Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Kinnaird College for Women University, Lahore, Pakistan.
GM Crops Food. 2021 Jan 2;12(1):292-302. doi: 10.1080/21645698.2021.1885288.
(Desi Cotton) holds a special place in cotton industry because of its inherent ability to withstand drought, salinity, and remarkable resistance to sucking pests and cotton leaf curl virus. However, it suffers yield losses due to weeds and bollworm infestation. Genetic modification of variety FBD-1 was attempted in the current study to combat insect and weedicide resistance by incorporating and genes under control of 35S promoter in two different cassettes using kanamycin and GUS as markers through -mediated shoot apex cut method of cotton transformation. The efficiency of transformation was found to be 1.57%. Amplification of 1700 bp for , 167 bp for and 111 bp for confirmed the presence of transgenes in cotton plants. The maximum mRNA expression of and was observed in transgenic cotton line L3 while minimum in transgenic cotton line L1. The maximum protein concentrations of Cry1Ac, Cry2A and Cp4-EPSPS of 3.534 µg g, 2.534 µg g and 3.58 µg-g respectively were observed for transgenic cotton line L3 as compared to control cotton line. On leaf-feed-based insect bioassay, almost 99% mortality was observed for on the transgenic cotton plant (L3). It completely survived the 1900 ml hectare glyphosate spray assay as compared to non-transgenic cotton plants. The necrotic spots appeared on the third day, leading to the complete death of control plants on the fifth day of assay. The successful multiple gene-stacking in FBD-1 variety could be further used for qualitative improvement of cotton fiber through plant breeding techniques.
(Desi Cotton)因其耐旱、耐盐和对吮吸性害虫和棉卷叶病毒的显著抗性而在棉花产业中占有特殊地位。然而，它因杂草和棉铃虫侵害而导致产量损失。本研究试图通过在两个不同的盒中，利用卡那霉素和 GUS 作为标记，在 35S 启动子的控制下，将 和 基因整合到 FBD-1 品种中，采用 -介导的棉花转化顶端切口法对其进行遗传改良，以对抗抗虫性和除草剂抗性。转化效率为 1.57%。1700 bp 的扩增证实了棉花植株中存在转基因，167 bp 的 ，111 bp 的 。在转基因棉花株系 L3 中观察到 和 的最大 mRNA 表达，而在转基因棉花株系 L1 中观察到的最小。转基因棉花株系 L3 中 Cry1Ac、Cry2A 和 Cp4-EPSPS 的最大蛋白浓度分别为 3.534µg g、2.534µg g 和 3.58µg g，而对照棉花株系的蛋白浓度分别为 3.534µg g、2.534µg g 和 3.58µg g。在基于叶片的昆虫生物测定中，转基因棉花植株（L3）上的 几乎有 99%的死亡率。与非转基因棉花植物相比，它完全耐受 1900 毫升公顷的草甘膦喷雾测定。第三天出现坏死斑，导致对照植物在测定的第五天完全死亡。成功地在 FBD-1 品种中进行了多个基因堆叠，可以进一步通过植物育种技术来提高棉花纤维的质量。
