The Malus baccata Receptor Like Protein MbRLP7 Negatively Regulates the Resistance of Pyrus betulifolia Bunge to Valsa Canker

doi:10.16420/j.issn.0513-353x.2022-0938

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (11): 2489-2498.doi: 10.16420/j.issn.0513-353x.2022-0938

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The Malus baccata Receptor Like Protein MbRLP7 Negatively Regulates the Resistance of Pyrus betulifolia Bunge to Valsa Canker

JIANG Daji¹, WEN Youfu², YU Hongqiang¹, GUO Zhigang⁵, ZUO Longgang¹, SUN E¹, LI Yanyan¹, ZUO Cunwu¹^,³^,^*(), ZHAO Junying⁴^,^*()

¹ College of Horticulture，Gansu Agricultural University，Lanzhou 730070，China
² Forestry and Grassland Bureau of Jingning County，Jingning，Gansu 743400，China
³ State Key Laboratory of Aridland Crop Science，Lanzhou 730070，China
⁴ Gansu Academy of Forestry，Lanzhou 730020，China
⁵ Tianshui Normal University，Tianshui，Gansu 741001，China

Received:2023-05-04 Revised:2023-08-17 Online:2023-11-25 Published:2023-11-28
Contact: ZUO Cunwu, ZHAO Junying

Abstract

Abstract:

Based on bioinformatics analysis，functional verification，and expression measurement，the role of the receptor-like protein MbRLP7 in Valsa pyri resistance，and its potential regulatory mechanisms were explored. The results showed that MbRLP7 had the highest sequence similarity with AtRLP6 and AtRLP7，with 43.0% and 42.8%，respectively. There are several cis-elements in the promoter region of MbRLP7 in response to stress-related signals and Valsa pyri signals. The MbRLP7 was transformed into suspension cells of Pyrus betulifoliaBunge，and three overexpressed cell lines were obtained. After inoculation with the pathogen Valsa pyri（Vp），the growth rate of mycelia on the overexpressing cells was significantly higher than that on the wild-type cells. After Valsa pyri metabolites treatment，the cell activity of all overexpressed cell lines was significantly lower than that of wild-type. In addition，the overexpression of gene MbRLP7 enhanced the induction of Valsa pyrimetabolites on PAMP-triggered immunity（PTI），reactive oxygen species related genes and pathogenesis-related protein PR1. In conclusion，MbRLP7 negatively regulates the resistance of Pyrus betulifoliaBunge to Valsa pyri，and PTI，reactive oxygen species and PR1 are involved in this regulatory process.

Key words: Pyrus betulifolia Bunge, receptor-like protein, Valsa pyri, PAMP-triggered immunity, ROS

JIANG Daji, WEN Youfu, YU Hongqiang, GUO Zhigang, ZUO Longgang, SUN E, LI Yanyan, ZUO Cunwu, ZHAO Junying. The Malus baccata Receptor Like Protein MbRLP7 Negatively Regulates the Resistance of Pyrus betulifolia Bunge to Valsa Canker[J]. Acta Horticulturae Sinica, 2023, 50(11): 2489-2498.

Figures/Tables 7

Table 1 Primers for PCR and quantitative real-time PCR

用途 Application	基因 Gene	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse prime
PCR	MbRLP7	CTGGCGCGCCATGAAAACTTTACAATCTTTTGGCC	TCCCTAGGTCAACGTCTCCACCTCTGATTTATG
qRT-PCR	MbRLP7	AACACATCTCGGGTGGCATT	AGCCGTGCAATAACCCTGAA
	PbeWRKY22	CATATCCAAGGGGATATTACAGATG	GTGACTATAAAAATATTCGGGTCGG
	PbeFRK1	TCGAGTCCAAAAAACGACAGTTTAC	TTGTCAAGTTTCTGTGATGAACTCT
	PbeOXI1	CAACTTGGAAAACTCCGAGAAGTC	TTGTCAGTATCCGATAAAACGTCTC
	TaNOX	TTGGAGTTGTTCGATGCGTTG	TCTTGTCCACCATGTCGAAGAAG
	PbePR1	AATCTTGTTCATTTTGGTGGGCC	AACAACCTGAGTATAATGCCCACAC
	PbePR4	GGTGAGAATTGTTGATCAGTGCAG	GTAGTTGACAATAAGGTGGCCTTG
	PbePR5	GACACACCCCAAGACTACCTCAAG	GTCACCAGTGCTCATGGCAAG

Table 2 Analysis of the main cis-elements in promoter region of MbRLP7

顺式元件名称 cis-Element name	序列 Sequence	位置 Site	响应信号 Signal of response
TC-rich repeats	ATTCTCTAAC	-2 699	防御和胁迫Defense and stress
CCAAT-box	CAACGG	-2 459	转录因子MYBv1结合位点MYBv1 binding site
CGTCA-motif	CGTCA	-2 414	茉莉酸甲酯MeJA
O2-site	GATGA(C/T)(A/G)TG(A/G)	-2 065	玉米素Zeatin
GARE-motif	TCTGTTG	-1 026	赤霉素GA
ABRE	ACGTG	-1 003	脱落酸ABA
TGA-element	AACGAC	-119	生长素IAA

Fig. 1 FPKM value（A）of MbRLP7 transcriptome data and qRT-PCR expression level（B）of MbRLP7 in Malus baccata suspension cells treated with Valsa pyri metabolites * P < 0.05；** P < 0.01. The same below.

Fig. 2 Overexpression of MbRLP7 promotes the spread of Valsa pyri on the Pyrus betulifolia Bunge cells surface A is the relative expression of MbRLP7 in wild type（WT）and overexpressed lines. B is the inoculation of cell surface Valsa pyri（Vp） and MTT staining test. C is the colony diameter statistics of Valsa pyri（Vp）.

Fig. 3 Analysis of cell viability of wild type（WT）and MbRLP7-OE7 after Valsa pyri metabolites（VpM）treatment

Fig. 4 Changes of active oxygen content of Pyrus betulifolia Bunge wild type（WT）and MbRLP7-OE7 after Valsa pyri metabolites treatment The relative fluorescence ratio represents the relative ROS level.

Fig. 5 Changes of defense gene expression in Pyrus betulifolia Bunge wild type（WT）and MbRLP7-OE7 overexpression line after Valsa pyri metabolites treatment

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The Malus baccata Receptor Like Protein MbRLP7 Negatively Regulates the Resistance of Pyrus betulifolia Bunge to Valsa Canker

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