Sun Zhongfeng, Song Jiabin, Xin Xi'an, Xie Xianan, Zhao Bin
State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, China.
Front Microbiol. 2018 Mar 5;9:91. doi: 10.3389/fmicb.2018.00091. eCollection 2018.
Arbuscular mycorrhizal (AM) fungi are soil-borne fungi belonging to the ancient phylum Glomeromycota and are important symbionts of the arbuscular mycorrhiza, enhancing plant nutrient acquisition and resistance to various abiotic stresses. In contrast to their significant physiological implications, the molecular basis involved is poorly understood, largely due to their obligate biotrophism and complicated genetics. Here, we identify and characterize three genes termed , and that encode 14-3-3-like proteins in the AM fungi and , respectively. The transcriptional levels of , and are strongly induced in the pre-symbiotic and symbiotic phases, including germinating spores, intraradical hyphae- and arbuscules-enriched roots. To functionally characterize the , and genes, we took advantage of a yeast heterologous system owing to the lack of AM fungal transformation systems. Our data suggest that all three genes can restore the lethal double mutant on galactose-containing media. Importantly, yeast one-hybrid analysis suggests that the transcription factor RiMsn2 is able to recognize the STRE (CCCCT/AGGGG) element present in the promoter region of gene. More importantly, Host-Induced Gene Silencing of both and in impairs the arbuscule formation in AM symbiosis and inhibits the expression of symbiotic and genes from plant and fungal partners, respectively. We further subjected the AM fungus- association system to drought or salinity stress. Accordingly, the expression profiles in both mycorrhizal roots and extraradical hyphae reveal that these three 14-3-3-like genes are involved in response to drought or salinity stress. Collectively, our results provide new insights into molecular functions of the AM fungal 14-3-3 proteins in abiotic stress responses and arbuscule formation during AM symbiosis.
丛枝菌根(AM)真菌是属于古老的球囊菌门的土壤传播真菌,是丛枝菌根的重要共生体,可增强植物对养分的获取以及对各种非生物胁迫的抗性。与其重要的生理意义形成对比的是,其中涉及的分子基础却知之甚少,这主要是由于它们的专性活体营养特性和复杂的遗传学。在此,我们分别在AM真菌和中鉴定并表征了三个名为、和的基因,它们编码类14-3-3蛋白。、和的转录水平在共生前期和共生阶段强烈诱导,包括萌发的孢子、富含根内菌丝和丛枝的根。为了从功能上表征、和基因,由于缺乏AM真菌转化系统,我们利用了酵母异源系统。我们的数据表明,所有这三个基因都可以在含半乳糖的培养基上恢复致死性双突变体。重要的是,酵母单杂交分析表明,转录因子RiMsn2能够识别基因启动子区域中存在的STRE(CCCCT/AGGGG)元件。更重要的是,在中对和进行宿主诱导的基因沉默会损害AM共生中的丛枝形成,并分别抑制来自植物和真菌伙伴的共生和基因的表达。我们进一步使AM真菌-共生系统遭受干旱或盐胁迫。相应地,菌根根和根外菌丝中的表达谱表明,这三个类14-3-3基因参与了对干旱或盐胁迫的响应。总体而言,我们的结果为AM真菌14-3-3蛋白在非生物胁迫响应和AM共生期间丛枝形成中的分子功能提供了新见解。