Application of airlift bioreactors to accelerate genetic transformation in American chestnut

Kong, Lisheng; Holtz, Christine T.; Nairn, Campbell J.; Houke, Haley; Powell, William A.; Baier, Kathleen; Merkle, S. A.

doi:10.1007/s11240-013-0418-8

Cited by 23 publications

(11 citation statements)

References 26 publications

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“…We have also performed experiments to determine whether or not overexpression of the gene increased the resistance of regenerated transgenic plants to the ink disease caused by Phytophthora cinnamomi Rand, although data are not yet available. Similarly, in American chestnut a Castanea mollissima thaumatin-like protein gene was also transformed into somatic embryos (Kong et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated transformation of European chestnut somatic embryos with a Castanea sativa (Mill.) endochitinase gene

et al. 2016

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Chestnut blight, caused by Cryphonectria parasitica, is a severe disease that has devastated chestnut stands in North America and Europe. Genes encoding hydrolytic enzymes such as chitinases, which can degrade fungal cell wall components, are attractive candidates for improving disease resistance. This report describes a reliable and efficient protocol for the Agrobacterium-mediated transformation of somatic embryos of European chestnut with the endogenous CsCh3 gene that codes for chitinase. The transformation efficiency, determined on the basis of the fluorescence of surviving explants, was genotype-dependent. Although somatic embryos of all three lines evaluated were transformed, the best results were obtained with somatic embryos derived from line CI-9 (20%). The addition of silver thiosulphate (20 or 40 μM) improved the transformation efficiency of somatic embryos derived from lines CI-3 and CI-9, although the differences were not significant. A total of 88 independent transformed lines were obtained. The presence of transgenes was confirmed by green fluorescent protein (GFP) expression, PCR and Southern blot analysis. Transgenic lines were maintained by secondary embryogenesis or cryopreservation following vitrification procedures. Maturation and germination of transformed somatic embryos yielded transgenic plants. Fluorescence indicating overexpression of the transgenes was observed in somatic embryos and also in shoots and leaves. No phenotypic differences were found relative to control plants, suggesting a lack of any cytotoxic effects of the GFP.

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Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated transformation of European chestnut somatic embryos with a Castanea sativa (Mill.) endochitinase gene

et al. 2016

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“…Among the procedures developed to prevent such problems, temporary immersion systems (TIS) are the most popular. These systems have been used to culture somatic embryos of different plant species, including woody plants such as Hevea brasiliensis (rubber tree), Coffea Arabica & N. Vidal nieves@iiag.csic.es 1 (coffee), and Theobroma cacao (cacao) (Etienne et al 1997; Etienne-Barry et al 1999;Niemenak et al 2008), besides members of Fagaceae family such as Quercus robur (pedunculate oak), Q. suber (cork oak), and Castanea dentata (American chestnut) (Mallón et al 2012(Mallón et al , 2013Pérez et al 2013;Kong et al 2014).…”

Section: Introductionmentioning

confidence: 99%

A temporary immersion system for micropropagation of axillary shoots of hybrid chestnut

Vidal

Blanco

Cuenca

2015

Plant Cell Tiss Organ Cult

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A protocol for culturing chestnut axillary shoots by temporary immersion in liquid medium was developed. The influence of type of explant, support material, bioreactor, and immersion was investigated for five artificial hybrids and five natural hybrids of Asian and European chestnut selected for resistance to ink disease. The type of explant influenced shoot quality and proliferation rates, and basal explants with callus produced more and longer shoots than apical and nodal segments. Use of rockwool cubes as support material prevented hyperhydricity and allowed proliferation of explants in Murashige and Skoog medium with half-strength nitrates supplemented with 0.22 lM BA and 3 % sucrose, cultured both in plantform TM and RITA Ò vessels with three or six immersions per day and additional aeration of 1 min per hour in the case of plantform TM bioreactors. Basal explants cultured in plantform TM for 5 weeks produced long shoots suitable for rooting, whereas apical and nodal explants cultured in plantform TM or RITA Ò produced shorter shoots that were suitable for maintenance of stock. For most of the clones, similar or higher proliferation rates were observed when cultured in liquid medium than in semisolid medium, with the additional benefit of cost-reduction of the former system. Shoots developed in liquid medium were submitted to ex vitro root induction by dipping in indole-3-butyric acid, and acclimatized under greenhouse conditions. This is the first demonstration of the production of chestnut plantlets from shoots cultured in liquid medium, and the protocol presented here shows good potential for application in large-scale propagation.

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“…Besides simple impeller system, further modification by using an airlift or simple tubing system for aeration could also be incorporated in both 2-in-1 MoSLIM and SLIM-FaTT systems. The application of airlift bioreactor system was proven effective in the multiplication of American chestnut cultures (Kong et al, 2014). Furthermore, this technology can be further exploited by semi or fully-automating the oil palm clonal production process.…”

Section: Resultsmentioning

confidence: 99%

Simple Impeller Systems for Maintenance of Oil Palm Culture Aggregates

Tarmizi¹

2016

JOPR

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Scaling up of liquid culture systems generally involves moving from the use of simple shake flasks to bioreactors or specialised vessels; this is costly. A new innovation called the Two-in-One MPOB Simple Impeller (2-in-1 MoSLIM) was developed using commonly available Schott bottles in the laboratory. This system provided simultaneous aeration and agitation (two-inone) in a single device for tissue propagation in liquid culture. The 2-in-1 MoSLIM produced cell aggregates with fresh weight increments of two-to six-fold over 30-40 days. This system was a convenient alternative compared to the conventional shake flask system. Multiplication of cultures in the 2-in-1 MoSLIM did not require any shaker or a big space area. This system with a working volume of 300-700 ml used a simple impeller and a pump for agitation and aeration purposes. However, with the 2-in-1 MoSLIM, media replenishment remained a tedious task. To overcome this, modifications were made to the system to enable media replenishment on-site without the need of a sterile hood. The adaptation of 2-in-1 MoSLIM with an earlier innovation, Fast Transfer Technique (MoFaTT) in Liquid Culture System, resulted in the development of the Simple Impeller with Fast Transfer Technique (SLIM-FaTT) system. This new system can be applied to the liquid culture system of any crop with a potential towards automation.

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Application of airlift bioreactors to accelerate genetic transformation in American chestnut

Cited by 23 publications

References 26 publications

Agrobacterium-mediated transformation of European chestnut somatic embryos with a Castanea sativa (Mill.) endochitinase gene

Agrobacterium-mediated transformation of European chestnut somatic embryos with a Castanea sativa (Mill.) endochitinase gene

A temporary immersion system for micropropagation of axillary shoots of hybrid chestnut

Simple Impeller Systems for Maintenance of Oil Palm Culture Aggregates

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