Showing papers in "Phytopathology in 2013"

A review of current knowledge of resistance aspects for the next-generation succinate dehydrogenase inhibitor fungicides.

Abstract: The new broad-spectrum fungicides from the succinate dehydrogenase inhibitor (SDHI) class have been quickly adopted by the market, which may lead to a high selection pressure on various pathogens. Cases of resistance have been observed in 14 fungal pathogens to date and are caused by different mutations in genes encoding the molecular target of SDHIs, which is the mitochondrial succinate dehydrogenase (SDH) enzyme. All of the 17 marketed SDHI fungicides bind to the same ubiquinone binding site of the SDH enzyme. Their primary biochemical mode of action is the blockage of the TCA cycle at the level of succinate to fumarate oxidation, leading to an inhibition of respiration. Homology models and docking simulations explain binding behaviors and some peculiarities of the cross-resistance profiles displayed by different members of this class of fungicides. Furthermore, cross-resistance patterns among SDHIs is complex because many mutations confer full cross resistance while others do not. The nature of the mutations found in pathogen populations varies with species and the selection compound used but cross resistance between all SDHIs has to be assumed at the population level. In most of the cases where resistance has been reported, the frequency is still too low to impact field performance. However, the Fungicide Resistance Action Committee has developed resistance management recommendations for pathogens of different crops in order to reduce the risk for resistance development to this class of fungicides. These recommendations include preventative usage, mixture with partner fungicides active against the current pathogen population, alternation in the mode of action of products used in a spray program, and limitations in the total number of applications per season or per crop.

Citrus huanglongbing: a newly relevant disease presents unprecedented challenges.

Abstract: Citrus huanglongbing (HLB) is one of the oldest citrus diseases and has been known for over a century. HLB is caused by ‘Candidatus Liberibacter’ spp. that are phloem-limited, fastidious α-proteobacteria and infect hosts in different Kingdoms (i.e., Animalia and Plantae). When compared with well-characterized, cultivatable plant-pathogenic Gram-negative bacteria, the interactions of uncultured insect-vectored plant-pathogenic bacteria, including ‘Ca. Liberibacter’ spp., with their hosts remain poorly understood. ‘Ca. Liberibacter’ spp. have been known to cause HLB, which has been rapidly spreading worldwide, resulting in dramatic economic losses. HLB presents an unprecedented challenge to citrus production. In this review, we focus on the most recent research on citrus, ‘Candidatus Liberibacter asiaticus’, and psyllid interactions, specifically considering the following topics: evolutionary relationships among ‘Ca. Liberibacter’ spp., genetic diversity, host range, genome analysis, transmission, virulence...

Major emerging problems with minor meloidogyne species.

Abstract: Elling, A. A. 2013. Major emerging problems with minor Meloidogyne species. Phytopathology 103:1092-1102. Root-knot nematodes (Meloidogyne spp.) represent one of the most polyphagous genera of plant-parasitic nematodes. To date, close to 100 valid species are recognized. In contrast to the size of the genus, the majority of past research focused on a small number of species, i.e., the so-called ‘major’ species M. arenaria, M. hapla, M. incognita, and M. javanica. This review highlights recent work aimed at ‘minor’ root-knot nematodes: M. chitwoodi, M. fallax, M. minor, M. enterolobii (=M. mayaguensis), M. exigua, and M. paranaensis. Some of these species have been described only recently. After a brief profile of each species, identification methods and their application in Meloidogyne spp. are summarized. Intraspecific variation and its impact on plant resistance breeding are discussed and interactions between M. enterolobii and Fusarium solani are highlighted as an example of synergistic interactions with other plant pathogens. Future research on Meloidogyne spp. is not only shaped by recent breakthroughs such as completing the genome sequences of M. hapla and M. incognita, but is also influenced by changes in agriculture. Taken together, the aim of this review is to draw attention to previously neglected and newly described Meloidogyne spp. that are developing into major problems for agriculture in tropical and temperate climates. Additional keywords: high-resolution melting curve, loop-mediated isothermal amplification, polymerase chain reaction, quarantine.

One Fungus, One Name: Defining the Genus Fusarium in a Scientifically Robust Way That Preserves Longstanding Use

Abstract: In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice.

