RESUMEN
Adventitious rooting is a process of postembryonic organogenesis strongly affected by endogenous and exogenous factors. Although adventitious rooting has been exploited in vegetative propagation programs for many plant species, it is a bottleneck for vegetative multiplication of difficult-to-root species, such as many woody species. The purpose of this research was to understand how N,N'-bis-(2,3-methylenedioxyphenyl)urea could exert its already reported adventitious rooting adjuvant activity, starting from the widely accepted knowledge that adventitious rooting is a hormonally tuned progressive process. Here, by using specific in vitro bioassays, histological analyses, molecular docking simulations and in vitro enzymatic bioassays, we have demonstrated that this urea derivative does not interfere with polar auxin transport; it inhibits cytokinin oxidase/dehydrogenase (CKX); and, possibly, it interacts with the apoplastic portion of the auxin receptor ABP1. As a consequence of this dual binding capacity, the lifespan of endogenous cytokinins could be locally increased and, at the same time, auxin signaling could be favored. This combination of effects could lead to a cell fate transition, which, in turn, could result in increased adventitious rooting.
RESUMEN
Phytochelatin synthases (PCSs) play pivotal roles in the detoxification of heavy metals and metalloids in plants; however, little information on the evolution of recently duplicated PCS genes in plant species is available. Here we characterize the evolution and functional differentiation of three PCS genes from the giant reed (Arundo donax L.), a biomass/bioenergy crop with remarkable resistance to cadmium and other heavy metals. Phylogenetic reconstruction with PCS genes from fully sequenced monocotyledonous genomes indicated that the three A. donax PCSs, namely AdPCS1-3, form a monophyletic clade. The AdPCS1-3 genes were expressed at low levels in many A. donax organs and displayed different levels of cadmium-responsive expression in roots. Overexpression of AdPCS1-3 in Arabidopsis thaliana and yeast reproduced the phenotype of functional PCS genes. Mass spectrometry analyses confirmed that AdPCS1-3 are all functional enzymes, but with significant differences in the amount of the phytochelatins synthesized. Moreover, heterogeneous evolutionary rates characterized the AdPCS1-3 genes, indicative of relaxed natural selection. These results highlight the elevated functional differentiation of A. donax PCS genes from both a transcriptional and an enzymatic point of view, providing evidence of the high evolvability of PCS genes and of plant responsiveness to heavy metal stress.
Asunto(s)
Aminoaciltransferasas/genética , Evolución Molecular , Proteínas de Plantas/genética , Poaceae/genética , Secuencia de Aminoácidos , Aminoaciltransferasas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Microorganismos Modificados Genéticamente/genética , Microorganismos Modificados Genéticamente/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Poaceae/enzimología , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Alineación de Secuencia/veterinariaRESUMEN
The foliar pathogen Pseudomonas syringae pv. tomato DC3000 (Pst) leads to consistent losses in tomato crops, urging to multiply investigations on the physiological bases for its infectiveness. As other P. syringae pathovars, Pst is equipped with photoreceptors for blue and red light, mimicking the photosensing ability of host plants. In this work we have investigated Pst strains lacking the genes for a blue-light sensing protein (PstLOV), for a bacteriophytochrome (PstBph1) or for heme-oxygenase-1. When grown in culturing medium, all deletion mutants presented a larger growth than wild-type (WT) Pst under all other light conditions, with the exception of blue light which, under our experimental conditions (photon fluence rate = 40 µmol m(-2) s(-1)), completely suppressed the growth of the deletion mutants. Each of the knockout mutants shows stronger virulence towards Arabidopsis thaliana than PstWT, as evidenced by macroscopic damages in the host tissues of infected leaves. Mutated bacteria were also identified in districts distant from the infection site using scanning electron microscopy. These results underscore the importance of Pst photoreceptors in responding to environmental light inputs and the partial protective role that they exert towards host plants during infection, diminishing virulence and invasiveness.
Asunto(s)
Arabidopsis/microbiología , Interacciones Huésped-Patógeno , Luz , Fotorreceptores Microbianos/metabolismo , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/crecimiento & desarrollo , Pseudomonas syringae/metabolismo , Eliminación de Gen , Fotorreceptores Microbianos/genética , Hojas de la Planta/microbiología , Pseudomonas syringae/genética , VirulenciaRESUMEN
Some years ago we demonstrated the cytokinin-like activity of the synthetic N-phenyl-N'-benzothiazol-6-ylurea (PBU) and a relevant adventitious rooting adjuvant activity of symmetric urea derivatives devoid of any cytokinin- or auxin-like activity per se. Here we report the synthesis and the biological activity evaluation of nine symmetric or asymmetric ureas/thioureas, structurally related to PBU. None of them show cytokinin-like activity, while we demonstrate for the first time that PBU interacts with Arabidopsis cytokinin receptor CRE1/AHK4 in a heterologous bioassay system. Among the PBU derivatives, all the symmetric ureas/thioureas show an adventitious rooting adjuvant activity in various bioassays, confirming that this activity is strictly dependent on their chemical structure.