Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici
Abstract
:1. Introduction
2. Results
2.1. Identification and Sequence Analysis of Tomato ShROP7
2.2. ShROP7 Cannot Induce Cell Death nor Suppress BAX-Induced Necrosis
2.3. ShROP7 Is Differentially Induced by On-Lz Infection
2.4. Silencing of ShROP7 Enhances the Susceptibility of Tomato to On-Lz
2.5. Histological Observation of LA1777 ShROP7-Silenced Plants
2.6. Screening and Identification of ShROP7 Interaction Proteins
3. Discussion
4. Materials and Methods
4.1. Plant and Pathogen Materials and Growth
4.2. Identification and Sequence Analysis of ShROP
4.3. Agrobacterium-Mediated Transient Overexpression in Tobacco (Nicotiana Benthamiana)
4.4. Real-Time Quantitative PCR (RT-qPCR) Analysis
4.5. TRV2-Mediated Silencing of ShROP7 in Tomato
4.6. Yeast Two-Hybrid and Bimolecular Fluorescence Complementation (BiFC) Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strain No. | Interacting Target Proteins | Accession No. | Description |
---|---|---|---|
1 | Calcium-binding EF-hand family protein | Solyc03g117470.2.1 | Ca2+ binding induces a conformational change in the EF-hand motif, leading to the activation or inactivation of target proteins. |
2 | 60S ribosomal protein L26-1 | Solyc02g064670.1.1 | Ribosomal large subunit biogenesis; cytoplasmic translation. |
3 | eukaryotic initiation factor 4A-2 | Solyc08g062800.2.1 | ATP-dependent RNA helicase activity; RNA secondary structure unwinding; translational initiation. |
4 | Uncharacterized protein | Solyc05g009220.2.1 | Glutamine-tRNA ligase; glutaminyl-tRNA synthetase; belongs to the class-I aminoacyl-tRNA synthetase family. |
5 | 40S ribosomal protein S20-2 | Solyc01g096580.2.1 | Structural constituent of ribosome; cytoplasmic translation. |
6 | PGH1 | Solyc09g009020.2.1 | Magnesium ion binding; phosphopyruvate hydratase activity; glycolytic process. |
7 | Uncharacterized protein | Solyc07g007680.2.1 | Membrane contacts sites (MCSs) domains are exclusively found at MCSs between different organelles; mediates lipid transfer between the two adjacent bilayers independently of membrane fusion and fission reactions. |
8 | proteasome subunit alpha type-3 | Solyc10g081130.1.1 | Proteasome subunit alpha type; the proteasome is a multicatalytic proteinase complex. |
9 | ranslocase of chloroplast34 | Solyc03g095220.2.1 | GTPase involved in protein precursor import into chloroplasts. Seems to recognize chloroplast-destined precursor proteins and regulate their presentation to the translocation channel through GTP hydrolysis. |
10 | U3 small nucleolar ribonucleoprotein | Solyc01g007250.2.1 | Some proteins in this family suggest a role in ribosome biogenesis and rRNA binding. |
11 | ERAD-associated E3 ubiquitin-protein ligase component HRD3A | Solyc03g118670.2.1 | It mediates protein-protein interactions and the assembly of multiprotein complexes. |
12 | wound-induced proteinase inhibitor 1 | Solyc09g084440.2.1 | The proteinase inhibitors inhibit peptidases of the S1 and S8 families; synthesis of the inhibitors throughout the plant is also induced by leaf damage. |
13 | exonuclease family protein | Solyc01g096570.2.1 | Ribonuclease T is responsible for the end-turnover of tRNA and removes the terminal AMP residue from uncharged tRNA. |
14 | Bap31 | Solyc02g032930.2.1 | Play a role in endoplasmic reticulum (ER) quality control and sorting |
15 | Fructose-bisphosphate aldolase | Solyc05g008600.2.1 | It is a glycolytic enzyme that catalyzes the reversible aldol cleavage or condensation of fructose-1,6-bisphosphate into dihydroxyacetone- phosphate and glyceraldehyde 3-phosphate. |
16 | Putative PTI1-like tyrosine-protein kinase 3 | Solyc12g098820.1.1 | Tyrosine-protein kinases can transfer a phosphate group from ATP to a tyrosine residue in a protein. |
17 | 5-oxoprolinase | Solyc09g010560.1.1 | Participate in glutathione metabolism. |
18 | EIL1 | Solyc06g073720.1.1 | DNA-binding transcription factor activity; cellular response to iron ion; ethylene-activated signaling pathway. |
