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標題: Development of transgenic watermelon resistant to Cucumber mosaic virus and Watermelon mosaic virus by using a single chimeric transgene construct
作者: Lin, Ching-Yi
Ku, Hsin-Mei
Chiang, Yi-Hua
Ho, Hsiu-Yin
Yu, Tsong-Ann
Jan, Fuh-Jyh
關鍵字: Watermelon;Transgenic plant;Viral resistance
Project: Transgenic Res., Volume 21, Page(s) 983–993.
Watermelon, an important fruit crop
worldwide, is prone to attack by several viruses that
often results in destructive yield loss. To develop a
transgenic watermelon resistant to multiple virus
infection, a single chimeric transgene comprising a
silencer DNA from the partial N gene of Watermelon
silver mottle virus (WSMoV) fused to the partial coat
protein (CP) gene sequences of Cucumber mosaic
virus (CMV), Cucumber green mottle mosaic virus
(CGMMV) and Watermelon mosaic virus (WMV)
was constructed and transformed into watermelon
(cv. Feeling) via Agrobacterium-mediated transformation.
Single or multiple transgene copies randomly inserted
into various locations in the genome were confirmed
by Southern blot analysis. Transgenic watermelon R0
plants were individually challenged with CMV,
CGMMV or WMV, or with a mixture of these three
viruses for resistance evaluation. Two lines were
identified to exhibit resistance to CMV, CGMMV,
WMV individually, and a mixed inoculation of the
three viruses. The R1 progeny of the two resistant R0
lines showed resistance to CMV and WMV, but not to
CGMMV. Low level accumulation of transgene
transcripts in resistant plants and small interfering
(si) RNAs specific to CMV and WMV were readily
detected in the resistant R1 plants by northern blot
analysis, indicating that the resistance was established
via RNA-mediated post-transcriptional gene silencing
(PTGS). Loss of the CGMMV CP-transgene fragment
in R1 progeny might be the reason for the failure to
resistant CGMMV infection, as shown by the absence
of a hybridization signal and no detectable siRNA
specific to CGMMV in Southern and northern blot
analyses. In summary, this study demonstrated that
fusion of different viral CP gene fragments in transgenic
watermelon contributed to multiple virus resistance
via PTGS. The construct and resistant
watermelon lines developed in this study could be
used in a watermelon breeding program for resistance
to multiple viruses.
DOI: 10.1007/s11248-011-9585-8
Appears in Collections:農藝學系

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