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Agronomy 2018, 8(1), 2; https://doi.org/10.3390/agronomy8010002

Generation of Transgenic Rootstock Plum ((Prunus pumila L. × P. salicina Lindl.) × (P. cerasifera Ehrh.)) Using Hairpin-RNA Construct for Resistance to the Plum pox virus

1
Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, 142290 Pushchino, Russia
2
Nikitsky Botanical Gardens, Yalta, 98648 Nikita, Russia
*
Author to whom correspondence should be addressed.
Received: 9 November 2017 / Revised: 4 December 2017 / Accepted: 15 December 2017 / Published: 22 December 2017
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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. View Full-Text
Keywords: sharka disease; stone fruits; genetic transformation; leaf explant; RNA interference sharka disease; stone fruits; genetic transformation; leaf explant; RNA interference
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Sidorova, T.; Pushin, A.; Miroshnichenko, D.; Dolgov, S. Generation of Transgenic Rootstock Plum ((Prunus pumila L. × P. salicina Lindl.) × (P. cerasifera Ehrh.)) Using Hairpin-RNA Construct for Resistance to the Plum pox virus. Agronomy 2018, 8, 2.

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