19 | OxaA/YidC-like membrane insertion protein | Solyc05g014050.2.1 | Required for the insertion of some light harvesting chlorophyll-binding proteins (LHCP) into the chloroplast thylakoid membrane. |
20 | Ferredoxin | Solyc10g075160.1.1 | Ferredoxins are iron-sulfur proteins that transfer electrons in a wide variety of metabolic reactions. |
21 | Thioredoxin | Solyc06g060290.2.1 | Thioredoxin serves as a general protein disulphide oxidoreductase. It interacts with a broad range of proteins by a redox mechanism based on reversible oxidation of two cysteine thiol groups to a disulphide. |
22 | Cellulose synthase | Solyc01g087210.2.1 | This protein is involved in the pathway plant cellulose biosynthesis. |
23 | Pectin acetylesterase (PAEs) | Solyc08g005800.2.1 | PAEs catalyze the deacetylation of pectin, a major compound of primary cell walls. |
24 | 60S ribosomal protein L24 | Solyc09g008800.2.1 | L24e/L24 appears to play a role in the kinetics of peptide synthesis and may be involved in interactions between the large and small subunits, either directly or through other factors. |
25 | Glutathione peroxidase (GSHPx) | Solyc08g006720.2.1 | GSHPx is an enzyme that catalyzes the reduction of hydroxyperoxides by glutathione. Its main function is to protect against the damaging effect of endogenously formed hydroxyperoxides. |
26 | 15.7 kDa heat shock protein | Solyc04g014480.2.1 | These seem to act as chaperones that can protect other proteins against heat-induced denaturation and aggregation. |
27 | vesicle transport v-SNARE 12 | Solyc05g013050.2.1 | SNAP receptor activity; vesicle-mediated transport; intracellular protein transport; V-SNARE proteins are required for protein traffic between eukaryotic organelles. |
28 | Serinethreonine-protein kinase sty46 | Solyc10g055720.1.1 | Protein kinases catalyze the transfer of the gamma phosphate from nucleotide triphosphates (often ATP) to one or more amino acid residues in a protein substrate side chain, resulting in a conformational change affecting protein function. |
29 | DAR GTPase 3 | Solyc06g084270.2.1 | GTP binding; GTPase activity |
30 | MAPK16 | Solyc12g040680.1.1 | Eukaryotic serine-threonine mitogen-activated protein (MAP) kinases are key regulators of cellular signal transduction systems and are conserved; MAPKs play important roles in the signaling of most plant hormones and in developmental processes |
31 | F-box protein At5g46170 | Solyc08g048430.2.1 | The F-box is a conserved domain that is present in numerous proteins with a bipartite structure. Through the F-box, these proteins are linked to the Skp1 protein and the core of SCFs (Skp1-cullin-F-box protein ligase) complexes. SCFs complexes constitute a new class of E3 ligases. |
32 | D111/G-patch domain-containing protein | Solyc08g081580.2.1 | It might be a previously undetected RNA-binding domain mediating a distinct type of RNA-protein interaction. |
33 | Uncharacterized protein | Solyc08g080110.2.1 | It is deubiquitinating (DUB) enzymes known as the MINDY family (MIU-containing novel DUB). |
34 | CHI3 | Solyc02g082920.3 | Defense against chitinase activity; cell wall macromolecule catabolic process; chitin catabolic process; defense response; polysaccharide catabolic process. |
35 | GATA transcription factor 5 | Solyc01g110310.2.1 | Transcriptional activator that specifically binds 5′-GATA-3′ or 5′-GAT-3′ motifs within gene promoters; positive regulation of transcription. |
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Meng, Y.; Zhang, A.; Ma, Q.; Xing, L. Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici. Int. J. Mol. Sci. 2022, 23, 8557. https://doi.org/10.3390/ijms23158557
Meng Y, Zhang A, Ma Q, Xing L. Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici. International Journal of Molecular Sciences. 2022; 23(15):8557. https://doi.org/10.3390/ijms23158557
Chicago/Turabian StyleMeng, Yanan, Ancheng Zhang, Qing Ma, and Lianxi Xing. 2022. "Functional Characterization of Tomato ShROP7 in Regulating Resistance against Oidium neolycopersici" International Journal of Molecular Sciences 23, no. 15: 8557. https://doi.org/10.3390/ijms23158557