Photocatalysis: effect of light-activated nanoscale formulations of TiO(2) on Xanthomonas perforans and control of bacterial spot of tomato

Abstract: Protection of crops from bacterial diseases presents a continuing challenge, mandating the development of novel agents and approaches. Photocatalysis is a process where chemically reactive oxygen species are catalytically generated by certain minerals in the presence of light. These reactive oxygen species have the capacity to destroy organic molecular structures critical to pathogen viability. In this study, the antibacterial potential of photocatalytic nanoscale titanium dioxide (TiO(2)), nanoscale TiO(2) doped (incorporation of other materials into the structure of TiO(2)) with silver (TiO(2)/Ag), and nanoscale TiO(2) doped with zinc (TiO(2)/Zn; AgriTitan) was evaluated against Xanthomonas perforans, the causal agent for bacterial spot disease of tomato. In vitro experiments on photocatalytic activity and dose dependency were conducted on glass cover slips coated with the nanoscale formulations by adding a known population of X. perforans strain Xp-F7 and illuminating the cover slips under a visible light source. TiO(2)/Ag and TiO(2)/Zn had high photocatalytic activity against X. perforans within 10 min of exposure to 3 × 10(4) lux. Greenhouse studies on naturally and artificially infected transplants treated with TiO(2)/Zn at ≈500 to 800 ppm significantly reduced bacterial spot severity compared with untreated and copper control. Protection was similar to the grower standard, copper + mancozeb. The use of TiO(2)/Zn at ≈500 to 800 ppm significantly reduced disease incidence in three of the four trials compared with untreated and copper control, and was comparable to or better than the grower standard. The treatments did not cause any adverse effects on tomato yield in any of the field trials.

Association of a DNA Virus with Grapevines Affected by Red Blotch Disease in California

Abstract: In the Napa Valley of California, vineyards of 'Cabernet Franc' (CF) clone 214, 'Cabernet Sauvignon' clone 337, and 'Zinfandel' clone 1A (Z1A) with grapevines exhibiting foliar symptoms of red blotches, marginal reddening, and red veins that were accompanied by reduced sugar accumulation in fruit at harvest were initially suspected to be infected with leafroll-associated viruses. However, reverse-transcription polymerase chain reaction (PCR) tests were negative for all known leafroll-associated viruses, with the exception of Grapevine leafroll-associated virus 2 in Z1A. Metagenomic analysis of cDNA libraries obtained from double-stranded RNA enriched nucleic acid (NA) preparations from bark scrapings of dormant canes on an Illumina platform revealed sequences having a distant relationship with members of the family Geminiviridae. Sequencing of products obtained by PCR assays using overlapping primers and rolling circle amplification (RCA) confirmed the presence of a single circular genome of 3,206 nucleotides which was nearly identical to the genome of a recently reported Grapevine cabernet franc-associated virus found in declining grapevines in New York. We propose to call this virus "Grapevine red blotch-associated virus" (GRBaV) to describe its association with grapevine red blotch disease. Primers specific to GRBaV amplified a product of expected size (557 bp) from NA preparations obtained from petioles of several diseased source vines. Chip bud inoculations successfully transmitted GRBaV to test plants of CF, as confirmed by PCR analysis. This is the first report of a DNA virus associated with red blotch disease of grapevines in California.

Identification of Eighteen Berberis Species as Alternate Hosts of Puccinia striiformis f. sp. tritici and Virulence Variation in the Pathogen Isolates from Natural Infection of Barberry Plants in China

Abstract: The wheat stripe rust pathogen (Puccinia striiformis f. sp. tritici) population in China has been reported to be a distinct genetic group with higher diversity than those in many other countries. Genetic recombination in the P. striiformis f. sp. tritici population has been identified with molecular markers but whether sexual reproduction occurs in China is unknown. In this study, we surveyed barberry plants for infection by rust fungi in the stripe rust “hotspot” regions in Gansu, Sichuan, and Shaanxi provinces; collected barberry plants and inoculated plants of 20 Berberis spp. with germinated teliospores under controlled greenhouse conditions for susceptibility to P. striiformis f. sp. tritici; and tested P. striiformis f. sp. tritici isolates obtained from aecia on naturally infected barberry plants on the wheat genotypes used to differentiate Chinese P. striiformis f. sp. tritici races to determine virulence variations. Different Berberis spp. were widely distributed and most surveyed plants...

Heat Treatment Eliminates ‘Candidatus Liberibacter asiaticus’ from Infected Citrus Trees Under Controlled Conditions

Abstract: Huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide. The three known causal agents of HLB are species of α-proteobacteria: ‘Candidatus Liberibacter asiaticus’, ‘Ca. L. africanus’, and ‘Ca. L. americanus’. Previous studies have found distinct variations in temperature sensitivity and tolerance among these species. Here, we describe the use of controlled heat treatments to cure HLB caused by ‘Ca. L. asiaticus’, the most prevalent and heat-tolerant species. Using temperature-controlled growth chambers, we evaluated the time duration and temperature required to suppress or eliminate the ‘Ca. L. asiaticus’ bacterium in citrus, using various temperature treatments for time periods ranging from 2 days to 4 months. Results of quantitative polymerase chain reaction (qPCR) after treatment illustrate significant decreases in the ‘Ca. L. asiaticus’ bacterial titer, combined with healthy vigorous growth by all surviving trees. Repeated qPCR testing confirmed that previously infecte...

