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      Mass spectrometry imaging as a potential technique for diagnostic of Huanglongbing disease using fast and simple sample preparation

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          Abstract

          Huanglongbing (HLB) is a disease of worldwide incidence that affects orange trees, among other commercial varieties, implicating in great losses to the citrus industry. The disease is transmitted through Diaphorina citri vector, which inoculates Candidatus Liberibacter spp. in the plant sap. HLB disease lead to blotchy mottle and fruit deformation, among other characteristic symptoms, which induce fruit drop and affect negatively the juice quality. Nowadays, the disease is controlled by eradication of sick, symptomatic plants, coupled with psyllid control. Polymerase chain reaction (PCR) is the technique most used to diagnose the disease; however, this methodology involves high cost and extensive sample preparation. Mass spectrometry imaging (MSI) technique is a fast and easily handled sample analysis that, in the case of Huanglongbing allows the detection of increased concentration of metabolites associated to the disease, including quinic acid, phenylalanine, nobiletin and sucrose. The metabolites abieta-8,11,13-trien-18-oic acid, suggested by global natural product social molecular networking (GNPS) analysis, and 4-acetyl-1-methylcyclohexene showed a higher distribution in symptomatic leaves and have been directly associated to HLB disease. Desorption electrospray ionization coupled to mass spectrometry imaging (DESI-MSI) allows the rapid and efficient detection of biomarkers in sweet oranges infected with Candidatus Liberibacter asiaticus and can be developed into a real-time, fast-diagnostic technique.

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          Most cited references 82

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          A method is presented for the rapid isolation of high molecular weight plant DNA (50,000 base pairs or more in length) which is free of contaminants which interfere with complete digestion by restriction endonucleases. The procedure yields total cellular DNA (i.e. nuclear, chloroplast, and mitochondrial DNA). The technique is ideal for the rapid isolation of small amounts of DNA from many different species and is also useful for large scale isolations.
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            Metabolomics--the link between genotypes and phenotypes.

             Oliver Fiehn (2001)
            Metabolites are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to genetic or environmental changes. In parallel to the terms 'transcriptome' and proteome', the set of metabolites synthesized by a biological system constitute its 'metabolome'. Yet, unlike other functional genomics approaches, the unbiased simultaneous identification and quantification of plant metabolomes has been largely neglected. Until recently, most analyses were restricted to profiling selected classes of compounds, or to fingerprinting metabolic changes without sufficient analytical resolution to determine metabolite levels and identities individually. As a prerequisite for metabolomic analysis, careful consideration of the methods employed for tissue extraction, sample preparation, data acquisition, and data mining must be taken. In this review, the differences among metabolite target analysis, metabolite profiling, and metabolic fingerprinting are clarified, and terms are defined. Current approaches are examined, and potential applications are summarized with a special emphasis on data mining and mathematical modelling of metabolism.
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              Mechanisms of plant defense against insect herbivores.

              Plants respond to herbivory through various morphological, biochemicals, and molecular mechanisms to counter/offset the effects of herbivore attack. The biochemical mechanisms of defense against the herbivores are wide-ranging, highly dynamic, and are mediated both by direct and indirect defenses. The defensive compounds are either produced constitutively or in response to plant damage, and affect feeding, growth, and survival of herbivores. In addition, plants also release volatile organic compounds that attract the natural enemies of the herbivores. These strategies either act independently or in conjunction with each other. However, our understanding of these defensive mechanisms is still limited. Induced resistance could be exploited as an important tool for the pest management to minimize the amounts of insecticides used for pest control. Host plant resistance to insects, particularly, induced resistance, can also be manipulated with the use of chemical elicitors of secondary metabolites, which confer resistance to insects. By understanding the mechanisms of induced resistance, we can predict the herbivores that are likely to be affected by induced responses. The elicitors of induced responses can be sprayed on crop plants to build up the natural defense system against damage caused by herbivores. The induced responses can also be engineered genetically, so that the defensive compounds are constitutively produced in plants against are challenged by the herbivory. Induced resistance can be exploited for developing crop cultivars, which readily produce the inducible response upon mild infestation, and can act as one of components of integrated pest management for sustainable crop production.
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                Author and article information

                Contributors
                taicia@unicamp.br
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                10 August 2020
                10 August 2020
                2020
                : 10
                Affiliations
                [1 ]Laboratório de Biologia Química Microbiana (LaBioQuiMi), IQ-UNICAMP, Campinas, SP Brazil
                [2 ]ThoMSon Mass Spectrometry Laboratory, IQ-UNICAMP, Campinas, SP Brazil
                [3 ]GRID grid.412403.0, ISNI 0000 0001 2359 5252, MackMass Laboratory, School of Engineering - PPGEMN, , Mackenzie Presbyterian University, ; São Paulo, SP Brazil
                [4 ]GRID grid.271762.7, ISNI 0000 0001 2116 9989, Programa de Pós-graduação Em Ciências Biológicas, , Universidade Estadual de Maringá, ; Maringá, PR Brazil
                [5 ]GRID grid.271762.7, ISNI 0000 0001 2116 9989, Departamento de Química, Laboratório de Biomoléculas E Espectrometria de Massas (LabioMass), , Universidade Estadual de Maringá, ; Maringá, PR Brazil
                [6 ]GRID grid.456716.7, ISNI 0000 0001 0379 8976, Departamento de Pesquisa & Desenvolvimento, , Fundo de Defesa da Citricultura (FUNDECITRUS), ; Araraquara, SP Brazil
                Article
                70385
                10.1038/s41598-020-70385-4
                7417563
                32778716
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100002322, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior;
                Award ID: Finance Code 001
                Award Recipient :
                Funded by: Fundação de Amparo à Pesquisa no Estado de São Paulo
                Award ID: 2017/24462-4
                Award Recipient :
                Funded by: L’Oréal Brazil, together with ABC and UNESCO
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized

                natural products, chemical biology

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