Describe in detail the procedure to develop a transgenic rice variety that is resistant to the brown planthopper, covering vector choice, gene insertion method, selectable marker, and evaluation of resistance.

Step-by-step solution

1. Identify a suitable insecticidal gene, e.g., *Bt* cry1Ab, that confers resistance to the brown planthopper.
2. Choose a binary vector (such as pCAMBIA) that contains a strong monocot promoter (e.g., maize ubiquitin promoter) and a selectable marker gene (e.g., hygromycin phosphotransferase).
3. Insert the *Bt* gene into the vector using restriction enzymes and ligase, confirming the construct by PCR and restriction analysis.
4. Introduce the recombinant vector into *Agrobacterium tumefaciens* by electroporation.
5. Prepare rice embryogenic calli and co‑cultivate them with the engineered *Agrobacterium* to facilitate T‑DNA transfer.
6. Wash calli and place them on medium containing hygromycin to select transformed cells; non‑transformed cells die.
7. Regenerate hygromycin‑resistant calli into whole plants using tissue‑culture hormones.
8. Confirm integration of the *Bt* gene in regenerated plants by PCR and Southern blotting.
9. Analyse expression of the toxin protein by ELISA or Western blot.
10. Conduct bioassays by exposing the transgenic plants to brown planthopper and record mortality or damage compared with wild‑type controls.
11. Select the best performing lines for further field trials and eventual release.
12. Summarise that the whole workflow combines molecular cloning, Agrobacterium‑mediated transformation, selection, regeneration, molecular verification, and phenotypic testing.

Concept tested: Creation of transgenic pest‑resistant rice

Published 17 September 2026 · How this solution was prepared and checked