Tissue culture and plant regeneration from immature embryo explants of Barley, Hordeum vulgare

Abstract

Immature embryo explants taken 8 days after anthesis were used to establish callus cultures of spring barley. Two types of calli were observed. A soft, watery callus produced a limited number of shoots and a harder, more compact, yellowish callus gave rise to numerous green primordia and shoots. Gamborg's B5 basal medium supplemented with either 2,4-D (2,4-dichlorophenoxyacetic acid) or Cl3 POP (2,4,5-trichlorophenoxypropionic acid) was found to give good callus growth and shoot initiation. Media containing 2,4-D at 1.0 mg L(-1) or Cl3 POP at 5.0 mg L(-1) produced numerous cultures resulting in regeneration of plants. Plantlets developed roots on basal medium with Cl3 POP at 1.0 mg L(-1) or on auxin-free medium. Twenty genetically diverse genotypes were screened to determine if these techniques were suitable for a wide range of spring barley cultivars. Regeneration of plantlets was obtained for 19 of the 20 genotypes approximately 4 months after culture initiation. Lines differed in the ability to develop vigorously growing calli and in the ability of calli to develop large numbers of shoots and regenerated plantlets.