ArticlePlant cell reports2026
Hormone-optimized hypocotyl-epicotyl-cotyledonary tri-complex explant system enables efficient soybean transformation with reduced genotype dependence.
Article in Plant cell reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
key messageA composite hypocotyl-epicotyl-cotyledonary tri-complex (HECC) explant significantly improves soybean regeneration and Agrobacterium-mediated transformation efficiency (40.3%) while reducing genotype dependence, providing a rapid and robust platform for functional genomics. Soybean genetic transformation is hindered by low regeneration efficiency and considerable genotype dependency. Here, we present a hormone-optimized Agrobacterium-mediated transformation system employing a composite hypocotyl-epicotyl-cotyledonary tri-complex (HECC) explant that integrates multiple meristematic regions within a single explant and significantly improves regeneration and transformation efficiency. Compared to conventional single-meristem explant systems such as cotyledonary node and half-seed explants, HECC explants exhibited substantially higher regeneration frequencies and achieved an average transformation efficiency of 40.3%, calculated as the number of PCR-positive shoots obtained per infected explant in the JS-335 cultivar. Optimization of hormone combinations, particularly trans-zeatin riboside in shoot induction and elongation and indole-3-butyric acid (IBA) during rooting, promoted robust organogenic responses and reduced tissue damage. The system supported efficient transformation (26-41%) across multiple soybean cultivars, including JS-335, JS-2034, JS-2069 and PUSA-9712, indicating reduced genotype dependence compared with conventional explant systems. To demonstrate the applicability of this system for genome engineering, a CRISPR/Cas9 construct targeting GmSPL9c (Glycine max SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9c), a transcription factor involved in regulating plant developmental transitions and shoot architecture, was introduced. Molecular analyses by PCR and qRT-PCR confirmed the integration and expression of cas9 and bar genes, while segregation analysis in T
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