scholarly journals Use of in vitro Cultures of Daphne cneorum L. for the Western Detection of Daphne Virus X

1992 ◽  
Vol 10 (3) ◽  
pp. 153-156
Author(s):  
C. Wei ◽  
M.J. Green ◽  
S.E. Godkin ◽  
P.L. Monette
Keyword(s):  
In Vitro Cultures ◽  
Early Screening ◽  
Virus Elimination ◽  
Viral Coat Protein ◽  
Western Analysis ◽  
Heat Therapy ◽  
Viral Coat ◽  
Ornamental Shrub ◽  
Commercial Micropropagation

Abstract In vitro cultures of Daphne cneorum L. were successfully used as test samples for the detection of the viral coat protein of daphne virus X (DVX) by Western analysis. This powerful serological technique could thus be used where viral antigen concentrations may be very low, such as in the early screening of in vitro-cultured “heat therapy” tips from virus elimination programs. Western analysis was conducted on randomly selected D. cneorum plants from a commercial nursery and on in vitro cultures from a commercial micropropagation lab. All the plants and cultures tested were DVX-infected, supporting the view that this ornamental shrub may be universally infected with DVX.

scholarly journals The elimination of Plum pox virus in plum cv. Bluefree and apricot cv. Hanita by chemotherapy of in vitro cultures

2011 ◽  
Vol 38 (No. 2) ◽  
pp. 49-53 ◽  
Author(s):  
A. Hauptmanová ◽  
J. Polák
Keyword(s):  
In Vitro Cultures ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Virus Elimination ◽  
Skoog Medium ◽  
Negative Results ◽  
Murashige Skoog ◽  
One Year

In vitro cultures of plum cv. Bluefree and apricot cv. Hanita infected with Plum pox virus (PPV) were used for the virus elimination by chemotherapy. Low ribavirin concentrations of 5 and 10 mg/l in Murashige-Skoog medium were applied in the treatment. Plum pox virus was completely eliminated by 5 mg/l of ribavirin in plum cv. Bluefree within twenty weeks and in apricot cv. Hanita in twelve weeks of the application. Plum pox virus was completely eliminated by 10 mg/l of ribavirin both in plum cv. Bluefree and apricot cv. Hanita within twelve weeks. The presence of PPV was not proved by RT-PCR. Clones of plum cv. Bluefree and apricot cv. Hanita were re-tested by RT-PCR one year after the termination of the ribavirin treatment and negative results confirmed the elimination of Plum pox virus.

scholarly journals Inconsistent transmission of banana bunchy top virus in micropropagated bananas and its implication for germplasm screening

1995 ◽  
Vol 46 (3) ◽  
pp. 663 ◽  
Author(s):  
JE Thomas ◽  
MK Smith ◽  
AF Kessling ◽  
SD Hamill
Keyword(s):  
Tissue Culture ◽  
Monoclonal Antibodies ◽  
Sandwich Elisa ◽  
In Vitro Cultures ◽  
Banana Bunchy Top Virus ◽  
Germplasm Screening ◽  
Musa Sp ◽  
Heat Therapy ◽  
Meristem Tip Culture

Banana bunchy top virus (BBTV) was readily transmitted through tissue culture in banana (Musa sp.) cv. Lady finger (AAB) and Cavendish cv. Williams (AAA). Lines derived from infected and healthy field plants had similar in vitro multiplication rates. BBTV-infected in vitro cultures displayed symptoms of stunting, leaf curling, chlorotic and green flecks, and poor root growth. Symptoms became milder with time, and were often difficult to discern in older, rapidly multiplying cultures. A triple antibody sandwich ELISA using polyclonal and monoclonal antibodies was very efficient for detecting BBTV in vitro. Symptomless, ELISA-negative plants arose in 10 out of 11 lines derived from BBTV-infected field plants and first appeared after 9 months continuous in vitro culture at a constant 28�C. Meristem tip culture or heat therapy was not used. These plants remained symptomless and ELISA-negative after planting out in the glasshouse (individual plants checked for up to 16 months). The implications of this inconsistent transmission of BBTV for germplasm indexing and exchange are discussed.

scholarly journals Mechanism of Assembly of Recombinant Murine Polyomavirus-Like Particles

2000 ◽  
Vol 74 (4) ◽  
pp. 1658-1662 ◽  
Author(s):  
Uli Schmidt ◽  
Rainer Rudolph ◽  
Gerald Böhm
Keyword(s):  
Coat Protein ◽  
Disulfide Bonds ◽  
Capsid Assembly ◽  
Particle Assembly ◽  
Viral Coat Protein ◽  
Virus Like Particles ◽  
Equilibrium Reaction ◽  
Viral Coat ◽  
Murine Polyomavirus

ABSTRACT VP1 is the major viral coat protein of murine polyomavirus and can be used for the generation of virus-like particles in vitro. Here, we demonstrate that capsid assembly is an equilibrium reaction followed by oxidation of intracapsomere disulfide bonds, which are not essential for the formation of virus-like particles but enable complete particle assembly and prevent capsid dissembly.

