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      An unconventionally secreted effector from the root knot nematode Meloidogyne incognita, Mi‐ISC‐1, promotes parasitism by disrupting salicylic acid biosynthesis in host plants

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          Abstract

          Plant‐parasitic nematodes need to deliver effectors that suppress host immunity for successful parasitism. We have characterized a novel isochorismatase effector from the root‐knot nematode Meloidogyne incognita, named Mi‐ISC‐1. The Mi‐isc‐1 gene is expressed in the subventral oesophageal glands and is up‐regulated in parasitic‐stage juveniles. Tobacco rattle virus‐induced gene silencing targeting Mi‐isc‐1 attenuated M. incognita parasitism. Enzyme activity assays confirmed that Mi‐ISC‐1 can catalyse hydrolysis of isochorismate into 2,3‐dihydro‐2,3‐dihydroxybenzoate in vitro. Although Mi‐ISC‐1 lacks a classical signal peptide for secretion at its N‐terminus, a yeast invertase secretion assay showed that this protein can be secreted from eukaryotic cells. However, the subcellular localization and plasmolysis assay revealed that the unconventional secretory signal present on the Mi‐ISC‐1 is not recognized by the plant secretory pathway and that the effector was localized within the cytoplasm of plant cells, but not apoplast, when transiently expressed in Nicotiana benthamiana leaves by agroinfiltration. Ectopic expression of Mi‐ISC‐1 in Nbenthamiana reduced expression of the PR1 gene and levels of salicylic acid (SA), and promoted infection by Phytophthora capsici. The cytoplasmic localization of Mi‐ISC‐1 is required for its function. Moreover, Mi‐ISC‐1 suppresses the production of SA following the reconstitution of the de novo SA biosynthesis via the isochorismate pathway in the cytoplasm of N. benthamiana leaves. These results demonstrate that M. incognita deploys a functional isochorismatase that suppresses SA‐mediated plant defences by disrupting the isochorismate synthase pathway for SA biosynthesis to promote parasitism.

          Abstract

          The nematode effector from the root‐knot nematode Meloidogyne incognita, Mi‐ISC‐1, diverts the cytoplasmic isochorismate away from conjugation to l‐glutamate, thus interrupting salicylic acid biosynthesis in the cytosol.

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          Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.

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          The BLAST programs are widely used tools for searching protein and DNA databases for sequence similarities. For protein comparisons, a variety of definitional, algorithmic and statistical refinements described here permits the execution time of the BLAST programs to be decreased substantially while enhancing their sensitivity to weak similarities. A new criterion for triggering the extension of word hits, combined with a new heuristic for generating gapped alignments, yields a gapped BLAST program that runs at approximately three times the speed of the original. In addition, a method is introduced for automatically combining statistically significant alignments produced by BLAST into a position-specific score matrix, and searching the database using this matrix. The resulting Position-Specific Iterated BLAST (PSI-BLAST) program runs at approximately the same speed per iteration as gapped BLAST, but in many cases is much more sensitive to weak but biologically relevant sequence similarities. PSI-BLAST is used to uncover several new and interesting members of the BRCT superfamily.
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            Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis. This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) were reached in LightCycler PCR using the established mathematical model.
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              Pfam: The protein families database in 2021

              Abstract The Pfam database is a widely used resource for classifying protein sequences into families and domains. Since Pfam was last described in this journal, over 350 new families have been added in Pfam 33.1 and numerous improvements have been made to existing entries. To facilitate research on COVID-19, we have revised the Pfam entries that cover the SARS-CoV-2 proteome, and built new entries for regions that were not covered by Pfam. We have reintroduced Pfam-B which provides an automatically generated supplement to Pfam and contains 136 730 novel clusters of sequences that are not yet matched by a Pfam family. The new Pfam-B is based on a clustering by the MMseqs2 software. We have compared all of the regions in the RepeatsDB to those in Pfam and have started to use the results to build and refine Pfam repeat families. Pfam is freely available for browsing and download at http://pfam.xfam.org/.
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                Author and article information

                Contributors
                xuanwang@njau.edu.cn
                Journal
                Mol Plant Pathol
                Mol Plant Pathol
                10.1111/(ISSN)1364-3703
                MPP
                Molecular Plant Pathology
                John Wiley and Sons Inc. (Hoboken )
                1464-6722
                1364-3703
                19 December 2021
                April 2022
                : 23
                : 4 ( doiID: 10.1111/mpp.v23.4 )
                : 516-529
                Affiliations
                [ 1 ] Key Laboratory of Integrated Management of Crop Disease and Pests Ministry of Education Nanjing Agricultural University Nanjing China
                [ 2 ] Key Laboratory of Plant Immunity Nanjing Agricultural University Nanjing China
                [ 3 ] School of Biology Biomedical Sciences Research Complex University of St Andrews St Andrews UK
                [ 4 ] Cell & Molecular Sciences Department The James Hutton Institute Dundee UK
                Author notes
                [*] [* ] Correspondence

                Xuan Wang, Key Laboratory of Integrated Management of Crop Disease and Pests, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, China.

                Email: xuanwang@ 123456njau.edu.cn

                Author information
                https://orcid.org/0000-0003-2074-0551
                https://orcid.org/0000-0001-7405-8377
                Article
                MPP13175
                10.1111/mpp.13175
                8916211
                34923729
                f1ff3a68-8b15-467c-8364-67b693b88df2
                © 2021 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                : 24 November 2021
                : 27 September 2021
                : 29 November 2021
                Page count
                Figures: 9, Tables: 0, Pages: 14, Words: 10385
                Funding
                Funded by: National Natural Science Foundation of China , doi 10.13039/501100001809;
                Award ID: 31872923
                Award ID: 31371922
                Funded by: Scottish Government Rural and Environmental Science and Analytical Services Division
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                April 2022
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.1.2 mode:remove_FC converted:11.03.2022

                Plant science & Botany
                effector,isochorismatase,meloidogyne incognita,parasitism,plant immunity,salicylic acid,secretion activity

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