Abstract
The use of Prunus rootstocks that are resistant to plum pox virus (PPV) is an important agronomic strategy to combat the spread of the Sharka disease in nurseries and orchards. Despite remarkable progress in developing stone fruit rootstocks to adapt to various stresses, breeding that ensures durable virus resistance has not yet been achieved. For this reason, the engineering of PPV resistant plants through genetic transformation is a very promising approach to control sharka disease. The aim of the present study is to produce transgenic plants of the clonal rootstock ‘Elita’, which is resistant to PPV using ribonucleic acid interference (RNAi) technology. The genetic construct containing the self-complementary fragments of the plum pox virus coat protein (PPV-CP) gene sequence were used to induce the mechanism of post-transcriptional gene silencing to ensure virus resistance. Transgenic plants have been produced after agrobacterium-mediated transformation of in vitro explanted leaves. The results of polymerase chain reaction (PCR) and Southern blotting analyses confirmed the stable genomic integration of the PPV-CP sense and antisense intron-hairpin-RNA sequence. The functionality of the introduced expression cassette was confirmed by the activity of including the uidA gene into the transferring T-DNA. To our knowledge, this is the first interspecific plum rootstock produced by genetic engineering to achieve PPV resistance.
Highlights
Plum pox virus (PPV), a member of the Potyvirus genus in the Potyviridae family, is a serious devastating disease affecting a number of the Prunus species
Since the spread of the PPV virus in nurseries or orchards can occur in an unpredictable manner, it is expected that economic losses in various regions will continue to grow [1]
Moderntheinterspecific rootstocks are oftenplum specially recalcitrance with in vitro organogenesis, successful generation of transgenic developed to withstand bacterial infection, the introduction of a transgene using plants is limited to a few genotypes [4]
Summary
Plum pox virus (PPV), a member of the Potyvirus genus in the Potyviridae family, is a serious devastating disease affecting a number of the Prunus species. The grafting of a non-genetically modified (non-GM) scion onto the transgenic rootstock is regarded as a very promising biotechnological tool, as it has fewer biosafety concerns [14] Such an approach opens up the possibility of producing non-transgenic fruit while ensuring the overall resistance to viral infection using a transgenic ribonucleic acid interference (RNAi) inducing rootstock. The success of this approach, has largely depended on the species, the level of expression and the interactions of the small interfering ribonucleic acid (siRNA) with the target gene [15]. Efforts have been directed to express the genetic construct containing the self-complementary sequences of fragments of PPV-CP for the induction of plum pox virus resistance in plum rootstock
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