Hyperhydricity in shoot cultures of Scrophularia yoshimurae can be effectively reduced by ventilation of culture vessels.
An effective procedure for obtaining healthy shoots from nodal segments of Scrophularia yoshimurae is described. Nodal segments cultured on Murashige and Skoog's (MS) basal medium supplemented with 1.0 mg L(-1) benzyladenine (BA) and 0.2 mg L(-1) alpha-naphthaleneacetic acid (NAA) induced multiple shoots in conical flasks that differed in the way they were closed and sealed. Hermitically sealed culture vessels resulted in high hyperhydricity/vitrification. High concentrations of ethylene and CO2 were found to accumulate in these vessels. The hyperhydricity of the shoot cultures could be decreased by progressively ventilating the vessels. Exchange of gases was achieved by removing the Parafilm sealing without affecting sterility. This reduced the hyperhydricity rate and gave a good recovery of vitrified shoots, but resulted in decreased proliferation and a dehydration of proliferating nodal segments and the culture medium. The best number of normal shoots was observed when the parafilm was removed for gaseous exchange after four weeks of culture incubation. The results show that hyperhydricity in shoot cultures of S. yoshimurae could be prevented by sufficient gas exchange during culture.
Lai
Chien-Chou
CC
Department of Bio-lndustrial Mechatronics Engineering, National Taiwan University, Taipei 106, Taiwan.
Lin
Hong-Ming
HM
Nalawade
Satish Manohar
SM
Fang
Wei
W
Tsay
Hsin-Sheng
HS
D000332:Aerobiosis; D002245:Carbon Dioxide / Q000032:analysis; D018929:Cell Culture Techniques; D003470:Culture Media; D035264:Flowers / Q000502:physiology; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D018520:Plant Shoots / Q000166:cytology / Q000502:physiology*; D032266:Scrophularia / Q000166:cytology / Q000502:physiology*
D003470:Culture Media; D009280:Naphthaleneacetic Acids; D002245:Carbon Dioxide; C034182:1-naphthaleneacetic acid