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Factors Influencing Somatic Embryo Induction from Orchardgrass Anther Cultures

Agronomy Dep., Univ. of Illinois, Urbana, IL 61801
Dep. of Plant and Soil Science, Univ of Tennessee, P. O. Box 1071, Knoxville, TN 37901-1071

^* Corresponding author.

Somatic embryos from orchardgrass (Ductylis glomerata L.) anther explants have been initiated in our lab but further research is needed to optimize this response. The objectives of this research were to: (i) determine the role of anther density/orientation and various medium supplements on embryogenesis; and (ii) examine chromosomes and nucleoli from regenerated plants to determine a gametic or somatic cell origin. Anthers were pretreated for 0, 3, or 6 wk at 4°C before planting on Schenk and Hildebrandt (SH) medium containing various levels of dicamba, sucrose, or the cytokinins, 6- furfurylaminopurine (kinetin) or benzylaminopurine (BAP). Incubation was in the dark at 25 °C. No differences in response were obtained from a flat vs. an on edge orientation but a density of 10 anthers/mL resulted in a higher frequency of embryogenesis than 100 anthers/mL. Optimum responses were obtained with 30 pmol L^–1 dicamba and 263 µmol L^–1 sucrose. Both cytokinins had a significant inhibitory effect at all concentrations tested. Somatic embryos were formed directly from somatic tissue of anthers receiving a 0- or 3-wk cold pretreatment whereas those receiving a 6-wk cold pretreatment initiated embryogenic calli. One of six plants regenerated from anthers exposed to the 6-wk cold pretreatment was a mixoploid containing cells with 14, 28,56, and more than 100 chromosomes and 1 to 8 nucleoli per nucleus. This work suggests that cytokinins hinder whereas sucrose, dicamba, and a cold pretreatment promote embryogenesis from orchardgrass anthers.

Received for publication December 30, 1987.