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Agriculture & Food, Volume 5, 2017

GENERATION OF TRANSGENIC CUCUMBER WITH RESISTANCE TO CUCURBIT CHLOROTIC YELLOWS VIRUS USING POLLEN ELECTROTRANSFORMATION
Hsiu-Fung Chao, Yi-Fang Hsu, Yung-Fu Yen
Pages: 16-25
Published: 17 Jul 2017
Views: 1,066
Downloads: 366
Abstract: The Cucurbit chlorotic yellows virus (CCYV) infects many Cucurbitaceae crops, resulting in chlorotic leaves, retarded growth, and a decreased yield. Transformed cucumber plants incorporating the coat protein (CP) gene of the CCYV were generated using the transformant of CP/pCAMBIA1302 vector. The plasmid DNA was introduced into the pollen through electroporation at voltage intensity of 25 μF × 1.5kV. The electropulsed pollen was used to pollinate emasculated flowers. The transformed T1 offspring were verified using PCR and about 22% of the T1 resistant offspring exhibited CP protein expression, as verified by Western blot analysis of the GFP. In the field evaluation, the transformed plants of the T3 generation showed a high level of resistance to the CCYV. Our results showed that these transgenic cucumber plants were resistance against the CCYV, which they obtained by incorporating the CP gene through a pollen electrotransformation approach.
Keywords: cucurbit chlorotic yellows virus, pollen electrotransformation, transgenic cucumber, coat protein
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