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<p>The increasing resistance of <i>Helicoverpa armigera</i> to currently deployed <i>Bacillus thuringiensis</i> (<i>Bt</i>) genes in transgenic crops necessitates the development of new insecticidal genes that can effectively control this pest. This study evaluates the <i>in-planta</i> efficacy of <i>cry1Ac34</i>, a truncated, plant codon-optimized gene derived from an Indian <i>Bt</i> isolate (SK-783). The <i>cry1Ac34</i> gene was successfully introduced into <i>Nicotiana tabacum</i> through Agrobacterium-mediated transformation. Following the transformation, PCR-positive T<sub>0</sub> transgenic plants expressing the Cry1Ac34 protein were identified. Subsequent analyses of T<sub>1</sub> progeny lines, using Southern blotting and qRT-PCR, confirmed the stable integration and expression of the gene. Insect bioassays conducted with detached leaves from transgenic lines demonstrated impressive mortality rates of up to 95% in line C12 of T<sub>1</sub> against first larval instars of <i>H. armigera</i>. Protein expression levels in T<sub>1</sub> transgenic plants ranged from 0.86&#xa0;µg/g to 1.36&#xa0;µg/g fresh leaf weight. Additionally, the study observed significant sub-lethal effects, including reduced weight gain and larval growth, delays in pupation, and pre-pupal death. Histopathological analysis revealed disintegration of brush border membranes in the midguts of fourth instar larvae that had been fed transgenic leaves. The successful expression and efficacy of the codon-optimized <i>cry1Ac34</i> gene highlight its potential as a viable solution for controlling <i>H. armigera</i>. This research suggests <i>cry1Ac34</i> as a promising candidate for deployment in the development of new insect-resistant transgenic crops, which may help combat the escalating resistance challenges currently faced in agricultural pest management.</p>

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Novel codon-optimized cry1Ac34 gene from Indian Bt isolate confers resistance against Helicoverpa armigera in Transgenic tobacco

  • 期刊论文

  • 发表日期：

    2026年1月6日

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  • Anupma Singh,
  • G. G. K. Gouthami,
  • V. Rakesh,
  • Mamta Gupta,
  • Harish Kumar,
  • Vinay Kalia,
  • Sarvjeet Kaur

摘要

The increasing resistance of Helicoverpa armigera to currently deployed Bacillus thuringiensis (Bt) genes in transgenic crops necessitates the development of new insecticidal genes that can effectively control this pest. This study evaluates the in-planta efficacy of cry1Ac34, a truncated, plant codon-optimized gene derived from an Indian Bt isolate (SK-783). The cry1Ac34 gene was successfully introduced into Nicotiana tabacum through Agrobacterium-mediated transformation. Following the transformation, PCR-positive T0 transgenic plants expressing the Cry1Ac34 protein were identified. Subsequent analyses of T1 progeny lines, using Southern blotting and qRT-PCR, confirmed the stable integration and expression of the gene. Insect bioassays conducted with detached leaves from transgenic lines demonstrated impressive mortality rates of up to 95% in line C12 of T1 against first larval instars of H. armigera. Protein expression levels in T1 transgenic plants ranged from 0.86 µg/g to 1.36 µg/g fresh leaf weight. Additionally, the study observed significant sub-lethal effects, including reduced weight gain and larval growth, delays in pupation, and pre-pupal death. Histopathological analysis revealed disintegration of brush border membranes in the midguts of fourth instar larvae that had been fed transgenic leaves. The successful expression and efficacy of the codon-optimized cry1Ac34 gene highlight its potential as a viable solution for controlling H. armigera. This research suggests cry1Ac34 as a promising candidate for deployment in the development of new insect-resistant transgenic crops, which may help combat the escalating resistance challenges currently faced in agricultural pest management.