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In this paper, micro-Raman mapping and conductive atomic force microscopy (C-AFM) were jointly applied to investigate the structural and electrical homogeneity of quasi-free-standing monolayer graphene (QFMLG), obtained by high temperature decomposition of 4H-SiC(0001) followed by hydrogen intercalation at 900 °C. Strain and doping maps, obtained by Raman data, showed the presence of sub-micron patches with reduced hole density correlated to regions with higher compressive strain, probably associated with a locally reduced hydrogen intercalation. Nanoscale resolution electrical maps by C-AFM also revealed the presence of patches with enhanced current injection through the QFMLG/SiC interface, indicating a locally reduced Schottky barrier height (ΦB). The ΦB values evaluated from local I–V curves by the thermionic emission model were in good agreement with the values calculated for the QFMLG/SiC interface using the Schottky–Mott rule and the graphene holes density from Raman maps. The demonstrated approach revealed a useful and non-invasive method to probe the structural and electrical homogeneity of QFMLG for future nano-electronics applications. [ABSTRACT FROM AUTHOR]

The combination of the unique physical properties of molybdenum disulfide (MoS 2 ) with those of gallium nitride (GaN) and related group-III nitride semiconductors have recently attracted increasing scientific interest for the realization of innovative electronic and optoelectronic devices. A deep understanding of MoS 2 /GaN interface properties represents the key to properly tailor the electronic and optical behavior of devices based on this heterostructure. In this study, monolayer (1L) MoS 2 was grown on GaN-on-sapphire substrates by chemical vapor deposition (CVD) at 700 °C. The structural, chemical, vibrational, and light emission properties of the MoS 2 /GaN heterostructure were investigated in detail by the combination of microscopic/spectroscopic techniques and ab initio calculations. XPS analyses on as-grown samples showed the formation of stoichiometric MoS 2 . According to micro-Raman spectroscopy, monolayer MoS 2 domains on GaN exhibit an average n -type doping of (0.11 ± 0.12) × 10 13 cm -2 and a small tensile strain (ε ≈ 0.25%), whereas an intense light emission at 1.87 eV was revealed by PL analyses. Furthermore, a gap at the interface was shown by cross-sectional TEM analysis, confirming the van der Waals (vdW) bond between MoS 2 and GaN. Finally, density functional theory (DFT) calculations of the heterostructure were carried out, considering three different configurations of the interface, i.e., (i) an ideal Ga-terminated GaN surface, (ii) the passivation of Ga surface by a monolayer of oxygen (O), and (iii) the presence of an ultrathin Ga 2 O 3 layer. This latter model predicts the formation of a vdW interface and a strong n -type doping of MoS 2 , in closer agreement with the experimental observations.

Metal-oxide-semiconductor (MOS) capacitors with Al 2 O 3 as a gate insulator are fabricated on cubic silicon carbide (3C-SiC). Al 2 O 3 is deposited both by thermal and plasma-enhanced Atomic Layer Deposition (ALD) on a thermally grown 5 nm SiO 2 interlayer to improve the ALD nucleation and guarantee a better band offset with the SiC. The deposited Al 2 O 3 /SiO 2 stacks show lower negative shifts of the flat band voltage V FB (in the range of about -3 V) compared with the conventional single SiO 2 layer (in the range of -9 V). This lower negative shift is due to the combined effect of the Al 2 O 3 higher permittivity (ε = 8) and to the reduced amount of carbon defects generated during the short thermal oxidation process for the thin SiO 2 . Moreover, the comparison between thermal and plasma-enhanced ALD suggests that this latter approach produces Al 2 O 3 layers possessing better insulating behavior in terms of distribution of the leakage current breakdown. In fact, despite both possessing a breakdown voltage of 26 V, the T-ALD Al 2 O 3 sample is characterised by a higher current density starting from 15 V. This can be attributable to the slightly inferior quality (in terms of density and defects) of Al 2 O 3 obtained by the thermal approach and, which also explains its non-uniform dC/dV distribution arising by SCM maps.