Salmonella interactions with plants and their associated microbiota.

Abstract: The increase in the incidence of gastroenteritis outbreaks linked to the consumption of foods of plant origin has ignited public concern and scientific interest in understanding interactions of human enteric pathogens with plants. Enteric disease caused by nontyphoidal Salmonella is a major public health burden, with the number of cases of illness linked to fresh produce, spices, and nuts surpassing those linked to foods of animal origin. Mounting evidence supports the hypothesis that colonization of plants is an important part of the life cycle of this human pathogen. Although plant responses to human pathogens are distinct from the more specific responses to phytopathogens, plants appear to recognize Salmonella, likely by detecting conserved microbial patterns, which subsequently activates basal defenses. Numerous Salmonella genes have been identified as playing a role in its colonization of plant surfaces and tissues, and in its various interactions with other members of the phyto-microbial community. Importantly, Salmonella utilizes diverse and overlapping strategies to interact with plants and their microflora, and to successfully colonize its vertebrate hosts. This review provides insight into the complex behavior of Salmonella on plants and the apparent remarkable adaptation of this human pathogen to a potentially secondary host.

Aggressiveness of Fusarium species and impact of root infection on growth and yield of soybeans.

Abstract: Fusarium spp. are commonly isolated from soybean roots but the pathogenic activity of most species is poorly documented. Aggressiveness and yield impact of nine species of Fusarium were determined on soybean in greenhouse (50 isolates) and field microplot (19 isolates) experiments. Root rot severity and shoot and root dry weights were compared at growth stages V3 or R1. Root systems were scanned and digital image analysis was conducted; yield was measured in microplots. Disease severity and root morphology impacts varied among and within species. Fusarium graminearum was highly aggressive (root rot severity >90%), followed by F. proliferatum and F. virguliforme. Significant variation in damping-off (20 to 75%) and root rot severity ( 60%) was observed among F. oxysporum isolates. In artificially-infested microplots, root rot severity was low (<25%) and mean yield was not significantly reduced. However, there were significant linear relationships between yield and root symptoms for some isolates. Root morphological characteristics were more consistent indicators of yield loss than root rot severity. This study provides the first characterization of aggressiveness and yield impact of Fusarium root rot species on soybean at different plant stages and introduces root image analysis to assess the impact of root pathogens on soybean.

Hidden host plant associations of soilborne fungal pathogens: An ecological perspective

Abstract: Much of the current knowledge on population biology and ecology of soilborne fungal pathogens has been derived from research based on populations recovered from plants displaying disease symptoms or soil associated with symptomatic plants. Many soilborne fungal pathogens are known to cause disease on a large number of crop plants, including a variety of important agronomical, horticultural, ornamental, and forest plants species. For instance, the fungus Verticillium dahliae causes disease on >400 host plants. From a phytopathological perspective, plants on which disease symptoms have not been yet observed are considered to be nonhosts for V. dahliae. This term may be misleading because it does not provide information regarding the nature of the plant–fungus association; that is, a nonhost plant may harbor the fungus as an endophyte. Yet, there are numerous instances in the literature where V. dahliae has been isolated from asymptomatic plants; thus, these plants should be considered hosts. In thi...

Evidence that the Biofungicide Serenade (Bacillus subtilis) Suppresses Clubroot on Canola via Antibiosis and Induced Host Resistance

Abstract: This study investigated how the timing of application of the biofungicide Serenade (Bacillus subtilis QST713) or it components (product filtrate and bacterial cell suspension) influenced infection of canola by Plasmodiophora brassicae under controlled conditions. The biofungicide and its components were applied as a soil drench at 5% concentration (vol/vol or equivalent CFU) to a planting mix infested with P. brassicae at seeding or at transplanting 7 or 14 days after seeding (DAS) to target primary and secondary zoospores of P. brassicae. Quantitative polymerase chain reaction (qPCR) was used to assess root colonization by B. subtilis as well as P. brassicae. The biofungicide was consistently more effective than the individual components in reducing infection by P. brassicae. Two applications were more effective than one, with the biofungicide suppressing infection completely and the individual components reducing clubroot severity by 62 to 83%. The biofungicide also reduced genomic DNA of P. brassicae in canola roots by 26 to 99% at 7 and 14 DAS, and the qPCR results were strongly correlated with root hair infection (%) assessed at the same time (r = 0.84 to 0.95). qPCR was also used to quantify the transcript activity of nine host-defense-related genes in inoculated plants treated with Serenade at 14 DAS for potential induced resistance. Genes encoding the jasmonic acid (BnOPR2), ethylene (BnACO), and phenylpropanoid (BnOPCL and BnCCR) pathways were upregulated by 2.2- to 23-fold in plants treated with the biofungicide relative to control plants. This induced defense response was translocated to the foliage (determined based on the inhibition of infection by Leptosphaeria maculans). It is possible that antibiosis and induced resistance are involved in clubroot suppression by Serenade. Activity against the infection from both primary and secondary zoospores of P. brassicae may be required for maximum efficacy against clubroot.

