Zinhari Zahra, Pourseyedi Shahram, Zolalo Jefar
Department of Biotechnology, Shahid Bahonar University of Kerman, Kerman, Iran.
3 Biotech. 2017 Oct;7(5):284. doi: 10.1007/s13205-017-0915-2. Epub 2017 Aug 16.
The present research is carried out to study gene transformation for the first time, using direct shoot explants. As a prerequisite for gene transformation, the regeneration conditions in were optimized. We achieved an efficient and reproducible protocol for successful direct shoot regeneration without intervening callus formation. The results indicate that is the insistent species of Brassicaceae in direct shoot regeneration. Various explants of were genetically transformed with different strains of viz., LBA4404, GV3850, GV3101, and EHA105, using the vector pBI121. Expression of GUS reporter protein was assayed by histochemical staining. In addition, using the PCR method with specific primers proved the integration of GUS gene into the plants. The highest transformation efficiency was achieved with strain GV3850. Moreover, we found that infected hypocotyl and root explants of interestingly yielded higher transformation efficiency, so that in hypocotyls on average exceeded 70% of the explants. This study showed that in addition to the numerous desirable traits, has a high potential for gene transfer.
本研究首次利用直接芽外植体进行基因转化研究。作为基因转化的前提条件，对再生条件进行了优化。我们成功建立了一种高效且可重复的方案，实现了直接芽再生，且无需经过愈伤组织形成阶段。结果表明，[具体物种名称未给出]是十字花科中直接芽再生的顽固物种。使用载体pBI121，用不同菌株（即LBA4404、GV3850、GV3101和EHA105）对[具体物种名称未给出]的各种外植体进行遗传转化。通过组织化学染色检测GUS报告蛋白的表达。此外，使用特异性引物的PCR方法证明了GUS基因已整合到植物中。菌株GV3850实现了最高的转化效率。此外，我们发现有趣的是，[具体物种名称未给出]受感染的下胚轴和根外植体产生了更高的转化效率，以至于在下胚轴中平均超过70%的外植体实现了转化。这项研究表明，[具体物种名称未给出]除了具有许多优良性状外，还具有很高的基因转移潜力。
