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.

Generation of Large Numbers of Independently Transformed Fertile Barley Plants

“…Callus response and the production of regenerable tissue from this explant is observed with many common cultivars (Goldstein and Kronstad, 1986;Lührs and Lorz, 1987;Bregitzer, 1991). Almost every IE of cv Golden Promise used in this study formed embryogenic callus.…”

Adventitious Shoots From Cotyledons of Immature Cherry and Apricot Embryos

“…1 (Goldstein and Kronstad 1986). Callus originating from immature embryos was also found to be a requirement for regeneration of peach plants (Hammerschlag et al 1985 …”

In vitro genetics of barley (Hordeum vulgare L.): Response of immature embryos to 2,4-dichlorophenoxyacetic acid

“…It must be noted, however, that this study differed from the present one in that the embryos were placed with the scutellum in contact with the medium. Embryo orientation has been reported to have a significant effect on the frequency and amount of callus produced from immature embryos (Dale and Deambrogio, 1979;Powell, Hayter and Wood, 1983), although this was not found to be so in the study of Goldstein and Kronstad (1986).…”

“…Furthermore, the relationship between callus initiation, callus proliferation and shoot differentiation will be investigated (Goldstein and Kronstad, 1986). This aspect of in vitro development is important since it will determine whether or not these responses are under separate genetic control and hence amenable to independent adjustment by suitable selection procedures.…”

“…These authors found differences in the amount of callus formation between the two cultivars Akka and Himalaya. In the most recent study (Goldstein and Kronstad, 1986), which involved 20 spring barley genotypes, callus fresh weight was used as an estimate of response in vitro.…”