Plant Pathogen Response Research Articles

Inter‐individual genetic variation in the temperature response of Leptosphaeria species pathogenic on oilseed rape

AbstractIt is important to understand the likely response of plant pathogens to increased temperatures due to anthropogenic climate change. This includes evolutionary change due to selection on genetically based variation in growth rate with temperature. We attempted to quantify this in two ways. First, radial mycelial growth rates in agar culture were determined for a collection of 44 English isolates of Leptosphaeria maculans and 17 isolates of L. biglobosa, at 14 temperatures. For L. maculans the genotypic variances in four parameters were measured: minimum temperature allowing growth, optimum temperature, growth rate at the optimum temperature, and growth rate at the highest usable temperature, 31.8°C. The standard deviations were 0.068°C, 1.28°C, 0.21 mm/day, and 0.31 mm⋅day−1⋅°C−1, respectively. For L. biglobosa, these figures were, respectively: immeasurably small, 1.31°C, 0.053 mm/day, and 0.53 mm⋅day−1⋅°C−1. In addition, the incidence and severity of phoma stem canker in planta over a natural growing cycle at four temperatures (16, 20, 24, and 28°C) around the average culture optimum were determined. There was no correlation between in vitro and in planta growth, and the decrease in pathogen measures either side of the optimum temperature was much less for in planta growth than for in vitro growth. We conclude that both pathogens have the capacity to evolve to adapt to changes in environmental conditions, but that predictions of the effect of this adaptation, or estimates of heritability in natural conditions, cannot be made from measurements in vitro.

Editor Profile: Pascal Genschik.

![Figure][1] Pascal Genschik Accounting for more than 1600 genes in Arabidopsis ( Arabidopsis thaliana ), the ubiquitin-proteasome system (UPS) plays a central role in nearly all facets of plant biology ranging from hormone biology and plant pathogen responses to development and

Salicylic Acid Biosynthesis in Plants.

Salicylic acid (SA) is an important plant hormone that is best known for mediating host responses upon pathogen infection. Its role in plant defense activation is well established, but its biosynthesis in plants is not fully understood. SA is considered to be derived from two possible pathways; the ICS and PAL pathway, both starting from chorismate. The importance of both pathways for biosynthesis differs between plant species, rendering it hard to make generalizations about SA production that cover the entire plant kingdom. Yet, understanding SA biosynthesis is important to gain insight into how plant pathogen responses function and how pathogens can interfere with them. In this review, we have taken a closer look at how SA is synthesized and the importance of both biosynthesis pathways in different plant species.

Effect of Exogenic Humic Substances on Various Growth Endpoints of Alternaria alternata and Trichoderma harzianum in the Experimental Conditions

Industrial humic products (HPs) manufactured from various organic matter resources, including fossils, peat, lake-bottom sediments and organic waste materials, have found multiple applications in soil environments. Fungi play a crucial role in the turnover of humic substances (HSs). However, the data on the biological effects of HSs on soil fungi are contradictory. Revealing mechanisms of how HSs interact with soil filamentous fungi is essential for understanding a sustainable soil system. Here we study the responses of phytopathogenic Alternaria alternata and the potential antagonist Trichoderma harzianum, to exogenic HPs (from leonardite and lignosulfonate) at two concentrations (0.1 and 0.02%) in growth media with varying sucrose concentrations (0, 3 and 30 g/L). We assessed the fungal growth endpoints including the mycelium biomass accumulation, fungal colony size and conidia production. Our results demonstrate the significant relationships between the extent of inhibitory/stimulation effect of HPs on the investigated fungi and the carbon status of the growth medium, some features of the fungal species, as well as the type and concentration of the introduced HPs. The responses of plant pathogens correlated specifically with various types of HPs. HP from lignosulfonate exhibited a more stimulating effect on A. alternata compared to the effect of HP from leonardite.

Lack of gene flow between Phytophthora infestans populations of two neighboring countries with the largest potato production.

Gene flow is an important evolutionary force that enables adaptive responses of plant pathogens in response to changes in the environment and plant disease management strategies. In this study, we made a direct inference concerning gene flow in the Irish famine pathogen Phytophthora infestans between two of its hosts (potato and tomato) as well as between China and India. This was done by comparing sequence characteristics of the eukaryotic translation elongation factor 1 alpha (eEF‐1α) gene, generated from 245 P. infestans isolates sampled from two countries and hosts. Consistent with previous results, we found that eEF‐1α gene was highly conserved and point mutation was the only mechanism generating any sequence variation. Higher genetic variation was found in the eEF‐1α sequences in the P. infestans populations sampled from tomato compared to those sampled from potato. We also found the P. infestans population from India displayed a higher genetic variation in the eEF‐1α sequences compared to China. No gene flow was detected between the pathogen populations from the two countries, which is possibly attributed to the geographic barrier caused by Himalaya Plateau and the minimum cross‐border trade of potato and tomato products. The implications of these results for a sustainable management of late blight diseases are discussed.

