Engineering broad-spectrum resistance against RNA viruses in potato |
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Authors: | M Arif U Azhar M Arshad Y Zafar S Mansoor S Asad |
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Institution: | (1) Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan;(2) Director General Agriculture and Biotechnology, PAEC, Islamabad, Pakistan; |
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Abstract: | RNA silencing technology has become the tool of choice for inducing resistance against viruses in plants. A significant discovery
of this technology is that double-stranded RNA (dsRNA), which is diced into small interfering RNAs (siRNAs), is a potent trigger
for RNA silencing. By exploiting this phenomenon in transgenic plants, it is possible to confer high level of virus resistance
by specific targeting of cognate viral RNA. In order to maximize the efficiency and versatility of the vector-based siRNA
approach, we have constructed a chimeric expression vector containing three partial gene sequences derived from the ORF2 gene
of Potato virus X, Helper Component Protease gene of Potato virus Y and Coat protein gene of Potato leaf roll virus. Solanum tuberosum cv. Desiree and Kuroda were transformed with this chimeric gene cassette via Agrobacterium tumefaciens-mediated transformation and transgenic status was confirmed by PCR, Southern and double antibody sandwich ELISA detection.
Due to simultaneous RNA silencing, as demonstrated by accumulation of specific siRNAs, the expression of partial triple-gene
sequence cassette depicted 20% of the transgenic plants are immune against all three viruses. Thus, expression of a single
transgene construct can effectively confer resistance to multiple viruses in transgenic plants. |
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