A protocol of homozygous haploid callus induction from endosperm of Taxus chinensis Rehd. var. mairei

Obtainment and characterization of the novel endosperm callus of Taxus chinensis Rehd. var. mairei are valuable for haploid breeding, genome, and functional genome in Taxus. Callus was obtained by hydropriming with sterile water for 3 days and suitable medium composition. The highest callus induction (70.89 %) and lower browning ratio (7.95 %) were obtained from Gamborg (B(5)) medium supplemented with 30 g l(−1) of sucrose, 2.5 mg l(−1) of 2,4-dichlorophenoxyacetic (2,4-D), 0.5 mg l(−1) of 6-benzylademine (6-BA) and 7 g l(−1) of agar under dark conditions. The auxin of 2,4-D had a better efficiency of callus induction than naphthylacetic acid, and over 1 mg l(−1) of 6-BA was inhibitory to the callogensis of endosperm. The endosperm callus was haploid which was detectable by the flow cytometry. The genome block of homozygosity of callus was homozygous which was indicated by PCR-based SNP marks. The homozygous haploid of endosperm callus in vitro culture may be useful tools for taxoid-metabolism of gene engineering and bio-fermentation engineering.