High-κ dielectrics are insulating materials with higher permittivity than silicon dioxide. These materials have already found application in microelectronics, mainly as gate insulators or passivating layers for silicon (Si) technology. However, since the last decade, the post-Si era began with the pervasive introduction of wide band gap (WBG) semiconductors, such as silicon carbide (SiC) and gallium nitride (GaN), which opened new perspectives for high-κ materials in these emerging technologies. In this context, aluminium and hafnium oxides (i.e., Al 2 O 3 , HfO 2 ) and some rare earth oxides (e.g., CeO 2 , Gd 2 O 3 , Sc 2 O 3 ) are promising high-κ binary oxides that can find application as gate dielectric layers in the next generation of high-power and high-frequency transistors based on SiC and GaN. This review paper gives a general overview of high-permittivity binary oxides thin films for post-Si electronic devices. In particular, focus is placed on high-κ binary oxides grown by atomic layer deposition on WBG semiconductors (silicon carbide and gallium nitride), as either amorphous or crystalline films. The impacts of deposition modes and pre- or postdeposition treatments are both discussed. Moreover, the dielectric behaviour of these films is also presented, and some examples of high-κ binary oxides applied to SiC and GaN transistors are reported. The potential advantages and the current limitations of these technologies are highlighted.

In this paper, we report a multiscale investigation of the compositional, morphological, structural, electrical, and optical emission properties of 2H-MoS 2 obtained by sulfurization at 800 °C of very thin MoO 3 films (with thickness ranging from ~2.8 nm to ~4.2 nm) on a SiO 2 /Si substrate. XPS analyses confirmed that the sulfurization was very effective in the reduction of the oxide to MoS 2, with only a small percentage of residual MoO 3 present in the final film. High-resolution TEM/STEM analyses revealed the formation of few (i.e., 2-3 layers) of MoS 2 nearly aligned with the SiO 2 surface in the case of the thinnest (~2.8 nm) MoO 3 film, whereas multilayers of MoS 2 partially standing up with respect to the substrate were observed for the ~4.2 nm one. Such different configurations indicate the prevalence of different mechanisms (i.e., vapour-solid surface reaction or S diffusion within the film) as a function of the thickness. The uniform thickness distribution of the few-layer and multilayer MoS 2 was confirmed by Raman mapping. Furthermore, the correlative plot of the characteristic A 1g -E 2g Raman modes revealed a compressive strain (ε ≈ -0.78 ± 0.18%) and the coexistence of n- and p-type doped areas in the few-layer MoS 2 on SiO 2 , where the p-type doping is probably due to the presence of residual MoO 3 . Nanoscale resolution current mapping by C-AFM showed local inhomogeneities in the conductivity of the few-layer MoS 2 , which are well correlated to the lateral changes in the strain detected by Raman. Finally, characteristic spectroscopic signatures of the defects/disorder in MoS 2 films produced by sulfurization were identified by a comparative analysis of Raman and photoluminescence (PL) spectra with CVD grown MoS 2 flakes.

This paper reports an investigation of the structural, chemical and electrical properties of ultra-thin (5 nm) aluminum nitride (AlN) films grown by plasma enhanced atomic layer deposition (PE-ALD) on gallium nitride (GaN). A uniform and conformal coverage of the GaN substrate was demonstrated by morphological analyses of as-deposited AlN films. Transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) analyses showed a sharp epitaxial interface with GaN for the first AlN atomic layers, while a deviation from the perfect wurtzite stacking and oxygen contamination were detected in the upper part of the film. This epitaxial interface resulted in the formation of a two-dimensional electron gas (2DEG) with a sheet charge density n s ≈ 1.45 × 10 12 cm -2 , revealed by Hg-probe capacitance-voltage (C-V) analyses. Nanoscale resolution current mapping and current-voltage (I-V) measurements by conductive atomic force microscopy (C-AFM) showed a highly homogeneous current transport through the 5 nm AlN barrier, while a uniform flat-band voltage (V FB ≈ 0.3 V) for the AlN/GaN heterostructure was demonstrated by scanning capacitance microscopy (SCM). Electron transport through the AlN film was shown to follow the Fowler-Nordheim (FN) tunneling mechanism with an average barrier height of <Φ B > = 2.08 eV, in good agreement with the expected AlN/GaN conduction band offset.

