scholarly journals Expression of several Phytophthora cinnamomi putative RxLRs provides evidence for virulence roles in avocado

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254645
Author(s):  
Melissa Joubert ◽  
Robert Backer ◽  
Juanita Engelbrecht ◽  
Noëlani van den Berg
Keyword(s):  
Phytophthora Cinnamomi ◽  
Expression Profiles ◽  
Functional Characterization ◽  
Effector Proteins ◽  
Virulence Determinants ◽  
In Planta ◽  
Phytophthora Root Rot ◽  
Host Immune Responses ◽  
Rxlr Effectors ◽  
Pathogen Fitness

Phytophthora cinnamomi is a plant pathogenic oomycete that causes Phytophthora root rot of avocado (PRR). Currently, there is a limited understanding of the molecular interactions underlying this disease. Other Phytophthora species employ an arsenal of effector proteins to manipulate host physiology, of which the RxLR effectors contribute to virulence by interfering with host immune responses. The aim of this study was to identify candidate RxLR effectors in P. cinnamomi that play a role in establishing PRR, and to infer possible functions for these effectors. We identified 61 candidate RxLR genes which were expressed during infection of a susceptible avocado rootstock. Several of these genes were present in multiple copies in the P. cinnamomi genome, suggesting that they may contribute to pathogen fitness. Phylogenetic analysis of the manually predicted RxLR protein sequences revealed 12 P. cinnamomi RxLRs that were related to characterised effectors in other Phytophthora spp., providing clues to their functions in planta. Expression profiles of nine more RxLRs point to possible virulence roles in avocado–highlighting a way forward for studies of this interaction. This study represents the first investigation of the expression of P. cinnamomi RxLR genes during the course of avocado infection, and puts forward a pipeline to pinpoint effector genes with potential as virulence determinants, providing a foundation for the future functional characterization of RxLRs that contribute to P. cinnamomi virulence in avocado.

scholarly journals Phytophthora parasitica Effector PpRxLR2 Suppresses Nicotiana benthamiana Immunity

2018 ◽  
Vol 31 (4) ◽  
pp. 481-493 ◽  
Author(s):  
R. J. D. Dalio ◽  
H. J. Maximo ◽  
T. S. Oliveira ◽  
R. O. Dias ◽  
M. C. Breton ◽  
...  
Keyword(s):  
Nicotiana Benthamiana ◽  
Control Strategies ◽  
Functional Characterization ◽  
Phytophthora Species ◽  
Effector Proteins ◽  
Phytophthora Parasitica ◽  
In Planta ◽  
Bioinformatics Pipeline ◽  
Rxlr Effectors

Phytophthora species secrete several classes of effector proteins during interaction with their hosts. These proteins can have multiple functions including modulation of host physiology and immunity. The RxLR effectors have the ability to enter plant cells using the plant machinery. Some of these effectors have been characterized as immunity suppressors; however, very little is known about their functions in the interaction between Phytophthora parasitica and its hosts. Using a bioinformatics pipeline, we have identified 172 candidate RxLR effectors (CREs) in the isolate IAC 01_95 of P. parasitica. Of these 172 CREs, 93 were found to be also present in eight other genomes of P. parasitica, isolated from different hosts and continents. After transcriptomics and gene expression analysis, we have found five CREs to be up-regulated in in-vitro and in-planta samples. Subsequently, we selected three CREs for functional characterization in the model plant Nicotiana benthamiana. We show that PpRxLR2 is able to completely suppress INF-1-induced cell death, whereas PpRxLR3 and PpRxLR5 moderately suppressed N. benthamiana immunity in a less-extensive manner. Moreover, we confirmed the effector-triggered susceptibility activity of these proteins after transient transformation and infection of N. benthamiana plants. All three CREs enhanced virulence of P. parasitica during the interaction with N. benthamiana. These effectors, in particular PpRxLR2, can be targeted for the development of biotechnology-based control strategies of P. parasitica diseases.

scholarly journals Global Gene Expression Profiles Suggest an Important Role for Nutrient Acquisition in Early Pathogenesis in a Plant Model of Pseudomonas aeruginosa Infection

2008 ◽  
Vol 74 (18) ◽  
pp. 5784-5791 ◽  
Author(s):  
Tiffany L. Weir ◽  
Valerie J. Stull ◽  
Dayakar Badri ◽  
Lily A. Trunck ◽  
Herbert P. Schweizer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Salicylic Acid ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Global Gene Expression ◽  
Defense Responses ◽  
N Gene ◽  
Virulence Determinants ◽  
In Planta

