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Improved Resistance to Cucumber mosaic virus in Petunia Transformed with Non‐Cytotoxic Pokeweed Antiviral Protein Gene
Author(s) -
Li Li,
Li Yu,
Chen Dinghu,
Feng Hui,
Wang Xifeng
Publication year - 2013
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12051
Subject(s) - biology , petunia , cucumber mosaic virus , virology , mutant , agrobacterium , transgene , gene , microbiology and biotechnology , southern blot , western blot , cauliflower mosaic virus , antiviral protein , virus , cytotoxic t cell , genetically modified crops , plant virus , genetics , rna , in vitro
Petunia hybrida is an important ornamental plant that can be seriously affected by cucumber mosaic virus ( CMV ). Pokeweed antiviral protein ( PAP ), a ribosome‐inactivating protein, has been recognized as a broad spectrum virus inhibitory agent. Mutant PAP efficiently inhibited viral gene expression at both the translational and transcriptional levels without causing host cell toxicity. We have transferred the non‐cytotoxic pokeweed antiviral protein (mutant PAP ) gene into petunia cells with Agrobacterium tumefacien s. Forty‐two putative transgenic regenerated lines were obtained from the selected explants. Fifty‐six plants immune to CMV infection were recovered from nine transgenic lines. Another 51 plants tolerant to CMV were obtained from 10 transgenic regenerated lines. Successful integration of the mutant PAP gene into the genome of transgenic petunia was confirmed by PCR and Southern blot analysis. Expression of the PAP gene was further confirmed by RT ‐ PCR and Western blot analysis. These results were consistent with the assay of resistance to CMV .