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<p>Potato virus Y (PVY) and potato tuber moth (PTM) pose significant threats to worldwide potato production. Gene pyramiding through genetic engineering has emerged as a valuable tool for introducing multiple genes into a single plant to enhance crop productivity by improving resistance to environmental stresses. This study focused on the simultaneous incorporation of RNAi-mediated PVY resistance and <i>cry</i>1Ab-mediated PTM resistance in double-transformed plants through the re-transformation of transgenic Bt-potato plants. The RNAi construct was introduced into Bt-transgenic potato plants <i>via</i> <i>Agrobacterium</i>-mediated transformation. The re-transformed potato plants were selected on phosphinothricin (PPT) selection media and confirmed by polymerase chain reaction (PCR) analysis. Double-transgenic plants were tested for resistance to PVY and PTM using virus and insect bioassays. Molecular analysis confirmed the presence of <i>cry</i>1Ab and PVY-RNAi cassette genes in transgenic plants. Bioassay analysis on re-transformed potato plants using PTM larvae demonstrated a significantly higher mortality rate in transgenic than in the non-transgenic potato plants. Moreover, enzyme-linked immunosorbent assay (ELISA) tests on the re-transformed lines demonstrated a 28- to 38-d delay in the appearance of PVY infection symptoms compared to the control lines. Herbicide resistance analysis revealed a range of full (100%) to relative resistance (50%) with visible drying symptoms on older leaves among the double-transformed potato plants. Double-transformed potato plants simultaneously showed resistance to PTM, PVY, and glufosinate herbicide. Consequently, the results suggest that the re-transformation strategy could serve as an efficient method for gene stacking, enabling the acquisition of multiple resistances in transgenic potato plants.</p>

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Gene stacking for resistance to potato virus Y (PVY) and potato tuber moth (PTM) in transgenic potato plants

  • 期刊论文

  • 发表日期：

    2025年8月22日

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  • Shahrzad Labaf,
  • Hassan Rahnama,
  • Masoumeh Emadpour,
  • Ziba Ghasimi Hagh

摘要

Potato virus Y (PVY) and potato tuber moth (PTM) pose significant threats to worldwide potato production. Gene pyramiding through genetic engineering has emerged as a valuable tool for introducing multiple genes into a single plant to enhance crop productivity by improving resistance to environmental stresses. This study focused on the simultaneous incorporation of RNAi-mediated PVY resistance and cry1Ab-mediated PTM resistance in double-transformed plants through the re-transformation of transgenic Bt-potato plants. The RNAi construct was introduced into Bt-transgenic potato plants via Agrobacterium-mediated transformation. The re-transformed potato plants were selected on phosphinothricin (PPT) selection media and confirmed by polymerase chain reaction (PCR) analysis. Double-transgenic plants were tested for resistance to PVY and PTM using virus and insect bioassays. Molecular analysis confirmed the presence of cry1Ab and PVY-RNAi cassette genes in transgenic plants. Bioassay analysis on re-transformed potato plants using PTM larvae demonstrated a significantly higher mortality rate in transgenic than in the non-transgenic potato plants. Moreover, enzyme-linked immunosorbent assay (ELISA) tests on the re-transformed lines demonstrated a 28- to 38-d delay in the appearance of PVY infection symptoms compared to the control lines. Herbicide resistance analysis revealed a range of full (100%) to relative resistance (50%) with visible drying symptoms on older leaves among the double-transformed potato plants. Double-transformed potato plants simultaneously showed resistance to PTM, PVY, and glufosinate herbicide. Consequently, the results suggest that the re-transformation strategy could serve as an efficient method for gene stacking, enabling the acquisition of multiple resistances in transgenic potato plants.