ABSTRACT Although Pseudomonas aeruginosa is an opportunistic pathogen that does not often naturally infect alternate hosts, such as plants, the plant-P. aeruginosa model has become a widely recognized system for identifying new virulence determinants and studying the pathogenesis of the organism. Here, we examine how both host factors and P. aeruginosa PAO1 gene expression are affected in planta after infiltration into incompatible and compatible cultivars of tobacco (Nicotiana tabacum L.). N. tabacum has a resistance gene (N) against tobacco mosaic virus, and although resistance to PAO1 infection is correlated with the presence of a dominant N gene, our data suggest that it is not a factor in resistance against PAO1. We did observe that the resistant tobacco cultivar had higher basal levels of salicylic acid and a stronger salicylic acid response upon infiltration of PAO1. Salicylic acid acts as a signal to activate defense responses in plants, limiting the spread of the pathogen and preventing access to nutrients. It has also been shown to have direct virulence-modulating effects on P. aeruginosa. We also examined host effects on the pathogen by analyzing global gene expression profiles of bacteria removed from the intracellular fluid of the two plant hosts. We discovered that the availability of micronutrients, particularly sulfate and phosphates, is important for in planta pathogenesis and that the amounts of these nutrients made available to the bacteria may in turn have an effect on virulence gene expression. Indeed, there are several reports suggesting that P. aeruginosa virulence is influenced in mammalian hosts by the availability of micronutrients, such as iron and nitrogen, and by levels of O2.

scholarly journals Transcriptional regulation of HSFA7 and post-transcriptional modulation of HSFB4a by miRNA4200 govern general and varietal thermotolerance in tomato

2021 ◽  
Author(s):  
Sombir Rao ◽  
Sonia Balyan ◽  
Jaishri Rubina Das ◽  
Radhika Verma ◽  
Saloni Mathur
Keyword(s):  
Heat Stress ◽  
Transcriptional Control ◽  
Expression Profiles ◽  
Functional Characterization ◽  
Promoter Sequence ◽  
Negative Regulator ◽  
Heat Shock Factors ◽  
In Planta ◽  
Transcriptional Modulation ◽  
Reporter Assays

AbstractHeat shock factors (HSFs) are at the core of heat stress (HS) response in plants. However, the contribution of HSFs governing the inherent thermotolerance mechanism in tomato from sub-tropical hot climates is poorly understood. With the above aim, comparative expression profiles of the HSF family in a HS tolerant (CLN1621L) and a sensitive cultivar (CA4) of tomato under HS revealed cultivar-biased regulation of an activator (HSFA7a) and repressor (HSFB4a) class HSF. Functional characterization of HSFA7a that was strongly up-regulated in the tolerant cultivar by VIGS-based silencing and overexpression established it as a positive regulator of HS-tolerance. While knock-down and overexpression analyses of HSFB4a that was down-regulated in CLN1621L in HS, showed it as a negative regulator of thermotolerance. Promoter:GUS reporter assays and promoter sequence analyses suggest heat-mediated transcriptional control of both the HSF genes in the contrasting cultivars. Moreover, we show HSFB4a is also regulated post-transcriptionally by microRNA Sly-miR4200 using degradome, short-tandem-target-mimic of Sly-miR4200 and transient in-planta Sly-miR4200-effector:HSFB4a-reporter assays. This miRNA is induced several folds upon HS in the tolerant variety thereby reducing HSFB4a levels. We thus propose that the alleviation of HSFB4a repressor governs thermotolerance in the tolerant cultivar by regulating downstream heat stress responsive genes.

scholarly journals The Pleiades are a cluster of fungal effectors that inhibit host defenses

2021 ◽  
Vol 17 (6) ◽  
pp. e1009641
Author(s):  
Fernando Navarrete ◽  
Nenad Grujic ◽  
Alexandra Stirnberg ◽  
Indira Saado ◽  
David Aleksza ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Functional Characterization ◽  
Rapid Evolution ◽  
Effector Proteins ◽  
In Planta ◽  
E3 Ligases ◽  
Effector Genes ◽  
Ros Burst ◽  
Molecular Patterns

Biotrophic plant pathogens secrete effector proteins to manipulate the host physiology. Effectors suppress defenses and induce an environment favorable to disease development. Sequence-based prediction of effector function is impeded by their rapid evolution rate. In the maize pathogen Ustilago maydis, effector-coding genes frequently organize in clusters. Here we describe the functional characterization of the pleiades, a cluster of ten effector genes, by analyzing the micro- and macroscopic phenotype of the cluster deletion and expressing these proteins in planta. Deletion of the pleiades leads to strongly impaired virulence and accumulation of reactive oxygen species (ROS) in infected tissue. Eight of the Pleiades suppress the production of ROS upon perception of pathogen associated molecular patterns (PAMPs). Although functionally redundant, the Pleiades target different host components. The paralogs Taygeta1 and Merope1 suppress ROS production in either the cytoplasm or nucleus, respectively. Merope1 targets and promotes the auto-ubiquitination activity of RFI2, a conserved family of E3 ligases that regulates the production of PAMP-triggered ROS burst in plants.

scholarly journals Analysis of Epichloë festucae small secreted proteins in the interaction with Lolium perenne

2018 ◽  
Author(s):  
Barry Scott ◽  
Berit Hassing ◽  
David Winter ◽  
Carl Mesarich ◽  
Carla Eaton ◽  
...  
Keyword(s):  
Expression Profiles ◽  
Vital Role ◽  
Effector Proteins ◽  
Secreted Proteins ◽  
In Planta ◽  
Dynamic Features ◽  
Symbiotic Interaction ◽  
Extracellular Medium ◽  
Epichloë Festucae ◽  
Small Secreted Proteins

Epichloë festucae is an endophyte of the agriculturally important perennial ryegrass. This species systemically colonises the aerial tissues of this host where its growth is tightly regulated thereby maintaining a mutualistic symbiotic interaction. Recent studies have suggested that small secreted proteins, termed effectors, play a vital role in the suppression of host defence responses. To date only a few effectors with important roles in mutualistic interactions have been described. Here we make use of the fully assembled E. festucae genome and EffectorP to generate a suite of 141 effector candidates. These were analysed with respect to their genome location and expression profiles in planta and in several symbiosis-defective mutants. We found an association between effector candidates and a class of transposable elements known as MITEs, but no correlation with other dynamic features of the E. festucae genome, such as transposable element-rich regions. Three effector candidates and a small GPI-anchored protein were chosen for functional analysis based on their high expression in planta compared to in culture and their differential regulation in symbiosis defective E. festucae mutants. All three candidate effector proteins were shown to possess a functional signal peptide and two could be detected in the extracellular medium by western blotting. Localization of the effector candidates in planta suggests that they are not translocated into the plant cell, but rather, are localized in the apoplastic space or are attached to the cell wall. Deletion and overexpression of the effector candidates, as well as the putative GPI-anchored protein, did not affect the plant growth phenotype or restrict growth of E. festucae mutants in planta. These results indicate that these proteins are either not required for the interaction at the observed life stages or that there is redundancy between effectors expressed by E. festucae.

scholarly journals Development of a Nested Quantitative Real-Time PCR for Detecting Phytophthora cinnamomi in Persea americana Rootstocks

Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1012-1017 ◽  
Author(s):  
J. Engelbrecht ◽  
T. A. Duong ◽  
N. van den Berg
Keyword(s):  
Real Time ◽  
Phytophthora Cinnamomi ◽  
Low Cost ◽  
Persea Americana ◽  
Economic Losses ◽  
In Planta ◽  
Phytophthora Root Rot ◽  
Disease Tolerance ◽  
Quantitative Manner ◽  
Pathogen Dna

Phytophthora cinnamomi causes Phytophthora root rot (PRR) in avocado (Persea americana), an important disease that causes severe economic losses to the avocado industry globally. To date, no PRR-resistant avocado rootstock variety has been discovered, although certain rootstock varieties have been shown to be more tolerant than others. In this study, we developed an accurate, low cost assay for in planta quantification of P. cinnamomi to evaluate disease tolerance. A nested real-time polymerase chain reaction assay was developed to sensitively detect pathogen DNA in plant tissues. Root samples from a highly tolerant (Dusa) and less tolerant (R0.12) rootstock were collected at 0, 3, 7, 14, and 21 days after inoculation with P. cinnamomi and used for pathogen quantification. Nested primers developed in this study were specific and sensitive and could detect P. cinnamomi in root tissues. The amount of P. cinnamomi quantified in roots was significantly higher in the less-tolerant R0.12 plants when compared with the highly tolerant Dusa plants at all time points. This study has confirmed the known status of disease tolerance of Dusa and R0.12 avocado rootstocks in a quantitative manner and provides a reliable molecular tool to assist with industry breeding programs for the selection of PRR-resistant avocado rootstock varieties.

