Application of airlift bioreactors to accelerate genetic transformation in American chestnut

• Published in: Plant cell, tissue and organ culture tissue and organ culture, 2014-04, Vol.117 (1), p.39-50
• Main Authors: Kong, Lisheng, Holtz, Christine T., Nairn, Campbell J., Houke, Haley, Powell, William A., Baier, Kathleen, Merkle, Scott A.
• Format: Article
• Language: English
• Subjects: Acceleration; Agrobacterium; Air flow; Biomedical and Life Sciences; Bioreactors; Blight; Castanea dentata; Castanea mollissima; Cell culture; Chestnut; Clumps; Cultivation; Dichlorophenoxyacetic acid; Flasks; Genes; Genetic transformation; Genotypes; Glutamine; Hybrids; Inoculation; Life Sciences; Original Paper; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant Sciences; Propagation; Sucrose; Sugar; Thaumatin; Tissue culture; Tissues; Woody plants
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-013-0418-8

Airlift bioreactors (ALBs) were constructed and tested for their potential to enhance chestnut embryogenic tissue proliferation for genetic transformation and mass propagation. Multiple genotypes of American chestnut ( Castanea dentata ) and backcross hybrids of American chestnut and Chinese chestnut ( C. dentata  ×  Castanea mollissima ) were cultured in ALBs. Medium for tissue proliferation in ALBs was woody plant medium supplemented with 3 g l −1 sucrose, 0.5 g l −1 l -glutamine and 2 mg l −1 2,4-dichlorophenoxyacetic acid. Optimum culture conditions for 1-l ALBs included 2 % (w/v) tissue inoculation density for culture initiation, 200 ml min −1 airflow rate, and weekly feeding with fresh medium (80–85 %, v/v). Compared with flasks, ALBs generated higher yields of tissue mass and larger fractions of small cell clumps (