scholarly journals Field Trials of Plum Clones Transformed with the Plum pox virus Coat Protein (PPV-CP) Gene

Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1012-1018 ◽  
Author(s):  
T. Malinowski ◽  
M. Cambra ◽  
N. Capote ◽  
B. Zawadzka ◽  
M. T. Gorris ◽  
...  

Transgenic clones C2, C3, C4, C5, C6, and PT-6, of plum (Prunus domestica L.) transformed with the coat protein (CP) gene of Plum pox virus (PPV), PT-23 transformed with marker genes only, and nontransgenic B70146 were evaluated for sharka resistance under high infection pressure in field trials in Poland and Spain. These sites differed in climatic conditions and virus isolates. Transgenic clone C5 showed high resistance to PPV at both sites. None of the C5 trees became naturally infected by aphids during seven (Spain) or eight (Poland) years of the test, although up to 100% of other plum trees (transgenic clones and nontransgenic control plants) grown in the same conditions showed disease symptoms and tested positively for PPV. Although highly resistant, C5 trees could be infected artificially by chip budding or via susceptible rootstock. Infected C5 trees showed only a few mild symptoms on single, isolated shoots, even up to 8 years post inoculation. These results clearly indicate the long-term nature and high level of resistance to PPV obtained through genetically engineered resistance.

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 532a-532
Author(s):  
Ralph Scorza ◽  
Laurene Levy ◽  
Vern Damsteegt ◽  
Ann Callahan ◽  
Kevin Webb ◽  
...  

Sharka or plum pox virus (PPV) is a major disease of stone fruit and causes severe economic losses in Europe. There is little resistance to PPV in most Prunus species, thus genetic engineering represents a potentially useful approach to obtain resistant germplasm. Transgenic plums containing the PPV coat protein (CP) or the related papaya ringspot virus (PRV)-CP gene were produced through Agrobacterium tumefaciens-mediated transformation. These transgenic plum clones were then evaluated for resistance to PPV infection in the greenhouse by graft or aphid inoculation with PPV. While symptoms of PPV appeared in most transgenic clones, all plants of PPV-CP transgenic clone C5 were symptomless and ELISA and immunocapture-reverse transcriptase PCR negative for over three years following inoculation with two strains of PPV (Ravelonandro et al., Plant Dis. 81:1231-1235, 1997). Clone C5, which contains multiple copies of the PPV-CP gene, was hybridized with PRV-CP transgenic plants or untransformed plum cultivars. Progeny were obtained containing no transgenes, only the PPV-CP, only the PRV-CP, or both the PRV-CP and PPV-CP transgenes. Seedlings were inoculated with PPV. At 5 and 11 months post-inoculation, seedlings containing the PPV-CP genes from C5 were symptomless and ELISA negative. Seedlings containing only PRV-CP transgenes or non-transformed controls showed symptoms of PPV infection and were ELISA positive. These results indicate that the PPV-CP transgenes can be transferred to progeny through hybridization and that these genes can impart resistance to PPV in transgenic seedlings. The inheritance of the multicopy inserts of the PPV-CP and PRV-CP transgenes is being analyzed. The combined effects of both transgenes on resistance to PPV and the stability of PPV resistance in the progeny of the resistant C5 transgenic line are currently under evaluation.


2012 ◽  
Vol 48 (Special Issue) ◽  
pp. S43-S48 ◽  
Author(s):  
J. Polák ◽  
J. Kumar ◽  
B. Krška ◽  
M. Ravelonandro

Commercialisation of Biotech/GM (Biotech) crops started in 1995. Not only field crops, but also horticultural transgenic crops are under development and are beginning to be commercialised. Genetic engineering has the potential to revolutionise fruit tree breeding. The development of transgenic fruit cultivars is in progress. Over the past 20 years an international public sector research team has collaborated in the development of HoneySweet plum which is highly resistant to Plum pox virus (PPV) the most devastating disease of plums and other stone fruits. HoneySweet was deregulated in the USA in 2010. HoneySweet (aka C5) has been evaluated for eleven years (2002–2012) in a regulated field trial in the CzechRepublic for the resistance to PPV, Prune dwarf virus (PDV), and Apple chlorotic leaf spot virus (ACLSV), all of them being serious diseases of plum. Even under the high and permanent infection pressure produced through grafting, PPV has only been detected in HoneySweet trees in several leaves and fruits situated close to the point of inoculum grafting. The lack of infection spread in HoneySweet demonstrates its high level of PPV resistance. Co-infections of PPV with PDV and/or ACLSV had practically no influence on the quantity and quality of HoneySweet fruit which are large, sweet, and of a high eating quality. In many respects, they are superior to the fruits of the well-known cultivar Stanley. Many fruit growers and fruit tree nurseries in the CzechRepublic are supportive of the deregulation of HoneySweet plum to help improve the plum production and control the spread of PPV.


