Premium
Somatic Embryogenesis and Plant Regeneration from Suspension Cultures of Red Fescue
Author(s) -
Zaghmout O. M. F.,
Torello W. A.
Publication year - 1989
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci1989.0011183x002900030055x
Subject(s) - somatic embryogenesis , biology , callus , sucrose , botany , tissue culture , agar , murashige and skoog medium , kinetin , subculture (biology) , percoll , explant culture , biochemistry , in vitro , genetics , bacteria
The development of embryogenic cell suspension cultures from which whole‐viable plants could be regenerated on a reliable basis would increase the range of techniques that may be applied by somatic approaches toward genetic modification. The objective of this study was to develop such a cultural system from embryogenic callus tissue of ‘Dawson’ red fescue ( Festuca rubra var trichophylla Gaud.). Embryogenic cell suspension cultures were initiated from highly embryogenic callus tissue that had been cultured for over 4 yr. The optimal basal medium for suspension cultures consisted of one‐half strength Murashige and Skoog (MS) salts supplemented with 30 g L −1 sucrose, Gamborgs B‐5 vitamins, 3 g L −1 casein hydrolysate (CH), and 4 mg L −1 of 2,4‐dichlorophenoxyacetic acid (2,4‐D). The addition of CH significantly enhanced culture growth rates as measured by packed cell volumes. Prefiltering newly initiated suspensions through a 31‐μm filter screen was critical in removing relatively faster growing non‐embryogenic (NE) cell types. The use of a discontinuous percoll gradient was as effective as filtering in removing NE cells but was more time consuming. Prolific regeneration of whole, viable plants was attained by transferring embryogenic cell clusters to a hormone‐free, half‐strength MS basal medium supplemented B‐5 vitamins and 8 g L −1 agar.