The present invention refers to the next-generation three genes cotton (Gossypium hirsutum) expressing chloroplast-targeted herbicide tolerant gene resistant to broad and narrow-leaved weedicide sprays and two B. thuringiensis insecticidal genes d-endotoxin Cry2A and vegetative insecticidal protein gene VIP3A. Both Cry2A and VIP3A genes have different modes of action in controlling a wide spectrum of lepidopteron insect pests, therefore, likely risk of pest resistance will be minimized which is a prevalent problem with single gene Bt cotton expressing Cry1Ac in Pakistan. The invention comprises novel nucleic acid segments encoding proteins comprising herbicide tolerance, Cry2A and VIP3A insecticidal toxins. The polynucleotide segments are revealed, as are Agro- bacterium-mediated transformation vectors holding the nucleic acid segments, plants transformed with claimed segments, methods for transforming plants, and methods for controlling plant infestation by pests.
