A C B D Figure 5. Epithelial cells of boll weevil guts analysed by confocal microscopy. Gut sections of insects fed on non-transformed plants (A and B) and TO-34 line (C and D). Conjugation reaction with biotinylated anti-Cry1la antibody revealed with avidin conjugated to Texas Red and DAPI. Magnification 10 μm. Arrow in (A), cell nuclei DAPI labelled.
A C B D Figure 5. Epithelial cells of boll weevil guts analysed by confocal microscopy. Gut sections of insects fed on non-transformed plants (A and B) and TO-34 line (C and D). Conjugation reaction with biotinylated anti-Cry1la antibody revealed with avidin conjugated to Texas Red and DAPI. Magnification 10 μm. Arrow in (A), cell nuclei DAPI labelled.
A C B D Figure 5. Epithelial cells of boll weevil guts analysed by confocal microscopy. Gut sections of insects fed on non-transformed plants (A and B) and TO-34 line (C and D). Conjugation reaction with biotinylated anti-Cry1la antibody revealed with avidin conjugated to Texas Red and DAPI. Magnification 10 μm. Arrow in (A), cell nuclei DAPI labelled.
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Background: Boll weevil is a serious pest of cotton crop. Effective control involves applications of chemical insecticides, increasing the cost of production and environmental pollution. The current genetically modified Bt crops have allowed great
benefits to farmers but show activity limited to lepidopteran pests. This work reports on procedures adopted for integration and expression of a cry transgene conferring resistance to boll weevil and fall armyworm by using molecular tools.
Four Brazilian cotton cultivars were microinjected with a minimal linear cassette generating 1248 putative lines.
Complete gene integration was found in only one line (TO-34) containing one copy of crylla detected by Southern blot. Protein was expressed in high concentration at 45 days after emergence (dae), decreasing by approximately 50% at 90 dae. Toxicity of the cry protein was demonstrated in feeding bioassays revealing 56.7% mortality to boll weevil fed buds and 88.1% mortality to fall armyworm fed leaves. A binding of crylla antibody was found in the midgut of boll weevils fed on TO-34 buds in an immunodetection assay.
The gene introduced into plants confers resistance to boll weevil and fall armyworm. Transmission of the transgene occurred normally to TI progeny. All plants showed phenotypically normal growth, with fertile flowers and abundant seeds.
Results relating to figure:
By confocal microscopy, the binding of anti-Cry1Ia antibody to midgut cells of boll weevil larvae was verified in samples of the T0-34 line (Figs 5B to D). The conjugation reaction of complex
protein × antibody that was revealed with avidin conjugated to Texas Red showed emission of fluorophore only in epithelial cells of larvae fed on artificial diet+young buds of the transgenic line. Disruptions of microvilli were also seen in T0-34 samples. No emission of Texas Red was seen in cells from non-transformed
samples (BRS 293) (Figs 5A to C).
Definition Definition Observable physical traits or characteristics of an organism, which are determined by its genetic makeup (genotype) as well as environmental factors. These traits can include anything from eye color or height to behavior or disease susceptibility. Understanding an organism's phenotype is important in fields such as genetics, medicine, and ecology.
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