YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora
Abstract
:1. Introduction
2. Results
2.1. The Induction Process, Histology, and Expression Profiling of the CcYUCs Transcribed during SE in C. canephora
2.2. Identification and Content of Free IAA and Conjugated IAA
2.3. Endogenous Free IAA Accumulation and Localization during the SE Induction Process
2.4. Effect of the Inhibition of Auxin IAA Biosynthesis by Yucasin during the SE Induction Process in C. canephora
2.5. Restoration of Somatic Embryogenesis by Exogenous Addition of IAA
3. Discussion
4. Materials and Methods
4.1. Plant Material and Growth Conditions
4.2. Induction of Somatic Embryogenesis in C. canephora
4.3. Extraction of Auxins and Their Conjugates
4.4. High-Performance Liquid Chromatography
4.5. Liquid Chromatography–Mass Spectrometry of Auxins
4.6. Preparation of Seedlings in the Presence of 3-14C-Trp
4.7. Yucasin Inhibition Assay
4.8. Plant Tissue Sampling
4.9. Real-Time Quantitative Analysis of Gene Expression
4.10. Histological Analyses
4.11. Immunolocalization Assays
4.12. Controls of IAA Immunolocalization
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
DAI | Days after induction |
IAA | Indole-3-acetic acid |
IPA | Indole-3-pyruvic acid |
Kin | Kinetin |
NAA | Naphthalenacetic acid |
SE | Somatic embryogenesis |
Trp | Tryptophan |
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Sample | Days | ||||
---|---|---|---|---|---|
−9 | −7 | −4 | 0 | ||
Leaf extracts | IAA | 154 | 174 | 274 | 1093 |
Trp | 0 | 0 | 34 | 40 | |
Leaves in medium | 287 | 764 | 755 | 1131 | |
Medium | 2,306,388 | 1,845,858 | 1,537,818 | 2,283,948 | |
Stem | - | - | - | 5860 | |
Root | - | - | - | - | |
Total counts per minute | 2,306,829 | 1,846,796 | 1,538,848 | 2,292,033 | |
Initial total cpm = 2,679,807 |
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Uc-Chuc, M.A.; Pérez-Hernández, C.; Galaz-Ávalos, R.M.; Brito-Argaez, L.; Aguilar-Hernández, V.; Loyola-Vargas, V.M. YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora. Int. J. Mol. Sci. 2020, 21, 4751. https://doi.org/10.3390/ijms21134751
Uc-Chuc MA, Pérez-Hernández C, Galaz-Ávalos RM, Brito-Argaez L, Aguilar-Hernández V, Loyola-Vargas VM. YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora. International Journal of Molecular Sciences. 2020; 21(13):4751. https://doi.org/10.3390/ijms21134751
Chicago/Turabian StyleUc-Chuc, Miguel A., Cleyre Pérez-Hernández, Rosa M. Galaz-Ávalos, Ligia Brito-Argaez, Víctor Aguilar-Hernández, and Víctor M. Loyola-Vargas. 2020. "YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora" International Journal of Molecular Sciences 21, no. 13: 4751. https://doi.org/10.3390/ijms21134751