Development of transgenic watermelon resistant to Cucumber mosaic virus and Watermelon mosaic virus by using a single chimeric transgene construct.
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 R(0) 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 R(1) progeny of the two resistant R(0) 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 R(1) 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.
Department of Plant Pathology, National Chung Hsing University, Taichung, 402, Taiwan.
Ching-Yi
CY
Lin
Hsin-Mei
HM
Ku
Yi-Hua
YH
Chiang
Hsiu-Yin
HY
Ho
Tsong-Ann
TA
Yu
Fuh-Jyh
FJ
Jan
D060054:Agrobacterium / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D015139:Blotting, Southern; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D029773:Citrullus / Q000235:genetics* / Q000276:immunology / Q000821:virology; D017799:Cucumovirus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D060467:Disease Resistance; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D018745:Genome, Plant; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000821:virology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D019076:Transgenes
D036022:Capsid Proteins; D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins