Production of fertile regenerants from protoplasts of Triticum tauschii (Coss.) Schmal.
Date
2000
Authors
Afshar-Sterle, S.
Kollmorgen, J.
Fincher, G.
Editors
Advisors
Journal Title
Journal ISSN
Volume Title
Type:
Journal article
Citation
Australian Journal of Botany, 2000; 48(4):501-506
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Abstract
<jats:p>
Four suspension cell lines generated from two accessions of
Triticum tauschii (Coss.) Schmal.
(Aegilops squarrosa, 2n =
2x = 14, DD genome) were used to develop an
efficient protocol for producing fertile regenerants from protoplasts.
Protoplasts were isolated from each cell line by incubating fine cell
aggregates (<500 µm in diameter) in a solution containing 3%
Cellulase ‘Onozuka’ RS, 0.5% Macerozyme R10 and 0.2%
Pectolyase Y23. Cell division occurred when the protoplasts were cultured at a
density of 1.0–1.5 x 106 protoplasts
mL–1 in half-strength MS medium supplemented with
1.1 mg L–1 2,4-dichlorophenoxyacetic acid (2,4-D),
0.6 M glucose and 1.2% agarose. The first cell divisions were observed
after 5–7 days. Cell colonies were observed after 14 days and these grew
quickly into large clumps when transferred to half-strength MS medium
supplemented with 2.2 mg L-1 2,4-D, 30 g
L–1 sucrose and solidified with 0.25%
Phytagel. The colonies produced somatic embryos within 21–28 days of
transfer to this medium. Somatic embryos were transferred to hormone-free MS
medium for regeneration into plantlets. Although many regenerants produced
shrivelled seeds, 9 of 16 were fertile and produced normal seeds.</jats:p>