCR	MdActin	F：TGACCGAATGAGCAAGGAAATTACT;R：TACTCAGCTTTGGCAATCCACATC
qRT-PCR	AtSOS1	F：GGAAGAGGAAGAGTGAGA;R：GGAAAGTAGGAGGGAGTAA
qRT-PCR	AtNHX1	F：TCTTGCTATTGGTGCCATAT;R：AGGTGTCTCGTCTTGATTC
qRT-PCR	AtActin	F：CGCTCTTTCTTTCCAAGCTC;R：AACAGCCCTGGGAGCATC
亚细胞定位 Subcellular localization	MdWRKY74	F：CAAGCTTGCATGCCTGCAGGTCGACATGGAGGAGGTTGAGGCAGC; R：CTCGCCCTTGCTCACCATGGATCCCCAGGCCGATTGTGAATGC

用途 Use	引物名称 Primer name	序列（5′-3′） Sequence
CDs	MdWRKY74	F：ATGGAGGAGGTTGAGGCAGC;R：CCAGGCCGATTGTGAATGC
qRT-PCR	MdWRKY74	F：GGAAGAGGAAGAGTGAGA;R：GGAAAGTAGGAGGGAGTAA
qRT-PCR	MdActin	F：TGACCGAATGAGCAAGGAAATTACT;R：TACTCAGCTTTGGCAATCCACATC
qRT-PCR	AtSOS1	F：GGAAGAGGAAGAGTGAGA;R：GGAAAGTAGGAGGGAGTAA
qRT-PCR	AtNHX1	F：TCTTGCTATTGGTGCCATAT;R：AGGTGTCTCGTCTTGATTC
qRT-PCR	AtActin	F：CGCTCTTTCTTTCCAAGCTC;R：AACAGCCCTGGGAGCATC
亚细胞定位 Subcellular localization	MdWRKY74	F：CAAGCTTGCATGCCTGCAGGTCGACATGGAGGAGGTTGAGGCAGC; R：CTCGCCCTTGCTCACCATGGATCCCCAGGCCGATTGTGAATGC

作用元件 Functional element	序列 Sequence	位点功能 Function of site	位置 Location
ABRE	ACGTG	参与ABA响应 cis-acting element involved in the abscisic acid responsiveness	+ 1 530
CGTCA-motif	CGTCA	参与MeJA响应 cis-acting regulatory element involved in the MeJA-responsiveness	-215
LTR	CCGAAA	低温响应 cis-acting element involved in low-temperature responsiveness	+ 485
MBS	CAACTG	与干旱诱导相关的MYB绑定位点 MYB binding site involved in drought-inducibility	-175
TC-rich repeats	ATTTTCTTCA	参与防御反应和胁迫响应 cis-acting element involved in defense and stress responsiveness	+ 927

作用元件 Functional element	序列 Sequence	位点功能 Function of site	位置 Location
ABRE	ACGTG	参与ABA响应 cis-acting element involved in the abscisic acid responsiveness	+ 1 530
CGTCA-motif	CGTCA	参与MeJA响应 cis-acting regulatory element involved in the MeJA-responsiveness	-215
LTR	CCGAAA	低温响应 cis-acting element involved in low-temperature responsiveness	+ 485
MBS	CAACTG	与干旱诱导相关的MYB绑定位点 MYB binding site involved in drought-inducibility	-175
TC-rich repeats	ATTTTCTTCA	参与防御反应和胁迫响应 cis-acting element involved in defense and stress responsiveness	+ 927

Cloning and Functional Analysis of MdWRKY74 in Apple

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