Soil Suppressiveness to Fusarium Disease: Shifts in Root Microbiome Associated with Reduction of Pathogen Root Colonization

Abstract: Soil suppressiveness to Fusarium disease was induced by incubating sandy soil with debris of wild rocket (WR; Diplotaxis tenuifolia) under field conditions. We studied microbial dynamics in the roots of cucumber seedlings following transplantation into WR-amended or nonamended soil, as influenced by inoculation with Fusarium oxysporum f. sp. radicis-cucumerinum. Disease symptoms initiated in nonamended soil 6 days after inoculation, compared with 14 days in WR-amended soil. Root infection by F. oxysporum f. sp. radicis-cucumerinum was quantified using real-time polymerase chain reaction (PCR). Target numbers were similar 3 days after inoculation for both WR-amended and nonamended soils, and were significantly lower (66%) 6 days after inoculation and transplanting into the suppressive (WR-amended) soil. This decrease in root colonization was correlated with a reduction in disease (60%) 21 days after inoculation and transplanting into the suppressive soil. Fungal community composition on cucumber roots was assessed using mass sequencing of fungal internal transcribed spacer gene fragments. Sequences related to F. oxysporum, Fusarium sp. 14005, Chaetomium sp. 15003, and an unclassified Ascomycota composed 96% of the total fungal sequences in all samples. The relative abundances of these major groups were highly affected by root inoculation with F. oxysporum f. sp. radicis-cucumerinum, with a 10-fold increase in F. oxysporum sequences, but were not affected by the WR amendment. Quantitative analysis and mass-sequencing methods indicated a qualitative shift in the root's bacterial community composition in suppressive soil, rather than a change in bacterial numbers. A sharp reduction in the size and root dominance of the Massilia population in suppressive soil was accompanied by a significant increase in the relative abundance of specific populations; namely, Rhizobium, Bacillus, Paenibacillus, and Streptomyces spp. Composition of the Streptomyces community shifted significantly, as determined by PCR denaturing gradient gel electrophoresis, resulting in an increase in the dominance of a specific population in suppressive soils after only 3 days. This shift was related mainly to the increase in Streptomyces humidus, a group previously described as antagonistic to phytopathogenic fungi. Thus, suitable soil amendment resulted in a shift in the root's bacterial communities, and infection by a virulent pathogen was contained by the root microbiome, leading to a reduced disease rate.

The 8D05 parasitism gene of Meloidogyne incognita is required for successful infection of host roots.

Abstract: Parasitism genes encode effector proteins that are secreted through the stylet of root-knot nematodes to dramatically modify selected plant cells into giant-cells for feeding. The Mi8D05 parasitism gene previously identified was confirmed to encode a novel protein of 382 amino acids that had only one database homolog identified on contig 2374 within the Meloidogyne hapla genome. Mi8D05 expression peaked in M. incognita parasitic second-stage juveniles within host roots and its encoded protein was limited to the subventral esophageal gland cells that produce proteins secreted from the stylet. Constitutive expression of Mi8D05 in transformed Arabidopsis thaliana plants induced accelerated shoot growth and early flowering but had no visible effects on root growth. Independent lines of transgenic Arabidopsis that expressed a double-stranded RNA complementary to Mi8D05 in host-derived RNA interference (RNAi) tests had up to 90% reduction in infection by M. incognita compared with wild-type control plants, suggesting that Mi8D05 plays a critical role in parasitism by the root-knot nematode. Yeast two-hybrid experiments confirmed the specific interaction of the Mi8D05 protein with plant aquaporin tonoplast intrinsic protein 2 (TIP2) and provided evidence that the Mi8D05 effector may help regulate solute and water transport within giant-cells to promote the parasitic interaction.

The endophytic lifestyle of Escherichia coli O157:H7: quantification and internal localization in roots.

Abstract: The foodborne pathogen Escherichia coli O157:H7 is increasingly associated with fresh produce (fruit and vegetables). Bacterial colonization of fresh produce plants can occur to high levels on the external tissue but bacteria have also been detected within plant tissue. However, questions remain about the extent of internalization, its molecular basis, and internal location of the bacteria. We have determined the extent of internalization of E. coli O157:H7 in live spinach and lettuce plants and used high-resolution microscopy to examine colony formation in roots and pathways to internalization. E. coli O157:H7 was found within internal tissue of both produce species. Colonization occurred within the apoplast between plant cells. Furthermore, colonies were detected inside the cell wall of epidermal and cortical cells of spinach and Nicotiana benthamiana roots. Internal colonization of epidermal cells resembled that of the phytopathogen Pectobacterium atrosepticum on potato. In contrast, only spor...

