11 research outputs found
Global, regional, and national burden of colorectal cancer and its risk factors, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019
Funding: F Carvalho and E Fernandes acknowledge support from Fundação para a Ciência e a Tecnologia, I.P. (FCT), in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy i4HB; FCT/MCTES through the project UIDB/50006/2020. J Conde acknowledges the European Research Council Starting Grant (ERC-StG-2019-848325). V M Costa acknowledges the grant SFRH/BHD/110001/2015, received by Portuguese national funds through Fundação para a Ciência e Tecnologia (FCT), IP, under the Norma Transitória DL57/2016/CP1334/CT0006.proofepub_ahead_of_prin
A SNARE-Like Protein and Biotin Are Implicated in Soybean Cyst Nematode Virulence.
Phytoparasitic nematodes that are able to infect and reproduce on plants that are considered resistant are referred to as virulent. The mechanism(s) that virulent nematodes employ to evade or suppress host plant defenses are not well understood. Here we report the use of a genetic strategy (allelic imbalance analysis) to associate single nucleotide polymorphisms (SNPs) with nematode virulence genes in Heterodera glycines, the soybean cyst nematode (SCN). To accomplish this analysis, a custom SCN SNP array was developed and used to genotype SCN F3-derived populations grown on resistant and susceptible soybean plants. Three SNPs reproducibly showed allele imbalances between nematodes grown on resistant and susceptible plants. Two candidate SCN virulence genes that were tightly linked to the SNPs were identified. One SCN gene encoded biotin synthase (HgBioB), and the other encoded a bacterial-like protein containing a putative SNARE domain (HgSLP-1). The two genes mapped to two different linkage groups. HgBioB contained sequence polymorphisms between avirulent and virulent nematodes. However, the gene encoding HgSLP-1 had reduced copy number in virulent nematode populations and appears to produce multiple forms of the protein via intron retention and alternative splicing. We show that HgSLP-1 is an esophageal-gland protein that is secreted by the nematode during plant parasitism. Furthermore, in bacterial co-expression experiments, HgSLP-1 co-purified with the SCN resistance protein Rhg1 α-SNAP, suggesting that these two proteins physically interact. Collectively our data suggest that multiple SCN genes are involved in SCN virulence, and that HgSLP-1 may function as an avirulence protein and when absent it helps SCN evade host defenses
Large read mapping of TN10 cDNA sequences to the HgSLP-1 genomic sequence.
<p>The Y-axis shows depth of mapped cDNA coverage (99% identical with 99%overlap of each read) and the X-axis indicates the base position along the <i>HgSLP-1</i> gene. The numbers mark the exons of <i>HgSLP-1</i>.</p
Immunolocalization of HgSNARE-like protein-1 (HgSLP-1).
<p>Panels A—D are 40 x light field images matched with corresponding epiflorescent images of sections of SCN in soybean roots stained using HgSLP-1 antibodies. Arrows point to the basal cell of a subventral esophageal gland in A, the median bulb and esophageal lumen in B and the stylet in C. Panel D shows negative control sections lacking HgSLP-1 antibody staining in the nematode. Arrows in D point to the basal cell of an esophageal gland and the stylet. Panel D is a composite of two sequential sections from the same nematode. For all light field images, 20 micron scale bars are shown.</p
Multiple sequence alignment of the HgSLP-1 SNARE domain to related t-SNARE proteins.
<p>The **marks the zero layer residue (red) critical for membrane fusion and * indicates conserved hydrophobic residues (green) in the flanking heptad repeat domains. The following sequences are in the alignment: 1). Locus: 2NPS_B; protein name: chain B; crystal structure of the early endosomal SNARE complex; accession: 2NPS_B; organism: <i>Rattus norvegicus</i> (Norway rat). 2). Locus: HgSLP; protein name: <i>Heterodera glycines</i> SNARE-like protein 1; accession: KM575849; organism: <i>Heterodera glycines</i> (soybean cyst nematode). 3). Locus: SYP24_ARATH, protein name: putative syntaxin-24; accession: Q9C615; organism: <i>Arabidopsis thaliana</i> (thale cress). 4. Locus: Q9SML5_CAPAN; protein name: syntaxin t-SNARE; accession: Q9SML5; organism: <i>Capsicum annuum</i> (peppers). 5). Locus: Q8S4W4_PORYE; protein name: Syntaxin PM. Accession: Q8S4W4; organism: <u><i>Pyropia yezoensis</i></u> (marine red alga). 6). Locus: SYP72_ARATH; protein name: Syntaxin-72; accession: Q94KK6; organism: <i>A</i>. <i>thaliana</i>. 7). Locus: BET1L_RAT; protein name: golgi SNARE 15 kDa; accession: O35152; organism: <i>R</i>. <i>norvegicus</i>. 8). Locus: SNP30_ARATH; putative SNAP25 homologous protein SNAP30; accession: Q9LMG8; organism: <i>A</i>. <i>thaliana</i>. 9). Locus: O44419_STRPU; Protein name: Synaptosomal-associated protein 25; accession: O44419; organism: <i>Strongylocentrotus purpuratus</i> (purple sea urchin). 10). Locus: O01389_HIRME; protein name: SNAP-25 homolog; accession: O01389; organism: <i>Hirudo medicinalis</i> (medicinal leech).</p
Alignment of paired SOLiD DNA sequencing reads from SCN inbred strain TN20 to the scaffold 385 reference sequence derived from TN10 genomic sequence.
<p>The Y-axis shows depth of coverage and the X-axis indicates the base position along the scaffold. The <i>P</i>. <i>dendritiformis</i>-like gene (<i>HgSLP-1</i>) spans bases 17816 to 21445.</p
Protein gel blot of HgSNARE-like protein (HgSLP-1) and soybean α-SNAP protein expressed in <i>E</i>. <i>coli</i>. Proteins in lanes 1–5 were detected using an antibody that binds to HgSLP-1.
<p>Proteins in lanes 6–9 were detected using an antibody that binds to soybean α-SNAP. Lanes 1 and 6 contain purified protein from <i>E</i>. <i>coli</i> co-expressing full size HgSLP-1 and soybean α-SNAP. Lane 2, 3, 7, and 8 contain independent replicates of proteins purified from <i>E</i>. <i>coli</i> co-expressing HgSLP-1 missing its signal peptide and soybean α-SNAP. Lanes 4 and 9 contain purified protein from <i>E</i>. <i>coli</i> that only expresses full sized HgSLP-1. Lane 5 contains total protein from <i>E</i>. <i>coli</i> that only expresses full sized HgSLP-1. Protein sizes are shown in kDa.</p
Quantitative PCR of <i>P</i>. <i>dendritiformis</i>-like gene (<i>HgSLP-1)</i> genomic copy number relative to <i>HgFAR-1</i> in inbred SCN strains, TN10, TN20 OP25, OP20 and OP50.
<p>Quantitative PCR of <i>P</i>. <i>dendritiformis</i>-like gene (<i>HgSLP-1)</i> genomic copy number relative to <i>HgFAR-1</i> in inbred SCN strains, TN10, TN20 OP25, OP20 and OP50.</p
SCN genetic linkage groups containing SCN SNPs linked to virulence.
<p>The left column has the map distance in centimorgans and the right column shows the SNP number. The SCN SNPs that show an allelic imbalance when grown on resistant and susceptible soybean plants are shown in red.</p