Abstract
We developed an innovative RNAi concept based on two gene constructs built from the capsid gene (CP) cistron of the Plum pox virus (PPV) genome. First, designated as amiCPRNA, a potential molecule interfering with PPV genome translation and the second one is the ami-siCPRNA to target viral genome translation and PPV RNA replication. Following the previous engineering of these constructs in an experimental herbaceous host, they were introduced into Prunus domestica (plum tree) genome. Previously propagated onto a susceptible rootstock, these clones were graft-inoculated with PPV. After four dormancy cycles, and consistent with our experience of PPV infection, some clones showed a common phenomenon of silencing that can differ between the detailed plant phenotypes. Three different phenotypes were developed by the amisiCPRNA clones. First, the high resistance character shown by the amisiCPRNA plum-7 that was similar to the resistance expressed by HoneySweet plum. Secondly, a recovery reaction was developed by the two other amisiCPRNA plum-3 and plum-4 that differed from the rest, characterized as susceptible clones, among these were the amiCPRNA plums. Having assessed the behavior of these plums versus the herbaceous host accumulating the similar form of RNAi: ami-, si-, and ami-siRNA, challenging assays in perennials consistently reflect the natural context of viral genome targeting.
Highlights
Plum pox virus (PPV) causes the severe disease known as sharka [1]
Jojo plum released by German researchers is an achievement [3], the phenotypes shown by this plum tree in field release raised a scientific debate about resistance and potential virus reservoirs [4]
In further analysis of RNAi accumulated in each clone, a variable amount of RNAi was detected in amisiCPRNA-plums (Figure 2B) and expectedly, all ami-RNA plums produced artificial miRNA (amiRNA)
Summary
PPV causes the severe disease known as sharka [1]. While control of local disease spread has been in many cases achieved through the harmonization of control measures, long distance virus spread has continued through human transport of propagative material in the last three decades [2].Throughout the European Union countries, researchers are seeking solutions to control the disease incidence but the economic imbalance between each and other state led to the lack of harmonization.Presently, no resistant plum cultivar is being extensively used by fruit-tree growers. PPV causes the severe disease known as sharka [1]. While control of local disease spread has been in many cases achieved through the harmonization of control measures, long distance virus spread has continued through human transport of propagative material in the last three decades [2]. Throughout the European Union countries, researchers are seeking solutions to control the disease incidence but the economic imbalance between each and other state led to the lack of harmonization. No resistant plum cultivar is being extensively used by fruit-tree growers. Jojo plum released by German researchers is an achievement [3], the phenotypes shown by this plum tree in field release raised a scientific debate about resistance and potential virus reservoirs [4]
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