THE DEVELOPMENT OF GENETIC RESISTANCE TO PLUM POX VIRUS IN TRANSGENIC NICOTIANA BENTHAMIANA AND PRUNUS DOMESTICA

Constructs with self-complementary sequences separated by an intron produce “hairpin” RNA [intron-hairpin-RNA (ihpRNA)] structures that efficiently elicit posttranscriptional gene silencing (PTGS). In the current study, the authors use this technology to confer resistance to plum pox virus (PPV) in herbaceous and woody perennial plants by silencing the PPV–coat protein (CP) gene. The authors confirmed the high capacity of ihpRNA constructs for inducing RNA silencing in Nicotiana benthamiana Domin., as more than 75% of the transformants displayed PTGS as evaluated by specific small interfering RNA (siRNA) production. The authors demonstrated that ihpRNA constructs provided PPV resistance, and they found a correlation between the length of the PPV sequence introduced in the ihpRNA constructs and the frequency of transgenic-resistant plants. Plants transformed with the full-length sequence produced a higher percentage of resistant lines. The authors further demonstrated for the first time that ihpRNA technology is applicable to a woody perennial species. A transgenic plum (Prunus domestica L.) PPV-CP ihpRNA line showed gene silencing characteristics (hypermethylation of the transgene sequence and specific siRNA production) and resistance to PPV infection 16 months after inoculation.

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Tracking the potyviral P1 protein in Nicotiana benthamiana plants during plum pox virus infection

Acta Virologica ◽

10.4149/av_2017_413 ◽

2017 ◽

Vol 61 (04) ◽

pp. 492-494 ◽

Cited By ~ 2

Author(s):

Z. VOZÁROVÁ ◽

M. GLASA ◽

Z. W. ŠUBR

Keyword(s):

Virus Infection ◽

Nicotiana Benthamiana ◽

Plum Pox Virus

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The phenotypically quantitative nature of hypersensitivity of European plum (Prunus domestica L.) against the Plum pox virus and its description using the hypersensitivity index

Horticultural Science ◽

10.17221/639-hortsci ◽

2008 ◽

Vol 35 (No. 2) ◽

pp. 50-64 ◽

Cited By ~ 14

Author(s):

M. Neumüller ◽

W. Hartmann

Keyword(s):

Quantitative Trait ◽

Leaf Blade ◽

Test System ◽

Artificial Inoculation ◽

Plum Pox Virus ◽

Prunus Domestica ◽

Individual Genotype ◽

Selection Tool ◽

Quantitative Nature

More than 1,300 seedlings of European plum originating from crossing combinations with at least one parent showing hypersensitivity resistance against PPV were analyzed for their reaction to artificial inoculation with PPV using the double grafting method with virus infected interstem. It was shown that the hypersensitivity resistance against the virus is a phenotypically quantitative trait. The different kinds of symptoms observed in the test system, which contribute to the hypersensitivity resistance, range from weak necrosis on the leaf blade and on the stem to the death of the whole young shoots. A hypersensitivity index was developed which helps to determine the degree of hypersensitivity resistance of an individual genotype. Its use is strongly recommended as selection tool in breeding for hypersensitivity resistance.

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Interactions of Plum pox virus strain Rec with Apple chlorotic leafspot virus and Prune dwarf viruses in field-grown transgenic plum Prunus domestica L., clone C5

Plant Protection Science ◽

10.17221/535-pps ◽

2008 ◽

Vol 44 (No. 1) ◽

pp. 1-5 ◽

Cited By ~ 1

Author(s):

J. Polák ◽

M. Ravelonandro ◽

J. Kumar-Kundu ◽

J. Pívalová ◽

R. Scorza

Keyword(s):

Sequence Analysis ◽

Transgenic Plants ◽

Open Field ◽

Recombinant Strain ◽

Dwarf Virus ◽

Plum Pox Virus ◽

Prunus Domestica ◽

Rt Pcr ◽

Antagonistic Effects ◽

Das Elisa

Transgenic plums, Prunus domestica L. clone C5, were inoculated by bud grafting with Plum pox virus (PPV-Rec, recombinant strain originated from plum), PPV-Rec + Apple chlorotic leafspot virus (ACLSV), PPV-Rec + Prune dwarf virus (PDV), and PPV-Rec + ACLSV + PDV. Non-inoculated transgenic plums served as controls. Plants were grown in an open field for 5 years. They were evaluated by visible symptoms, by DAS-ELISA and RT-PCR. Mild PPV symptoms, diffuse spots or rings appeared two years after inoculation in some leaves of plants artificially inoculated with PPV-Rec, PPV-Rec + ACLSV, PPV-Rec + PDV, and PPV-Rec + ACLSV + PDV. Severe PPV symptoms appeared in leaves of shoots growing from infected buds used for inoculation. During the following three years, further weakening of PPV symptoms was observed in transgenic plants. In 2007, very mild PPV symptoms were found in only a few leaves, and over 60%, resp. 70% of the C5 trees showed no PPV symptoms. The presence of PPV was confirmed by ELISA, ISEM and RT-PCR. No difference in PPV symptoms was observed between PPV-Rec and combinations PPV-Rec + ACLSV, PPV-Rec + PDV, PPV-Rec + ACLSV + PDV. No symptoms of ACLSV appeared in combinations of ACLSV with PPV-Rec and PPV-Rec + PDV during 2004–2007, but the presence of ACLSV in leaves of transgenic plants clone C5 was proved by ELISA and RT-PCR. Neither synergistic nor antagonistic effects of ACLSV on PPV-Rec were observed. No symptoms of PDV appeared in combinations of viruses with PDV during 2004–2007. PDV was not detected by ELISA, and the presence of PDV was uncertain by RT-PCR in most of inoculated trees in 2006 and 2007. The results of RT-PCR will be further confirmed by sequence analysis and discussed. These results suggest a possible antagonistic interaction between PPV-Rec and PDV in plum clone C5.

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