The endophytic microbiome of X. fastidiosa susceptible and resistant olives

A multi-factorial strategy is required to co-exist with X. fastidiosa infections, which are devastating olive trees in the southern area of Apulia (Italy). Observations in the outbreak area can provide information on potential approaches for containment. Olive cvs Leccino and FS17 show lessened symptoms and host lower bacterial populations (1,2) than cvs Ogliarola salentina, Cellina di Nardò and Kalamata. We are evaluating whether microbial communities inhabiting the xylem vessels of olive cvs showing different susceptibilities to X. fastidiosa -infection play a role in resistance. To explore these endophytic microbiomes, a whole-metagenome shotgun analysis is currently ongoing. X. fastidiosa -infected and healthy olive plants of the cultivars FS17, Leccino and Kalamata, were selected from the same plot to limit the influence of diverse soil composition and crop management. Shotgun sequencing of DNA extracted from the xylem tissues will be used to investigate the microbiome community by bio-informatic analysis. Moreover, efforts to isolate culturable microorganisms to be used in antagonistic assays against X. fastidiosa, will be performed. Concurrently, the X. fastidiosa-biocontrol potency of Paraburkholderia phytofirmans PsJN strain, whose beneficial effects in the reduction of symptoms in Pierce’s Disease (3) have been recently described, are under evaluation. We are testing the ability of P. phytofirmans to colonise xylem vessels and interact with X. fastidiosa in tobacco and olive

The rapid moving Capriglio earth flow (Parma Province, North Italy): multi-temporal mapping and GB-InSAR monitoring

This research presents the main findings of the multi-temporal mapping and of the long-term, real-time monitoring of the Capriglio landslide in the Emilian Apennines (Northern Italy). The landslide, triggered by prolonged rainfall and rapid snowmelt, activated of April 6th 2013. It is constituted by two main adjacent enlarging bodies with a roto-translational kinematics. They activated in sequence and subsequently joined into a large fast moving earth flow, channelizing downstream the Bardea Creek, for a total length of about 3600 meters. The landslide completely destroyed a 450 m sector of the provincial roadway S.P. 101, and its retrogression tendency put at high risk the Capriglio and Pianestolla villages, located in the upper watershed area of the Bardea River. Furthermore, the advancing toe seriously threatened the Antria bridge, representing the "Massese" provincial roadway S.P. 665R transect over the Bardea Creek, the only strategic roadway left able to connect the above-mentioned villages. With the final aim of supporting local authorities in the hazard assessment and risk management during the emergency phase, on May 5th 2013 aerial optical surveys were conducted to accurately map the landslide extension and evolution. Moreover, a GB-InSAR monitoring campaign was started in order to assess displacements of the whole landslide area. The versatility and flexibility of the GB-InSAR sensors allowed acquiring data with two different configurations, designed and set up to continuously retrieve information on the landslide movements rates (both in its upper slow-moving sectors and in its fast-moving toe). The first acquisition mode revealed that the Capriglio and Pianestolla villages were affected by minor displacements (order of magnitude of few millimetres per month). The second acquisition mode allowed to acquire data every 28'', reaching very high temporal resolution values by applying GB-InSAR technique (Monserrat et al., 2014; Caduff et al., 2015)

Transcriptome profiling of two olive cultivars in response to infection by the CoDiRO strain of Xylella fastidiosa subsp. pauca

Background: The recent Xylella fastidiosa subsp. pauca (Xfp) outbreak in olive (Olea europaea) groves in southern Italy is causing a destructive disease denoted Olive Quick Decline Syndrome (OQDS). Field observations disclosed that Xfp-infected plants of cv. Leccino show much milder symptoms, than the more widely grown and highly susceptible cv. Ogliarola salentina. To determine whether these field observations underlie a tolerant condition of cv. Leccino, which could be exploited for lessening the economic impact of the disease on the local olive industry, transcriptional changes occurring in plants of the two cultivars affected by Xfp were investigated. Results: A global quantitative transcriptome profiling comparing susceptible (Ogliarola salentina) and tolerant (Leccino) olive cultivars, infected or not by Xfp, was done on messenger RNA (mRNAs) extracted from xylem tissues. The study revealed that 659 and 447 genes were differentially regulated in cvs Leccino and Ogliarola upon Xfp infection, respectively, whereas 512 genes were altered when the transcriptome of both infected cultivars was compared. Analysis of these differentially expressed genes (DEGs) shows that the presence of Xfp is perceived by the plants of both cultivars, in which it triggers a differential response strongly involving the cell wall. Up-regulation of genes encoding receptor-like kinases (RLK) and receptor-like proteins (RLP) is the predominant response of cv. Leccino, which is missing in cv. Ogliarola salentina. Moreover, both cultivars react with a strong re-modelling of cell wall proteins. These data suggest that Xfp elicits a different transcriptome response in the two cultivars, which determines a lower pathogen concentration in cv. Leccino and indicates that this cultivar may harbor genetic constituents and/or regulatory elements which counteract Xfp infection. Conclusions: Collectively these findings suggest that cv. Leccino is endowed with an intrinsic tolerance to Xfp, which makes it eligible for further studies aiming at investigating molecular basis and pathways modulating its different defense response