Gold-assisted mechanical exfoliation currently represents a promising method to separate ultralarge (centimeter scale) transition metal dichalcogenide (TMD) monolayers (1L) with excellent electronic and optical properties from the parent van der Waals (vdW) crystals. The strong interaction between Au and chalcogen atoms is key to achieving this nearly perfect 1L exfoliation yield. On the other hand, it may significantly affect the doping and strain of 1L TMDs in contact with Au. In this paper, we systematically investigated the morphology, strain, doping, and electrical properties of large area 1L MoS 2 exfoliated on ultraflat Au films (0.16-0.21 nm roughness) and finally transferred to an insulating Al 2 O 3 substrate. Raman mapping and correlative analysis of the E' and A 1 ' peak positions revealed a moderate tensile strain (ε ≈ 0.2%) and p-type doping ( n ≈ -0.25 × 10 13 cm -2 ) of 1L MoS 2 in contact with Au. Nanoscale resolution current mapping and current-voltage ( I - V ) measurements by conductive atomic force microscopy (C-AFM) showed direct tunneling across the 1L MoS 2 on Au, with a broad distribution of tunneling barrier values (Φ B from 0.7 to 1.7 eV) consistent with p-type doping of MoS 2 . After the final transfer of 1L MoS 2 on Al 2 O 3 /Si, the strain was converted to compressive strain (ε ≈ -0.25%). Furthermore, an n-type doping ( n ≈ 0.5 × 10 13 cm -2 ) was deduced by Raman mapping and confirmed by electrical measurements of an Al 2 O 3 /Si back-gated 1L MoS 2 transistor. These results provide a deeper understanding of the Au-assisted exfoliation mechanism and can contribute to its widespread application for the realization of novel devices and artificial vdW heterostructures.

The integration of graphene (Gr) with nitride semiconductors is highly interesting for applications in high-power/high-frequency electronics and optoelectronics. In this work, we demonstrated the direct growth of Gr on Al 0.5 Ga 0.5 N/sapphire templates by propane (C 3 H 8 ) chemical vapor deposition at a temperature of 1350 °C. After optimization of the C 3 H 8 flow rate, a uniform and conformal Gr coverage was achieved, which proved beneficial to prevent degradation of AlGaN morphology. X-ray photoemission spectroscopy revealed Ga loss and partial oxidation of Al in the near-surface AlGaN region. Such chemical modification of a ∼2 nm thick AlGaN surface region was confirmed by cross-sectional scanning transmission electron microscopy combined with electron energy loss spectroscopy, which also showed the presence of a bilayer of Gr with partial sp 2 /sp 3 hybridization. Raman spectra indicated that the deposited Gr is nanocrystalline (with domain size ∼7 nm) and compressively strained. A Gr sheet resistance of ∼15.8 kΩ sq -1 was evaluated by four-point-probe measurements, consistently with the nanocrystalline nature of these films. Furthermore, nanoscale resolution current mapping by conductive atomic force microscopy indicated local variations of the Gr carrier density at a mesoscopic scale, which can be ascribed to changes in the charge transfer from the substrate due to local oxidation of AlGaN or to the presence of Gr wrinkles.

