SlMAPKKK43 Regulates Tomato Resistance to Gray Mold

doi:10.16420/j.issn.0513-353x.2023-0757

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (2): 309-320.doi: 10.16420/j.issn.0513-353x.2023-0757

• Genetic&Breeding?Germplasm Resources?Molecular Biology • Previous Articles Next Articles

SlMAPKKK43 Regulates Tomato Resistance to Gray Mold

DONG Xiaonan¹, LÜ Hongmei¹, ZHAO Liqun³, HE Bingqing⁴, ZHANG Jiaojiao¹, ZHAO Bing¹, GUO Yangdong¹^,²^,^*(), ZHANG Na¹^,²^,^*()

¹ Beijing Key Laboratory of Growth and Development Regulation of Protected Vegetables，Department of Vegetable，College of Horticulture，China Agricultural University，Beijing 100193，China
² Sanya Institute of China Agricultural University，Sanya，Hainan 572025，China
³ Beijing Agricultural Technology Extension Station，Beijing 100029，China
⁴ Beijing Changping District Agricultural Technology Extension Station，Beijing 102200，China

Received:2023-10-07 Revised:2024-01-07 Online:2024-02-25 Published:2024-02-26
Contact: GUO Yangdong, ZHANG Na

Abstract

Abstract:

The mitogen-activated protein kinase（MAPK）cascade plays an important role in multiple developmental and physiological processes in plants and in response to various biotic and abiotic stresses. In this study，SlMAPKKK43 overexpressed plants and CRISPR-Cas9-mediated SlMAPKKK43 knockout mutant using Agrobacterium infection genetic transformation were obtained. Through the inoculation of Botrytis cinerea with transgenic lines，we found that SlMAPKKK43 positively regulated the tolerance of tomato to Botrytis cinerea. In order to further elucidate the molecular mechanism of SlMAPKKK43 regulating the resistance of gray mold，the known gray mold regulators SlMKK2 and SlMKK4 were screened by pull down-MS technology，and in vitro phosphorylation assay was used to preliminarily verify that SlMAPKKK43 could phosphorylate SlMKK2 and SlMKK4.

Key words: tomato, Botrytis cinerea, MAPK cascade, SlMAPKKK43

DONG Xiaonan, LÜ Hongmei, ZHAO Liqun, HE Bingqing, ZHANG Jiaojiao, ZHAO Bing, GUO Yangdong, ZHANG Na. SlMAPKKK43 Regulates Tomato Resistance to Gray Mold[J]. Acta Horticulturae Sinica, 2024, 51(2): 309-320.

Figures/Tables 9

References 38

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用途 Function	引物名称 Primer name	序列（5′-3′） Sequence
扩增基因全长 Full length amplification	SlMAPKKK43	F：ATGGATAGTCCCACAATGACCAA R：TTTGACATTCATTGCCGCCG
构建过表达载体 Construct overexpression vector	1305-SlMAPKKK43	F：AGCTCGGTACCCGGGGATCCATGGATAGTCCCACAATGACCAA R：ATGGTCTTTGTAGTCAAGCTTTTTGACATTCATTGCCGCCG
构建基因编辑载体 Construct gene editing vector	PB-SlMAPKKK43	BsF：ATATATGGTCTCGATTGAAGTATCAGCAGCCTGACGGTT BsR：ATTATTGGTCTCGAAACGAAGAGTACAGTCGTCTTCCAA F0：TGAAGTATCAGCAGCCTGACGGTTTTAGAGCTAGAAATAGC R0：AACGAAGAGTACAGTCGTCTTCCAATCTCTTAGTCGACTCTAC
qRT-PCR	qSlMAPKKK43	F：CACATTCGAACTGGACGGGA R：CATGGGCTGATTTCCCTGGT
	qSlUbi3	F：TCCATCTCGTGCTCCGTCT R：CTGAACCTTTCCAGTGTCATCAA
	qSlEFα	F：GGAACTTGAGAAGGAGCCTAAG R：CAACACCAACAGCAACAGTCT
过表达植株鉴定 Identification of overexpressing lines	OE- SlMAPKKK43	F：GATGACGCACAATCCCACTATCC R：CCATCACCAGAATCCAACTTATCG
基因编辑植株鉴定 Identification of gene edited lines	CR- SlMAPKKK43	F1：ATGGATAGTCCCACAATGACCAA R1：TCCACATACCTTCTCTCATTTTCCC F2：ACTCTTGATGGTTGTCTCCACAA R2：TTTCCTGTATCGAGAAGGGAATGC