Plant pathogen responses to Late Pleistocene and Holocene climate change in the central Atacama Desert, Chile

Future climate change has the potential to alter the distribution and prevalence of plant pathogens, which may have significant implications for both agricultural crops and natural plant communities. However, there are few long-term datasets against which modelled predictions of pathogen responses to climate change can be tested. Here, we use 18S metabarcoding of 28 rodent middens (solidified deposits of rodent coprolites and nesting material) from the Central Atacama, spanning the last ca. 49 ka, to provide the first long-term late Quaternary record of change in plant pathogen communities in response to changing climate. Plant pathogen richness was significantly greater in middens deposited during the Central Andean Pluvial Event (CAPE); a period of increased precipitation between 17.5–8.5 ka. Moreover, the occurrence frequency of Pucciniaceae (rust fungi) was significantly greater during the CAPE, and the highest relative abundances for five additional potentially pathogenic taxa also occurred during this period. The results demonstrate the promising potential for ancient DNA analysis of late Quaternary samples to reveal insights into how plant pathogens responded to past climatic and environmental change, which could help predict how pathogens may responded to future change.

Transcriptomic response of Ralstonia solanacearum to antimicrobial Pseudomonas fluorescens SN15-2 metabolites.

To develop efficient biocontrol agents, it is essential to investigate the response of soil-borne plant pathogens to such agents. For example, the response of Ralstonia solanacearum, the tomato wilt pathogen, to antimicrobial metabolites of Pseudomonas fluorescens is unknown. Thus, we assessed the effects of P. fluorescens SN15-2 fermentation broth on R. solanacearum by transmission electron microscopy and transcriptome technology. RNA sequencing identified 109 and 155 genes that are significantly upregulated and downregulated, respectively, in response to P. fluorescens metabolites, many of which are associated with the cell membrane and cell wall, and with nucleotide acid metabolism, iron absorption, and response to oxidative stress. This study highlights the effectiveness of P. fluorescens metabolites against the tomato wilt pathogen and helps clarify the underlying molecular mechanisms.

The use of fungal entomopathogens as endophytes in biological control: a review

ABSTRACTFungal entomopathogens have been proposed as environmentally friendly alternatives to chemical control. Unfortunately, their effectiveness continues to be limited by their susceptibility to ultraviolet (UV) light and low moisture. A relatively recent development, the use of fungal entomopathogens as endophytes, might overcome the traditional obstacles impeding the widespread adoption of fungal entomopathogens and also provide a novel alternative for management of insect pests and plant pathogens. In addition, some fungal entomopathogens could also function as biofertilizers. Eighty-five papers covering 109 individual fungal entomopathogen studies involving 12 species in six genera are reviewed. Thirty-eight plant species in 19 families were studied, with maize, common bean, and tomato being the most investigated. Of the 85 papers, 39 (46%) examined the effects of fungal entomopathogen endophytism on 33 insect species in 17 families and eight orders. Thirty-four (40%) examined plant response to endophytism, corresponding to 20 plant species. Various inoculation techniques (e.g., foliar sprays, soil drenching, seed soaking, injections, etc.) are effective in introducing fungal entomopathogens as endophytes, but colonization appears to be localized and ephemeral. The field of insect pathology will not substantially profit from dozens of additional studies attempting to introduce fungal entomopathogens into a wider array of plants, without attempting to understand the mechanisms underlying endophytism, the responses of the plant to such endophytism, and the consequent responses of insect pests and plant pathogens. This review presents several areas that should receive focused attention to increase the probability of success for making this technology an effective alternative to chemical control.

Investigating Plant Pathogen Responses: Using a Common Moss and a Soil Pathogen to Demonstrate Plant Defense Mechanisms

Plant–pathogen interactions are often omitted as a topic in most introductory and upper-level biology courses. The infection process of the plant pathogen Pythium irregulare on the moss Mnium cuspidatum can be observed and exploited to provide lessons on host–pathogen responses, as well as introduce other biological topics such as microscopy, spectrophotometry, and enzymes. Students can qualitatively analyze plant responses to pathogen infection using microscopy and observe quantitative enzyme responses to draw conclusions. Students are also encouraged to generate hypotheses and test them using this biological system as a method to develop scientific skills.

