Studies on the Resistance of Apple MdMYB116 to Powdery Mildew

doi:10.16420/j.issn.0513-353x.2021-0060

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (11): 2133-2145.doi: 10.16420/j.issn.0513-353x.2021-0060

• Original article • Previous Articles Next Articles

Studies on the Resistance of Apple MdMYB116 to Powdery Mildew

JI Miaomiao¹, WAN Ye¹, ZHANG Yiping¹, MA Hai¹, WANG Xiping¹^,²^,^*(), GAO Hua¹^,²^,^*()

¹College of Horticulture,State Key Laboratory of Crop Stress Biology in Arid Areas,Northwest A & F University,Yangling,Shaanxi 712100,China
²Key Laboratory of Horticultural Crop Biology and Germplasm Development in Northwest China,Ministry of Agriculture and Rural Affairs,Yangling,Shaanxi 712100,China

Received:2021-05-21 Revised:2021-07-20 Published:2021-12-02
Contact: WANG Xiping,GAO Hua E-mail:wangxiping@nwsuaf.edu.cn;gaohua2378@163.com

Abstract

Abstract:

The full-length cDNA sequence of MdMYB116 was cloned from the powdery mildew resistant‘Gala’apple. The open reading frame of this gene was 900 bp,encoded 299 amino acids and contained two conserved domains. The phylogenetic tree results showed that MdMYB116 shared close relationship with PbMYB108-like in pear,with a identity of 93.38%. Subcellular localization assay evidenced that MdMYB116 was targeted to the nucleus. The expression level of MdMYB116 increased gradually from 6 h to 12 h and it was induced to a peak at 12 h after inoculation with powdery mildew. Yeast transcriptional activation verification revealed that the full length of MdMYB116 exhibited self-activating activity. MdMYB116 was transformed into Arabidopsis thaliana heterologously,and transgenic plant was obtained. After being inoculated with powdery mildew,it was found that the resistance of the transgenic lines was significantly higher than that of the wild type and pad4 mutant plants. Histochemical staining assays displayed that compared with wild type and pad4 mutant plants,the transgenic lines had a higher level of reactive oxygen species（ROS）burst,and more dead cells and calluses accumulation,indicating that the transgenic lines had a serious allergic lethal situation,which proved that MdMYB116 participated in the plant^’s powdery mildew immunomodulatory response. Expression analysis of some disease related genes speculated that MdMYB116 may enhanced defense responses though SA and/or MeJA signal pathways.

Key words: apple, MdMYB116, powdery mildew, functional analysis

CLC Number:

S661.1

JI Miaomiao, WAN Ye, ZHANG Yiping, MA Hai, WANG Xiping, GAO Hua. Studies on the Resistance of Apple MdMYB116 to Powdery Mildew[J]. Acta Horticulturae Sinica, 2021, 48(11): 2133-2145.

Figures/Tables 11

References 40

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引物用途	引物名称	序列（5′-3′）
Useage	Primer name	Sequence
基因克隆 Gene cloning	MdMYB116-Clone	F:ATGTCGACTAAGACTAAAACCCTAA
基因克隆 Gene cloning	MdMYB116-Clone	R:CTACATTTCGTCCGTGTTCCATATG
过表达/亚细胞定位 Overexpression/ subcellular localization	MdMYB116-OE	F:GGTACCATGTCGACTAAGACTAAAACCCTAA
过表达/亚细胞定位 Overexpression/ subcellular localization	MdMYB116-OE	R:GGATCCCATTTCGTCCGTGTTCCATATG
酵母双杂交 Yeast two-hybrid	MdMYB116-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-BD	R:CCGCTGCAGGTCGACGGATCCCTACATTTCGTCCGTGTTCCATATG
	MdMYB116-D1-BD	F:ATGGCCATGGAGGCCGAATTCTGGACTCTTGAAGAAGACTCTCTTC
	MdMYB116-D1-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D2-BD	F:ATGGCCATGGAGGCCGAATTCTGGTCGAAAATCGCGCAATA
	MdMYB116-D2-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D3-BD	F:ATGGCCATGGAGGCCGAATTCTGGATGCCAAGGTTGCAGCA
	MdMYB116-D3-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D4-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-D4-BD	R:CCGCTGCAGGTCGACGGATCCCGATATTGTGGCTGTAAAATGTGGG
	MdMYB116-D5-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-D5-BD	R:CCGCTGCAGGTCGACGGATCCTTTCTGCACCCGTGTTCTCCAGTAG
实时荧光定量分析 qRT-PCR	MdMYB116-RT	F:TGATAAGGTGCTATTGGATGC
	MdMYB116-RT	R:TGTAAAATGTGGGAAGGTGTT
	AtICS1	F:CTTCCGTGACCTTGATCCTTTCT
	AtICS1	R:CAGCGATCTTGCCATTAGGATC
	AtPR1	F:AACTACGCTGCGAACACGTG
	AtPR1	R:TCACTTTGGCACATCCGAGTC
	AtLOX3	F:TCTCCGTACAACAAGCGTTGG
	AtLOX3	R:GCGTCCGTCTAGCGCATTAAT
	AtMYC2	F:TCATACGACGGTTGCCAGAA
	AtMYC2	R:AGCAACGTTTACAAGCTTTGATTG
内参基因 Reference gene	AtActin1	F:TCAATCCAGGAGATGTTTAGG
	AtActin1	R:ACTGCTGGTACTCTGCGACA
	MdTubulin	F:AGGATGCTACAGCCGATGAG
	MdTubulin	R:GCCGAAGAACTGACGAGAATC

