Ferraz Ricardo, Casimiro Bruno, Cordeiro Daniela, Canhoto Jorge, Correia Sandra
Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal.
Laqv Requimte, Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre s/n, 4169-007 Porto, Portugal.
Plants (Basel). 2025 Mar 26;14(7):1028. doi: 10.3390/plants14071028.
Cav. (tamarillo) has a strong biotechnological potential given the ease of obtaining cell lines from it that can be genetically transformed. However, genetic transformation of tamarillo cell suspension cultures has not yet been described. This study presents a simple method for -mediated transformation of these cells and demonstrates the successful insertion of the β-glucuronidase gene () and the yellow fluorescent protein gene () in their genome. For the success of this protocol, the selection of actively growing sub-cultured callus as explant and isolation of bacterial colonies with a cell density OD of 0.6-0.8 were key steps. Also, the inoculation of the callus in a bacteria liquid culture, the use of sonication, and the addition of antioxidants were essential. The transient expression of the gene in tamarillo callus was confirmed and quantified, and no significant differences were observed between using LBA4404 or EHA105 strains. Finally, the insertion of the gene in the tamarillo genome enabled the in vivo confirmation of the transformation success. The present study showed that tamarillo cell suspension cultures can be genetically modified, opening the way for metabolite production in transformed cells and future scaling-up in bioreactors.
番茄树（蛋树）具有强大的生物技术潜力，因为可以很容易地从它获得能够进行遗传转化的细胞系。然而，尚未有关于番茄树细胞悬浮培养物遗传转化的描述。本研究提出了一种用于介导这些细胞转化的简单方法，并证明了β-葡萄糖醛酸酶基因（GUS）和黄色荧光蛋白基因（YFP）成功插入其基因组。对于该方案的成功，选择活跃生长的继代培养愈伤组织作为外植体以及分离细胞密度OD为0.6 - 0.8的细菌菌落是关键步骤。此外，将愈伤组织接种到细菌液体培养物中、使用超声处理以及添加抗氧化剂也至关重要。证实并定量了番茄树愈伤组织中GUS基因的瞬时表达，并且在使用LBA4404或EHA105菌株之间未观察到显著差异。最后，GUS基因在番茄树基因组中的插入使得能够在体内确认转化成功。本研究表明番茄树细胞悬浮培养物可以进行基因改造，为转化细胞中的代谢产物生产以及未来在生物反应器中的扩大规模开辟了道路。
