In vitro propagation of plant virus using different forms of plant tissue culture and modes of culture operation

Shih, Sharon; Doran, Pauline M.

doi:10.1016/j.jbiotec.2009.07.007

Cited by 11 publications

(9 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The recombinant virus failed to replicate or lead to significant GFP accumulation in hairy roots due to foreign gene deletion that occurred during serial passaging of the recombinant TMV stocks in plants [46]. Similarly, very limited uptake and replication was observed for infectious recombinant TMV particles co-incubated with cell suspensions [47]. In an alternative approach, hairy roots of N. benthamiana were established from the leaves of plants previously infected with the same recombinant TMV vector.…”

Section: Discussionmentioning

confidence: 99%

RNA viral vectors for improved Agrobacterium-mediated transient expression of heterologous proteins in Nicotiana benthamiana cell suspensions and hairy roots

Larsen

Curtis

2012

BMC Biotechnol

View full text Add to dashboard Cite

BackgroundPlant cell suspensions and hairy root cultures represent scalable protein expression platforms. Low protein product titers have thus far limited the application of transient protein expression in these hosts. The objective of this work was to overcome this limitation by harnessing A. tumefaciens to deliver replicating and non-replicating RNA viral vectors in plant tissue co-cultures.ResultsReplicating vectors derived from Potato virus X (PVX) and Tobacco rattle virus (TRV) were modified to contain the reporter gene β-glucuronidase (GUS) with a plant intron to prevent bacterial expression. In cell suspensions, a minimal PVX vector retaining only the viral RNA polymerase gene yielded 6.6-fold more GUS than an analogous full-length PVX vector. Transient co-expression of the minimal PVX vector with P19 of Tomato bushy stunt virus or HC-Pro of Tobacco etch virus to suppress post-transcriptional gene silencing increased GUS expression by 44 and 83%, respectively. A non-replicating vector containing a leader sequence from Cowpea mosaic virus (CPMV-HT) modified for enhanced translation led to 70% higher transient GUS expression than a control treatment. In hairy roots, a TRV vector capable of systemic movement increased GUS accumulation by 150-fold relative to the analogous PVX vector. Histochemical staining for GUS in TRV-infected hairy roots revealed the capacity for achieving even higher productivity per unit biomass.ConclusionsFor the first time, replicating PVX vectors and a non-replicating CPMV-HT vector were successfully applied toward transient heterologous protein expression in cell suspensions. A replicating TRV vector achieved transient GUS expression levels in hairy roots more than an order of magnitude higher than the highest level previously reported with a viral vector delivered by A. tumefaciens.

show abstract

Section: Discussionmentioning

confidence: 99%

RNA viral vectors for improved Agrobacterium-mediated transient expression of heterologous proteins in Nicotiana benthamiana cell suspensions and hairy roots

Larsen

Curtis

2012

BMC Biotechnol

View full text Add to dashboard Cite

show abstract

“…With this respect, Pelah et al [25] reported the establishment of callus cultures from TYLCV-infected tomato plants that were suitable for in vitro storage of TYLCV-infected callus up to 8 months. Similar tissue culture approaches were developed for the purpose of Tobacco mosaic virus (TMV) propagation in hairy root cultures of Nicotiana benthamiana where the hairy root cultures were directly inoculated by the addition of the virus to the culture medium [26]. Therefore, the described in vitro -inoculation method can be used for prolonged storage of infected material and can be used in exchanging infected plant materials between locations.…”

Section: Discussionmentioning

confidence: 99%

An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Al‐Abdallat

Debei

Asmar

et al. 2010

Virol J

View full text Add to dashboard Cite

BackgroundTomato yellow leaf curl virus (TYLCV) is a member of the family Geminiviridae, genus Begomovirus. To test the infectivity of TYLCV in tomato plants, an improved protocol for inoculation of in vitro-cultured tomato plants was developed.ResultsA TYLCV isolate was cloned, sequenced and used to construct a 1.8-mer infectious clone. Three weeks old microshoots of TYLCV-susceptible tomato plants were inoculated with Agrobacterium tumefaciens harboring the infectious clone for the TYLCV isolate. After two weeks, the TYLCV symptoms started to appear on the in vitro-inoculated plants and the symptoms became more severe and pronounced eight weeks post-inoculation. The method was used efficiently to uncover the resistance mechanism against TYLCV in Solanum habrochaites accession LA 1777, a wild tomato known for its high resistance to whitefly and TYLCV.ConclusionsThe reported in vitro-inoculation method can be used to screen tomato genotypes for their responses to TYLCV under controlled conditions and it will be a useful tool for better understanding of the TYLCV biology in tomato plants.

show abstract

“…Several systems have been developed for transient expression of foreign proteins in plant suspension and hairy root cultures based on viral vectors or agrobacterial co-culture [29,39,[50][51][52]. This approach restricts transformation and protein expression to only certain stages of cell culture, thereby avoiding the reductions in expression levels that occur during multiple cell divisions due to mutations or genetic re-arrangements.…”

Section: Culture and Transgene Stability And Gene Silencingmentioning

confidence: 99%

“…Because chemical agents are applied easily to plant cultures by direct addition to the medium, a wide variety of inducible promoters activated by compounds such as ethanol, estradiol, salicylic acid, dexamethasone and tetracycline [61] are also suitable for in vitro application. In principle, transient transformation and expression systems [29,39,[50][51][52] coupled with knowledge of the timecourse of protease production and/or secretion could also be used to allow higher levels of foreign protein to accumulate in the absence of major protease effects.…”

Section: Inducible Promoters and Transient Expressionmentioning

confidence: 99%

Therapeutically Important Proteins From In Vitro Plant Tissue Culture Systems

Doran¹

2013

Current Medicinal Chemistry

View full text Add to dashboard Cite

Plant cells cultured in liquid medium in bioreactors are now being used commercially to produce biopharmaceutical proteins. The emergence of in vitro plant cell culture as a production vehicle reflects the importance of key biosafety and biocontainment concerns affecting the competitiveness of alternative systems such as mammalian cell culture and agriculture. Food plant species are particularly attractive as hosts for in vitro protein production: the risk of transgene escape and food chain contamination is eliminated using containment facilities, while regulatory approval for oral delivery of drugs may be easier than if non-edible species were used. As in whole plants, proteolysis in cultured plant cells can lead to significant degradation of foreign proteins after synthesis; however, substantial progress has been made to counter the destructive effects of proteases in plant systems. Although protein secretion into the culture medium is advantageous for product recovery and purification, measures are often required to minimise extracellular protease activity and product losses due to irreversible surface adsorption. Disposable plastic bioreactors, which are being used increasingly in mammalian cell bioprocessing, are also being adopted for plant cell culture to allow rapid scale-up and generation of saleable product. This review examines a range of technical and regulatory issues affecting the choice of industrial production platform for foreign proteins, and assesses progress in the development of in vitro plant systems for biopharmaceutical production.

show abstract

In vitro propagation of plant virus using different forms of plant tissue culture and modes of culture operation

Cited by 11 publications

References 33 publications

RNA viral vectors for improved Agrobacterium-mediated transient expression of heterologous proteins in Nicotiana benthamiana cell suspensions and hairy roots

RNA viral vectors for improved Agrobacterium-mediated transient expression of heterologous proteins in Nicotiana benthamiana cell suspensions and hairy roots

An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Therapeutically Important Proteins From In Vitro Plant Tissue Culture Systems

Contact Info

Product

Resources

About