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. 2023 Dec 27;25(1):375.
doi: 10.3390/ijms25010375.

Technology Invention and Mechanism Analysis of Rapid Rooting of Taxus × media Rehder Branches Induced by Agrobacterium rhizogenes

Ying Wang  1   2 , Xiumei Luo  1 , Haotian Su  1   3 , Ge Guan  1   3 , Shuang Liu  1   3 , Maozhi Ren  1   2   3
Affiliations

Technology Invention and Mechanism Analysis of Rapid Rooting of Taxus × media Rehder Branches Induced by Agrobacterium rhizogenes

Ying Wang et al. Int J Mol Sci. .

Abstract

Taxus, a vital source of the anticancer drug paclitaxel, grapples with a pronounced supply-demand gap. Current efforts to alleviate the paclitaxel shortage involve expanding Taxus cultivation through cutting propagation. However, traditional cutting propagation of Taxus is difficult to root and time-consuming. Obtaining the roots with high paclitaxel content will cause tree death and resource destruction, which is not conducive to the development of the Taxus industry. To address this, establishing rapid and efficient stem rooting systems emerges as a key solution for Taxus propagation, facilitating direct and continuous root utilization. In this study, Agrobacterium rhizogenes were induced in the 1-3-year-old branches of Taxus × media Rehder, which has the highest paclitaxel content. The research delves into the rooting efficiency induced by different A. rhizogenes strains, with MSU440 and C58 exhibiting superior effects. Transcriptome and metabolome analyses revealed A. rhizogenes' impact on hormone signal transduction, amino acid metabolism, zeatin synthesis, and secondary metabolite synthesis pathways in roots. LC-MS-targeted quantitative detection showed no significant difference in paclitaxel and baccatin III content between naturally formed and induced roots. These findings underpin the theoretical framework for T. media rapid propagation, contributing to the sustainable advancement of the Taxus industry.

Keywords: Agrobacterium rhizogenes; Taxus × media Rehder; branch rooting; paclitaxel.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
The technical process of branch rooting of T. media. (a) Girdling treatment of 1–3-year-old branches. (b) Plant high-pressure propagation box fixing at the girdling site. (c) Swelling and formation of root apical meristem at the upper girdling site. (d) Formation of young roots at the upper girdling site. (e) Elongation and growth of branch adventitious roots.
Figure 2
Figure 2
Demonstration of branch rooting. (a) Morphology of roots after opening the high-pressure propagation box, with new shoots emerging from the branch. (b) The morphology of roots formed under the treatment of different A. rhizogenes strains, showing their growth from the girdling site. CK means the normally formed roots of T. media branches without the treatment of A. rhizogenes. (c) The rooting rate of T. media branches under the treatment of different A. rhizogenes strains that, without the treatment of A. rhizogenes, was as a control (CK). (d) The number of adventitious roots in T. media branches under treatments of different A. rhizogenes strains that, without the treatment of A. rhizogenes, was as a control (CK). (e) The growth status of the rooting branches induced by A. rhizogenes strains MSU440 and C58 after being transplanted into pots. Each experiment was repeated three times. Each replicate contains at least 35 branch rooting treatments. Asterisks indicate significant differences (*** p < 0.001).
Figure 3
Figure 3
Analysis of differentially expressed genes (DEGs) in TR vs. WR; gene ontology (GO); and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs in TR vs. WR. (a) Differentially expressed genes in TR vs. WR. (b) GO enrichment analysis showed that biological processes, cellular anatomical entity, and oxidoreductase activity were involved in the biological process, cellular component, and molecular function, respectively. (c) KEGG enrichment analysis showed that metabolic pathways, biosynthesis of secondary metabolites, and phenylpropanoid biosynthesis were the top three significant enrichment pathways.
Figure 4
Figure 4
Analysis of differentially expressed metabolites (DEMs) in TR vs. WR. (a) Differentially expressed metabolites in TR vs. WR. (b) Principal component analysis revealed clear differences between the metabolites in TR and WR. (c) Cluster analysis of DEMs. (d) KEGG enrichment analysis showed that DEMs were the main components for the biosynthesis of amino acids and their derivatives.
Figure 5
Figure 5
Combined analysis of transcriptome and metabolome (DEGs/DEMs) in TR vs. WR. (a) Top ten KEGG pathways correlated with DEGs/DEMs in TR vs. WR. (b) Plant hormone signal transduction pathway in the correlated DEGs/DEMs in TR vs. WR. (c) Zeatin biosynthesis pathway in the correlated DEGs/DEMs in TR vs. WR. In (b,c), the red rectangles indicate the DEGs encoded in the protein were all upregulated, the blue rectangles indicate the DEGs encoded in the protein were all downregulated and the yellow rectangle indicates the genes encoded in the protein contain both upregulated and downregulated DEGs.
Figure 6
Figure 6
The quantitative detection of paclitaxel and baccatin III. (a) The LC-MS/MS analysis of paclitaxel and baccatin III in 20-year-old roots, WR, and TR. The retention time (RT) of paclitaxel is 3.08 min and that of baccatin III is 2.86 min. (b) The content of paclitaxel in 20-year-old roots, WR, and TR. (c) The content of baccatin III in 20-year-old roots, WR, and TR. Asterisks indicate significant differences (*** p < 0.001).
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