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      Infections of the Xylella fastidiosa subsp. pauca Strain “De Donno” in Alfalfa ( Medicago sativa) Elicits an Overactive Immune Response

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          Abstract

          Diseases caused by Xylella fastidiosa are among the most destructive for several agricultural productions. A deadly disease of olive, termed olive quick decline syndrome, is one of the most recent examples of the severe impacts caused by the introduction and spread of this bacterium in new ecosystems with favorable epidemiological conditions. Deciphering the cascade of events leading to the development of severe alterations in the susceptible host plants is a priority of several research programs investigating strategies to mitigate the detrimental impacts of the infections. However, in the case of olives, the long latent period (>1 year) makes this pathosystem not amenable for such studies. We have inoculated alfalfa ( Medicago sativa) with the olive-infecting strain “De Donno” isolated from a symptomatic olive in Apulia (Italy), and we demonstrated that this highly pathogenic strain causes an overactive reaction that ends up with the necrosis of the inoculated stem, a reaction that differs from the notoriously Alfalfa Dwarf disease, caused by X. fastidiosa strains isolated from grapes and almonds. RNASeq analysis showed that major plant immunity pathways are activated, in particular, several calcium transmembrane transporters and enzymes responsible for the production of reactive oxygen species (ROS). Signs of the necrotic reaction are anticipated by the upregulation of genes responsible for plant cell death and the hypersensitive reaction. Overall the whole infection process takes four months in alfalfa, which makes this pathosystem suitable for studies involving either the plant response to the infection or the role of Xylella genes in the expression of symptoms.

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          Regulation of pattern recognition receptor signalling in plants.

          Recognition of pathogen-derived molecules by pattern recognition receptors (PRRs) is a common feature of both animal and plant innate immune systems. In plants, PRR signalling is initiated at the cell surface by kinase complexes, resulting in the activation of immune responses that ward off microorganisms. However, the activation and amplitude of innate immune responses must be tightly controlled. In this Review, we summarize our knowledge of the early signalling events that follow PRR activation and describe the mechanisms that fine-tune immune signalling to maintain immune homeostasis. We also illustrate the mechanisms used by pathogens to inhibit innate immune signalling and discuss how the innate ability of plant cells to monitor the integrity of key immune components can lead to autoimmune phenotypes following genetic or pathogen-induced perturbations of these components.
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            An improved genome release (version Mt4.0) for the model legume Medicago truncatula

            Background Medicago truncatula, a close relative of alfalfa, is a preeminent model for studying nitrogen fixation, symbiosis, and legume genomics. The Medicago sequencing project began in 2003 with the goal to decipher sequences originated from the euchromatic portion of the genome. The initial sequencing approach was based on a BAC tiling path, culminating in a BAC-based assembly (Mt3.5) as well as an in-depth analysis of the genome published in 2011. Results Here we describe a further improved and refined version of the M. truncatula genome (Mt4.0) based on de novo whole genome shotgun assembly of a majority of Illumina and 454 reads using ALLPATHS-LG. The ALLPATHS-LG scaffolds were anchored onto the pseudomolecules on the basis of alignments to both the optical map and the genotyping-by-sequencing (GBS) map. The Mt4.0 pseudomolecules encompass ~360 Mb of actual sequences spanning 390 Mb of which ~330 Mb align perfectly with the optical map, presenting a drastic improvement over the BAC-based Mt3.5 which only contained 70% sequences (~250 Mb) of the current version. Most of the sequences and genes that previously resided on the unanchored portion of Mt3.5 have now been incorporated into the Mt4.0 pseudomolecules, with the exception of ~28 Mb of unplaced sequences. With regard to gene annotation, the genome has been re-annotated through our gene prediction pipeline, which integrates EST, RNA-seq, protein and gene prediction evidences. A total of 50,894 genes (31,661 high confidence and 19,233 low confidence) are included in Mt4.0 which overlapped with ~82% of the gene loci annotated in Mt3.5. Of the remaining genes, 14% of the Mt3.5 genes have been deprecated to an “unsupported” status and 4% are absent from the Mt4.0 predictions. Conclusions Mt4.0 and its associated resources, such as genome browsers, BLAST-able datasets and gene information pages, can be found on the JCVI Medicago web site (http://www.jcvi.org/medicago). The assembly and annotation has been deposited in GenBank (BioProject: PRJNA10791). The heavily curated chromosomal sequences and associated gene models of Medicago will serve as a better reference for legume biology and comparative genomics.
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              The Evolution of Calcium-Based Signalling in Plants.

              The calcium-based intracellular signalling system is used ubiquitously to couple extracellular stimuli to their characteristic intracellular responses. It is becoming clear from genomic and physiological investigations that while the basic elements in the toolkit are common between plants and animals, evolution has acted in such a way that, in plants, some components have diversified with respect to their animal counterparts, while others have either been lost or have never evolved in the plant lineages. In comparison with animals, in plants there appears to have been a loss of diversity in calcium-influx mechanisms at the plasma membrane. However, the evolution of the calcium-storing vacuole may provide plants with additional possibilities for regulating calcium influx into the cytosol. Among the proteins that are involved in sensing and responding to increases in calcium, plants possess specific decoder proteins that are absent from the animal lineage. In seeking to understand the selection pressures that shaped the plant calcium-signalling toolkit, we consider the evolution of fast electrical signalling. We also note that, in contrast to animals, plants apparently do not make extensive use of cyclic-nucleotide-based signalling. It is possible that reliance on a single intracellular second-messenger-based system, coupled with the requirement to adapt to changing environmental conditions, has helped to define the diversity of components found in the extant plant calcium-signalling toolkit.
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                Author and article information

                Journal
                Plants (Basel)
                Plants (Basel)
                plants
                Plants
                MDPI
                2223-7747
                07 September 2019
                September 2019
                : 8
                : 9
                : 335
                Affiliations
                [1 ]CNR, Institute for Sustainable Plant Protection, via Amendola 122/D, 70126 Bari, Italy
                [2 ]Department of Soil Science, Plants and Foods, University of Bari, via Amendola 165/A, 70126 Bari, Italy
                Author notes
                [* ]Correspondence: pasquale.saldarelli@ 123456ipsp.cnr.it ; Tel.: +39-080-5443065
                [†]

                Equally contributed to the work.

                Author information
                https://orcid.org/0000-0002-8633-7813
                Article
                plants-08-00335
                10.3390/plants8090335
                6784145
                31500293
                c00d57a7-6374-4baa-b678-1e0453ceb39c
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 29 July 2019
                : 05 September 2019
                Categories
                Article

                xylella,alfalfa,medicago,transcriptome,olive,pauca,immune response,necrosis

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