The Rice Transcription Factor, OsbHLH046, Contributes to Salt Stress Tolerance at Seedling Stage

Rice basic-Helix-Loop-Helix (OsbHLH) transcription factors (TFs) belong to a multiple gene family and are known to involve in rice growth, development, regulating abiotic stress-responsive gene expression and tolerance of rice. However, because functional redundancy within the super gene family and the pleiotropic effect of individual TF, the specific function for corresponding OsbHLH is not easy to be determined. In this study, we identified an OsbHLH046 TF which is down-regulated by salt and cold treatments from previous microarray result and further confirmed the gene expression by reverse transcription-PCR (RT-PCR). The OsbHLH046-GFP fusion protein showed OsbHLH046 is located in nucleus. Based on the predication of rice eFP browser and the GUS-histochemical staining indicated that OsbHLH046 is expressed in the embryo, seedling root, lateral roots, shoots and anthers. The gene expression was higher at the early germination stage then declined to lower level within 3rd leaf stage. Compared with WT, the Osbhlh046, a Tos17 knocked-down rice mutant, displayed late seed germination, growth retardation, and unable to recover from salt stress at seedling stage. The q-PCR data indicated that salt tolerance related genes in such as OsABI5, OsNHX2 were differential expressed between two plants during recovery stage. The MDA was overaccumulated in Osbhlh046 mutant compared to wild type after recover from salt stress. The ICP-OES elemental analysis also revealed that the Osbhlh046 plant have lower K+ content level, which may not able to maintain the Na+ homeostasis. Meanwhile, the callus of OsbHLH046 overexpression line showed a relative poor regeneration rate. Taken together, these results suggested that OsbHLH046 plays as a positive regulator in seed germination, seedling growth, and contributes to the ability of rice to recover from salt stress.