Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jun 20:14:1189499.
doi: 10.3389/fpls.2023.1189499. eCollection 2023.

Integrated physicochemical, hormonal, and transcriptomic analysis reveals the underlying mechanism of callus formation in Pinellia ternata hydroponic cuttings

Xiaoqing Duan  1 , Lin Chen  1 , Youping Liu  1 , Hongping Chen  1 , Fu Wang  1 , Yuan Hu  1
Affiliations

Integrated physicochemical, hormonal, and transcriptomic analysis reveals the underlying mechanism of callus formation in Pinellia ternata hydroponic cuttings

Xiaoqing Duan et al. Front Plant Sci. .

Abstract

Introduction: P. ternata is a perennial herb of the family Araceae that grows in China and has various medicinal properties and applications. At present, the artificial cultivation of P. ternata is constrained by seedling propagation. To address the problems of low seedling breeding propagation efficiency and high cost, our group has developed a highly efficient cultivation technology for "hydroponic cuttings of P. ternata "for the first time. P. ternata is used as the source material and is grown in a hydroponic system, increasing the seedling production rate 10-fold compared with the traditional cultivation mode. However, the callus formation mechanism in cuttings from hydroponic cultivation is still remains unclear.

Methods: In order to better understand the biological process of callus formation in cuttings from hydroponic P. ternata, anatomical characterization, endogenous hormone content determination and transcriptome sequencing were performed on five callus stages from early growth to early senescence.

Results: Regarding the four major hormones during the callus developmental stages of P. ternata hydroponic cuttings, cytokinins showed an increasing trend during callus formation. IAA(indole-3-acetic acid) and abscisic acid contents increased at 8d and then decreased, while jasmonic acid content gradually decreased. A total of 254137 unigenes were identified by transcriptome sequencing in five callus formation stages. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed genes (DEGs) that differentially expressed unigenes were involved in various plant hormone signaling and hormone synthesis-related pathways. The expression patterns of 7 genes were validated using quantitative real-time PCR.

Discussion: This study presented integrated transcriptomic and metabolic analysis approach to obtain insights into the underlying biosynthetic mechanisms and function of key hormones involved in the callus formation process from hydroponic P. ternata cuttings.

Keywords: P. ternate; histomorphological; phytohormone; plant hormone signaling and synthesis pathways; transcriptome.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Morphological and histological characteristics of callus from P. ternata hydroponic cuttings cultured in water at different induction stages (0d, 8d, 15d, 25d, and 30d). (A) External morphology of different callus in order from top to bottom. (B) PAS staining of transverse callus sections at different points in order from top to bottom. (C) Staining of callus cross sections at different periods of time in order from top to bottom. (D) Local organization of saffron green in cross sections of callus at different induction stages periods of time in order from top to bottom, with green arrows indicating the process of bud formation at different induction stages. The scale bars are 2 cm, 200 μm, 1000 μm, and 500 μm, in that order.
Figure 2
Figure 2
Changes in endogenous cytokinins (CKs) content at five stages of callus formation from hydroponic cuttings. (A) trans-zeatin (tZ) type; (B) cis-zeatin (cZ) type; (C) N6-(Δ2-prenyl) adenine (iP) type; (D) methyl mercaptan type (2MeScZR, 2MeSiP); (E) dihydrozeatin; (F) 6- benzylaminopurine (BAP) type; (G) 3-[(9H-purin-6-ylamino) methyl] phenol (mT) type; (H) 2-[(9H-purin-6-ylamino) methyl] phenol (pT); (I) 4-[(9H-purin-6-ylamino) methyl] phenol (oT) type. The bars with different letters are significantly different (p < 0.05). Values are means of three replicates ± SE.
Figure 3
Figure 3
Changes in endogenous auxin, jasmonic acid, and abscisic acid contents in five stages of callus formation in P. ternata hydroponic cuttings. (A) IAA active compound; (B) tryptophan, a key compound for IAA synthesis; (C) tryptamine; (D) amino acid coupling substance of IAA; (E) JA; (F) amino acid conjugates of JA. The bars with different letters are significantly different from each treatment (p < 0.05). Values are means of three replicates ± SE.
Figure 4
Figure 4
Changes in the expression levels of differentially expressed genes (DEGs) involved in auxin, cytokinin, jasmonic acid, and abscisic acid signaling pathways during the five stages of P. ternata callus formation. (A) Expression levels of DEGs in the auxin signaling pathway. (B) Expression levels of DEGs involved in cytokinin signaling. (C) Expression levels of DEGs of the abscisic acid signaling pathway. (D) Expression levels of DEGs of the jasmonic acid signaling pathway.
Figure 5
Figure 5
(A) Changes in expression of genes and metabolites of the auxin biosynthesis pathway. (B) Changes in the expression of genes and metabolites of the cytokinin biosynthetic pathway.
Figure 6
Figure 6
(A) Changes in expression of genes and metabolites of the auxin biosynthesis pathway. (B) Changes in the expression of genes and metabolites of the cytokinin biosynthetic pathway.
Figure 7
Figure 7
Changes in gene expression in phytohormone signaling in developing callus. (A) Expression levels of the bHLH TFs family differentially expressed genes (DEGs). (B) Expression levels of the AP2/ERF-ERF TFs family DEGs. (C) Expression levels of the C2H2 TFs family DEGs. (D) Expression levels of the bZIP TFs family DEGs. (E) Expression levels of the AUX/IAA TFs family DEGs.
Figure 8
Figure 8
Proposed model of callus proliferation regulation in P. ternata hydroponic cuttings.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Ahuja M. R. (1965). Genetic control of tumor formation in higher plants. Q. Rev. Biol. 40, 329–340. doi: 10.1086/404744 - DOI
    1. Aleman F., Yazaki J., Lee M., Takahashi Y., Kim A. Y., Li Z., et al. . (2016). An ABA-increased interaction of the PYL6 ABA receptor with MYC2 transcription factor: a putative link of ABA and JA signaling. Sci. Rep. 6, 28941. doi: 10.1038/srep28941 - DOI - PMC - PubMed
    1. Bai J., Qi J., Yang L., Wang Z., Wang R., Shi Y. (2022). A comprehensive review on ethnopharmacological, phytochemical, pharmacological and toxicological evaluation, and quality control of pinellia ternata (Thunb.) breit. J. Ethnopharmacol. 298, 115650. doi: 10.1016/j.jep.2022.115650 - DOI - PubMed
    1. Birnbaum K. D., Alvarado A. S. (2008). Slicing across kingdoms: regeneration in plants and animals. Cell 132, 697–710. doi: 10.1016/j.cell.2008.01.040 - DOI - PMC - PubMed
    1. Buechel S., Leibfried A., To J. P. C., Zhao Z., Andersen S. U., Kieber J. J., et al. . (2010). Role of a-type ARABIDOPSIS RESPONSE REGULATORS in meristem maintenance and regeneration. Eur. J. Cell Biol. 89, 279–284. doi: 10.1016/j.ejcb.2009.11.016 - DOI - PubMed

LinkOut - more resources