Abstract
MicroRNAs play an important role in abiotic stress responses in higher plants and animals, but their role in stress adaptation in algae remains unknown. In this study, the expression of identified and putative miRNAs in Chlamydomonas reinhardtii was assessed using quantitative polymerase chain reaction; some of the miRNAs (Cre-miR906-3p) were up-regulated, whereas others (Cre-miR910) were down-regulated when the species was subjected to multiple abiotic stresses. With degradome sequencing data, we also identified ATP4 (the d-subunit of ATP synthase) and NCR2 (NADPH: cytochrome P450 reductase) as one of the several targets of Cre-miR906-3p and Cre-miR910, respectively. Q-PCR data indicated that ATP4, which was expressed inversely in relation to Cre-miR906-3p under stress conditions. Overexpressing of Cre-miR906-3p enhanced resistance to multiple stresses; conversely, overexpressing of ATP4 produced the opposite effect. These data of Q-PCR, degradome sequencing and adaptation of overexpressing lines indicated that Cre-miR906-3p and its target ATP4 were a part of the same pathway for stress adaptation. We found that Cre-miR910 and its target NCR2 were also a part of this pathway. Overexpressing of Cre-miR910 decreased, whereas that of NCR2 increased the adaption to multiple stresses. Our findings suggest that the two classes of miRNAs synergistically mediate stress adaptation in algae.
Highlights
Epigenetic regulation, especially by means of small RNAs, is involved in the adaptation of plants to environmental stress[1]
200 miRNAs have been found in Chlamydomonas[33,34]; it is possible to select miRNAs of interest and analyzing their expression pattern under multiple stresses by using quantitative polymerase chain reaction (q-PCR)
We performed q-PCR screening and found that Cre-miR906-3p was up-regulated under multiple stresses, suggesting that it might play an essential role in a common adaptation mechanism to multiple stresses
Summary
Epigenetic regulation, especially by means of small RNAs (sRNAs), is involved in the adaptation of plants to environmental stress[1]. Previous studies have shown a limited, modulatory role for miRNA-mediated gene regulation in Chlamydomonas under standard conditions[6], but numerous sRNAs are known to be up-regulated under nitrogen or sulfur starvation conditions in wild-type Chlamydomonas[33,34,35] The findings of these studies suggest that miRNAs might potentially be involved in regulating specific responses to environmental stress rather than in controlling normal cellular functions[36]. This study focused on the regulation mechanism of miRNAs in stress adaptation, which is important for stress biology research in plants and algae and might be used to improve algal growth during biofuel[18] and high-value bio-product[37] production
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