Semiconducting transition metal dichalcogenides (TMDs) are promising materials for future electronic and optoelectronic applications. However, their electronic properties are strongly affected by peculiar nanoscale defects/inhomogeneities (point or complex defects, thickness fluctuations, grain boundaries, etc.), which are intrinsic of these materials or introduced during device fabrication processes. This paper reviews recent applications of conductive atomic force microscopy (C-AFM) to the investigation of nanoscale transport properties in TMDs, discussing the implications of the local phenomena in the overall behavior of TMD-based devices. Nanoscale resolution current spectroscopy and mapping by C-AFM provided information on the Schottky barrier uniformity and shed light on the mechanisms responsible for the Fermi level pinning commonly observed at metal/TMD interfaces. Methods for nanoscale tailoring of the Schottky barrier in MoS 2 for the realization of ambipolar transistors are also illustrated. Experiments on local conductivity mapping in monolayer MoS 2 grown by chemical vapor deposition (CVD) on SiO 2 substrates are discussed, providing a direct evidence of the resistance associated to the grain boundaries (GBs) between MoS 2 domains. Finally, C-AFM provided an insight into the current transport phenomena in TMD-based heterostructures, including lateral heterojunctions observed within Mo x W 1-x Se 2 alloys, and vertical heterostructures made by van der Waals stacking of different TMDs (e.g., MoS 2 /WSe 2 ) or by CVD growth of TMDs on bulk semiconductors., Competing Interests: The authors declare no conflict of interest.

Graphene (Gr)-a single layer of two-dimensional sp 2 carbon atoms-and Carbon Dots (CDs)-a novel class of carbon nanoparticles-are two outstanding nanomaterials, renowned for their peculiar properties: Gr for its excellent charge-transport, and CDs for their impressive emission properties. Such features, coupled with a strong sensitivity to the environment, originate the interest in bringing together these two nanomaterials in order to combine their complementary properties. In this work, the investigation of a solid-phase composite of CDs deposited on Gr is reported. The CD emission efficiency is reduced by the contact of Gr. At the same time, the Raman analysis of Gr demonstrates the increase of Fermi energy when it is in contact with CDs under certain conditions. The interaction between CDs and Gr is modeled in terms of an electron-transfer from photoexcited CDs to Gr, wherein an electron is first transferred from the carbon core to the surface states of CDs, and from there to Gr. There, the accumulated electrons determine a dynamical n-doping effect modulated by photoexcitation. The CD-graphene interaction unveiled herein is a step forward in the understanding of the mutual influence between carbon-based nanomaterials, with potential prospects in light conversion applications.

In this paper, micro-Raman mapping and conductive atomic force microscopy (C-AFM) were jointly applied to investigate the structural and electrical homogeneity of quasi-free-standing monolayer graphene (QFMLG), obtained by high temperature decomposition of 4H-SiC(0001) followed by hydrogen intercalation at 900 °C. Strain and doping maps, obtained by Raman data, showed the presence of sub-micron patches with reduced hole density correlated to regions with higher compressive strain, probably associated with a locally reduced hydrogen intercalation. Nanoscale resolution electrical maps by C-AFM also revealed the presence of patches with enhanced current injection through the QFMLG/SiC interface, indicating a locally reduced Schottky barrier height (Φ B ). The Φ B values evaluated from local I-V curves by the thermionic emission model were in good agreement with the values calculated for the QFMLG/SiC interface using the Schottky-Mott rule and the graphene holes density from Raman maps. The demonstrated approach revealed a useful and non-invasive method to probe the structural and electrical homogeneity of QFMLG for future nano-electronics applications.

