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Regeneration of Ryegrass Plants in Tissue Culture 1
Author(s) -
Ahloowalia B. S.
Publication year - 1975
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci1975.0011183x001500040001x
Subject(s) - kinetin , callus , biology , botany , zeatin , murashige and skoog medium , tissue culture , agar , shoot , horticulture , cytokinin , auxin , biochemistry , genetics , gene , bacteria , in vitro
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.