Genome-wide identification of CRF gene family members in four rice subspecies and expression analysis of OsCRF members in response to cold stress at seedling stage.

Cytokinin Response Factors (CRFs) play a crucial role in plant growth and development, hormone signaling, and responses to biotic and abiotic stresses. However, there have been no reports on CRF genes in rice until now. We analyzed the CRF families in four rice subspecies: cultivated rice Oryza sativa Japonica Group, Oryza sativa Indica Group, and Oryza sativa (circum-Aus1 var. N22), as well as wild rice Oryza rufipogon. We identified 7, 6, 6, and 7 CRF in their genomes, respectively, distributed across different chromosomes. The protein motifs and gene structures of CRF in these four types of rice show high conservation. Cis-regulatory element analysis revealed that the promoter regions of the CRF contain numerous hormone and stress-related elements. The number of CRF in these four types of rice is not influenced by gene duplication. The expression pattern showed that OsCRF exhibit significant tissue-specific expression. The qRT-PCR results showed that OsCRF strongly responded to low-temperature stress and can be induced by melatonin and cytokinin to increase expression levels. In addition, the nuclear localisation of OsCRF4/5 was confirmed to be as predicted. The results above will provide a foundation for further and deeper investigation of CRFs.