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© 1995 Oxford University Press

RESEARCH-ARTICLE

Establishment and physiological analyses of photoautotrophic callus cultures of the fern Platycerium coronarium (Koenig) Desv. under CO2 enrichmen

Siew-Hwa Kwa1, Yeow-Chin Wee1, Tit-Meng Lim2 and Prakash P. Kumar1,3

1 Department of Botany, National University of Singapore, Lower Kent Ridge Road, Singapore 0511, Republic of Singapore
2 Department of Zoology, National University of Singapore, Lower Kent Ridge Road, Singapore 0511, Republic of Singapore

3To whom correspondence should be addressed. Fax: +65 779 5671. E-mail: botprakh{at}leonis.nus.sg

Gametophyte-derived callus cultures of Platycerium coronarium could be maintained under photoautotrophic conditions on Murashige and Skoog medium supplemented with 2µM 2,4-dichlorophenoxyacetic acid (2,4-D) and with CO2 enrichment. Progressive reduction of sucrose from the medium resulted in a reduction in growth, but an increase in total chlorophyll content. When subculturing was delayed beyond 2 weeks, callus cells differentiated into gametophytes on the medium with 0.2≤ sucrose and no CO2 enrichment. Enriching the photoautotrophic cultures on 2µM 2, 4-D with 1% CO2 resulted in about 1.7-fold increase in fresh weight within 42 d. Total chlorophyll content was generally higher with 1% CO2 enrichment than with 10%. Fv/Fm ratio was higher for callus on low levels of sucrose (>0.5%) than that on sucrose 1.0%. An increase in autofluorescence of chloroplasts, but not the size, was observed with decreasing sucrose levels in the medium. Autofluorescence decreased with increase in CO2 from 0.03%. Our data are in agreement with the view that long-term exposure to high levels of decrease in photosynthetic capacity.

Key words: Platycerium coronarium, stag's horn fern, autofluorescence of chloroplasts, confocal laser scanning microscope, Fv/Fm ratio, photoautotrophic callus


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