Olive cultivation is affected by a wide range of biotic constraints. Verticillium wilt of olive is one of the most devastating diseases affecting this woody crop, inflicting major economic losses in many areas, particularly within the Mediterranean Basin. Little is known about gene-expression changes during plant infection by of woody plants such as olive. A complete RNA-seq transcriptomic analysis of olive tree roots was made. Trinity assembler proved to be the best option to assemble the olive and transcriptomes. The olive transcriptome (Oleup) consisted of 68,259 unigenes (254,252 isoforms/transcripts), and the transcriptome (Vedah) consisted of 37,425 unigenes (52,119 isoforms/transcripts). Most unigenes of the Oleup transcriptome corresponded to cellular processes (12,339), metabolic processes (10,974), single-organism processes (7263), and responses to stimuli (5114). As for the Vedah transcriptome, most unigenes correspond to metabolic processes (25,372), cellular processes (23,718), localization (6385), and biological regulation (4801). Differential gene-expression analysis of both transcriptomes was made at 2 and 7 d post-infection. The induced genes of both organisms during the plant-pathogen interaction were clustered in six subclusters, depending on the expression patterns during the infection. Subclusters A to C correspond to plant genes, and subcluster D to F correspond to genes. A relevant finding was that the differentially expressed gene (DEGs) included in subclusters B and C were highly enriched in proteolysis as well as protein-folding and biosynthesis genes. In addition, a reactive oxygen species (ROS) defense was induced first in the pathogen and later in the plant roots., (Copyright © 2017 Crop Science Society of America.)

High-quality thin insulating films on graphene (Gr) are essential for field-effect transistors (FETs) and other electronics applications of this material. Atomic layer deposition (ALD) is the method of choice to deposit high-κ dielectrics with excellent thickness uniformity and conformal coverage. However, to start the growth on the sp 2 Gr surface, a chemical prefunctionalization or the physical deposition of a seed layer are required, which can effect, to some extent, the electrical properties of Gr. In this paper, we report a detailed morphological, structural, and electrical investigation of Al 2 O 3 thin films grown by a two-steps ALD process on a large area Gr membrane residing on an Al 2 O 3 -Si substrate. This process consists of the H 2 O-activated deposition of a Al 2 O 3 seed layer a few nanometers in thickness, performed in situ at 100 °C, followed by ALD thermal growth of Al 2 O 3 at 250 °C. The optimization of the low-temperature seed layer allowed us to obtain a uniform, conformal, and pinhole-free Al 2 O 3 film on Gr by the second ALD step. Nanoscale-resolution mapping of the current through the dielectric by conductive atomic force microscopy (CAFM) demonstrated an excellent laterally uniformity of the film. Raman spectroscopy measurements indicated that the ALD process does not introduce defects in Gr, whereas it produces a partial compensation of Gr unintentional p-type doping, as confirmed by the increase of Gr sheet resistance (from ∼300 Ω/sq in pristine Gr to ∼1100 Ω/sq after Al 2 O 3 deposition). Analysis of the transfer characteristics of Gr field-effect transistors (GFETs) allowed us to evaluate the relative dielectric permittivity (ε = 7.45) and the breakdown electric field (E BD = 7.4 MV/cm) of the Al 2 O 3 film as well as the transconductance and the holes field-effect mobility (∼1200 cm 2 V -1 s -1 ). A special focus has been given to the electrical characterization of the Al 2 O 3 -Gr interface by the analysis of high frequency capacitance-voltage measurements, which allowed us to elucidate the charge trapping and detrapping phenomena due to near-interface and interface oxide traps.

Graphene is an ideal candidate for next generation applications as a transparent electrode for electronics on plastic due to its flexibility and the conservation of electrical properties upon deformation. More importantly, its field-effect tunable carrier density, high mobility and saturation velocity make it an appealing choice as a channel material for field-effect transistors (FETs) for several potential applications. As an example, properly designed and scaled graphene FETs (Gr-FETs) can be used for flexible high frequency (RF) electronics or for high sensitivity chemical sensors. Miniaturized and flexible Gr-FET sensors would be highly advantageous for current sensors technology for in vivo and in situ applications. In this paper, we report a wafer-scale processing strategy to fabricate arrays of back-gated Gr-FETs on poly(ethylene naphthalate) (PEN) substrates. These devices present a large-area graphene channel fully exposed to the external environment, in order to be suitable for sensing applications, and the channel conductivity is efficiently modulated by a buried gate contact under a thin Al 2 O 3 insulating film. In order to be compatible with the use of the PEN substrate, optimized deposition conditions of the Al 2 O 3 film by plasma-enhanced atomic layer deposition (PE-ALD) at a low temperature (100 °C) have been developed without any relevant degradation of the final dielectric performance.

