title: A protocol of homozygous haploid callus induction from endosperm of Taxus chinensis Rehd. var. mairei.
abstract: 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 (B5) 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.
authors.affiliation: Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha, 410128 Hunan People's Republic of China ; College of Horticulture and Landscape, Hunan Agricultural University, Changsha, 410128 Hunan People's Republic of China.
authors.forename: Yan-Lin
authors.initials: YL
authors.lastname: Li
authors.affiliation: Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha, 410128 Hunan People's Republic of China ; The Institute of Vegetables and Flowers, Chinese Academy of Agriculture Sciences, Beijing, 100081 People's Republic of China.
authors.forename: San-Wen
authors.initials: SW
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Subhealth Intervention Technology, Changsha, 410128 Hunan People's Republic of China.
authors.forename: Jia-Yin
authors.initials: JY
authors.lastname: Zhang
authors.affiliation: The Institute of Vegetables and Flowers, Chinese Academy of Agriculture Sciences, Beijing, 100081 People's Republic of China.
authors.forename: Feng-Jiao
authors.initials: FJ
authors.lastname: Bu
authors.affiliation: The Institute of Vegetables and Flowers, Chinese Academy of Agriculture Sciences, Beijing, 100081 People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.affiliation: The Institute of Vegetables and Flowers, Chinese Academy of Agriculture Sciences, Beijing, 100081 People's Republic of China.
authors.forename: Zhong-Hua
authors.initials: ZH
authors.lastname: Zhang
authors.affiliation: Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha, 410128 Hunan People's Republic of China ; The Institute of Vegetables and Flowers, Chinese Academy of Agriculture Sciences, Beijing, 100081 People's Republic of China.
authors.forename: Xing-Yao
authors.initials: XY
authors.lastname: Xiong
keywords: Callus initiation; Endosperm; Haploid; Homozygote; SNP mark; Taxus