scholarly journals 454 Genome Sequencing of Pseudoperonospora cubensis Reveals Effector Proteins with a QXLR Translocation Motif

2011 ◽  
Vol 24 (5) ◽  
pp. 543-553 ◽  
Author(s):  
Miaoying Tian ◽  
Joe Win ◽  
Elizabeth Savory ◽  
Alyssa Burkhardt ◽  
Michael Held ◽  
...  
Keyword(s):  
Genomic Sequence ◽  
Effector Proteins ◽  
Pseudoperonospora Cubensis ◽  
Sequence Information ◽  
Rxlr Effectors ◽  
Rxlr Effector ◽  
Pathogen Fitness ◽  
Oomycete Pathogen ◽  
Bona Fide ◽  
Secreted Peptides

Pseudoperonospora cubensis is a biotrophic oomycete pathogen that causes downy mildew of cucurbits, a devastating foliar disease threatening cucurbit production worldwide. We sequenced P. cubensis genomic DNA using 454 pyrosequencing and obtained random genomic sequences covering approximately 14% of the genome, thus providing the first set of useful genomic sequence information for P. cubensis. Using bioinformatics approaches, we identified 32 putative RXLR effector proteins. Interestingly, we also identified 29 secreted peptides with high similarity to RXLR effectors at the N-terminal translocation domain, yet containing an R-to-Q substitution in the first residue of the translocation motif. Among these, a family of QXLR-containing proteins, designated as PcQNE, was confirmed to have a functional signal peptide and was further characterized as being localized in the plant nucleus. Internalization of secreted PcQNE into plant cells requires the QXLR-EER motif. This family has a large number of near-identical copies within the P. cubensis genome, is under diversifying selection at the C-terminal domain, and is upregulated during infection of plants, all of which are common characteristics of characterized oomycete effectors. Taken together, the data suggest that PcQNE are bona fide effector proteins with a QXLR translocation motif, and QXLR effectors are prevalent in P. cubensis. Furthermore, the massive duplication of PcQNE suggests that they might play pivotal roles in pathogen fitness and pathogenicity.

scholarly journals Cellulase Activity as a Mechanism for Suppression of Phytophthora Root Rot in Mulches

2011 ◽  
Vol 101 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Brantlee Spakes Richter ◽  
Kelly Ivors ◽  
Wei Shi ◽  
D. M. Benson
Keyword(s):  
Root Rot ◽  
Phytophthora Cinnamomi ◽  
Cellulase Activity ◽  
Enzyme Concentration ◽  
Disease Suppression ◽  
In Vitro Assays ◽  
Infested Soil ◽  
Phytophthora Root Rot ◽  
Standard Curve

Wood-based mulches are used in avocado production and are being tested on Fraser fir for reduction of Phytophthora root rot, caused by Phytophthora cinnamomi. Research with avocado has suggested a role of microbial cellulase enzymes in pathogen suppression through effects on the cellulosic cell walls of Phytophthora. This work was conducted to determine whether cellulase activity could account for disease suppression in mulch systems. A standard curve was developed to correlate cellulase activity in mulches with concentrations of a cellulase product. Based on this curve, cellulase activity in mulch samples was equivalent to a cellulase enzyme concentration of 25 U ml–1 or greater of product. Sustained exposure of P. cinnamomi to cellulase at 10 to 50 U ml–1 significantly reduced sporangia production, but biomass was only reduced with concentrations over 100 U ml–1. In a lupine bioassay, cellulase was applied to infested soil at 100 or 1,000 U ml–1 with three timings. Cellulase activity diminished by 47% between 1 and 15 days after application. Cellulase applied at 100 U ml–1 2 weeks before planting yielded activity of 20.08 μmol glucose equivalents per gram of soil water (GE g–1 aq) at planting, a level equivalent to mulch samples. Cellulase activity at planting ranged from 3.35 to 48.67 μmol GE g–1 aq, but no treatment significantly affected disease progress. Based on in vitro assays, cellulase activity in mulch was sufficient to impair sporangia production of P. cinnamomi, but not always sufficient to impact vegetative biomass.

scholarly journals Identification of the specific long-noncoding RNAs involved in night-break mediated flowering retardation in Chenopodium quinoa

