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Cloning and Characterization of miRNAs and Their Targets, Including a Novel miRNA-Targeted nbs-lrr protein Class Gene in Apple （Golden Delicious）

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Molecular Plant 2014年第1期7卷 218-230页

作者： Chao Ma You Lu Songlin Bai Wennan Zhang Xuwei Duan Dong Meng Zhigang Wang Aide Wang Zongshan Zhou Tianzhong Li Laboratory of Fruit Cell and Molecular Breeding China Agricultural University Beijing 100193 China Research Institute of Pomology Chinese Academy of Agricultural Sciences Xingcheng Liaoning 125100 China College of Horticulture Shenyang Agricultural University Shenyang 110866 China

MicroRNA （miRNA） has emerged as an important regulator of gene expression in plants. 146 miRNAs were identified from apple （Malus domestica cv. Golden Delicious） by bioinformatic analysis and RNA library sequencing. From these, 135 were conserved and 11 were novel miRNAs. Target analysis predicted one of the novel miRNAs, Md-miRLn11 （Malus domestica microRNA Ln11）, targeted an apple nucleotide-binding site （nbs）-Ieucine-rich repeat （lrr） class protein coding gene （Md-nbs）. 5/ RACE assay confirmed the ability of Md-miRLn11 to cleave Md-nbs at the 11-12-nt position. Analysis of the expression of Md-miRLn11 and Md-nbs during the optimum invasion period in 40 apple varieties showed that the expression of Md-nbs gene in resistant varieties is higher than in susceptible varieties, with an inverse pattern for Md-miRLn11. Seedlings from the resistant apple variety ＇JiGuan＇ were used to carry out an Agrobacterium infiltration assay, and then inoculated with the apple leaf spot disease. The result showed a clear decline of disease resistance in JiGuan apples. In contrast, the susceptible variety ＇FuJi＇ infiltrated with the Md-nbs gene showed a significant increase in disease resistance. Based on the above results, we propose that Md-miRLn11 regulates Md-nbs gene expression in particular under the condition of pathogen infection, and that the Md-miRLn11 targeting P-loop site may regulate many nbs-lrr protein class genes in woody plants.

关键词： apple microRNA nbs-lrr protein bioinformatics analysis gene function analysis

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nbs-lrr proteins and Their Partners:Molecular Switches of Plant Defense

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Journal of Northeast Agricultural University(English Edition) 2008年第4期15卷 49-57页

作者： LIU Chunyan QIU Hongmei WANG Jialin WANG Jing CHEN Qingshan HU Guohua Crop Research and Breeding Center of Land-Reclamation Harbin 150090 China Soybean Research Institute Northeast Agricultural University Harbin 150030 China

Specificity of the plant innate immune system is often conferred by resistance（R）*** plant disease resistance （R）proteins contain a series of leucine-rich repeats（lrrs）,a nucleotide-binding site（nbs）,and a putative amino-terminal signaling *** are termed nbs-lrr *** lrrs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the nbs domain presumably functions as a molecular *** the past years,the most important discoveries are the role of partners in nbs-lrr gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within nbs- lrr proteins as a mode of signaling *** outcome of these interactions determines whether a plant activates its defense responses.

关键词： plant defense nbs-lrr protein intramolecular interaction intermolecular interactions

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