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Development of grapevine somatic embryogenesis using an air-lift bioreactor as an efficient tool in the generation of transgenic plants

Paper ID Volume ID Publish Year Pages File Format Full-Text
24762 43535 2009 7 PDF Available
Title
Development of grapevine somatic embryogenesis using an air-lift bioreactor as an efficient tool in the generation of transgenic plants
Abstract

The grapevine genetic transformation programs have relayed on the use of solid media-based somatic embryogenesis. To reach a high throughput of candidate gene evaluation in ‘Thompson Seedless’, a semi-automatic system allowing viable transformation of explants was designed. An intermediate procedure using liquid media and agitated flasks was first characterized, leading to reduction in the biomass duplication time of pro-embryogenic (PE) cells from 30 d in dishes to 14 d. The oxygen transfer coefficient value in this system was 213 h−1 at 120 rpm and 25 °C with a 16/8-h (light/darkness) photoperiod. The scaling-up to the air-lift bioreactor decreased the biomass duplication time of PE cells up to 5.3 d post-inoculation (pi) and an average volumetric productivity of 1.6 g/(d × L). Although slight browning was seen in the explants during the phase of 8–14 d pi, no losses in their viability and regenerative capability were observed. Cultured cells showed normal elongation in the transition from heart- to the torpedo-shape and finally to advanced developmental stages, with radicle emergence and whole plant generation. Agrobacterium-mediated transformation of cells was efficiently incorporated after this multiplication process by use of conventional procedures in dishes, allowing the generation of transgenic plantlets confirmed by PCR.

Keywords
SE, somatic embryogenesis; PE, pro-embryonic; E, embryonic; μ, specific growth rate; kLa, volumetric oxygen transfer coefficient; Qx, volumetric productivities; LPM, liters per minute; ɛ, gas hold-up; tc, circulation time; td, biomass duplication time; tm
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Development of grapevine somatic embryogenesis using an air-lift bioreactor as an efficient tool in the generation of transgenic plants
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 139, Issue 1, 1 January 2009, Pages 95–101
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us