用途 Function	引物名称 Primer name	序列（5′-3′） Sequence
扩增基因全长 Full length amplification	SlMAPKKK43	F：ATGGATAGTCCCACAATGACCAA R：TTTGACATTCATTGCCGCCG
构建过表达载体 Construct overexpression vector	1305-SlMAPKKK43	F：AGCTCGGTACCCGGGGATCCATGGATAGTCCCACAATGACCAA R：ATGGTCTTTGTAGTCAAGCTTTTTGACATTCATTGCCGCCG
构建基因编辑载体 Construct gene editing vector	PB-SlMAPKKK43	BsF：ATATATGGTCTCGATTGAAGTATCAGCAGCCTGACGGTT BsR：ATTATTGGTCTCGAAACGAAGAGTACAGTCGTCTTCCAA F0：TGAAGTATCAGCAGCCTGACGGTTTTAGAGCTAGAAATAGC R0：AACGAAGAGTACAGTCGTCTTCCAATCTCTTAGTCGACTCTAC
qRT-PCR	qSlMAPKKK43	F：CACATTCGAACTGGACGGGA R：CATGGGCTGATTTCCCTGGT
	qSlUbi3	F：TCCATCTCGTGCTCCGTCT R：CTGAACCTTTCCAGTGTCATCAA
	qSlEFα	F：GGAACTTGAGAAGGAGCCTAAG R：CAACACCAACAGCAACAGTCT
过表达植株鉴定 Identification of overexpressing lines	OE- SlMAPKKK43	F：GATGACGCACAATCCCACTATCC R：CCATCACCAGAATCCAACTTATCG
基因编辑植株鉴定 Identification of gene edited lines	CR- SlMAPKKK43	F1：ATGGATAGTCCCACAATGACCAA R1：TCCACATACCTTCTCTCATTTTCCC F2：ACTCTTGATGGTTGTCTCCACAA R2：TTTCCTGTATCGAGAAGGGAATGC

基因ID Gene ID	基因注释 Description
Solyc04g071615	脱落酸应激成熟蛋白 Abscicic acid stress ripening protein 4
Solyc04g074410	油菜素类固醇调节基因 Protein EXORDIUM-like
Solyc03g123800	SlMKK2 Mitogen-activated protein kinase kinase 4/5
Solyc02g080670	L-抗坏血酸氧化酶同源物 L-ascorbate oxidase homolog
Solyc06g062370	磷酸酶 Acid phosphatase 1
Solyc03g097920	SlMKK4 SF344-mitogen-activated kinase kinase kinase
Solyc02g086880	脱氢酶 Formate dehydrogenase
Solyc01g091330	谷胱甘肽转移酶 Glutathione S-transferase zeta class-like isoform X1
Solyc01g007260	F-box 蛋白 MORE AXILLARY GROWTH2 Protein SMAX1-LIKE 3

基因ID Gene ID	基因注释 Description
Solyc04g071615	脱落酸应激成熟蛋白 Abscicic acid stress ripening protein 4
Solyc04g074410	油菜素类固醇调节基因 Protein EXORDIUM-like
Solyc03g123800	SlMKK2 Mitogen-activated protein kinase kinase 4/5
Solyc02g080670	L-抗坏血酸氧化酶同源物 L-ascorbate oxidase homolog
Solyc06g062370	磷酸酶 Acid phosphatase 1
Solyc03g097920	SlMKK4 SF344-mitogen-activated kinase kinase kinase
Solyc02g086880	脱氢酶 Formate dehydrogenase
Solyc01g091330	谷胱甘肽转移酶 Glutathione S-transferase zeta class-like isoform X1
Solyc01g007260	F-box 蛋白 MORE AXILLARY GROWTH2 Protein SMAX1-LIKE 3

SlMAPKKK43 Regulates Tomato Resistance to Gray Mold

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