Association of Little cherry virus 1 (LChV1) with the Shirofugen stunt disease and characterization of the genome of a divergent LChV1 isolate.

Abstract: Double-stranded RNAs purified from the V2356 (‘Successa’) sour cherry source of the Shirofugen stunt disease (SSD) were sequenced using a 454 pyrosequencing multiplex approach. The 15,646 reads obtained were assembled into 279 contigs, 5 of which, totaling almost 16.9 kbp and 5,332 reads (34% of sample reads), showed high Blast scores and homology to Little cherry virus 1 (LChV1). The five contigs were further assembled manually into three supercontigs spanning the full LChV1 genome with only two small gaps (17 and 55 bases). Completion of the sequencing of the viral genome was performed using targeted polymerase chain reaction and primers designed from the contigs. No evidence for the presence of other viral agents in the V2356 source could be obtained in the remaining contigs or singletons. The V2356 LChV1 isolate is only ≈76% identical with the reference complete LChV1 sequences and, in particular, with the ITMAR isolate associated with the Kwanzan stunting syndrome. However, it is highly homo...

Brassinosteroid Enhances Resistance to Fusarium Diseases of Barley

Abstract: Fusarium pathogens are among the most damaging pathogens of cereals. These pathogens have the ability to attack the roots, seedlings, and flowering heads of barley and wheat plants with disease, resulting in yield loss and head blight disease and also resulting in the contamination of grain with mycotoxins harmful to human and animal health. There is increasing evidence that brassinosteroid (BR) hormones play an important role in plant defense against both biotic and abiotic stress agents and this study set out to determine if and how BR might affect Fusarium diseases of barley. Application of the epibrassinolide (epiBL) to heads of 'Lux' barley reduced the severity of Fusarium head blight (FHB) caused by Fusarium culmorum by 86% and reduced the FHB-associated loss in grain weight by 33%. Growth of plants in soil amended with epiBL resulted in a 28 and 35% reduction in Fusarium seedling blight (FSB) symptoms on the Lux and 'Akashinriki' barley, respectively. Microarray analysis was used to determine whether growth in epiBL-amended soil changed the transcriptional profile in stem base tissue during the early stages of FSB development. At 24 and 48 h post F. culmorum inoculation, there were 146 epiBL-responsive transcripts, the majority being from the 48-h time point (n = 118). Real-time reverse-transcription polymerase chain reaction analysis validated the results for eight transcripts, including five defense genes. The results of gene expression studies show that chromatin remodeling, hormonal signaling, photosynthesis, and pathogenesis-related genes are activated in plants as a result of growth in epiBL.

Escherichia coli O157:H7 induces stronger plant immunity than Salmonella enterica Typhimurium SL1344.

Abstract: Consumption of fresh produce contaminated with bacterial human pathogens has resulted in various, sometimes deadly, disease outbreaks. In this study, we assessed plant defense responses induced by the fully pathogenic bacteria Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium SL1344 in both Arabidopsis thaliana and lettuce (Lactuca sativa). Unlike SL1344, O157:H7 induced strong plant immunity at both pre-invasion and post-invasion steps of infection. For instance, O157:H7 triggered stomatal closure even under high relative humidity, an environmental condition that generally weakens plant defenses against bacteria in the field and laboratory conditions. SL1344 instead induced a transient stomatal immunity. We also observed that PR1 gene expression was significantly higher in Arabidopsis leaves infected with O157:H7 compared with SL1344. These results suggest that plants may recognize and respond to some human pathogens more effectively than others. Furthermore, stomatal immunity...

A novel virus of the genus Cilevirus causing symptoms similar to citrus leprosis.

Abstract: Citrus leprosis in Colombia was previously shown to be caused by cytoplasmic Citrus leprosis virus (CiLV-C). In 2011, enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction (RT-PCR)-based diagnostic methods failed to identify CiLV-C from citrus samples with symptoms similar to citrus leprosis; however, virions similar to CiLV-C were observed in the cytoplasm of the symptomatic leaves by transmission electron microscopy. Furthermore, the causal organism was transmitted by the false spider mite, Brevipalpus phoenicis, to healthy citrus seedlings. A library of small RNAs was constructed from symptomatic leaves and used as the template for Illumina high-throughput parallel sequencing. The complete genome sequence and structure of a new bipartite RNA virus was determined. RNA1 (8,717 nucleotides [nt]) contained two open reading frames (ORFs). ORF1 encoded the replication module, consisting of five domains: namely, methyltransferase (MTR), cysteine protease-like, FtsJ-MTR, helicase (Hel), and RNA-dependent RNA polymerase (RdRp); whereas ORF2 encoded the putative coat protein. RNA2 (4,989 nt) contained five ORFs that encode the movement protein (MP) and four hypothetical proteins (p7, p15, p24, and p61). The structure of this virus genome resembled that of CiLV-C except that it contained a long 3' untranslated terminal region and an extra ORF (p7) in RNA2. Both the RNA1 and RNA2 of the new virus had only 58 and 50% nucleotide identities, respectively, with known CiLV-C sequences and, thus, it appears to be a novel virus infecting citrus. Phylogenetic analyses of the MTR, Hel, RdRp, and MP domains also indicated that the new virus was closely related to CiLV-C. We suggest that the virus be called Citrus leprosis virus cytoplasmic type 2 (CiLV-C2) and it should be unambiguously classified as a definitive member of the genus Cilevirus. A pair of CiLV-C2 genome-specific RT-PCR primers was designed and validated to detect its presence in citrus leprosis samples collected from the Casanare and Meta states in Colombia.