Proteomics offers insight to the mechanism behind Pisum sativum L. response to pea seed-borne mosaic virus (PSbMV)

Pea seed-borne mosaic virus (PSbMV) significantly reduces yields in a broad spectra of legumes. The eukaryotic translation initiation factor has been shown to confer resistance to this pathogen, thus implying that translation and proteome dynamics play a role in resistance. This study presents the results of a proteome-wide analysis of Pisum sativum L. response to PSbMV infection. LC–MS profiling of two contrasting pea cultivars, resistant (B99) and susceptible (Raman) to PSbMV infection, detected >2300 proteins, 116 of which responded to PSbMV ten and/or twenty days post-inoculation. These differentially abundant proteins are involved in number of processes that have previously been reported in the plant-pathogen response, including protein and amino acid metabolism, stress signaling, redox homeostasis, carbohydrate metabolism, and lipid metabolism. We complemented our proteome-wide analysis work with targeted analyses of free amino acids and selected small molecules, fatty acid profiling, and enzyme activity assays. Data from these additional experiments support our findings and validate the biological relevance of the observed proteome changes. We found surprising similarities in the resistant and susceptible cultivars, which implies that a seemingly unaffected plant, with no detectable levels of PSbMV, actively suppresses viral replication. Biological significancePlant resistance to PSbMV is connected to translation initiation factors, yet the processes involved are still poorly understood at the proteome level. To the best of our knowledge, this is the first survey of the global proteomic response to PSbMV in plants. The combination of label-free LC–MS profiling and two contrasting cultivars (resistant and susceptible) provided highly sensitive snapshots of protein abundance in response to PSbMV infection. PSbMV is a member of the largest family of plant viruses and our results are in accordance with previously characterized potyvirus-responsive proteomes. Hence, the results of this study can further extend our knowledge about these pathogens. We also show that even though no viral replication is detected in the PSbMV-resistant cultivar B99, it is still significantly affected by PSbMV inoculation.

Response of tomato wilt pathogen Ralstonia solanacearum to the volatile organic compounds produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9.

It is important to study the response of plant pathogens to the antibiosis traits of biocontrol microbes to design the efficient biocontrol strategies. In this study, we evaluated the role of volatile organic compounds (VOCs) produced by a biocontrol strain Bacillus amyloliquefaciens SQR-9 on the growth and virulence traits of tomato wilt pathogen Ralstonia solanacearum (RS). The VOCs of SQR-9 significantly inhibited the growth of RS on agar medium and in soil. In addition, the VOCs significantly inhibited the motility traits, production of antioxidant enzymes and exopolysaccharides, biofilm formation and tomato root colonization by RS. The strain SQR-9 produced 22 VOCs, but only nine VOCs showed 1–11% antibacterial activity against RS in their corresponding amounts; however, the consortium of all VOCs showed 70% growth inhibition of RS. The proteomics analysis showed that the VOCs of SQR-9 downregulated RS proteins related to the antioxidant activity, virulence, carbohydrate and amino acid metabolism, protein folding and translation, while the proteins involved in the ABC transporter system, amino acid synthesis, detoxification of aldehydes and ketones, methylation, protein translation and folding, and energy transfer were upregulated. This study describes the significance and effectiveness of VOCs produced by a biocontrol strain against tomato wilt pathogen.

LOB Domain Proteins: Beyond Lateral Organ Boundaries.

LATERAL ORGAN BOUNDARIES DOMAIN (LBD) proteins defined by a conserved LATERAL ORGAN BOUNDARIES (LOB) domain are key regulators of plant organ development. Recent studies have expanded their functional diversity beyond the definition of lateral organ boundaries to pollen development, plant regeneration, photomorphogenesis, pathogen response, and specific developmental functions in non-model plants, such as poplar and legumes. The identification of a range of upstream regulators, protein partners, and downstream targets of LBD family members has unraveled the molecular networks of LBD-dependent processes. Moreover, it has been demonstrated that LBD proteins have essential roles in integrating developmental changes in response to phytohormone signaling or environmental cues. As we discuss here, these novel discoveries of LBD functions and their molecular contexts promote a better understanding of this plant-specific transcription factor family.

Transcriptome responses to Ralstonia solanacearum infection in the roots of the wild potato Solanum commersonii.

