Graduate Thesis Or Dissertation
 

Tissue culture and plant regeneration from immature embryo explants of twenty-two genotypes of barley, Hordeum vulgare L.

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  • Immature embryo explants taken eight days after anthesis were used to establish callus cultures of spring barley. Two types of calli were observed. A soft watery callus which produced a limited number of shoots and a harder yellowish callus that gave rise to numerous green primordia and shoots. Gamborg's B5 basal medium supplemented with either 2,4-D 2,4-dichlorophenoxyacetic acid) or C1₃ POP (2,4,5- trichlorophenoxypropionic acid) was found to support the growth and formation of callus capable of shoot and root development. Among the hormone concentrations tested, 2,4-D at 1.0 mg L⁻¹ and C1₃, POP at 5.0 mg L⁻¹ gave rise to numerous callus cultures capable of plantlet regeneration. Twenty genetically diverse genotypes were screened to determine if the methods developed were suitable for tissue culture propagation of spring barley cultivars. Regeneration of plantlets was achieved for 19 of the 20 genotypes approximately 4 months after culture initiation. Differences were detected in the ability of these genotypes to develop into vigorously growing callus cultures capable of shoot and root initiation. Specific cultivars and advanced breeding program lines have been identified for their ability to produce many shoots and several plantlets from a single embryo explant. Evidence for heterosis in tissue culture was found for callus production, root and shoot development, and regeneration of plantlets in four spring barley crosses. A heterotic effect was detected in all F₁ tissue culture populations for plantlet regeneration and gain in fresh weight 9 weeks after culture initiation. The results clearly demonstrate that heterosis was present in F₁ tissue culture populations from spring barley crosses. The increase in numbers of plants regenerated suggests that F₁ materials are well suited for tissue culture studies directed towards the genetic improvement of cereal crops.
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