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Qi Wu ◽  
Yiming Luo ◽  
Xiaoyong Wu ◽  
Xue Bai ◽  
Xueling Ye ◽  
...  
Keyword(s):  
Expression Profiles ◽  
Functional Characterization ◽  
Chenopodium Quinoa ◽  
Noncoding Rnas ◽  
Long Noncoding Rnas ◽  
Rna Seq ◽  
Short Day ◽  
Homologous Genes ◽  
The Relationship ◽  
Encoding Genes

Abstract Background Night-break (NB) has been proven to repress flowering of short-day plants (SDPs). Long-noncoding RNAs (lncRNAs) play key roles in plant flowering. However, investigation of the relationship between lncRNAs and NB responses is still limited, especially in Chenopodium quinoa, an important short-day coarse cereal. Results In this study, we performed strand-specific RNA-seq of leaf samples collected from quinoa seedlings treated by SD and NB. A total of 4914 high-confidence lncRNAs were identified, out of which 91 lncRNAs showed specific responses to SD and NB. Based on the expression profiles, we identified 17 positive- and 7 negative-flowering lncRNAs. Co-expression network analysis indicated that 1653 mRNAs were the common targets of both types of flowering lncRNAs. By mapping these targets to the known flowering pathways in model plants, we found some pivotal flowering homologs, including 2 florigen encoding genes (FT (FLOWERING LOCUS T) and TSF (TWIN SISTER of FT) homologs), 3 circadian clock related genes (EARLY FLOWERING 3 (ELF3), LATE ELONGATED HYPOCOTYL (LHY) and ELONGATED HYPOCOTYL 5 (HY5) homologs), 2 photoreceptor genes (PHYTOCHROME A (PHYA) and CRYPTOCHROME1 (CRY1) homologs), 1 B-BOX type CONSTANS (CO) homolog and 1 RELATED TO ABI3/VP1 (RAV1) homolog, were specifically affected by NB and competed by the positive and negative-flowering lncRNAs. We speculated that these potential flowering lncRNAs may mediate quinoa NB responses by modifying the expression of the floral homologous genes. Conclusions Together, the findings in this study will deepen our understanding of the roles of lncRNAs in NB responses, and provide valuable information for functional characterization in future.

scholarly journals Transcriptome analysis reveals potential function of long non-coding RNAs in 20-hydroxyecdysone regulated autophagy in Bombyx mori

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Huili Qiao ◽  
Jingya Wang ◽  
Yuanzhuo Wang ◽  
Juanjuan Yang ◽  
Bofan Wei ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Target Gene ◽  
Fat Body ◽  
Expression Profiles ◽  
Functional Characterization ◽  
Differentially Expressed ◽  
Post Injection ◽  
Biological Processes ◽  
Potential Target ◽  
Non Coding Rnas

Abstract Background 20-hydroxyecdysone (20E) plays important roles in insect molting and metamorphosis. 20E-induced autophagy has been detected during the larval–pupal transition in different insects. In Bombyx mori, autophagy is induced by 20E in the larval fat body. Long non-coding RNAs (lncRNAs) function in various biological processes in many organisms, including insects. Many lncRNAs have been reported to be potential for autophagy occurrence in mammals, but it has not been investigated in insects. Results RNA libraries from the fat body of B. mori dissected at 2 and 6 h post-injection with 20E were constructed and sequenced, and comprehensive analysis of lncRNAs and mRNAs was performed. A total of 1035 lncRNAs were identified, including 905 lincRNAs and 130 antisense lncRNAs. Compared with mRNAs, lncRNAs had longer transcript length and fewer exons. 132 lncRNAs were found differentially expressed at 2 h post injection, compared with 64 lncRNAs at 6 h post injection. Thirty differentially expressed lncRNAs were common at 2 and 6 h post-injection, and were hypothesized to be associated with the 20E response. Target gene analysis predicted 6493 lncRNA-mRNA cis pairs and 42,797 lncRNA-mRNA trans pairs. The expression profiles of LNC_000560 were highly consistent with its potential target genes, Atg4B, and RNAi of LNC_000560 significantly decreased the expression of LNC_000560 and Atg4B. These results indicated that LNC_000560 was potentially involved in the 20E-induced autophagy of the fat body by regulating Atg4B. Conclusions This study provides the genome-wide identification and functional characterization of lncRNAs associated with 20E-induced autophagy in the fat body of B. mori. LNC_000560 and its potential target gene were identified to be related to 20-regulated autophagy in B. mori. These results will be helpful for further studying the regulatory mechanisms of lncRNAs in autophagy and other biological processes in this insect model.

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