The Apple Receptor Like Kinase Gene MdLYK1 Positively Regulates Valsa Canker Resistance

doi:10.16420/j.issn.0513-353x.2022-0922

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (11): 2499-2508.doi: 10.16420/j.issn.0513-353x.2022-0922

• Plant Protection • Previous Articles Next Articles

The Apple Receptor Like Kinase Gene MdLYK1 Positively Regulates Valsa Canker Resistance

LI Yanyan¹, GUO Shoujun², CAI Minrui¹, SUN E¹, YU Hongqiang¹, JIANG Daji¹, ZUO Cunwu¹^,³^,^*(), WANG Hai¹^,^*()

¹ College of Horticulture，Gansu Agricultural University，Lanzhou 730070，China
² Gansu Yasheng Industrial（Group） Co.，Ltd. Tiaoshan Agricultural and Industrial Development Branch，Jingtai，Gansu 730400，China
³ State Key Laboratory of Aridland Crop Science，Lanzhou 730070，China

Received:2023-04-23 Revised:2023-08-17 Online:2023-11-25 Published:2023-11-28
Contact: ZUO Cunwu, WANG Hai

Abstract

Abstract:

The effect and potential mechanism of receptor like kinase gene MdLYK1（MD09G1111800）on the resistance to Valsa canker were investigated by bioinformatics analysis，functional verification and expression analysis. The results showed that MdLYK1 was homologous to Arabidopsis receptor kinase gene AtLYK1，with 68.1% amino acid sequence similarity. The transient expression of this gene in‘Yanfu 6’fruit significantly improved the fruit Valsa canker resistance. Three overexpressed cell lines were obtained by introducing them into the suspension cells of Pyrus betulifolia Bunge. After inoculation with Valsa pyri，the growth rate of colonies on all overexpressing cell lines was significantly lower than that of wild-type cells. All overexpression cell lines were significantly less sensitive to Valsa pyri metabolites compared to the wild type. In addition，MdLYK1 overexpression significantly enhanced the up-regulated expression of key genes related to signaling such as pathogen-associated molecular patterns triggered immunity，reactive oxygen species，jasmonic acid and other signal-related key genes were up-regulated in the suspension cells of Pyrus betulifolia Bunge. In conclusion，MdLYK1 positively regulates the resistance to apple and pear Valsa canker，and pathogen-associated molecular pattern-triggered immunity，reactive oxygen species and jasmonic acid signals are involved in the regulation of resistance.

Key words: apple, Valsa canker, receptor-like kinase gene, LysM-RLK, pattern recognition receptor

LI Yanyan, GUO Shoujun, CAI Minrui, SUN E, YU Hongqiang, JIANG Daji, ZUO Cunwu, WANG Hai. The Apple Receptor Like Kinase Gene MdLYK1 Positively Regulates Valsa Canker Resistance[J]. Acta Horticulturae Sinica, 2023, 50(11): 2499-2508.

Figures/Tables 6

Table 1 Primers for PCR and quantitative real-time PCR

用途 Usage	基因 Gene	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer
RT-PCR	MdLYK1	CTGGCGCGCCATGGGATTTCGAATCGGGTTAGGGTTCC	TCTCTAGACTGCCTTCCTGACATCAGATTGACAA
qRT-PCR	MdLYK1	TTGAGGAGGCTTTATTTTCAATTTT	GTATCACCTGAGCTCGACTTCAAA
	PbeWRKY22	CATATCCAAGGGGATATTACAGATG	GTGACTATAAAAATATTCGGGTCGG
	PbeFRK1	TCGAGTCCAAAAAACGACAGTTTAC	TTGTCAAGTTTCTGTGATGAACTCT
	PbeOXI1	CAACTTGGAAAACTCCGAGAAGTC	TTGTCAGTATCCGATAAAACGTCTC
	PbeTaNOX	TTGGAGTTGTTCGATGCGTTG	TCTTGTCCACCATGTCGAAGAAG
	PbePR1	AATCTTGTTCATTTTGGTGGGCC	AACAACCTGAGTATAATGCCCACAC
	PbePR4	GGTGAGAATTGTTGATCAGTGCAG	GTAGTTGACAATAAGGTGGCCTTG
	PbePDF1.2	GTCAGGGTTTTGTGCAAACACC	TGAAGTAGCAGAAGCAAGCAACTC
	PbeLOX1	GCTTATGTGGCTGTAAATGACTCTG	GAGGATGCAGAAGTTTGTAAATTGG

Fig. 1 Evolutionary characteristics and tissue specific expressional patterns of apple MdLYK1 The black triangle in A represents the genes contained in this branch of the evolutionary tree. The numbers on the left side of each branch are bootstrap values used to evaluate the reliability of the branch. Ruler represents the unit length of the difference value between sequences. Os：Oryza sativa；At：Arabidopsis thaliana；Sl：Solanum lycopersicum.

Fig. 2 MdLYK1 positive regulate the resistance of apple fruits to Valsa mali *P < 0.05；** P < 0.01. The same below.

Fig. 3 MdLYK1 positive regulates the resistance of Pyrus betulifolia Bunge suspension cells to Valsa pyri

Fig. 4 Activity of suspension cells of wild-type（WT）and MdLYK1 overexpressing strains（OE11）of Pyrus betulifolia Bunge under different concentrations of Valsa pyri metabolites（VpM）treatment

Fig. 5 Expression of defense genes in Pyrus betulifolia Bunge MdLYK1 overexpressing cells（OE11） and wild-type（WT）after 20% Valsa pyri metabolites（VpM）treatment

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The Apple Receptor Like Kinase Gene MdLYK1 Positively Regulates Valsa Canker Resistance

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