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This article in CS

  1. Vol. 15 No. 4, p. 449-452
     
    Received: Oct 23, 1974
    Published: July, 1975


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doi:10.2135/cropsci1975.0011183X001500040001x

Regeneration of Ryegrass Plants in Tissue Culture1

  1. B. S. Ahloowalia2

Abstract

Abstract

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.

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