Both symbiosis between legumes and rhizobia and nitrogen fixation in functional nodules are dramatically affected by salt stress. Better understanding of the molecular mechanisms that regulate the salt tolerance of functional nodules is essential for genetic improvement of nitrogen fixation efficiency. microRNAs (miRNAs) have been implicated in stress responses in many plants and in symbiotic nitrogen fixation (SNF) in soybean. However, the dynamic regulation of miRNAs in functioning nodules during salt stress response remains unknown. We performed deep sequencing of miRNAs to understand the miRNA expression profile in normal or salt stressed-soybean mature nodules. We identified 110 known miRNAs belonging to 61 miRNA families and 128 novel miRNAs belonging to 64 miRNA families. Among them, 104 miRNAs were dramatically differentially expressed (>2-fold or detected only in one library) during salt stress. qRT-PCR analysis of eight miRNAs confirmed that these miRNAs were dynamically regulated in response to salt stress in functional soybean nodules. These data significantly increase the number of miRNAs known to be expressed in soybean nodules, and revealed for the first time a dynamic regulation of miRNAs during salt stress in functional nodules. The findings suggest great potential for miRNAs in functional soybean nodules during salt stress.

Background: Low temperature severely limits the growth, yield, and geographic distributions of soybean. Soybean plants respond to cold stress by reprogramming the expression of a series of cold-responsive genes. However, the intrinsic mechanism underlying cold-stress tolerance in soybean remains unclear. A. thaliana tolerant to chilling and freezing 1 (AtTCF1) is a regulator of chromosome condensation 1 (RCC1) family protein and regulates freezing tolerance through an independent C-repeat binding transcription factor (CBF) signaling pathway. Results: In this study, we identified a homologous gene of AtTCF1 in soybean (named GmTCF1a), which mediates plant tolerance to low temperature. Like AtTCF1, GmTCF1a contains five RCC1 domains and is located in the nucleus. GmTCF1a is strongly and specifically induced by cold stress. Interestingly, ectopic overexpression of GmTCF1a in Arabidopsis greatly increased plant survival rate and decreased electrolyte leakage under freezing stress. A cold-responsive gene, COR15a, was highly induced in the GmTCF1a-overexpressing transgenic lines. Conclusions: GmTCF1a responded specifically to cold stress, and ectopic expression of GmTCF1a enhanced cold tolerance and upregulated COR15a levels. These results indicate that GmTCF1a positively regulates cold tolerance in soybean and may provide novel insights into genetic improvement of cold tolerance in crops.