Plant Disease ◽  
1997 ◽  
Vol 81 (11) ◽  
pp. 1231-1235 ◽  
Author(s):  
M. Ravelonandro ◽  
R. Scorza ◽  
J. C. Bachelier ◽  
G. Labonne ◽  
L. Levy ◽  
...  

Transgenic plum trees (Prunus domestica) containing the plum pox potyvirus coat protein (PPV-CP) gene were inoculated with PPV by aphid feeding or chip budding. Infection was monitored by evaluation of virus symptoms, DAS-ELISA, and immunoblot assays. Based on observations and analyses over 3 years including two dormancy cycles, one out of five transgenic clones (C-5), was found to be resistant to infection whether inoculated by aphids or by chip budding. PPV could not be detected in any inoculated plants of the C-5 clone by immunoblot or immunocap-ture-reverse transcriptase-polymerase chain reaction assays. To our knowledge, this is the first P. domestica clone resistant to PPV infection produced by genetic engineering.


HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 862D-862
Author(s):  
Jean-Michel Hily ◽  
Ralph Scorza* ◽  
Michel Ravelonandro

We have shown that high-level resistance to plum pox virus (PPV) in transgenic plum clone C5 is based on post-transcriptional gene silencing (PTGS), otherwise termed RNA silencing (Scorza et al. Transgenic Res. 10:201-209, 2001). In order to more fully characterize RNA silencing in woody perennial crops, we investigated the production of short interfering RNA (siRNA) in transgenic plum clones C3 and C5, both of which harbor the capsid protein (CP) gene of PPV. We used as a control, plum PT-23, a clone only transformed with the two marker genes, NPTII and GUS. We show in the current report that C5 constitutively produces two classes of siRNA, the short (21-22 nucleotides) and long (≈27 nucleotides) species in the absence of PPV inoculation. Transgenic susceptible clone C3 and the control clone PT-23, when healthy, produce no siRNA. Upon infection, these clones produce only the short siRNA (21-22 nt). This siRNA production suggests that plum trees naturally respond to virus infection by initiating PTGS or PTGS-like mechanisms. This study also suggests that high-level virus resistance in woody perennials may require the production of both the short and long size classes of siRNA, as are produced by the resistant C5 plum clone.


2009 ◽  
Vol 22 (10) ◽  
pp. 1302-1311 ◽  
Author(s):  
V. Decroocq ◽  
B. Salvador ◽  
O. Sicard ◽  
M. Glasa ◽  
P. Cosson ◽  
...  

In Arabidopsis thaliana Columbia (Col-0) plants, the restriction of Tobacco etch virus (TEV) long-distance movement involves at least three dominant RTM (restricted TEV movement) genes named RTM1, RTM2, and RTM3. Previous work has established that, while the RTM-mediated resistance is also effective against other potyviruses, such as Plum pox virus (PPV) and Lettuce mosaic virus (LMV), some isolates of these viruses are able to overcome the RTM mechanism. In order to identify the viral determinant of this RTM-resistance breaking, the biological properties of recombinants between PPV-R, which systemically infects Col-0, and PPV-PSes, restricted by the RTM resistance, were evaluated. Recombinants that contain the PPV-R coat protein (CP) sequence in an RTM-restricted background are able to systemically infect Col-0. The use of recombinants carrying chimeric CP genes indicated that one or more PPV resistance-breaking determinants map to the 5′ half of the CP gene. In the case of LMV, sequencing of independent RTM-breaking variants recovered after serial passages of the LMV AF199 isolate on Col-0 plants revealed, in each case, amino acid changes in the CP N-terminal region, close to the DAG motif. Taken together, these findings demonstrate that the potyvirus CP N-terminal region determines the outcome of the interaction with the RTM-mediated resistance.


2007 ◽  
Vol 37 (3) ◽  
pp. 515-522 ◽  
Author(s):  
Tore Skrøppa ◽  
Ketil Kohmann ◽  
Øystein Johnsen ◽  
Arne Steffenrem ◽  
Øyvind M. Edvardsen

We present results from early tests and field trials of offspring from two Norway spruce ( Picea abies (L.) Karst.) seed orchards containing clones that have been transferred from high altitudes to sea level and from northern to southern latitudes. Seedlings from seeds produced in the low-altitude seed orchard developed frost hardiness later at the end of the growth season, flushed later in field trials, and grew taller than seedlings from seeds produced in natural stands. They had the lowest mortality rate and the lowest frequency of injuries in the field trials. Similar results were observed in seedlings from seeds produced in the southern seed orchard. We found no adverse effects of the changed growth rhythm. Seedlings from two seed crops in the southern orchard, produced in years with a warm and a cold summer, had different annual growth rhythms. The results are explained mainly by the effects of the climatic conditions during the reproductive phase. Seed crops from different years in the same seed orchard may produce seedlings that perform as if they were from different provenances. It is argued that the effects of the climatic conditions during seed production must contribute to the variation among provenances of Norway spruce.


