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

Yan-Lin Li,Feng-Jiao Bu,Xing-Yao Xiong,San-Wen Huang,Tao Lin,Zhong-Hua Zhang,Jia-Yin Zhang

doi:10.1186/s40064-016-2320-4

Yan-Lin Li, Feng-Jiao Bu + Show 5 more

Open Access

https://doi.org/10.1186/s40064-016-2320-4

Copy DOI

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.

Highlights

Taxus plants are well-known by their ornamental values, cancer-inhibitory alkaloid paclitaxel (Taxol), and timber uses (Xi et al 2014)
We investigated the effects of hydropriming, media compositions, auxins and cytokinins on endosperm callus induction of Taxus chinensis Rehd. var. mairei, and fully dissect the ploidy level and the genomic homozygosity of them
These inhibitors could be removed by hydropriming with sterile distilled water for endosperm callus induction, and optimal soaking time was 3 days (Fig. 1a)

Summary

Introduction

Taxus plants are well-known by their ornamental values, cancer-inhibitory alkaloid paclitaxel (Taxol), and timber uses (Xi et al 2014). As an effective curing cancer of chemical compounds for 50 years (Foa et al 1994; Bi et al 2014), the same difficulties with other supply of active compounds: limited quantity of active compounds in tissues, low growth rate, limited localization of active ingredients in the specific organs, and destruction of the nature resource (Kazmi et al 1991; Frense 2007; Hussain et al 2013). Other seed plants (gymnosperms) of endosperm, as a nutritive tissue supporting the embryo growth, arises from the haploid (1C) cells of the female gametophyte (Wang et al 2009). We investigated the effects of hydropriming, media compositions, auxins and cytokinins on endosperm callus induction of Taxus chinensis Rehd. var. mairei, and fully dissect the ploidy level and the genomic homozygosity of them

Methods

Results

Conclusion