Antagonistic Peptide Technology for Functional Dissection of CLE Genes in Arabidopsis
作 者：Xiu-Fen Song、Peng Guo、Shi-Chao Ren、Ting-Ting Xu、Chun-Ming Liu
影响因子：6.535
刊物名称：Plant Physiology
出版年份：2013
卷： 期： 页码：DOI:10.1104/pp.112.211029
论文摘要：
In recent years, peptide hormones have been recognized as important signal molecules in plants. Genetic characterization of such peptides is challenging since they are usually encoded by small genes. As a proof of concept, we used the well-characterized stem cell-restricting CLV3 to develop an antagonistic peptide technology by transformations of wildtype Arabidopsis with constructs carrying the full-length CLV3 with every residues in the peptide-coding region replaced, one at a time, by Ala. Analyses of transgenic plants allowed us to identify one line exhibited a dominant-negative clv3-like phenotype, with enlarged shoot apical meristems and increased numbers of floral organs. We then performed second dimensional amino acid substitutions to replace the Gly residue individually with the other 18 possible proteinacious amino acids. Examination of transgenic plants showed that a Gly to Thr substitution gave the strongest antagonistic effect in the wildtype, in which over 70% of transgenic lines showed the clv3-like phenotype. Among these substitutions, a negative correlation was observed between the antagonistic effects in the wildtype and the complementation efficiencies in clv3. We also demonstrated that such an antagonistic peptide technology is applicable to other CLV3/ESR (CLE) genes, CLE8 and CLE22, as well as in vitro treatments. We believe this technology provides a powerful tool for functional dissection of widely occurring CLE genes in plants.
