Journal of Integrative Agriculture ›› 2018, Vol. 17 ›› Issue (12): 2694-2703.DOI: 10.1016/S2095-3119(18)62009-X

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  • 收稿日期:2018-01-11 出版日期:2018-12-01 发布日期:2018-12-03

Molecular cloning and functional identification of an apple flagellin receptor MdFLS2 gene

QI Chen-hui1, ZHAO Xian-yan1, JIANG Han1, LIU Hai-tao2, WANG Yong-xu1, HU Da-gang1, HAO Yu-jin1   

  1. 1 National Key Laboratory of Crop Biology/Key Laboratory of Horticultural Crop Biology and Germplasm Innovation, Ministry of Agriculture and Rural Affairs of China/College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an 271018, P.R.China
    2 Shandong Yihui Detection Technology Co., Ltd., Tai’an 271000, P.R.China
  • Received:2018-01-11 Online:2018-12-01 Published:2018-12-03
  • Contact: CorrespondenceHUD a-gang, Tel/Fax: +86-538-8246151,E-mail:fap_296566@163.com; HAO Yu-jin, E-mail: haoyujin@sdau.edu.cn
  • About author:QI Chen-hui, E-mail: 18763823286@163.com;
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (31601728 and 31430074), the Ministry of Education of China (IRT15R42), the Natural Science Foundation of Shandong Province, China (ZR2016CQ13 and SDAIT-06-03), the Young Scientists Funds of Shandong Agricultural University, China (564024 and 24024).

Abstract:

The leucine-rich repeat receptor kinase flagellin-sensing 2 gene (MdFLS2; Gene ID: MDP0000254112) was cloned from Royal Gala apple (Malus×domestica Borkh.).  This gene contained a complete open reading frame of 3 474 bp that encoded 1 158 amino acids.  The phylogenetic tree indicated that Prunus persica FLS2 exhibited the highest sequence similarity to MdFLS2.  The PlantCare database suggests that the promoter sequence of MdFLS2 contains several typical cis-acting elements, including ethylene-, gibberellin-, salicylic acid-, and drought-responsive elements.  Quantitative real-time PCR analysis showed that MdFLS2 was widely expressed in the different tissues of the apple and most highly expressed in the leaves.  Furthermore, MdFLS2 was significantly induced by the flagellin elicitor peptide flg22.  Treatment of the apple seedling leaves with flg22 resulted in an increase in leaf callose levels with increased treatment duration.  An increase in the production of O2 along with the expression of disease-related genes was also observed.  An oxidative burst was detected in the treated seedlings, but not in the control seedlings, indicating that flg22 had stimulated the expression of the MdFLS2 gene and its downstream target genes.  Furthermore, the ectopic expression of MdFLS2 complemented the function of the Arabidopsis fls2 mutant and conferred enhanced flg22 tolerance to the transgenic Arabidopsis, suggesting that MdFLS2 acts as a positive regulator in the response to pathogens in apple.

Key words: apple ,  flagellin receptor ,  flagellin elicitor peptide ,  MdFLS2 ,  pathogen infection