Peony is an excellent ornamental, medicinal, and oily plant. Its traditional seed propagation methods have the disadvantages of low propagation coefficient, long seedling cycle, and low seedling emergence rate, which severely restrict the supply of seedlings for the peony industry. Efficient tissue culture technology is an important basis for accelerating its breeding and reproduction, and in vitro seed embryo culturing into seedlings can also effectively avoid the above problems. However, the browning phenomenon caused by man-made damage in the process of seed embryo stripping leads to problems such as low induction rate and difficulty in rooting, and the relationship between anti-browning agents and seed embryo root formation is still unclear. This study intends to improve the induction rate of peony seedlings by using different anti-browning agents and different combinations and to clarify the relationship between anti-browning agents and seedling rooting using transcriptome sequencing methods. The results show that both anti-browning agents, activated carbon (AC) and polyvinyl pyrrolidone (PVP), can increase the germination rate of seed embryos. Testing with 0.9 g/L of AC showed excellent performance of peony rooting rate and seedling growth, but only AC and the combination of AC and PVP can further promote rooting development. Through transcriptome analysis, we found that the AC vs. control check (CK), AC vs. PVP, and PVP vs. AC and PVP groups have significantly more differentially expressed genes than the AC vs. AC and PVP groups. Pathway enrichment analysis shows that “phenylpropanoid biosynthesis”/“cutin, suberin, and wax biosynthesis” is significantly enriched in these groups, while the AC vs. AC and PVP groups are mainly enriched in “cytochrome P450,” indicating that AC may promote the further development of roots into seedlings by stimulating “phenylpropanoid biosynthesis” and biosynthesis of stratum cutin and suberin. This study can lay the foundation for understanding the potential molecular mechanism of the anti-browning agent promoting the rooting of seed embryo seedlings and also provide a theoretical basis for perfecting the construction of the peony tissue culture and rapid propagation system.
The efficient induction of peony embryogenic callus is of great significance to the improvement and establishment of its regeneration technology system. In this study, the in vitro embryos of ‘Fengdanbai’ at different developmental stages were selected as explants, the effects of different concentrations and types of plant growth regulator combinations on the induction and proliferation of embryonic callus at different developmental stages were investigated, and comparative transcriptome analysis of callus with different differentiation potentials were performed to explore the molecular mechanisms affecting callus differentiation. The results showed that the germination rate of 90d seed embryo was the best, which was 94.17%; the 70d and 80d cotyledon callus induction effect was the best, both reaching 100%, but the 80d callus proliferation rate was higher, the proliferation rate reached 5.31, and the optimal induction medium was MS+0.1 mg·L–1NAA+0.3 mg·L–1TDZ+3 mg·L–12,4-D, the callus proliferation multiple was 4.77. Based on the comparative transcriptomic analysis, we identified 3470 differentially expressed genes (DEGs) in the callus with high differentiation rate and low differentiation rate, including 1767 up-regulated genes and 1703 down-regulated genes. Pathway enrichment analysis showed that the “Phenylpropanoid biosynthesis” metabolic pathway was significantly enriched, which is associated with promoting further development of callus shoots and roots. This study can provide reference for genetic improvement and the improvement of regeneration technology system of peony.
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