Comprehensive genomic analysis and expression profiling of Argonaute gene family and examination of their regulatory roles in water-use efficiency and abiotic stress responses in apple

Argonaute proteins are key players in small RNA-guided gene-silencing, which plays an important role in plant stress tolerance. However, little is known about how Argonaute genes affect the water-use efficiency (WUE) of apple (Malus domestica) or its responses to different abiotic stresses. We identified and characterized MdAGOs in apple and analyzed their chromosome locations, exon/intron structures, phylogeny, and the distribution of conserved motifs. We also examined the expression profiling of responses to abiotic stress and conducted molecular cloning of MdAGO4.1. In all, 16 MdAGOs were identified and characterized, then grouped into three separate clusters. Our qRT-PCR data demonstrated that these genes are induced by drought, salt, cold, and ABA treatments, indicating that they are good candidates for further analysis of their activities and functions. We have previously shown that, during long-term moderate drought, the abundance of MdAGO4.1 protein is increased in 'Qinguan' apple (a cultivar with high WUE) leaves, but not in 'Naganofuji No. 2'. These changes are in accordance with alterations in their WUE and expression of MdAGO4.1 under the same test conditions as those used in the current study. Therefore, MdAGO4.1 is a putative gene that positively regulates WUE. Our findings provide evidence that MdAGOs have a role in plant adaptations to abiotic stress and can be exploited to improve WUE. These results will serve as a framework for future functional studies of that gene family in apple.