2020 ◽  
Vol 15 (1) ◽  
pp. 711-720
Author(s):  
Janetta Niemann ◽  
Justyna Szwarc ◽  
Jan Bocianowski ◽  
Dorota Weigt ◽  
Marek Mrówczyński

AbstractRapeseed (Brassica napus) can be attacked by a wide range of pests, for example, cabbage root fly (Delia radicum) and cabbage aphid (Brevicoryne brassicae). One of the best methods of pest management is breeding for insect resistance in rapeseed. Wild genotypes of Brassicaceae and rapeseed cultivars can be used as a source of resistance. In 2017, 2018, and 2019, field trials were performed to assess the level of resistance to D. radicum and B. brassicae within 53 registered rapeseed cultivars and 31 interspecific hybrid combinations originating from the resources of the Department of Genetics and Plant Breeding of Poznań University of Life Sciences (PULS). The level of resistance varied among genotypes and years. Only one hybrid combination and two B. napus cultivars maintained high level of resistance in all tested years, i.e., B. napus cv. Jet Neuf × B. carinata – PI 649096, Galileus, and Markolo. The results of this research indicate that resistance to insects is present in Brassicaceae family and can be transferred to rapeseed cultivars. The importance of continuous improvement of rapeseed pest resistance and the search for new sources of resistance is discussed; furthermore, plans for future investigations are presented.


Author(s):  
K. Saratbabu ◽  
K. Vemana ◽  
A.K. Patibanda ◽  
B. Sreekanth ◽  
V. Srinivasa Rao

Background: Peanut stem necrosis disease (PSND) caused by Tobacco streak virus (TSV) is a major constraint for groundnut production in Andhra Pradesh (A.P.). However, studies on prevalence and spread of the disease confined to only few districts of A.P. with this background current study focused on incidence and spread of the disease in entire state of A.P. Further an isolate of TSV occurring in A.P. characterized on the basis of genetic features by comparing with other TSV isolates originated from different hosts and locations from world.Methods: Roving survey was conducted during kharif 2017-18 in groundnut growing districts of Andhra Pradesh (A.P.) for peanut stem necrosis disease incidence. Groundnut plants showing PSND symptoms were collected and tested with direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). Groundnut samples found positive by ELISA once again tested by reverse transcription polymerase chain reaction (RT-PCR). The representative TSV-GN-INDVP groundnut isolate from Prakasham district was maintained on cowpea seedlings by standard sap inoculation method in glasshouse for further molecular characterization. The Phylogenetic tree for coat protein (CP) gene was constructed using aligned sequences with 1000 bootstrap replicates following neighbor-joining phylogeny.Result: Thirty-eight (52.7%) of seventy-two groundnut samples collected from different locations in A.P were given positive reaction to TSV by DAC-ELISA. For the first time, PSND incidence observed in coastal districts (Krishna, Guntur, Sri Pottisriramulu Nellore, Prakasham) of A.P. Maximum PSND incidence recorded from Bathalapalli (22.2%) and the minimum incidence in Mulakalacheruvu (4.1%). The coat protein (CP) gene of TSV-GN-INDVP groundnut isolate was amplified by RT-PCR and it shared maximum per cent nucleotide identity (97.51-98.62%) with TSV isolates from groundnut and other different crops reported in India. All Indian isolates cluster together irrespective of crop and location based on the phylogenetic analysis.


Author(s):  
E.N. Tyukhtina ◽  

The article deals with one of the most relevant topics of the modern city and ecology - the problem of landscaping in Volgograd. The city is characterized by an industrial urbanism, with a high concentration of population, saturation of production facilities and vehicles, which contributes to a high level of negative impact on the environment. The pace of greening of the city lags behind the pace of construction of the residential sector, and the existing green spaces do not meet the sanitary and hygienic, aesthetic requirements. The prospects of this problem are determined, the existing green spaces are considered, data on the climatic conditions of the city and the ecological situation are given. Improvement in the field of landscaping is associated with the scientifically sound placement of green spaces, taking into account all the factors that affect the growth and development of plants in the urban environment. Unfortunately, due to the uneven distribution of the assortment of trees and shrubs, the species composition of the central streets and squares of the city is extremely poor. The study focuses on the role of the botanical garden in urban landscaping and aims to create recommendations for the sorting of tree and shrub plants for urban landscaping. As a result of the analysis of the assortment of ornamental plants of the open ground of the botanical garden, the article proposed species and varieties of woody and shrubby plants that have passed the introduction tests for more than 5-10 years. Recommendations for sorting are aimed at improving the ecological situation of the city and greening the recreation areas of citizens. The proposed list of tree and shrub species cannot fully solve the problem of greening the city, but it will help to significantly enrich the composition of tree and shrub plants.


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