Verticillium wilt of olive (VWO) is caused by the vascular pathogen Verticillium dahliae. One of the best VWO management measures is the use of tolerant cultivars; however, our knowledge on VWO tolerance/resistance genetics is very limited. A transcriptomic analysis was conducted to (i) identify systemic defense responses induced/repressed in aerial tissues of the tolerant cultivar Frantoio upon root colonization by V. dahliae, and (ii) determine the expression pattern of selected defense genes in olive cultivars showing differential susceptibility to VWO. Two suppression subtractive hybridization cDNA libraries, enriched in up-regulated (FU) and down-regulated (FD) genes respectively, were generated from "Frantoio" aerial tissues. Results showed that broad systemic transcriptomic changes are taking place during V. dahliae-"Frantoio" interaction. A total of 585 FU and 381 FD unigenes were identified, many of them involved in defense response to (a)biotic stresses. Selected genes were then used to validate libraries and evaluate their temporal expression pattern in "Frantoio." Four defense genes were analyzed in cultivars Changlot Real (tolerant) and Picual (susceptible). An association between GRAS1 and DRR2 gene expression patterns and susceptibility to VWO was observed, suggesting that these transcripts could be further evaluated as markers of the tolerance level of olive cultivars to V. dahliae.

Pseudomonas fluorescens PICF7 is an indigenous inhabitant of olive (Olea europaea L.) rhizosphere, able to display endophytic lifestyle in roots, to induce a wide range of defence responses upon colonization of this organ and to exert effective biological control against Verticillium wilt of olive (VWO) (Verticillium dahliae). We aimed to evaluate the involvement of specific PICF7 phenotypes in olive root colonization and VWO biocontrol effectiveness by generating mutants impaired in swimming motility (fliI) or siderophore pyoverdine production (pvdI). Besides, the performance of mutants with diminished in vitro growth in potato dextrose agar medium (gltA) and cysteine (Cys) auxotrophy was also assessed. Results showed that olive root colonization and VWO biocontrol ability of the fliI, pvdI and gltA mutants did not significantly differ from that displayed by the parental strain PICF7. Consequently, altered in vitro growth, swimming motility and pyoverdine production contribute neither to PICF7 VWO suppressive effect nor to its colonization ability. In contrast, the Cys auxotroph mutant showed reduced olive root colonization capacity and lost full biocontrol efficacy. Moreover, confocal laser scanning microscopy revealed that all mutants tested were able to endophytically colonize root tissue to the same extent as wild-type PICF7, discarding these traits as relevant for its endophytic lifestyle., (© 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Calcium-doped praseodymium manganite films (Pr0.7Ca0.3MnO3, PCMO) were prepared by metal-organic chemical vapor deposition (MOCVD) on SrTiO3 (001) and SrTiO3 (110) single-crystal substrates. Structural characterization through X-ray diffraction (XRD) measurements and transmission electron microscopy (TEM) analyses confirmed the formation of epitaxial PCMO phase films. Energy dispersive X-ray (EDX) and X-ray photoelectron spectroscopy (XPS) characterization was used to confirm lateral and vertical composition and the purity of the deposited films. Magnetic measurements, obtained in zero-field-cooling (ZFC) and field-cooling (FC) modes, provided evidence of the presence of a ferromagnetic (FM) transition temperature, which was correlated to the transport properties of the film. The functional properties of the deposited films, combined with the structural and chemical characterization collected data, indicate that the MOCVD approach represents a suitable route for the growth of pure, good quality PCMO for the fabrication of novel spintronic devices.

