Hajiahmadi Zahra, Shirzadian-Khorramabad Reza, Kazemzad Mahmood, Sohani Mohammad Mehdi, Khajehali Jahangir
Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, 4199613776 Iran.
Department of Energy, Materials and Energy Research Center, Tehran, 14155-477 Iran.
3 Biotech. 2020 Aug;10(8):370. doi: 10.1007/s13205-020-02359-2. Epub 2020 Aug 3.
In this study, a novel and stable gene transformation system was developed under control of () as an inducible promoter using the Mesoporous Silica Nanoparticles (MSNs). The functionalized MSNs with a proper particle size were synthesized and attached to a recombinant construct (pDNA) containing gene under the control of promoter (pPZP122:::MSN [pDNA: MSN]) following transformation of tomato plants through injection of the pDNA: MSN complex into tomato red fruit at early ripening stage and then, putative transgenic seeds were collected. As an initial selection, gentamicin-resistant seedlings of T (24.24%) and T (61.37%) plants were identified. The transgene integration and expression were confirmed through the PCR, RT-PCR, and western blot approaches in the selected seedlings. PCR analysis showed that transformation frequency was equal to 10.71% in T plants. Semi-quantitative RT-PCR analysis confirmed the transcript expression of in all the T and T PCR-positive plants. Western blot analysis confirmed the existence of CryIAb protein in the leaves of T putative transgenic plants. Accordingly, the results demonstrated that the transgene has more likely integrated into the tomato genome through homologous recombination. Bioassay was carried out for further assessment of the plant responses to resulting in an enhanced tolerance of the plant. In conclusion, the MSN-mediated stable transformation system under the as an inducible promoter can be used as a suitable alternative for conventional genetic transformation methods due to its biodegradability, biocompatibility, cost and time-effectiveness, and positive effect on the plant defense against pathogens and pests.
在本研究中,利用介孔二氧化硅纳米颗粒(MSNs),在()作为诱导型启动子的控制下,开发了一种新型稳定的基因转化系统。合成了具有合适粒径的功能化MSNs,并将其连接到含有在启动子(pPZP122:::MSN [pDNA: MSN])控制下基因的重组构建体(pDNA)上。在番茄果实早熟期将pDNA: MSN复合物注射到番茄红果中,对番茄植株进行转化,然后收集推定的转基因种子。作为初步筛选,鉴定出T代(24.24%)和T代(61.37%)植株中对庆大霉素具有抗性的幼苗。通过PCR、RT-PCR和western blot方法在所选幼苗中确认了转基因的整合和表达。PCR分析表明,T代植株的转化频率为10.71%。半定量RT-PCR分析证实了在所有T代和T代PCR阳性植株中均有转录表达。western blot分析证实了在T代推定转基因植株的叶片中存在CryIAb蛋白。因此,结果表明转基因更有可能通过同源重组整合到番茄基因组中。进行了生物测定以进一步评估植株对()的反应,结果表明植株的耐受性增强。总之,以()作为诱导型启动子的MSN介导的稳定转化系统,由于其生物可降解性、生物相容性、成本和时间效益以及对植物抵御病原体和害虫的积极作用,可作为传统遗传转化方法的合适替代方法。