A study of weeds as potential inoculum sources for a tomato-infecting begomovirus in central Brazil.

Abstract: Tomato severe rugose virus (ToSRV) is the most important begomovirus species in Brazilian tomato production. Many weeds are associated with tomato, and some are hosts of begomoviruses. Only one species of weed, Nicandra physaloides, has been found to be infected with ToSRV. In this study, four weed species were investigated for their capacity to be infected by ToSRV and serve as a potential source of inoculum for tomato. Begomoviruses from naturally infected Crotalaria spp., Euphorbia heterophylla, N. physaloides, and Sida spp. were successfully transferred to tomato plants by biolistic inoculation. ToSRV was the major virus transferred to tomato. In contrast, other begomoviruses were transferred to weeds, such as Sida micrantha mosaic virus and Euphorbia yellow mosaic virus. Furthermore, a new strain of Sida micrantha mosaic virus is reported. We also confirmed that Crotalaria spp., E. heterophylla, and Sida spp. are infected with ToSRV but at low viral titers and in mixed infections with weed-infecting begomoviruses. Thus, it was demonstrated that weeds are potential sources of ToSRV for tomato in central Brazil.

Resident Bacteria on Leaves Enhance Survival of Immigrant Cells of Salmonella enterica

Abstract: Although Salmonella enterica apparently has comparatively low epiphytic fitness on plants, external factors that would influence its ability to survive on plants after contamination would be of significance in the epidemiology of human diseases caused by this human pathogen. Viable population sizes of S. enterica applied to plants preinoculated with Pseudomonas syringae or either of two Erwinia herbicola strains was ≥10-fold higher than that on control plants that were not precolonized by such indigenous bacteria when assessed 24 to 72 h after the imposition of desiccation stress. The protective effect of P. fluorescens, which exhibited antibiosis toward S. enterica in vitro, was only ≈50% that conferred by other bacterial strains. Although S. enterica could produce small cellular aggregates after incubation on wet leaves for several days, and the cells in such aggregates were less susceptible to death upon acute dehydration than solitary cells (as determined by propidium iodide staining), most S...

Transfer of Escherichia coli O157:H7 to Spinach by House Flies, Musca domestica (Diptera: Muscidae)

Abstract: Filth flies are known mechanical vectors of pathogenic bacteria in hospital and restaurant settings, but their role as vectors for disseminating microbes to plants has not been demonstrated. Escherichia coli O157:H7 deposition by flies onto spinach was studied using molecular, microbiological, and microscopy techniques. Relative quantitative polymerase chain reaction studies showed that bacteria acquired by flies from contaminated cattle manure and deposited in regurgitation spots on leaves survived and multiplied. Scanning electron microscopy of the regurgitation spots of flies exposed to manure inoculated with E. coli suggested the multiplication of bacteria-like organisms within the spots. This finding implies that the bacteria were active and is consistent with a hypothesis that regurgitation spots serve as a nutrition source allowing E. coli O157:H7 to survive on the spinach phylloplane. E. coli O157:H7 persisted on fly body surfaces up to 13 days after exposure to acquisition sources, suggesting that fly cuticular surfaces are conducive to the growth of this pathogen. These results are consistent with the hypothesis of bioenhanced transmission of human pathogens by house flies and suggest that filth flies may affect the microbial safety of fresh produce.

Dynamics of Southern rice black-streaked dwarf virus in rice and implication for virus acquisition.

