Rna Interference for Sharka Disease Resistance

Ilardi, Vincenza; Nicola-Negri, E. Di; Brunetti, Angela; Gentile, A.; Monticelli, S.; Damiano, C.

doi:10.17660/actahortic.2007.738.77

Cited by 3 publications

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Plums

Callahan

2008

Compendium of Transgenic Crop Plants

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Plums arose in three distinct regions, European plums ( Prunus domestica and others) from southern Europe and Asia Minor, Japanese plums ( Prunus salicina and others) from China, and the American plums ( Prunus Americana and others) from North America. The two major commercial species of plum are P. domestica (primarily for the dried fruit industry) and P. salicina (primarily for the fresh fruit industry). Breeding efforts have resulted in many high quality fruit cultivars that have been adapted for various local environmental and disease pressures. At present though, there have been few successes in breeding for resistance to plum pox potyvirus, a virus that not only affects the marketability of the fruit crop but the health of the tree. A number of research groups have begun to develop the techniques necessary to introduce resistance to plum pox virus through genetic engineering. These efforts have resulted in “HoneySweet”, a prune plum ( P. domestica ) that has been found to be highly resistant to plum pox virus in field trials. Work is continuing on improving the techniques used to engineer a plum pox virus‐resistant plum as well as producing more cultivars of plum with the resistance.

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Plums

Callahan

2008

Compendium of Transgenic Crop Plants

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Silencing of Plum pox virus 5′UTR/P1 sequence confers resistance to a wide range of PPV strains

et al. 2010

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An effective disease-control strategy should protect the host from the major economically important and geographically widespread variants of a pathogen. Plum pox virus (PPV) is the causal agent of sharka, the most devastating viral disease of Prunus species. We have shown previously that the hairpin RNA expression driven by h-UTR/P1, h-P1/HCPro, h-HCPro and h-HCPro/P3 constructs, derived from the PPV-M ISPaVe44 isolate, confers resistance to the homologous virus in Nicotiana benthamiana plants. Since the production of transgenic stone fruits and their evaluation for PPV resistance would take several years, the ISPaVe44-resistant plant lines were used to evaluate which construct would be the best candidate to be transferred to Prunus elite cultivars. To do that, nine PPV isolates of the D, M, Rec, EA and C strains originally collected from five Prunus species in different geographical areas, were typed by sequencing and used to challenge the transgenic N. benthamiana lines; 464 out of 464 virus-inoculated plants of lines h-UTR/P1, h-HCPro and h-HCPro/P3 showed complete and long-lasting resistance to the seven PPV isolates of D, M and Rec strains. Moreover, the h-UTR/P1 plants were also fully resistant to PPV-C and -EA isolates. Our data suggest that the h-UTR/P1 construct is of particular practical interest to obtain stone fruit plants resistant to the sharka disease.

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Production and in vitro assessment of transgenic plums for resistance to Plum pox virus: a feasible, environmental risk‐free, cost‐effective approach

Monticelli¹,

Nicola-Negri²,

Gentile³

et al. 2012

Annals of Applied Biology

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During the production and assessment of transgenic plants resistant to quarantine viruses, the need to contain genetically modified plants (GMPs) and pathogens severely limits working options. Moreover, in the case of fruit trees, acclimatisation and viral inoculation are very time‐consuming, thus a quick and safe method to assess the resistance to quarantine viruses, such as Plum pox virus (PPV), is desirable. This article focuses on the production of transgenic plums together with a contained and rapid evaluation in vitro for PPV resistance. The plum ‘Stanley’ was transformed by Rhizobium radiobacter (syn. Agrobacterium tumefaciens) using a PPV‐M derived hairpin construct (h‐UTR/P1) that had previously been shown to confer high and broad‐spectrum PPV resistance in model plant. Two transgenic clones, St24 and St28, were obtained. To assess their ability to resist PPV infection, micropropagated shoots of the transgenic clones were micrografted in vitro onto PPV‐D infected ‘GF305’ rootstock. Following successful grafting, the transgenic scions were analysed by immunocapture‐reverse transcription‐polymerase chain reaction (IC‐RT‐PCR) for PPV detection. A total of 97% (47/48) of St24 and 73% (17/23) of St28 tested plants were resistant to the heterologous strain of PPV. In line with an RNA silencing mediated resistance mechanism, the St24 clone was shown to accumulate higher concentration of PPV UTR/P1‐specific small interfering RNAs (siRNAs) than St28 one. The results are of practical interest not only developing plum clones that are highly resistant to PPV, but also for setting up quick and contained inoculation test procedure.

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Rna Interference for Sharka Disease Resistance

Cited by 3 publications

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Plums

Plums

Silencing of Plum pox virus 5′UTR/P1 sequence confers resistance to a wide range of PPV strains

Production and in vitro assessment of transgenic plums for resistance to Plum pox virus: a feasible, environmental risk‐free, cost‐effective approach

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