BackgroundSolanum commersonii is a wild potato species that exhibits high tolerance to both biotic and abiotic stresses and has been used as a source of genes for introgression into cultivated potato. Among the interesting features of S. commersonii is resistance to the bacterial wilt caused by Ralstonia solanacearum, one of the most devastating bacterial diseases of crops.ResultsIn this study, we used deep sequencing of S. commersonii RNA (RNA-seq) to analyze the below-ground plant transcriptional responses to R. solanacearum. While a majority of S. commersonii RNA-seq reads could be aligned to the Solanum tuberosum Group Phureja DM reference genome sequence, we identified 2,978 S. commersonii novel transcripts through assembly of unaligned S. commersonii RNA-seq reads. We also used RNA-seq to study gene expression in pathogen-challenged roots of S. commersonii accessions resistant (F118) and susceptible (F97) to the pathogen. Expression profiles obtained from read mapping to the S. tuberosum reference genome and the S. commersonii novel transcripts revealed a differential response to the pathogen in the two accessions, with 221 (F118) and 644 (F97) differentially expressed genes including S. commersonii novel transcripts in the resistant and susceptible genotypes. Interestingly, 22.6% of the F118 and 12.8% of the F97 differentially expressed genes had been previously identified as responsive to biotic stresses and half of those up-regulated in both accessions had been involved in plant pathogen responses. Finally, we compared two different methods to eliminate ribosomal RNA from the plant RNA samples in order to allow dual mapping of RNAseq reads to the host and pathogen genomes and provide insights on the advantages and limitations of each technique.ConclusionsOur work catalogues the S. commersonii transcriptome and strengthens the notion that this species encodes specific genes that are differentially expressed to respond to bacterial wilt. In addition, a high proportion of S. commersonii-specific transcripts were altered by R. solanacearum only in F118 accession, while phythormone-related genes were highly induced in F97, suggesting a markedly different response to the pathogen in the two plant accessions studied.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1460-1) contains supplementary material, which is available to authorized users.

MicroRNAs in fruit trees: discovery, diversity and future research directions.

Since the first description of microRNAs (miRNAs) 20 years ago, the number of miRNAs identified in different eukaryotic organisms has exploded, largely due to the recent advances in DNA sequencing technologies. Functional studies, mostly from model species, have revealed that miRNAs are major post-transcriptional regulators of gene expression in eukaryotes. In plants, they are implicated in fundamental biological processes, from plant development and morphogenesis, to regulation of plant pathogen and abiotic stress responses. Although a substantial number of miRNAs have been identified in fruit trees to date, their functions remain largely uncharacterised. The present review aims to summarise the progress made in miRNA research in fruit trees, focusing specifically on the economically important species Prunus persica, Malus domestica, Citrus spp, and Vitis vinifera. We also discuss future miRNA research prospects in these plants and highlight potential applications of miRNAs in the on-going improvement of fruit trees.

RsERF1 derived from wild radish (Raphanus sativus) confers salt stress tolerance in Arabidopsis

The change in environmental parameters affects normal growth of plants, eventually reduces agricultural production. Ethylene plays vital roles in plant stress responses, germination, fruit ripening, organ abscission, pathogen response, and senescence. Expression of an ethylene-responsive transcription factor (ERF) was induced in Korean halophyte, Raphanus sativus var. hortensis f. raphanistroides (wild radish) by 200-mM sodium chloride (NaCl). Raphanus sativus ethylene-responsive transcription factor 1 (RsERF1) is also localized to nucleus, similar to other transcription factors. In yeast, RsERF1 showed transcriptional activation property, by expressing the reporter gene. Being a TF, RsERF1 specifically bound to the cis-acting elements, GCC box and DRE/CRT in vitro, to initiate transcription. Homozygous T3 transgenic Arabidopsis, overexpressing RsERF1, showed significant tolerance against salt stress in soil-grown conditions. The tolerance was also marked by an increased germination rate of RsERF1 transgenics in salt-containing media. In RsERF1 overexpression lines, abiotic stress-related genes such as ABF3, ABF4, ADH, Rab18, and SUS1 were upregulated by 200-mM NaCl. ERFs have been studied and proven for their tolerance potential against various abiotic stresses, but RsERF1 belongs to an ERF subgroup called ethylene-responsive transcription factor related to AP2 (ERF-RAP2). Thus, this is a first report for ERF-RAP2 from Korean halophyte cDNA library. We believe that extensive posttranslational modification studies will reveal the role and location of RsERF1 in stress tolerance pathway.

