Preliminary Study on Mechanisms of Resistance to Citrus Canker of Kumquat FcRGA1

doi:10.16420/j.issn.0513-353x.2021-0940

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (11): 2325-233.doi: 10.16420/j.issn.0513-353x.2021-0940

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Preliminary Study on Mechanisms of Resistance to Citrus Canker of Kumquat FcRGA1

DAI Wenshan, WU Yue, WANG Min()

National Navel Orange Engineering Research Center，College of Navel Orange，Gannan Normal University，Ganzhou，Jiangxi 341000，China

Received:2022-03-30 Revised:2022-08-25 Online:2022-11-25 Published:2022-11-25
Contact: WANG Min E-mail:wangmin0624@foxmail.com

Abstract

Abstract:

NBS-LRR resistant proteins are the most abundant R protein members in plants. This family contains typical NB-ARC domains and LRR domains，which are mainly involved in the defense process of plants against pathogen’s invasion. In this study，an R gene containing the NB-ARC domain was cloned from kumquat（Fortunella crassifolia）and named FcRGA1. This gene contained a complete open reading frame（ORF）of 3 954 bp and encoded 1 317 amino acids. qRT-PCR analysis showed that the FcRGA1 gene was expressed in the roots，stems and leaves of kumquat，with the highest expression level in the leaves and the lowest in the roots. Furthermore，the expression of FcRGA1 gene was up-regulated by infection with canker bacteria and reached the highest value at 12 h，which was 12.3 times higher than that of the control. After silencing the FcRGA1 gene by VIGS（Virus-induced Gene Silencing），kumquat leaves in the silenced group showed more severe chlorosis symptom than those in control group after inoculated by Xcc（Xanthomonas citri subsp. citri），concurrent with significantly higher lesion area，bacterial growth and relative electrolytic leakage. Whereas，the chlorophyll content and hydrogen peroxide content were significantly lower in comparison with the control group，indicating that FcRGA1 might affect the resistance of kumquat to citrus canker through the regulation of reactive oxygen species.

Key words: kumquat, citrus canker, NBS-LRR, FcRGA1, gene silencing, resistance mechanism

CLC Number:

S666.1

DAI Wenshan, WU Yue, WANG Min. Preliminary Study on Mechanisms of Resistance to Citrus Canker of Kumquat FcRGA1[J]. Acta Horticulturae Sinica, 2022, 49(11): 2325-233.

Figures/Tables 6

Fig. 1 Cloning and gene structure analysis of FcRGA1

Fig. 2 Amino acid sequences phylogenetic tree of FcRGA1 and Arabidopsis（At）R protein containing NB-ARC domain The abscissa is a distance scale，representing the unit length of difference between sequences，corresponding a distance of 10 changes per 100 amino acid positions，and the numbers at node represent the distance of the kinship.

Fig. 3 Expression pattern analysis of FcRAG1 Asterisks indicate significant difference levels，**P < 0.01.

Fig. 4 Expression analysis of the FcRGA1 in the FcRGA1-silencing kumquat leaves The gene expression at each time of the control was taken as 1 for normalization. Asterisks indicate significant difference levels，**P < 0.01.

Fig. 5 The phenotype（a），fluorescence observation（b）and bacterial count（c）of control and FcRGA1-silencing kumquat leaves inoculated with Xcc C：Control;R：FcRGA1-VIGS;dpi：Days post inoculation. Asterisks indicate significant difference between FcRGA1-VIGS and control at the same time point. *P < 0.05，**P < 0.01.

Fig. 6 Electrolyte leakage，chlorophyll content and H2O2 content of control and FcRGA1-silencing kumquat leaves after inoculated with Xcc Asterisks indicate significant difference levels，**P < 0.01.

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Preliminary Study on Mechanisms of Resistance to Citrus Canker of Kumquat FcRGA1

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