Friable embryogenic callus (FEC) currently forms the tissue of choice for gene transfer to cassava tissues. However, this technology has not been sufficiently adopted in most laboratories, partly because of the challenging protocols involved in FEC production. Therefore, the development of transformation systems targeting readily accessible explants could enable the rapid improvement of this essential root tuber. This study developed a simple, efficient, and rapid transformation protocol for three elite cassava cultivars (TMS 60444, 08/354, KME 1) using immature leaf lobes (ILLs). Factors affecting callus and somatic embryo formation, such as auxin concentration and culture photoperiod, were optimized. Cassava transformation was achieved by co-cultivation of ILLs and callus with Agrobacterium strain EHA101 harboring pTF102 plasmid. We report a positive interactive effect between dark treatment and an increasing auxin concentration. Exposure to light was found to negatively affect the formation and viability of SEs by promoting tissue browning. The results further show that cassava transformation success depends on the cultivar, explant type, and age. Using this simple protocol, transgenic plants of TMS 60444, 08/354, and KME 1 with an average transformation frequency of 0.4% (0.5, 0.4, and 0.4%, respectively) were obtained in 6 months. Reverse transcription PCR, Southern blot, and histochemical GUS staining confirmed the stable integration of the gusA gene into the regenerated plantlets. This optimized transformation system for cassava is rapid, efficient, and simple to operate and will facilitate more genetic applications in cassava research.

易碎的胚性愈伤组织（FEC）当前形成基因转移到木薯组织的选择组织。但是，该技术尚未在大多数实验室中得到充分采用，部分原因是FEC生产涉及的挑战性协议。因此，针对易获得的外植体的转化系统的开发可以使这种必不可少的块根得以快速改善。这项研究使用未成熟的叶瓣（ILLs）为三个优良木薯品种（TMS 60444，08/354，KME 1）开发了一种简单，高效且快速的转化方案。优化了影响愈伤组织和体细胞胚形成的因素，例如植物生长素的浓度和培养的光周期。木薯转化是通过将ILLs和愈伤组织与农杆菌共培养而实现的带有pTF102质粒的EHA101菌株。我们报道了黑暗处理和生长素浓度增加之间的积极相互作用。发现暴露于光通过促进组织褐变对SE的形成和生存能力具有负面影响。结果进一步表明，木薯转化成功与否取决于品种，外植体类型和年龄。使用此简单协议，在6个月内获得了平均转化频率为0.4％（分别为0.5、0.4和0.4％）的TMS 60444、08 / 354和KME 1转基因植物。逆转录PCR，Southern印迹和组织化学GUS染色证实了gusA的稳定整合基因进入再生苗。这种针对木薯的优化转化系统快速，高效且易于操作，将促进木薯研究中更多的遗传应用。