- In silico identification of effector proteins from generalist herbivore Spodoptera litura. - The current study aims to identify genes encoding effector proteins from salivary glands of S. - Eight hundred eight proteins from the head and 267 from salivary gland proteins were predicted to be potential effector proteins.. - litura is responsible for 10–30% reduction in the yield of various crops due to vigorous defoliation and fruit feeding [3]. - Variety of HAMPs have been identified and analyzed in the oral secretion of chewing insects, including fatty acid-amino acid conju- gates (e.g., volicitin), long-chain α,ω-diols (e.g., bru- chins), and plant-derived peptides named inceptins [5].. - The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. - If material is not included in the article's Creative Commons licence 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. - NlSEF1 and endo β 1–4 endoglucanase effector proteins were identified in the oral secretion of brown plant hop- per, N. - Endo β 1–4 endoglucanase effector degrades cellulose in the rice cell wall, resulting in decreased cell wall defense [17]. - In the head, the maximum length of a transcript was 33.54 Kbp, and the minimum was 201 bp. - In the head and salivary gland, the length of transcripts var- ied from ≥200 bp to ≥10 Kbp, with maximum transcript of ≥200 bp (Fig. - From the salivary gland 127 families were. - From this assembly, 595 and 271 poten- tial effector proteins were predicted in head and SG (Additional file 3: Table S2). - Functional annotation of potential effector proteins using public databases. - Sequence similarity search of potential effector proteins were performed for functional annotation and taxo- nomic profiling. - 4, the number of proteins anno- tated to each public database in the head (Fig. - Annotation sum- mary of all the potential effector proteins in this study is provided in Additional file 2: Table S1. - The similarity distribution of potential effector proteins that have significant BLASTp hits against the Nr-NCBI protein database is shown in Fig. - GO terms were assigned to and potential effector proteins in the head and SG, respectively. - In the head, the three most abundant categories of. - In the ‘molecular func- tion’, proteins were mainly distributed amongst ‘struc- tural constituent of cuticle’(9.44%) and ‘serine-type endopeptidase activity’ (6.66%) (Fig. - Whereas, in the salivary gland, the two most abundant categories in the ‘biological process’ are. - In the ‘molecular function’, proteins were mainly distributed amongst ‘serine-type endopeptidase activity’ (5.49%) and ‘calcium ion binding’ (4.81%) (Fig.. - In the Pie Chart most of the categories were repeated at similar hierarchical positon, such as proteolysis in the head as well as in the SG (Additional File 1: Figure S3A &B).. - In the head, the group ‘function unknown’ had the most number of proteins (43.32%) followed by ‘post- translational modification, protein turnover’. - In the saliv- ary gland similar pattern was observed, where the group. - All of the sequences were further assigned to the refer- ence canonical pathways in the Kyoto Encyclopaedia of Genes and Genomes (KEGG), to study which biological pathways and molecular interactions are active in the salivary glands [30]. - Such a characteristic feature was shown in a study [31] by detecting the dense cytoplasm and organized whorls of rough endoplasmic reticulum in the salivary-gland cells. - Consequently, “metabolism” is important among these pathways in the salivary glands (26 head proteins and 17 salivary gland proteins) (Fig.. - This search identified 168 unique domains in the head sample and 107 unique domains in the SG sample (Additional file 5: Table S4). - The top three most abun- dant domains in the head sample are ‘Insect cuticle protein’, ‘Trypsin’ and ‘Lectin C-type domain’. - In the salivary gland, the top three most abundant domains. - are some of the other interesting domains of predicted effector proteins (Additional file 6: Table S5). - 2 Overview of in silico pipeline to identify potential effector proteins from generalist herbivore, Spodoptera litura. - Numbers shown here are predicted peptides remaining after each step in the pipeline. - Number of potential effector proteins 808 267. - A total of 288 potential effector proteins were predicted in SG in fed condition (Additional file 8: Table S7).. - salivary gland (SG), head (H), whole body (WB) and whole body excluded of head and saliv- ary gland WB-(H + S). - Expression analysis in the head (a proxy for mandibu- lar glands) (Fig. - Hence, these genes could have additional func- tions apart from being potential effector proteins.. - b The number of potential effector proteins predicted in the head and salivary gland of Spodoptera litura using de novo based assembly and reference based assembly. - of the insect. - Many of the predicted effector proteins are required by the insects to digest and assimi- late plant material during feeding . - We briefly describe the key potential effector proteins predicted from our transcriptome data with S. - Previ- ously they were also identified in the salivary gland of Helicoverpa armigera [42] pea aphid (Acyrthosiphon pisum) [43], vetch aphid (Megoura viciae) and M. - 4 Potential effector proteins annotated with six public databases. - In total, 808 proteins from the head and 267 proteins from the salivary gland were predicted to be potential effector proteins. - Annotation of head effector proteins (a) and SG effector proteins (b). - Previous studies revealed OBPs and CSPs in the salivary gland transcriptome of N.. - OBPs are found in the salivary gland of mosquito, Anopheles