Abstract
The current study evaluated the effect of (i) genotypes on somatic embryogenesis initiation of Pinus elliottii x Pinus caribaea var. hondurensis and (ii) 5-azacytidine (5-azaC), a potential agent for embryogenic competence reacquisition. For the establishment of embryogenic cell lines (ECLs), immature megagametophytes of thirteen families were cultured on an initiation medium. After 17 months, two aged ECLs (ECL02 and ECL05) were exposed for 10 days to 5-azaC (0, 50 and 150 μM). Gene expression analysis of Somatic Embryogenesis Receptor Kinase 1 (SERK1) and Leafy Cotyledon 1 (LEC1) were carried out by RT-qPCR in both ECLs. Metabolomic analysis via GC–MS were performed in the aged ECL05 and in a young ECL. ECLs initiation was genotype-dependent. The aged ECL05 exposed to 150 µM 5-azaC showed an increase in SERK1 and LEC1 expressions in comparison to the control and a similar metabolic profile to the young ECL. It suggests that 5-azaC has an effect on embryogenic morphology of the aged ECL05. On the other hand, it seems that the response to 5-azaC is also affected by the genotype, as the expression of both genes showed an opposite pattern in the ECL02.
Key message
The 5-azaC affect the proliferation and morphology of Pinus aged pro-embryogenic masses which might be associated to embryogenic competence, although is a genotype-dependent response.
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Data availability
The data are available from the corresponding author on reasonable request.
Abbreviations
- 5-azaC:
-
Azacytidine
- cDNA:
-
Complementary DNA
- EC:
-
Embryogenic competence
- CG-MS:
-
Gas chromatography-mass spectrometry
- ECL:
-
Embryogenic cell line
- ROS:
-
Reactive oxygen species
- SE:
-
Somatic embryogenesis
- LEC1 :
-
Leafy Cotyledon 1
- mLV:
-
Modified Litvay medium
- PEM:
-
Pro-embryogenic mass
- PAs:
-
Polyamines
- PCA:
-
Principal components analysis
- RT-qPCR:
-
Quantitative reverse transcription polymerase chain reaction
- Put:
-
Putrescine
- SERK1 :
-
Somatic Embryogenesis Receptor Kinase 1
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Acknowledgements
The authors would like to thank the Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA Forestry) for funding and for the facilities assigned to carry out this project. Also, we thank Daniela Rodrigues Pereira for the photos taken from the cytochemical analysis.
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This work was supported by Fundo Cooperativo para Melhoramento de Pinus (FUNPINUS).
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ELS and JD have substantial contributions to the conception and design of the work; ELS, FAH and JD conceived and conceptualized the manuscript; Material preparation, data collection and analysis were performed by ELS and JATS. JCBF, CGH, NS contributed to reviewing and critically revising the manuscript. JD and JCBF visualized and carefully supervised the work and the manuscript. All authors read and approved the final manuscript.
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Dos Santos, E.L., Filho, J.C.B., Hansel, F.A. et al. 5-Azacytidine affects gene expression and metabolic profile of Pinus elliottii x Pinus caribaea var. hondurensis embryogenic cell lines. Plant Cell Tiss Organ Cult 155, 637–651 (2023). https://doi.org/10.1007/s11240-023-02584-6
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DOI: https://doi.org/10.1007/s11240-023-02584-6