Yu Jing-Jing, Deng Shiling, Mo Jinhui, Huang Han, Xu Zeng-Fu, Wang Yi
Key Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry, Guangxi University, Nanning, 530000, China.
State Key Laboratory for Conservation and Utilization of Subtropical Agro‑Bioresources, College of Forestry, Guangxi University, Nanning, 530000, China.
Plant Cell Rep. 2025 May 3;44(5):112. doi: 10.1007/s00299-025-03493-y.
Identified a robust regeneration-related genomic sequence of PvoB3-69 from the B3 superfamily in Plukenetia volubilis. gPvoB3-69-assisted transformation of P. volubilis obtained transgenic shoots for the first time. The regenerative capacity of host cells is critical for the genetic transformation efficiency of woody plants. The B3 superfamily is particularly involved in early embryo morphogenesis and late-stage embryo maturation. In this study, 74 PvoB3 members were identified in the genome of P. volubilis, and classified into four subfamilies: LAV, RAV, ARF, and REM. RNA-seq and RT-qPCR analyses revealed that PvoB3-69, a member of the LAV subfamily, has specific expression patterns similar with LEC2. Overexpression of gPvoB3-69 enhanced the bud regeneration capacity of transgenic Nicotiana benthamiana organs during in vitro culture. Additionally, overexpression of gPvoB3-69 significantly improved somatic embryogenesis in transgenic Arabidopsis plants, especially with the aid of 2,4-D. Transcriptome analysis in Arabidopsis thaliana revealed that PvoB3-69 may enhance somatic embryo induction efficiency by activating developmental regulators such as WUS and LEC1, while also modulating salicylic acid, ABA, and ethylene metabolism. Furthermore, overexpression of gPvoB3-69 in the cotyledons of P. volubilis increased the regeneration ability of host cells, and broke the genetic transformation barrier through the Agrobacterium-mediated method, allowing the regenerated transgenic shoots obtained for the first time. This study provides the first systematic analysis of the B3 superfamily in P. volubilis, identifying PvoB3-69 as a key regulator of regeneration. These findings establish a foundation for further comprehensive studies of PvoB3 genes and deepen our understanding of the regulatory mechanism of shoot regeneration in P. volubilis.
从南美油藤的B3超家族中鉴定出一个与再生相关的强大基因组序列PvoB3-69。gPvoB3-69介导的南美油藤转化首次获得了转基因芽。宿主细胞的再生能力对木本植物的遗传转化效率至关重要。B3超家族尤其参与早期胚胎形态发生和后期胚胎成熟。在本研究中，在南美油藤基因组中鉴定出74个PvoB3成员，并分为四个亚家族：LAV、RAV、ARF和REM。RNA测序和RT-qPCR分析表明，LAV亚家族成员PvoB3-69具有与LEC2相似的特异性表达模式。gPvoB3-69的过表达增强了转基因本氏烟草器官在体外培养期间的芽再生能力。此外，gPvoB3-69的过表达显著提高了转基因拟南芥植株的体细胞胚胎发生，尤其是在2,4-D的作用下。拟南芥的转录组分析表明，PvoB3-69可能通过激活WUS和LEC1等发育调节因子来提高体细胞胚胎诱导效率，同时还调节水杨酸、脱落酸和乙烯代谢。此外，gPvoB3-69在南美油藤子叶中的过表达提高了宿主细胞的再生能力，并通过农杆菌介导的方法打破了遗传转化障碍，首次获得了再生的转基因芽。本研究首次对南美油藤中的B3超家族进行了系统分析，确定PvoB3-69为再生的关键调节因子。这些发现为进一步全面研究PvoB3基因奠定了基础，并加深了我们对南美油藤芽再生调控机制的理解。
