The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica |
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Authors: | Shinjiro Ogita Hamako Sasamoto Edward C Yeung Trevor A Thorpe |
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Institution: | (1) Cell Manipulation Laboratory, Bio-Resources Technology Division, Forestry and Forest Products Research Institute, 305-8687 Ibaraki, Japan;(2) Department of Biological Sciences, University of Calgary, T2N 1N4 Calgary, Alberta, Canada;(3) Present address: Research Association for Biotechnology, Technology Department, 8916-5, Takayama, Ikoma, 630-0101 Nara, Japan |
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Abstract: | Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l−1 glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic
and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic
aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation
of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing
(2400 mg l−1) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic
aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level
of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell
masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues. |
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Keywords: | amino acid Cryptomeria japonica embryogenic tissue glutamine HPLC somatic embryogenesis |
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