Regeneration of Ryegrass Plants in Tissue Culture1
- B. S. Ahloowalia2
Ryegrass plants (Lolium spp.) were regenerated in callus cultures induced from triploid (2n = 21) embryos. Developing seeds from diploid Χ tetraploid and reciprocal crosses were cultured on i) Bacto-Orchid agar and ii) modified medium of Niizeki and Oono containing 1.5 mg 2,4-dichlorophenoxyacefic acid (2,4-D); 6.5 mg indoleacetic acid (IAA); and 0.25 mg zeatin/liter.
Of the 219 seeds cultured, 10% formed triploid callus and 35% produced triploid and near-triploid (2n = 19, 20, 22) plants. The calli differentiated root and shoot primordia on modified Murashige and Skoog RM medium containing 1.5 mg 2,4-D; 6.5 mg IAA; and 2.15 mg kinetin/liter. Addition of 1 ml coconut milk, with or without 2 mg IAA and 2 mg zeatin/liter, to the medium stimulated chlorophyll development in the calli. Subsequent culture of these calli on half-strength Murashige and Skoog RM medium containing 0.75 mg 2,4-D; 3.25 mg IAA; and 1.075 mg kinetin/liter produced numerous normal and a few albino plants. The calli maintained for 18 months stayed totipotent through eight subcultures. Callus suspensions formed roots in liquid medium conraining 0.05 mg/liter each of IAA and kinetin. Cell suspensions plated on half-strength RM medium with 2,4-D; IAA; and kinetin, developed colonies which included elongate and spherical parenchymatous cells, cells with bipolar axis, vacuolated hair-like cells, etc., of varying size.
The established procedures permitted a rapid regeneration of xyegrass plants in large numbers through tissue culture and hold potential for chromosome manipulation and selection at cell level.Please view the pdf by using the Full Text (PDF) link under 'View' to the left.
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