Optimization of anther culture of awnless triticale

无芒小黑麦花药培养的优化

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Abstract

BACKGROUND: Compared with traditional breeding methods, anther culture is an effective method for quickly obtaining homozygotes within one generation. The method of cultivating doubled haploid plants derived from the anthers of awnless triticale was studied and optimized. METHODS: Young awnless triticale spikes were pretreated at 4 °C for 5, 10, 15, 20, or 25 days, respectively, and then the anthers of different treatment days were cultured on cysteine heart blood (CHB) with antibiotics media with four different hormone concentrations, respectively. RESULTS: Overall, 15 days was the best low-temperature treatment, and CHB medium containing 1.5 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) and 1.5 mg/L Kinetin (KT) was the best hormone concentration treatment. The callus induction rate (CIR) was highest (22.20%) for anthers pretreated for 15-days and inoculated on CHB medium containing 1.5 mg/L 2,4-D and 1.5 mg/L KT. The green plantlet differentiation frequency (DFG) was highest (30.20%) for anthers pretreated for 25-days and inoculated on CHB medium containing 0.5 mg/L 2,4-D+0.5 mg/L KT. Green plantlet production (PRG) was highest (4.58%) for anthers which were pretreated for 10-days and inoculated on CHB medium containing 0.5 mg/L 2,4-D+0.5 mg/L KT. The success rate of chromosome doubling for regenerated green plantlets was 52.8%. Nine of the thirteen DH1 plants (the first generation of double haploid plants) had tip and side awns shorter than 5 mm, implying that they may be used for cultivating awnless triticale. CONCLUSION: Triticale anther culture technology was optimized in this work, enabling the rapid breeding of homozygous varieties of awnless triticale and accelerating the breeding of new varieties of awnless triticale.

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