The Establishments of Tissue 4

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Abstract
Callus cultures of 12 temperate grasses were established, some for the first time, by incubating detached roots or whole seedlings on a Linsmaier and Skoog basal medium which contained 2, 4-dichlorophenoxyacetic acid (2, 4-D) at 1.0 mg 1−1 as the only growth hormone. The callus, which developed in the pericycle of the roots and on the embryo of the seeds, was subcultured on the same medium; further growth of the callus varied from good in the case of Dactylis glomeraia, Agrostis tennis, Cynosurus cristatus, and Poa trivialis, to poor in several Lolium species and varieties. Most cultures developed root primordia which sometimes grew into visible roots, but shoot primordia, none of which grew into shoots, were found only in the callus of Lolium multiflorum var. westerwoldicum. Cell suspension cultures were also readily established and maintained using the same culture medium. Most cultures contained a high proportion of round or oval cells, which ranged from 18 to 77 μm in diameter or length, while many also had a significant proportion of larger, more elongated cells which varied in length from 46 to 182 μm. The cells of Dactylis glomerata were characteristically larger and more convoluted than the cells of other grass species that were examined. The addition of kinetin at 0.1 mg 1−1 to the 2, 4-D-containing culture medium increased the proportion of irregular-shaped cells and reduced the dispersion of the cells, perhaps by improving cellular contact and adhesion; in some species, such as Agrostis tenuis and Phleum pratense, the presence of kinetin promoted the deposition of starch granules in cells.

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