A Microfluidic Assay for Identifying Differential Responses of Plant and Human Fungal Pathogens to Tobacco Phylloplanins

Phylloplanins are defensive glycoproteins secreted onto leaf surfaces by trichome-bearing plants such as tobacco (Nicotiana tabacum). They are of interest because of their antimicrobial properties, but like other natural product bioactives, the assessment and screening of phylloplanins biological activity is impeded by limited availabilities of active compounds. Here we report an inexpensive microfluidic approach that requires ≤ 20 microliters of tobacco phylloplanins to assess spore germination inhibition of plant and human fungal pathogens. Spores of Colletotrichum coccodes and Aspergillus fumigatus suspended in solutions containing tobacco phylloplanins did not germinate at 48 and 30 h post-treatment, respectively. Tobacco phylloplanins transiently inhibited spore germination of Fusarium sambucinum, but had no detectable activity against Alternaria solani or Verticillium albo-atrum at the concentrations tested, demonstrating differential sensitivity of fungi to tobacco phylloplanins. 4 August 2014. 14 August 2014.

Local adaptation and evolutionary potential along a temperature gradient in the fungal pathogen Rhynchosporium commune

To predict the response of plant pathogens to climate warming, data are needed on current thermal adaptation, the pathogen's evolutionary potential, and the link between them. We conducted a common garden experiment using isolates of the fungal pathogen Rhynchosporium commune from nine barley populations representing climatically diverse locations. Clonal replicates of 126 genetically distinct isolates were assessed for their growth rate at 12°C, 18°C, and 22°C. Populations originating from climates with higher monthly temperature variation had higher growth rate at all three temperatures compared with populations from climates with less temperature fluctuation. Population differentiation in growth rate (QST) was significantly higher at 22°C than population differentiation for neutral microsatellite loci (GST), consistent with local adaptation for growth at higher temperatures. At 18°C, we found evidence for stabilizing selection for growth rate as QST was significantly lower than GST. Heritability of growth rate under the three temperatures was substantial in all populations (0.58–0.76). Genetic variation was lower in populations with higher growth rate at the three temperatures and evolvability increased under heat stress in seven of nine populations. Our findings imply that the distribution of this pathogen is unlikely to be genetically limited under climate warming, due to its high genetic variation and plasticity for thermal tolerance.

Plant Stress and Biotechnology

Plant Stress and Biotechnology

Unique secreted–surface protein complex of Lactobacillus rhamnosus, identified by phage display

Proteins are the most diverse structures on bacterial surfaces; hence, they are candidates for species- and strain-specific interactions of bacteria with the host, environment, and other microorganisms. Genomics has decoded thousands of bacterial surface and secreted proteins, yet the function of most cannot be predicted because of the enormous variability and a lack of experimental data that would allow deduction of function through homology. Here, we used phage display to identify a pair of interacting extracellular proteins in the probiotic bacterium Lactobacillus rhamnosus HN001. A secreted protein, SpcA, containing two bacterial immunoglobulin-like domains type 3 (Big-3) and a domain distantly related to plant pathogen response domain 1 (PR-1-like) was identified by screening of an L. rhamnosus HN001 library using HN001 cells as bait. The SpcA-“docking” protein, SpcB, was in turn detected by another phage display library screening, using purified SpcA as bait. SpcB is a 3275-residue cell-surface protein that contains general features of large glycosylated Serine-rich adhesins/fibrils from gram-positive bacteria, including the hallmark signal sequence motif KxYKxGKxW. Both proteins are encoded by genes within a L. rhamnosus-unique gene cluster that distinguishes this species from other lactobacilli. To our knowledge, this is the first example of a secreted-docking protein pair identified in lactobacilli.

Two Homologous Putative Protein Tyrosine Phosphatases, OsPFA-DSP2 and AtPFA-DSP4, Negatively Regulate the Pathogen Response in Transgenic Plants

Protein phosphatases, together with protein kinases, regulate protein phosphorylation and dephosphorylation, and play critical roles in plant growth and biotic stress responses. However, little is known about the biological functions of plant protein tyrosine dual-specificity phosphatase (PFA-DSP) in biotic stresses. Here, we found that OsPFA-DSP2 was mainly expressed in calli, seedlings, roots, and young panicles, and localized in cytoplasm and nucleus. Ectopic overexpression of OsPFA-DSP2 in rice increased sensitivity to Magnaporthe grisea (M. grisea Z1 strain), inhibited the accumulation of hydrogen peroxide (H2O2) and suppressed the expression of pathogenesis-related (PR) genes after fungal infection. Interestingly, transgenic Arabidopsis plants overexpressing AtPFA-DSP4, which is homologous to OsPFA-DSP2, also exhibited sensitivity to Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), reduced accumulation of H2O2 and decreased photosynthesic capacity after infection compared with Col-0. These results indicate that OsPFA-DSP2 and AtPFA-DSP4 act as negative regulators of the pathogen response in transgenic plants.