Procedures were developed to determine if live, adult two-spotted spidermites (Tetranychus urticae Koch) could be surface disinfested before being introduced into in vitro cultures of torenia (Torenia fournieri L.). Three time periods (5, 10, and 15 minutes) and five levels of sodium hypochlorite (0.05% to 0.25%) were evaluated. Surface disinfestation was accomplished by agitating 2 × 3 cm pieces of infested bean leaves in sodium hypochlorite solutions and then drying in a mite drier apparatus. All sodium hypochlorite concentrations disinfested the mites completely, however high concentration levels were lethal to the mites. Exposure periods of 10 and 15 minutes also significantly increased mortality. For optimum disinfestation of two-spotted spidermites with minimum mortality, a concentration of 0.05% sodium hypochlorite and 0.05% Tween-20 for 5 minutes should be used.
Masood Z. Hadi, Mark P. Bridgen, and John P. Sanderson
Mark P. Bridgen, Masood Z. Hadi, and Madeleine Spencer-Barreto
A laboratory exercise on direct and indirect organogenesis from leaf explants is presented for students of plant tissue culture or plant propagation. Torenia fournieri, the wishbone flower, is used for this laboratory exercise because the in vitro production of adventitious shoots from Torenia is easy to control, seeds are easy to obtain, and plants are easy to grow. Direct shoot organogenesis results from leaf explants without an intervening callus phase, and indirect shoot organogenesis is possible after 4 to 6 weeks of callus production from leaf explants. The basal medium for all forms of organogenesis contains Murashige and Skoog (MS) salts and vitamins, 30 g sucrose/liter, and 7 g agar/liter at pH 5.7. To obtain direct shoot organogenesis, leaf explants should be placed on the MS basal medium with 1.1 μM (0.25 mg·liter-1) 6-benzylaminopurine (BAP) and 0.25 μM (0.05 mg·liter-1) indole-3-butyric acid (IBA). If leaf explants are placed on MS medium with 2.3 μM (0.5 mg·liter-1) 2,4-dichlorophenoxyacetic acid (2,4-D), callus formation will occur. Callus can be subcultured onto a MS medium with 8.88 μM BAP (2.0 mg·liter-1) plus 2.5 μM IBA (0.5 mg·liter-1) for indirect shoot organogenesis to occur.