Abstract: A novel viral disease of rice caused by Southern rice black-streaked dwarf virus (SRBSDV) has spread throughout East and Southeast Asia since the mid-2000s. Outbreaks of this viral disease occur yearly in southern parts of Japan concurrently with overseas migration of the planthopper vector Sogatella furcifera from southern China during the rainy season (from late June to early July). We examined the dynamics (changes in titer and localization) of SRBSDV on rice using reverse-transcription real-time polymerase chain reaction and determined the relationship between virus titer in plants and virus acquisition by S. furcifera. Under a constant temperature of 27°C, a substantial increase of SRBSDV titer in the leaf sheath together with typical symptoms (stunted growth and twisting of leaf tips) was observed at 20 days after the end of a 7-day exposure of viruliferous S. furcifera. Approximately 40% of S. furcifera acquired SRBSDV through feeding for 5 days on rice plants that were infected following exposure to viruliferous vectors for 10 to 15 days. These results suggest that rice infected by S. furcifera can be a source of SRBSDV before the next generation of S. furcifera emerges.

Functional analysis of Pid3-A4, an ortholog of rice blast resistance gene Pid3 revealed by allele mining in common wild rice.

Abstract: The rice blast resistance gene Pid3 encodes a nucleotide-binding-site leucine-rich repeat (NBS-LRR) protein. This gene was cloned from the rice 'Digu' (indica) by performing a genome-wide comparison of the NBS-LRR gene family between two genome-sequenced varieties, '9311' (indica) and 'Nipponbare' (japonica). In this study, we performed functional analysis of Pid3-A4, an ortholog of Pid3 revealed by allele mining in the common wild rice A4 (Oryza rufipogon). The predicted protein encoded by Pid3-A4 shares 99.03% sequence identity with Pid3, with only nine amino-acid substitutions. In wild rice plants, Pid3-A4 is constitutively expressed, and its expression is not induced by Magnaporthe oryzae isolate Zhong-10-8-14 infection. Importantly, in transgenic plants, Pid3-A4, as compared with Pid3, displays a distinct resistance spectrum to a set of M. oryzae isolates, including those that prevail in the rice fields of Sichuan Province. Therefore, Pid3-A4 should be quite useful for the breeding of rice blast resistance, especially in southwestern China.

Evidence for acquisition of copper resistance genes from different sources in citrus-associated xanthomonads.

Abstract: Behlau, F., Hong, J. C., Jones, J. B., and Graham, J. H. 2013. Evidence for acquisition of copper resistance genes from different sources in citrusassociated xanthomonads. Phytopathology 103:409-418. We determined that multiple and independent introductions of copper resistance genes have taken place for strains of Xanthomonas citri subsp. citri from Argentina and strains of X. alfalfae subsp. citrumelonis from Florida. This study compared the partial nucleotide sequences of principal copper resistance genes copL, copA, and copB from X. citri subsp. citri and X. alfalfae subsp. citrumelonis to strains of other Xanthomonas spp. resistant to copper that were isolated from 12 different countries or territories. The survey confirmed that the copLAB gene cluster is present in many species of Xanthomonas from different parts of the world. Alignment of partial nucleotide sequences of copper resistance genes among the copper-resistant (CuR) strains of Xanthomonas detected homology of ≥92, ≥96, and ≥91% for copL, copA, and copB, respectively. Grouping of strains based on branching patterns of phylogenetic trees was similar for copL and copA but differed for copB. When the three genes were concatenated and analyzed using various phylogenetic methods, it appeared that the plasmid had been horizontally transferred and various populations were mutating based on selection pressure unique to geographic regions. Although high homology of the genes among the strains indicated that the copper resistance in xanthomonads has a common origin, the slight differences in nucleotide sequences within groups of strains indicated that CuR genes have been independently exchanged among species of Xanthomonas throughout the world.

Organically Managed Soils Reduce Internal Colonization of Tomato Plants by Salmonella enterica Serovar Typhimurium

Abstract: A two-phase experiment was conducted twice to investigate the effects of soil management on movement of Salmonella enterica Typhimurium in tomato plants. In the first phase, individual leaflets of 84 tomato plants grown in conventional or organic soils were dip inoculated two to four times before fruiting with either of two Salmonella Typhimurium strains (10(9) CFU/ml; 0.025% [vol/vol] Silwet L-77). Inoculated and adjacent leaflets were tested for Salmonella spp. densities for 30 days after each inoculation. Endophytic bacterial communities were characterized by polymerase chain reaction denaturing gradient gel electrophoresis before and after inoculation. Fruit and seed were examined for Salmonella spp. incidence. In phase 2, extracted seed were planted in conventional soil, and contamination of leaves and fruit of the second generation was checked. More Salmonella spp. survived in inoculated leaves on plants grown in conventional than in organic soil. The soil management effect on Salmonella spp. survival was confirmed for tomato plants grown in two additional pairs of soils. Endophytic bacterial diversities of tomato plants grown in conventional soils were significantly lower than those in organic soils. All contaminated fruit (1%) were from tomato plants grown in conventional soil. Approximately 5% of the seed from infested fruit were internally contaminated. No Salmonella sp. was detected in plants grown from contaminated seed.