gambiae and is predicted to manipulate the host physiology by scavenging host amines [46]. - CSPs are known as abun- dant proteins in the mandibular gland of painted lady butterfly (Vanessa cardui) and New Zealand red admiral butterfly (Vanessa gonerilla) [47] and play an important role in host plant recognition. - 5 Phylogenetic Relationship and similarity percentage of all the potential effector proteins in the head and salivary gland. - A and B are for species distribution of the BLAST result for each protein in the head and salivary gland, respectively. - The Ca 2+ binding calumenin, calreticulin and EF-hand domain proteins are also reported as effectors in the salivary transcriptome of Sitobion avenae [15] and A. - They were reported in the salivary glands of different insects, including Sitobion avenae [15], Empoasca fabae [12], seed feeding bug, Oncopeltus fas- ciatus, [52] Mayetiola destructor [53] and tobacco. - 6 The enriched Gene Ontology categories (biological process and molecular function) for all predicted potential effector proteins. - In total, 693 of 808 proteins in the head and 194 of 267 in salivary gland were assigned GO term. - 7 a The predicted KEGG pathways for all potential effector proteins. - 142 out of 808 in the head and 80 out of 267 in the salivary gland were annotated. - The proteins in the second hierarchy of the KEGG pathway were assigned to 5 categories A: Metabolism, B: Genetic Information Processing, C: Environmental Information Processing, D: Cellular Processes, E:. - b Predicted Pfam domains in the transcriptome of the head and Salivary gland of S. - In Pfam database, 558 out of 808 proteins in the head and 178 out of 267 proteins in the salivary gland were annotated. - Pfam domains were mapped on potential effector proteins via Pfam standalone software using the default parameters. - Numerical values at the top of each bar represent the number of times that domain has occurred in the annotated proteins. - The domains which have occurred ≥ 5 times in the annotated proteins are shown here. - Peroxidase was previously reported in the saliv- ary gland of M. - Spodoptera litura growth and salivary gland isolation An inbred strain of Spodoptera litura (Fabricius strain) was used in the study. - 8 a Potential effector proteins from salivary glands present in two different environment conditions i.e. - Venn diagram depicting the number of potential effector proteins in artificial diet fed and Arabidopsis fed. - 110 potential effector proteins are common under both conditions. - b Predicted Pfam domains in the common effector proteins . - Numerical values at the top of each bar represent the number of times that domain has occurred in the common proteins.. - The domains that occurred ≥ 5 times in the common proteins are shown here. - The GTF output file from StringTie was used in the TransDecoder program to identify the ORFs within the transcripts. - Identification of in silico secretory proteins and potential effector proteins. - Annotation of potential effector proteins. - Potential effector proteins were annotated to six differ- ent public databases (Nr, UniProt, Pfam, KO, GO, and COG) for taxonomic profiling and functional annotation.. - Homology search of the potential effector proteins was performed using BLASTp at an E- value cut-off of 1e-5 against the constructed database.. - Gene ontology (GO) analysis was performed to identify the putative function of the potential effector proteins. - Species distribution and similarity percentage of all the potential effector proteins in head and salivary gland against UniProt (Order: Lepidoptera). - Enriched GO terms of potential effector proteins assembled by referenced genome of S. - The enriched COG functions for predicted potential effector proteins in head and salivary gland of S. - The predicted Pfam domains of potential effector proteins that only present in plant fed salivary gland (SGF) of S. - Annotation summary of potential effector proteins.. - Annotation summary of potential effector proteins (based of reference genome).. - The Pfam domains found in the head and salivary gland transcriptome.. - Annotation summary of potential effector proteins (based of Arabidopsis fed S. - SG: Salivary Gland. - The raw datasets generated during the sequencing of current study are available in the BioProject-NCBI repository under the accession PRJNA544612.. - Transcriptional profiling analysis of Spodoptera litura larvae challenged with Vip3Aa toxin and possible involvement of trypsin in the toxin activation. - Predicted effector molecules in the salivary secretome of the pea aphid (Acyrthosiphon pisum): a dual transcriptomic/proteomic approach. - Analysis of transcripts and proteins expressed in the salivary glands of hessian fly (Mayetiola destructor) larvae. - Transcriptome analysis of the salivary glands of potato leafhopper, Empoasca fabae. - Transcriptome analysis of the salivary glands of Nephotettix cincticeps (Uhler). - Transcriptome analysis of the salivary glands of the grain aphid, Sitobion avenae. - Comparative analysis of the development of the mandibular salivary glands and the labial silk glands in the mulberry silkworm, Bombyx mori . - Genome evolution following host jumps in the Irish potato famine pathogen lineage. - Effector proteins that modulate plant--insect interactions. - A massive expansion of effector genes underlies gall-formation in the wheat pest Mayetiola destructor.. - Characterization and expression analysis of a gene encoding a secreted lipase-like protein expressed in the salivary glands of the larval hessian fly, Mayetiola destructor (say)
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