Cadmium (Cd) is a non-essential heavy metal that is toxic to plants. microRNAs (miRNAs) are 21-nucleotide RNAs that are ubiquitous regulators of gene expression at the post-transcriptional level. Several plant miRNAs, such as miR390, have vital roles in plant growth, development and responses to environmental stresses including heavy metal stress. In this study, the expression of mature miR390 was significantly downregulated under Cd stress in rice. Consequently, the target gene of miR390, OsSRK was dramatically induced by Cd treatment. Transgenic rice plants overexpressing miR390 displayed reduced Cd tolerance and higher Cd accumulation compared with wild-type plants. Simultaneously, expression of OsSRK was less pronounced in 35S:MIR390 plants than in wild-type. These results indicate that miR390 was a negative regulator involved in Cd stress tolerance in rice.Keywords: cadmium, microRNA, miR390, OsSRK, rice
INTRODUCTIONCadmium is an important environmental pollutant that is highly toxic to plants (Schützendübel et al., 2001). It is widely released into soil, air, and water mainly by effluent from industrial sources, mining, and phosphate fertilization (Pinto et al., 2004). Cd is readily taken up by plants, resulting in toxic symptoms. It damages the photosynthetic apparatus, decreases carbon assimilation and chlorophyll content, and eventually leads to wilting and plant death (Hsu and Kao, 2003;Rodriguez-Serrano et al., 2009). Plants have evolved various molecular mechanisms to respond to heavy metal stress, one of which involves microRNA-guided gene regulation at the post-transcriptional level (Huang et al., , 2010Zhou et al., 2008Zhou et al., , 2012. miRNAs are a class of endogenous non-coding small RNAs that can base pair their target mRNAs to induce their degradation or repress their translation in eukaryotic organisms (Bartel, 2004). In plants, increasing reports have demonstrated that miRNAs have vital regulatory roles in growth, development and plant resistance to abiotic and biotic stresses, including heavy metal stress (Sunkar et al., 2006;Ding and Zhu, 2009;Khraiwesh et al., 2012). microRNA390 (miR390) is conserved in rice (Oryza sativa), maize (Zea mays), and Arabidopsis thaliana. In rice, it is represented by one member with one locus, whereas in Arabidopsis and poplar (Populus trichocarpa), it is represented by two members with three and six loci, respectively (Sunkar et al., 2005). In Arabidopsis, miR390 exerts its action through the biogenesis of ta-siRNAs that, in turn, leads to the degradation of ARFs (auxin response
MicroRNA390 Involved in Cadmium Tolerancefactors) that play critical roles in lateral root development (Marin et al., 2010;Meng et al., 2010). Recent studies indicate that it also plays an important role in plant stress tolerance because it was up-regulated under drought stress in cowpea (Vigna unguiculata; Barrera-Figueroa et al., 2011) and Brachypodium distachyon (Budak and Akpinar, 2011), while down-regulated under aluminum (Al) treatment in Medicago truncatula (Chen et al., 2...