Identification of a new enamovirus associated with citrus vein enation disease by deep sequencing of small RNAs.

Abstract: To identify the causal agent of citrus vein enation disease, we examined by deep sequencing (Solexa-Illumina) the small RNA (sRNA) fraction from infected and healthy Etrog citron plants. Our results showed that virus-derived sRNAs (vsRNAs): (i) represent about 14.21% of the total sRNA population, (ii) are predominantly of 21 and 24 nucleotides with a biased distribution of their 5′ nucleotide and with a clear prevalence of those of (+) polarity, and (iii) derive from all the viral genome, although a prominent hotspot is present at a 5′-proximal region. Contigs assembled from vsRNAs showed similarity with luteovirus sequences, particularly with Pea enation mosaic virus, the type member of the genus Enamovirus. The genomic RNA (gRNA) sequence of a new virus, provisionally named Citrus vein enation virus (CVEV), was completed and characterized. The CVEV gRNA was found to be single-stranded, positive-sense, with a size of 5,983 nucleotides and five open reading frames. Phylogenetic comparisons based ...

The usefulness of fungicide mixtures and alternation for delaying the selection for resistance in populations of Mycosphaerella graminicola on winter wheat: a modeling analysis.

Abstract: A fungicide resistance model (reported and tested previously) was amended to describe the development of resistance in Mycosphaerella graminicola populations in winter wheat (Triticum aestivum) crops in two sets of fields, connected by spore dispersal. The model was used to evaluate the usefulness of concurrent, alternating, or mixture use of two high-resistance-risk fungicides as resistance management strategies. We determined the effect on the usefulness of each strategy of (i) fitness costs of resistance, (ii) partial resistance to fungicides, (iii) differences in the dose-response curves and decay rates between fungicides, and (iv) different frequencies of the double-resistant strain at the start of a treatment strategy. Parameter values for the quinine outside inhibitor pyraclostrobin were used to represent two fungicides with differing modes of action. The effectiveness of each strategy was quantified as the maximum number of growing seasons that disease was effectively controlled in both sets of fields. For all scenarios, the maximum effective lives achieved by the use of the strategies were in the order mixtures ≥ alternation ≥ concurrent use. Mixtures were of particular benefit where the pathogen strain resistant to both modes of action incurred a fitness penalty or was present at a low initial frequency.

Phenazine production by Pseudomonas sp. LBUM223 contributes to the biological control of potato common scab.

Abstract: Arseneault, T., Goyer, C., and Filion, M. 2013. Phenazine production by Pseudomonas sp. LBUM223 contributes to the biological control of potato common scab. Phytopathology 103:995-1000. Common scab of potato is mainly caused by Streptomyces scabies. Currently, no method can efficiently control this economically important disease. We have previously determined that Pseudomonas sp. LBUM223 exhibits antagonistic properties toward S. scabies under in vitro conditions. Inhibition was mainly attributed to phenazine-1-carboxylic acid (PCA) production because an isogenic mutant of LBUM223 (phzC–), not producing PCA, was incapable of significantly reducing S. scabies growth. In order to understand the impact of PCA production by LBUM223 in controlling common scab under soil conditions, pot experiments were performed to determine its effect on (i) reducing scab symptoms development, (ii) S. scabies population dynamics, and (iii) txtA expression in S. scabies, a key gene involved in thaxtomin A biosynthesis and required for pathogenesis. Symptoms were significantly reduced following inoculation with LBUM223 but not its mutant. Surprisingly, pathogen populations increased in the geocaulosphere in the presence of both wild-type and mutant strains of LBUM223; however, significant repression of txtA expression in S. scabies was only observed in the presence of PCA-producing LBUM223, not its mutant. These results suggest that, under soil conditions, PCA production by LBUM223 does not control common scab development by antibiosis but, instead, reduces S. scabies thaxtomin A production in the geocaulosphere, leading to reduced virulence.

Sclerotinia sclerotiorum populations infecting canola from China and the United States are genetically and phenotypically distinct.

Abstract: Genetic and phenotypic diversity and population differentiation of Sclerotinia sclerotiorum isolates infecting canola from China and the United States were investigated. Genetic diversity was assessed with eight microsatellite markers and mycelial compatibility groups (MCGs). Phenotypic diversity was assessed with sensitivity to three fungicides, production of oxalate and sclerotia, growth rate, and virulence on two canola cultivars. No shared MCGs or multilocus haplotypes were detected between the two populations, and populations differed significantly (P < 0.001). Recombination was detected in both populations but was greater in the Chinese population. A polymerase chain reaction detection assay showed that ~60% of the isolates were inversion-plus at the mating type locus. The two populations differed significantly (P < 0.05) for all of the phenotypic traits except for sensitivity to fungicide fluazinam and virulence. Isolates in the Chinese population were unique in several aspects. Despite th...