Antioxidant enzyme activity and microRNA are associated with growth ofPoa pratensiscallus under salt stress

Kentucky bluegrass (Poa pratensisL.) is an important species of turfgrass that is commonly planted on golf courses and landscapes all over the world. It is sensitive to salt stress; however, details relating to its molecular mechanisms of salt resistance are not available. We, therefore, analyzed the changes in growth, antioxidant enzyme activity, and microRNA expression in the callus 1 week after treatment with 200 mM NaCl for 12 h, 24 h, 48 h, 96 h, and 144 h. The results demonstrated that callus growth declined and thiobarbituric acid-reactive substance production and cell membrane permeability increased. Treatment with salt increased ascorbate peroxidase, glutathione reductase, superoxide dismutase, catalase, peroxidase, and polyphenol oxidase activity. Changes in the expression levels of microRNAs were observed under salt treatment. The expression of miR162, miR173, miR391, miR408, miR773, and miR857 increased by 70% after 24 h of salt treatment, after which it declined to a level similar to that of the control. The expression level of miR775 and miR827 decreased by 20% after 24 h, and then further decreased by 80% after 144 h. The expression level of miR841 increased by 50% after 24 h of salt treatment, and then stabilized. In contrast, salt treatment increased the expression of the auxin response factors ARF6, ARF8, ARF10, and ARF16 in the callus from 12 to 144 h of salt treatment, during which the expression increased twofold. Gene expression analysis indicated that salt-responsive gene families were regulated by microRNAs in the callus under salinity stress. The activity of antioxidant enzymes is also changing. MiR841 is considered to be a positive regulator of antioxidant enzyme biosynthesis. The present investigation elucidates the manner in whichP. pratensisresponds to salt stress in the callus, and could be used to inform further studies on the molecular mechanisms of stress tolerance.