Background Somatic embryogenesis (SE) is a useful biotechnological tool to study the morpho-physiological, biochemical and molecular processes during the development of Coffea canephora. Plant growth regulators (PGR) play a key role during cell differentiation in SE. The Auxin-response-factor (ARF) and Auxin/Indole-3-acetic acid (Aux/IAA) are fundamental components involved in the signaling of the IAA. The IAA signaling pathway activates or represses the expression of genes responsive to auxins during the embryogenic transition of the somatic cells. The growing development of new generation sequencing technologies (NGS), as well as bioinformatics tools, has allowed us to broaden the landscape of SE study of various plant species and identify the genes directly involved. Methods Analysis of transcriptome expression profiles of the C. canephora genome and the identification of a particular set of differentially expressed genes (DEG) during SE are described in this study. Results A total of eight ARF and seven Aux/IAA differentially expressed genes were identified during the different stages of the SE induction process. The quantitative expression analysis showed that ARF18 and ARF5 genes are highly expressed after 21 days of the SE induction, while Aux/IAA7 and Aux/IAA12 genes are repressed. Discussion The results of this study allow a better understanding of the genes involved in the auxin signaling pathway as well as their expression profiles during the SE process.
Cytokinins (CK) are plant growth regulators involved in multiple physiological processes in plants. One less studied aspect is CK homeostasis (HM). The primary genes related to HM are involved in biosynthesis (IPT), degradation (CKX), and signaling (ARR). This paper demonstrates the effect of auxin (Aux) and CK and their cross talk in a Coffea canephora embryogenic system. The transcriptome and RT-qPCR suggest that Aux in pre-treatment represses biosynthesis, degradation, and signal CK genes. However, in the induction, there is an increase of genes implicated in the CK perception/signal, indicating perhaps, as in other species, Aux is repressing CK, and CK are inducing per se genes involved in its HM. This is reflected in the endogenous concentration of CK; pharmacology experiments helped study the effect of each plant growth regulator in our SE system. We conclude that the Aux–CK balance is crucial to directing somatic embryogenesis in C. canephora.
El objetivo del presente trabajo fue identi car el grado de tolerancia a sequía en diferentes genotipos élite de Coffea canephora P. var. Robusta. El estudio se realizó del 2013 al 2015 en Tuxtla Chico, Chiapas, México. De cada genotipo (INIFAP P9, INIFAP P4, INIFAP P5, INIFAP P7, INIFAP P8) se utilizaron veinte plantas, excepto para el INIFAP P9 con doce; un grupo con la mitad de plantas de cada genotipo se mantuvo en condiciones de capacidad de campo (CC) y la otra mitad en punto de marchitez permanente (PMP). Al nalizar la etapa de estrés y después del riego de recuperación, se evaluaron variables morfológicas y siológicas, tales como parámetros de crecimiento, contenido relativo de agua, acumulación de prolina, índice y densidad estomática e índice de tolerancia a sequía. Los genotipos de C. canephora presentaron entre ellos diferente grado de tolerancia al dé cit hídrico, pero compartieron entre sí la detención de sus funciones y activación de diferentes rutas para tolerar la sequía, como disminución en el contenido relativo de agua y aumento de prolina durante el periodo de estrés. Al determinar el índice de tolerancia a sequía, el genotipo más tolerante fue el INIFAP P5 de acuerdo con las variables número de hojas retenidas y totales, tasa de crecimiento relativo, número de pares de ramas plagiotrópicas y nudos totales; mientras que el INIFAP P4 se identi có como uno de los más susceptibles de acuerdo con la tasa de crecimiento relativo y el número de nudos totales, en las cuales presentó los valores más bajos.
Coffea arabica is one of the most important crops worldwide. In vitro culture is an alternative for achieving Coffea regeneration, propagation, conservation, genetic improvement, and genome editing. The aim of this work was to identify proteins involved in auxin homeostasis by isobaric tandem mass tag (TMT) and the synchronous precursor selection (SPS)-based MS3 technology on the Orbitrap Fusion™ Tribrid mass spectrometer™ in three types of biological materials corresponding to C. arabica: plantlet leaves, calli, and suspension cultures. Proteins included in the β-oxidation of indole butyric acid and in the signaling, transport, and conjugation of indole-3-acetic acid were identified, such as the indole butyric response (IBR), the auxin binding protein (ABP), the ATP-binding cassette transporters (ABC), the Gretchen-Hagen 3 proteins (GH3), and the indole-3-acetic-leucine-resistant proteins (ILR). A more significant accumulation of proteins involved in auxin homeostasis was found in the suspension cultures vs. the plantlet, followed by callus vs. plantlet and suspension culture vs. callus, suggesting important roles of these proteins in the cell differentiation process.
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