Liu Chao, Huang Renwei, Wang Lingli, Liang Guolu
College of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Sichuan Provincial Key Laboratory for Development and Utilization of Characteristic Horticultural Biological Resources, College of Chemistry and Life Sciences, Chengdu Normal University, Chengdu, China.
Front Plant Sci. 2021 Jan 12;11:612055. doi: 10.3389/fpls.2020.612055. eCollection 2020.
Although several results have been obtained in triploid loquat heterosis (i.e., leaf size of triploid loquat) studies in the past years, the underlying mechanisms of the heterosis are still largely unknown, especially the regulation effects of one specific gene on the corresponding morphology heterosis. In this study, we sought to further illustrate the regulatory mechanisms of one specific gene on the leaf size heterosis of triploid loquats. A leaf size development-related gene () and its promoter were successfully cloned. Ectopic expression of in showed that the leaf size of transgenic plantlets was larger than that of WTs, and the transgenic plantlets had more leaves than WTs. Quantitative Reverse Transcription PCR (qRT-PCR) showed that the expression level of showed an AHP expression pattern in most of the hybrids, and this was consistent with our previous phenotype observations. Structure analysis of promoter showed that there were significantly more light-responsive elements than other elements. To further ascertain the regulatory mechanisms of on triploid loquat heterosis, the methylation levels of promoter in different ploidy loquats were analyzed by using bisulfite sequencing. Surprisingly, the total methylation levels of promoter in triploid showed a decreasing trend compared with the mid-parent value (MPV), and this was also consistent with the qRT-PCR results of . Taken together, our results suggested that played an important role in promoting leaf size development of loquat, and demethylation of promoter in triploid loquats caused to exhibit over-dominance expression pattern and then further to promote leaf heterosis formation. In conclusion, played an important role in the formation of triploid loquat leaf size heterosis.
尽管在过去几年的三倍体枇杷杂种优势(即三倍体枇杷的叶片大小)研究中已取得了一些成果,但杂种优势的潜在机制仍大多未知,尤其是一个特定基因对相应形态杂种优势的调控作用。在本研究中,我们试图进一步阐明一个特定基因对三倍体枇杷叶片大小杂种优势的调控机制。成功克隆了一个与叶片大小发育相关的基因()及其启动子。在中的异位表达表明,转基因植株的叶片大小比野生型大,且转基因植株的叶片比野生型多。定量逆转录聚合酶链反应(qRT-PCR)表明,在大多数杂种中,的表达水平呈现AHP表达模式,这与我们之前的表型观察结果一致。启动子的结构分析表明,光响应元件明显多于其他元件。为了进一步确定对三倍体枇杷杂种优势的调控机制,利用亚硫酸氢盐测序分析了不同倍性枇杷中启动子的甲基化水平。令人惊讶的是,三倍体中启动子的总甲基化水平与中亲值(MPV)相比呈下降趋势,这也与的qRT-PCR结果一致。综上所述,我们的结果表明在促进枇杷叶片大小发育中起重要作用,三倍体枇杷中启动子的去甲基化导致呈现超显性表达模式,进而进一步促进叶片杂种优势的形成。总之,在三倍体枇杷叶片大小杂种优势的形成中起重要作用。