Effect of chemical and heat therapy on virus concentrations in in vitro potato plantlets

1990 ◽  
Vol 68 (7) ◽  
pp. 1515-1521 ◽  
Author(s):  
Helen M. Griffiths ◽  
Steven A. Slack ◽  
John H. Dodds
Keyword(s):  
Virus Titer ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Elimination ◽  
Potato Viruses ◽  
Host Interaction ◽  
Virus Assay ◽  
Heat Therapy ◽  
Nodal Cuttings ◽  
The Impact

Potato (Solanum tuberosum) plantlets were established in vitro to evaluate the effectiveness of chemicals and heat therapy for antiviral activity. Ribavirin was shown to be the most active chemical. Further studies were performed in which plantlets established from nodal cuttings were exposed to an alternating 4-h cycle of 35–31 °C and ribavirin (20 mg/L) therapy for 4 weeks, and then tested quantitatively for virus titer by the enzyme-linked immunosorbent assay. Plantlets were further propagated via nodal cuttings at room temperature without exposure to ribavirin. Virus assays were performed and virus-free plantlets were grown out as mature plants for a final virus assay. Ribavirin alone or in combination with heat therapy was effective in reducing potato virus M, S, and X titers (between 10- and 60-fold). Virus-free plants were obtained from both treatments. For potato viruses Y and leafroll, about a fourfold reduction in titer was observed after the ribavirin–heat treatments. When multiple viruses were present, quantitative (10- to 20-fold) reductions in potato viruses M, S, X, and leafroll were observed and resulted in plants free of potato viruses M, S, and X. This protocol enables the investigator to quantitatively monitor the impact of a single variable on the virus–host interaction. We have also been able to demonstrate that virus elimination can be obtained without a requisite meristem-tip excision step.

scholarly journals 052 In Vitro Culture Initiation and Proliferation of Exochorda racemosa

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 397C-397
Author(s):  
Guochen Yang ◽  
Marihelen Kamp-Glass
Keyword(s):  
In Vitro Culture ◽  
Woody Plant ◽  
Shoot Proliferation ◽  
Deionized Water ◽  
In Vitro Cultures ◽  
Culture Initiation ◽  
Woody Plant Medium ◽  
Ornamental Shrub ◽  
Ornamental Species

Exochorda racemosa is an ornamental shrub with white flowers that is spiraea-like, deciduous, and hardy. The buds resemble pearls. Normally it is propagated by seeds, layers, and cuttings of softwood. However, it is a slow process that takes a few years to produce a reasonable size plant for the demanding market. Our objective was to establish a successful in vitro culture and to rapidly multiply this ornamental species. Softwood explant materials were collected from a local nursery and were disinfested with 15% bleach solution and rinsed three times with sterile distilled and deionized water. In vitro cultures were established and maintained in woody plant medium (WPM) supplemented with BA at 0.1 mg·L-1, 3% sucrose, and 0.7% agar with the pH adjusted to 5.8. Then shoots were transferred to the multiplication medium containing BA, CPPU, or thidiazuron (TDZ) at various concentrations. Preliminary results show that explants cultured on medium containing TDZ produced the best shoot proliferation.

scholarly journals Infectious transcripts from cloned cDNA of potato leafroll luteovirus.

1998 ◽  
Vol 45 (2) ◽  
pp. 611-619 ◽  
Author(s):  
E Sadowy ◽  
K Pluta ◽  
B Gronenborn ◽  
D Hulanicka
Keyword(s):  
Plant Viruses ◽  
Cdna Clones ◽  
Viral Coat Protein ◽  
Restriction Sites ◽  
Viral Coat ◽  
Cloned Cdna ◽  
Potato Leafroll Luteovirus ◽  
Leafroll Virus ◽  
Rna Polymerase Promoter

Infectious transcripts play a key role in the research on plant viruses at the molecular level. A number of cDNA clones covering the whole genome of the Polish isolate of potato leafroll virus were constructed. Four overlapping clones were selected and assembled using restriction sites. The full copy was positioned between T7 RNA polymerase promoter and unique ScaI site. The full-length capped transcripts of the sequence of the viral genome synthesised in vitro were able to replicate in protoplasts and to produce the viral coat protein.

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