In vitro culture of isolated microspores and plant regeneration in six-row barley (Hordeum vulgare L.)
1995
Xiang Youbin | Liu Jing (Zhejiang Provincial Academy of Agricultural Sciences, Hangzhou (China). Biotechnology Research Center)
Microspores were isolated from the spike of the six-row barley (Hordeum vulgare L.) cultivar Frances at the mid-unicellular pollen stage, pretreated at 4 C degrees for 4 weeks, by directly cutting the spike into small segments and homogenizing. The isolated microspores were incubated in MSm medium supplemented with 60 g/L maltose, 1.0 mg/L BAP with slowly shaking at 25 C degrees in darkness. Cell aggregates from microspores were observed after 7 days from initial culturing and the number of them reached to the highest point at a frequency of 2.36 percent after about 3 weeks of culture. The cell aggregates developed embryo-like structure and callus at the rate of 80 percent and 20 percent, respectively. The embryo-like structures on 1/2 MS containing 1.0 mg/L BAP, 0.2 mg/L NAA and 0.2 mg/L ZT regenerated green plantlets (39 percent) and albino plantlets (28 percent). The calli improved on L3 medium with 2.0 mg/L 2,4-D, 0.1 mg/L ABA and 0.2 mg/L ZT, regenerated green plantlets (18 percent)and albino plantlets (44 percent), on 1/2 MS medium by adding 4.0 mg/L BAP , 0.5 mg/L NAA and 0.5 mg/L IAA
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