Activity of the SNARE Protein SNAP29 at the Endoplasmic Reticulum and Golgi Apparatus

Snap29 is a conserved regulator of membrane fusion essential to complete autophagy and to support other cellular processes, including cell division. In humans, inactivating SNAP29 mutations causes CEDNIK syndrome, a rare multi-systemic disorder characterized by congenital neuro-cutaneous alterations. The fibroblasts of CEDNIK patients show alterations of the Golgi apparatus (GA). However, whether and how Snap29 acts at the GA is unclear. Here we investigate SNAP29 function at the GA and endoplasmic reticulum (ER). As part of the elongated structures in proximity to these membrane compartments, a pool of SNAP29 forms a complex with Syntaxin18, or with Syntaxin5, which we find is required to engage SEC22B-loaded vesicles. Consistent with this, in HeLa cells, in neuroepithelial stem cells, and in vivo, decreased SNAP29 activity alters GA architecture and reduces ER to GA trafficking. Our data reveal a new regulatory function of Snap29 in promoting secretory trafficking

Xylella fastidiosa in Olive: A Review of Control Attempts and Current Management

Since 2013, Xylella fastidiosa Wells et al. has been reported to infect several hosts and to be present in different areas of Europe. The main damage has been inflicted on the olive orchards of southern Apulia (Italy), where a severe disease associated with X. fastidiosa subspecies pauca strain De Donno has led to the death of millions of trees. This dramatic and continuously evolving situation has led to European and national (Italian and Spanish) measures being implemented to reduce the spread of the pathogen and the associated olive quick decline syndrome (OQDS). Research has been also carried out to find solutions to better and directly fight the bacterium and its main insect vector, Philaenus spumarius L. In the course of this frantic effort, several treatments based on chemical or biological substances have been tested, in addition to plant breeding techniques and integrated pest management approaches. This review aims to summarize the attempts made so far and describe the prospects for better management of this serious threat, which poses alarming questions for the future of olive cultivation in the Mediterranean basin and beyond

Conserved genetic defense response against X. fastidiosa subsp. pauca in olive and citrus

X. fastidiosa subsp. pauca causes diseases in citrus and olive plants. Fortunately, there are citrus species and olive varieties more tolerant to X. fastidiosa and therefore good genotypes to search for genetic sources of resistance. Following this approach, global gene expression analyses were recently achieved using Citrus reticulata cv. Ponkan and Olea europaea cv. Leccino allowing the identification of potential genes involved in plant defense response. Thus, the objective of this work was to identify key genes involved in common genetic defense responses that could be further explored to get resistant varieties. Overall we identified two main mechanisms for both plant species: i. Bacteria recognition and ii. Cell wall fortification. The former involve the expression of patternrecognition receptors, which recognize pathogen molecular patterns and trigger cell defense responses. Some of these receptors belong to the LRR-XII group which contains cell surface immune receptors. The latter involves downregulation of genes in tolerant host such as expansin, pectate lyases and polygalacturonases, related with cell wall expansion and degradation. This suggests that in tolerant hosts, plant cell recognizes X. fastidiosa and reprograms the cell wall development to impair its colonization through the xylem vessels. Therefore these genes represent good candidates to be explored aiming their use in breeding and/or genetic engineering program. Giampetruzzi A., Morelli M., Saponari M., Loconsole G., Chiumenti M., Boscia D., Savino V.N., Martelli G.P. & Saldarelli P. 2016. Transcriptome profiling of two olive cultivars in response to infection by the CoDiRO strain of Xylella fastidiosa subsp. pauca. BMC Genomics 17:475. Rodrigues C.M., De Souza A.A., Takita M.A., Kishi L.T. & Machado M.A. 2013. RNA-Seq analysis of Citrus reticulata in the early stages of Xylella fastidiosa infection reveals auxin-related genes as a defense response. BMC Genomics 14: 676

Differences in the endophytic microbiome of olive cultivars infected by xylella fastidiosa across seasons

The dynamics of Xylella fastidiosa infections in the context of the endophytic microbiome was studied in field-grown plants of the susceptible and resistant olive cultivars Kalamata and FS17. Whole metagenome shotgun sequencing (WMSS) coupled with 16S/ITS rRNA gene sequencing was carried out on the same trees at two different stages of the infections: In Spring 2017 when plants were almost symptomless and in Autumn 2018 when the trees of the susceptible cultivar clearly showed desiccations. The progression of the infections detected in both cultivars clearly unraveled that Xylella tends to occupy the whole ecological niche and suppresses the diversity of the endophytic microbiome. However, this trend was mitigated in the resistant cultivar FS17, harboring lower population sizes and therefore lower Xylella average abundance ratio over total bacteria, and a higher α-diversity. Host cultivar had a negligible effect on the community composition and no clear associations of a single taxon or microbial consortia with the resistance cultivar were found with both sequencing approaches, suggesting that the mechanisms of resistance likely reside on factors that are independent of the microbiome structure. Overall, Proteobacteria, Actinobacteria, Firmicutes, and Bacteriodetes dominated the bacterial microbiome while Ascomycota and Basidiomycota those of Fungi.CIMO -Kansainvälisen Liikkuvuuden ja Yhteistyön Keskus(UIDB/04046/2020