National Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Mol Biol Rep. 2022 Jun;49(6):5315-5323. doi: 10.1007/s11033-021-06972-z. Epub 2021 Nov 27.
The efficacy of Bt crystal proteins has been compromised due to their extensive utilization in the field. The second-generation Bt vegetative insecticidal proteins could be the best-suited alternative to combat resistance build-up due to their broad range affinity with midgut receptors of insects.
The codon-optimized synthetic vegetative insecticidal proteins (Vip3Aa) gene under the control of CaMV35S promoter was transformed into a locally developed transgenic cotton variety (CKC-01) expressing cry1Ac and cry2A genes. Transformation efficiency of 1.63% was recorded. The highest Vip3Aa expression (51.98-fold) was found in MS3 transgenic cotton plant. Maximum Vip3Aa protein concentration (4.23 µg/mL) was calculated in transgenic cotton plant MS3 through ELISA. The transgenic cotton plant (MS3) showed one copy number on both chromatids in the homozygous form at chromosome 8 at the telophase stage. Almost 99% mortality of H. armigera was recorded in transgenic cotton plants expressing double crystal proteins pyramided with Vip3Aa gene as contrasted to transgenic cotton plant expressing only double crystal protein with 70% mortality.
The results obtained during this study suggest that the combination of Bt cry1Ac, cry2A, and Vip3Aa toxins is the best possible alternative approach to combat chewing insects.
由于在田间的广泛应用,Bt 晶体蛋白的功效已经受到影响。第二代 Bt 营养期杀虫蛋白可能是应对抗药性产生的最佳选择,因为它们与昆虫中肠受体具有广泛的亲和力。
在 CaMV35S 启动子的控制下,经密码子优化的合成营养期杀虫蛋白(Vip3Aa)基因被转化到一个表达 cry1Ac 和 cry2A 基因的本地开发的转基因棉花品种(CKC-01)中。转化效率为 1.63%。在 MS3 转基因棉花植物中发现 Vip3Aa 表达最高(51.98 倍)。通过 ELISA 计算,MS3 转基因棉花植物中的 Vip3Aa 蛋白浓度最高(4.23µg/mL)。在有丝分裂末期,MS3 转基因棉花植物在同源染色体的两条染色单体上都显示出一个拷贝数,位于第 8 号染色体上。与仅表达双晶体蛋白的转基因棉花植物 70%的死亡率相比,表达双晶体蛋白与 Vip3Aa 基因的转基因棉花植物对 H. armigera 的死亡率几乎达到 99%。
本研究获得的结果表明,Bt cry1Ac、cry2A 和 Vip3Aa 毒素的组合是应对咀嚼性昆虫的最佳选择。