Pseudomonas fluorescens strain PICF7 is a native endophyte of olive roots. Previous studies have shown this motile, Gram-negative, non-sporulating bacterium is an effective biocontrol agent against the soil-borne fungus Verticillium dahliae, the causal agent of one of the most devastating diseases for olive (Olea europaea L.) cultivation. Here, we announce and describe the complete genome sequence of Pseudomonas fluorescens strain PICF7 consisting of a circular chromosome of 6,136,735 bp that encodes 5,567 protein-coding genes and 88 RNA-only encoding genes. Genome analysis revealed genes predicting factors such as secretion systems, siderophores, detoxifying compounds or volatile components. Further analysis of the genome sequence of PICF7 will help in gaining insights into biocontrol and endophytism.

Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA) against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH) cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets), many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR) experiments aiming to: (1) validate the induction of these genes, and (2) shed light on their expression pattern along time (from 1 to 15 days). Induction of olive genes potentially coding for lipoxygenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase, and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e., JERF, bHLH, WRKY), as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mounts a wide array of systemic defense responses in distant tissues (stems, leaves). This sheds light on how olive plants respond to the "non-hostile" colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

Knowledge on the genetic basis underlying interactions between beneficial bacteria and woody plants is still very limited, and totally absent in the case of olive. We aimed to elucidate genetic responses taking place during the colonization of olive roots by the native endophyte Pseudomonas fluorescens PICF7, an effective biocontrol agent against Verticillium wilt of olive. Roots of olive plants grown under non-gnotobiotic conditions were collected at different time points after PICF7 inoculation. A Suppression Subtractive Hybridization cDNA library enriched in induced genes was generated. Quantitative real time PCR (qRT-PCR) analysis validated the induction of selected olive genes. Computational analysis of 445 olive ESTs showed that plant defence and response to different stresses represented nearly 45% of genes induced in PICF7-colonized olive roots. Moreover, quantitative real-time PCR (qRT-PCR) analysis confirmed induction of lipoxygenase, phenylpropanoid, terpenoids and plant hormones biosynthesis transcripts. Different classes of transcription factors (i.e., bHLH, WRKYs, GRAS1) were also induced. This work highlights for the first time the ability of an endophytic Pseudomonas spp. strain to mount a wide array of defence responses in an economically-relevant woody crop such as olive, helping to explain its biocontrol activity.

The use of indigenous bacterial root endophytes with biocontrol activity against soil-borne phytopathogens is an environmentally-friendly and ecologically-efficient action within an integrated disease management framework. The earliest steps of olive root colonization by Pseudomonas fluorescens PICF7 and Pseudomonas putida PICP2, effective biocontrol agents (BCAs) against Verticillium wilt of olive (Olea europaea L.) caused by the fungus Verticillium dahliae Kleb., are here described. A gnotobiotic study system using in vitro propagated olive plants, differential fluorescent-protein tagging of bacteria, and confocal laser scanning microscopy analysis have been successfully used to examine olive roots-Pseudomonas spp. interactions at the single-cell level. In vivo simultaneous visualization of PICF7 and PICP2 cells on/in root tissues enabled to discard competition between the two bacterial strains during root colonization. Results demonstrated that both BCAs are able to endophytically colonized olive root tissues. Moreover, results suggest a pivotal role of root hairs in root colonization by both biocontrol Pseudomonas spp. However, colonization of root hairs appeared to be a highly specific event, and only a very low number of root hairs were effectively colonized by introduced bacteria. Strains PICF7 and PICP2 can simultaneously colonize the same root hair, demonstrating that early colonization of a given root hair by one strain did not hinder subsequent attachment and penetration by the other. Since many environmental factors can affect the number, anatomy, development, and physiology of root hairs, colonization competence and biocontrol effectiveness of BCAs may be greatly influenced by root hair's fitness. Finally, the in vitro study system here reported has shown to be a suitable tool to investigate colonization processes of woody plant roots by microorganisms with biocontrol potential.