National Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, 53700, Lahore, Pakistan.
School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
Mol Biol Rep. 2022 Nov;49(11):10557-10564. doi: 10.1007/s11033-022-07824-0. Epub 2022 Sep 28.
Cotton is continuously exposed to sucking and chewing insect pest pressure since emergence to harvesting. Pink bollworm (Pectinophora gossypiella) has become major chewing insect pest to reduce the cotton yield and results in bad lint quality even in transgenic crops. The efficiency of insecticidal genes has been compromised due to extensive utilization of transgenic crops.
The present study was conducted to evaluate the efficacy of an alternate cry1Ia12 insecticidal gene against pink bollworm (PBW) in cotton. Agrobacterium tumefaciens strain LBA4404 harboring pCAMBIA2300 expression vector containing cry1Ia12 gene under the control of 35S CaMV was used to transform a local cotton cultivar GS-01. The various molecular analyses revealed the transgene integration and expression in primary transformants. Among five selected transgenic plants, tcL-08 showed maximum (16.06-fold) mRNA expression of cry1Ia12 gene whereas tcL-03 showed minimum (2.33-fold) expression. Feeding bioassays of 2nd and 3rd instar pink bollworm (PBW) larvae on immature cotton bolls, flowers and cotton squares revealed up to 33.33% mortality on tcL-08 while lowest mortality (13.33%) was observed in tcL-03 and tcL-15. Furthermore, the average weight and size of survived larvae fed on transgenic plants was significantly lesser than the average weight of larvae survived on non-transgenic plants.
The present study suggests the cry1Ia12 gene as an alternate insecticidal gene for the resistance management of cotton bollworms, especially PBW.
棉花从出苗到收获,一直受到吮吸和咀嚼害虫的压力。粉红棉铃虫(Pectinophora gossypiella)已成为主要的咀嚼害虫,降低了棉花的产量,并导致棉花品质下降,即使在转基因作物中也是如此。由于广泛利用转基因作物,杀虫基因的效率已经受到影响。
本研究旨在评估Cry1Ia12 杀虫基因对棉花中粉红棉铃虫(PBW)的防治效果。携带 Cry1Ia12 基因的 pCAMBIA2300 表达载体的根癌农杆菌菌株 LBA4404 在 35S CaMV 的控制下转化当地棉花品种 GS-01。各种分子分析表明,转基因在原代转化体中整合和表达。在五个选择的转基因植物中,tcL-08 显示出最高的 cry1Ia12 基因(16.06 倍)mRNA 表达,而 tcL-03 显示出最低的(2.33 倍)表达。对第 2 龄和第 3 龄粉红棉铃虫(PBW)幼虫在未成熟棉铃、花朵和棉铃上的取食生物测定表明,tcL-08 上的死亡率高达 33.33%,而在 tcL-03 和 tcL-15 上的死亡率最低(13.33%)。此外,在转基因植物上取食的存活幼虫的平均体重和大小明显小于在非转基因植物上取食的幼虫的平均体重。
本研究表明,Cry1Ia12 基因是棉花棉铃虫,特别是 PBW 的一种替代杀虫基因,可用于抗虫管理。