Efficient genetic transformation of common bean: system development and optimization
摘要
Common bean (Phaseolus vulgaris L.) is an important leguminous crop with high nutritional and economic value. However, the low efficiency of genetic transformation in common bean has limited gene function research and molecular breeding. This study aims to establish an efficient and stable genetic transformation system for common bean. We first screened the germplasm resources. Among ten common bean germplasms evaluated, Pv174 was identified as optimal due to its high germination rate (96%) and compact seed size. For Agrobacterium-mediated transformation, the best conditions were an OD600 of 0.3 combined with a 3-day co-culture, which enhanced GFP (Green Fluorescent Protein) fluorescence while minimizing explant browning and contamination. In terms of regeneration, the hormone combination of 2 mg/L 6-BA (6-Benzylaminopurine) and 0.1 mg/L IAA (Indole-3-acetic acid) most effectively promoted adventitious shoot elongation without yellowing. The optimized system was validated through overexpression of PvNIP6;1 (Phvul.005G182400), achieving a 6.5% transformation efficiency. By performing preliminary treatment on the cotyledon nodes of common bean, introducing exogenous genes via vacuum transfer, and enhancing visual screening with GFP tag, the transformation efficiency was significantly improved. Furthermore, the hairy root system in common bean demonstrated high efficiency in validating CRISPR/Cas9 target site editing. This system thereby offers a reliable technical foundation for the efficient generation of transgenic common bean plants for functional genomics and breeding applications.