引物用途	引物名称	序列（5′-3′）
Useage	Primer name	Sequence
基因克隆 Gene cloning	MdMYB116-Clone	F:ATGTCGACTAAGACTAAAACCCTAA
基因克隆 Gene cloning	MdMYB116-Clone	R:CTACATTTCGTCCGTGTTCCATATG
过表达/亚细胞定位 Overexpression/ subcellular localization	MdMYB116-OE	F:GGTACCATGTCGACTAAGACTAAAACCCTAA
过表达/亚细胞定位 Overexpression/ subcellular localization	MdMYB116-OE	R:GGATCCCATTTCGTCCGTGTTCCATATG
酵母双杂交 Yeast two-hybrid	MdMYB116-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-BD	R:CCGCTGCAGGTCGACGGATCCCTACATTTCGTCCGTGTTCCATATG
	MdMYB116-D1-BD	F:ATGGCCATGGAGGCCGAATTCTGGACTCTTGAAGAAGACTCTCTTC
	MdMYB116-D1-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D2-BD	F:ATGGCCATGGAGGCCGAATTCTGGTCGAAAATCGCGCAATA
	MdMYB116-D2-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D3-BD	F:ATGGCCATGGAGGCCGAATTCTGGATGCCAAGGTTGCAGCA
	MdMYB116-D3-BD	R:CCGCTGCAGGTCGACGGATCCCATTTCGTCCGTGTTCCATATGCTA
	MdMYB116-D4-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-D4-BD	R:CCGCTGCAGGTCGACGGATCCCGATATTGTGGCTGTAAAATGTGGG
	MdMYB116-D5-BD	F:ATGGCCATGGAGGCCGAATTCATGTCGACTAAGACTAAAACCCTAA
	MdMYB116-D5-BD	R:CCGCTGCAGGTCGACGGATCCTTTCTGCACCCGTGTTCTCCAGTAG
实时荧光定量分析 qRT-PCR	MdMYB116-RT	F:TGATAAGGTGCTATTGGATGC
	MdMYB116-RT	R:TGTAAAATGTGGGAAGGTGTT
	AtICS1	F:CTTCCGTGACCTTGATCCTTTCT
	AtICS1	R:CAGCGATCTTGCCATTAGGATC
	AtPR1	F:AACTACGCTGCGAACACGTG
	AtPR1	R:TCACTTTGGCACATCCGAGTC
	AtLOX3	F:TCTCCGTACAACAAGCGTTGG
	AtLOX3	R:GCGTCCGTCTAGCGCATTAAT
	AtMYC2	F:TCATACGACGGTTGCCAGAA
	AtMYC2	R:AGCAACGTTTACAAGCTTTGATTG
内参基因 Reference gene	AtActin1	F:TCAATCCAGGAGATGTTTAGG
	AtActin1	R:ACTGCTGGTACTCTGCGACA
	MdTubulin	F:AGGATGCTACAGCCGATGAG
	MdTubulin	R:GCCGAAGAACTGACGAGAATC

Studies on the Resistance of Apple MdMYB116 to Powdery Mildew

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