
%0 Journal Article
%A Afshar-Sterle, Shoukat 
%A Kollmorgen, James F. 
%A Fincher, Geoffrey B. 
%T Reduction of fertile regenerants from protoplasts of <emph type="2">Triticum tauschii</emph> (Coss.) Schmal.
%J Australian Journal of Botany
%D 2000/08/29
%V 48
%N 4
%P 501-506
%K 
%R https://doi.org/10.1071/BT99014
%U https://www.publish.csiro.au/paper/BT99014 
%X 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 (&#60;500 &#181;m in diameter) in a solution containing 3% Cellulase &#8216;Onozuka&#8217; RS, 0.5% Macerozyme R10 and 0.2% Pectolyase Y23. Cell division occurred when the protoplasts were cultured at a density of 1.0&#8211;1.5 x 106 protoplasts mL&#8211;1 in half-strength MS medium supplemented with 1.1 mg L&#8211;1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.6 M glucose and 1.2% agarose. The first cell divisions were observed after 5&#8211;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&#8211;1 sucrose and solidified with 0.25% Phytagel. The colonies produced somatic embryos within 21&#8211;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. 
