Symbiotic nitrogen fixation boosts legume growth and production in nitrogen-poor soils. It has long been assumed that fixed nitrogen increases reproductive success, but until now, the regulatory mechanism was unknown. Here, we report a symbiotic flowering pathway that couples symbiotic and nutrient signals to the flowering induction pathway in legumes. We show that the symbiotic microRNA–microRNA172c (miR172c) and fixed nitrogen systemically and synergistically convey symbiotic and nutritional cues from roots to leaves to promote soybean ( Glycine max ) flowering. The combinations of symbiotic miR172c and local miR172c elicited by fixed nitrogen and development in leaves activate florigen-encoding FLOWERING LOCUS T ( FT ) homologs ( GmFT2a/5a ) by repressing TARGET OF EAT1-like 4a ( GmTOE4a ). Thus, FTs trigger reproductive development, which allows legumes to survive and reproduce under low-nitrogen conditions.

Soybean (Glycine max (L.) Merr.) oil is an important source of vegetable oil for supporting the human diet. However, the high level of polyunsaturated fatty acids in natural soybean oil renders the oil unstable and thus susceptible to the development of unpalatable flavors and trans fatty acids. Therefore, reducing the content of polyunsaturated fatty acids and increasing the content of monounsaturated fatty acids is a longstanding and important target for soybean breeding. However, soybean varieties with a high oleic acid content are rare in soybean germplasm resources, which introduces substantial difficulties in the cultivation of high-oleic-acid soybeans. In this study, CRISPR/Cas9-mediated gene-editing technology was used to create targeted knockout of the soybean fatty acid desaturase encoding genes GmFAD2-1A and GmFAD2-1B that contribute to the formation of polyunsaturated fatty acids. We obtained fad2-1a, fad2-1b, and fad2-1a/fad2-1b homozygous mutants using two sgRNAs. We found that the oleic acid content increased from 11% to 40-50% in the fad2-1a and fad2-1b mutants and to 85% in the fad2-1a/fad2-1b mutants. We also generated transgene-free double mutants that conferred higher oleic acid, and the fad2-1a/fad2-1b mutant had no adverse phenotyping compared with the wild type. Our study provided new materials for the selection and breeding of high-oleic-acid soybean varieties.

Ambient light and the endogenous circadian clock play key roles in regulating Arabidopsis (Arabidopsis thaliana) seedling photomorphogenesis. PHYTOCHROME-INTERACTING FACTOR 4 (PIF4) acts downstream of both light and the circadian clock to promote hypocotyl elongation. Several members of the R2R3-MYB transcription factor (TF) family, the most common type of MYB TF family in Arabidopsis, have been shown to be involved in regulating photomorphogenesis. Nonetheless, whether R2R3-MYB TFs are involved in connecting the light and clock signaling pathways during seedling photomorphogenesis remains unknown. Here, we report that MYB112, a member of the R2R3-MYB family, acts as a negative regulator of seedling photomorphogenesis in Arabidopsis. The light signal promotes the transcription and protein accumulation of MYB112. myb112 mutants exhibit short hypocotyls in both constant light and diurnal cycles. MYB112 physically interacts with PIF4 to enhance the transcription of PIF4 target genes involved in the auxin pathway, including YUCCA8 (YUC8), INDOLE-3-ACETIC ACID INDUCIBLE 19 (IAA19) and IAA29. Furthermore, MYB112 directly binds to the promoter of LUX ARRHYTHMO (LUX), the central component of clock oscillators, to repress its expression mainly in the afternoon and relieve LUX-inhibited expression of PIF4. Genetic evidence confirms that LUX acts downstream of MYB112 in regulating hypocotyl elongation. Thus, the enhanced transcript accumulation and transcriptional activation activity of PIF4 by MYB112 additively promotes the expression of auxin-related genes, thereby increasing auxin synthesis and signaling and fine-tuning hypocotyl growth under diurnal cycles.

Flowering time and growth period are key agronomic traits which directly affect soybean (Glycine max (L.) Merr.) adaptation to diverse latitudes and farming systems. The FLOWERING LOCUS T (FT) homologs GmFT2a and GmFT5a integrate multiple flowering regulation pathways and significantly advance flowering and maturity in soybean. Pinpointing the genes responsible for regulating GmFT2a and GmFT5a will improve our understanding of the molecular mechanisms governing growth period in soybean. In this study, we identified the Nuclear Factor Y‐C (NFY‐C) protein GmNF‐YC4 as a novel flowering suppressor in soybean under long‐day (LD) conditions. GmNF‐YC4 delays flowering and maturation by directly repressing the expression of GmFT2a and GmFT5a. In addition, we found that a strong selective sweep event occurred in the chromosomal region harboring the GmNF‐YC4 gene during soybean domestication. The GmNF‐YC4^Hap3 allele was mainly found in wild soybean (Glycine soja Siebold & Zucc.) and has been eliminated from G. max landraces and improved cultivars, which predominantly contain the GmNF‐YC4^Hap1 allele. Furthermore, the Gmnf‐yc4 mutants displayed notably accelerated flowering and maturation under LD conditions. These alleles may prove to be valuable genetic resources for enhancing soybean adaptability to higher latitudes.

Soybean (Glycine max) is a globally important crop; however, its productivity is severely impacted by phosphorus (P) deficiency. Understanding the transcriptional regulation of low P (LP) response mechanisms is essential for enhancing soybean P use efficiency. In this study, we found that the Nuclear Factor-Y (NF-Y) transcription factor GmNF-YC4, in addition to its previously discovered role in regulating flowering time, possesses another functions in modulating root morphology and P uptake. Knockout of GmNF-YC4 notably boosted root proliferation and P uptake while also influencing the expression of genes related to LP stress. GmNF-YC4 acts as a specific DNA-binding transcriptional repressor, modulating the expression of the soybean α-EXPANSIN 7 (GmEXPA7) gene, which encodes a cell wall-loosening factor, through direct binding to its promoter region. Further investigation revealed that GmEXPA7 expression is predominantly root-specific and induced by LP. Moreover, overexpression of GmEXPA7 in soybean hairy roots enhanced LP tolerance by stimulating root growth and P uptake. We further screened and obtained more potential target genes of GmNF-YC4 via DNA affinity purification sequencing, including those related to LP stress. These findings underscore the pivotal role of the GmNF-YC4-GmEXPA7 module as a key regulator in mitigating LP stress in soybean.