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Soybean is the primary source of vegetable protein and is used for various purposes, mainly to feed animals. This crop can have diverse seed coat colors, varying from yellow, black, brown, and green to bicolor. Black seed coat cultivars have already been assigned as favorable for both seed and grain production. Thus, this work aimed to identify genes associated with soybean seed quality by comparing the transcriptomes of soybean seeds with contrasting seed coat colors. The results from RNA-seq analyses were validated with real-time PCR using the cultivar BRS 715A (black seed coat) and the cultivars BRS 413 RR and DM 6563 IPRO (yellow seed coat). We found 318 genes differentially expressed in all cultivars (freshly harvested seeds and seeds stored in cold chamber). From the in silico analysis of the transcriptomes, the following genes were selected and validated with RT-qPCR: ACS1, ACSF3, CYP90A1, CYP710A1, HCT, CBL, and SAHH. These genes are genes induced in the black seed coat cultivar and are part of pathways responsible for ethylene, lipid, brassinosteroid, lignin, and sulfur amino acid biosynthesis. The BRSMG 715A gene has almost 4times more lignin than the yellow seed coat cultivars. These attributes are related to the BRSMG 715A cultivar’s higher seed quality, which translates to more longevity and resistance to moisture and mechanical damage. Future silencing studies may evaluate the knockout of these genes to better understand the biology of soybean seeds with black seed coat.

BackgroundThe drought condition is responsible for considerable losses in soybean production, which in turn may result in billionaire losses. After drought perception, plants activate a cascade of protecting genes against water deficit (WD), many of which are responsive to abscisic acid, the most important phytohormone to plants’ adaptation. This work aimed to recover abscisic acid (ABA)-responsive differentially expressed genes (DEG) from an RNA-Seq, carried out from leaves and roots of drought-sensitive (BR16) and tolerant (Embrapa 48) soybean cultivars under mild (MiWD), moderate (MoWD), and severe (SWD) water-deficit treatments. Enriched ABA-responsive pathways important to drought tolerance in soybean were also identified.ResultsIn drought-sensitive and tolerant soybean cultivars, approximately 75% of genes were identified as ABA-responsive by containing more than two ABRE (ABA-responsive elements) in the promoter region. Most of these genes were positively regulated. Roots were the tissue with more ABA-responsive genes and pathways triggered in response to WD in both cultivars, although, on the tolerant cultivar, these pathways were higher expressed. The most important enriched ABA pathways observed in the roots of both cultivars were involved in sugar and sulfur amino acid biosynthesis, osmoregulation, and crosstalk among ABA and ethylene, jasmonate, auxin, and cytokinin. Other pathways enriched were involved in phytoalexin production, ROS homeostasis, and membrane stability by glycerolipid and glycerophospholipid production. ABA-responsive genes were also ordered based on their expression profile in tissue and cultivar, and nine confidence groups could be observed. More than 80% of these clustered genes showed the same regulation profile under MiWd, MoWD, and SWD treatments. Activation of ABA biosynthesis under water deficit was validated by RT-qPCR by increasing the expression level of NCED3, an important enzyme in this pathway, and GOLS, a known ABA-responsive gene.ConclusionsA robust catalog of ABA-responsive genes was made available in this work. Considering ABA’s role in drought-response mechanisms, the genes in the groups pointed out in this study would be reliable candidates to be used in strategies to develop soybean lines more tolerant to drought. This paper, presented for the first time, ABA-responsive genes and ABA-enriched pathways in contrasting soybean cultivars for drought tolerance.