Production of thaxtomin a by two species of Streptomyces causing potato scab

A total of nine isolates of streptomycetes were isolated from scab lesions on potato tubers. Five out of them were pathogenic on potato minitubers and four of the pathogenic isolates produced thaxtomin A in infected tubers tissues. The lesion surface areas induced by thaxtomin A were highest in treatment of the minitubers with extract of OMB inoculated with S-6 and S-7, intermediate with that inoculated with S-4 and lowest with S-3. The pathogenic isolates were identified by their colour of aerial mycelia, melanin pigment productivity (+ or -), the type of spore chains morphology and carbon utilization as either S. scabies strains S-3, S-4 and S-8, or S. acidiscabies strains S-6 and S-7. S-3 and S-4 produced 0.65 and 1.60 micrograms thaxtomin A per milliliter of OMB, respectively, whereas S-6 and S-7 produced similar amounts of thaxtomin A, 2.36 and 2.10 micrograms per ml of OMB, respectively. The optimal temperature for production of thaxtomin A by S. scabies and S. acidiscabies was 28 degrees C. Production of thaxtomin A by S. scabies strain S-4 and S. acidiscabies strain S-6 was suppressed at least 50-fold at 0.5 and 0.3% of glucose, respectively. Fructose enhanced the production of thaxtomin A by both S. scabies and S. acidiscabies.     

Production of thaxtomin a by two species ofStreptomyces causing potato scab

A total of nine isolates of streptomycetes were isolated from scab lesions on potato tubers. Five of nine isolates were pathogenic on potato minitubers. Four of the pathogenic isolates produced thaxtomin A (ThxA) in infected tuber tissues. The lesion surface areas inducing ThxA were highest in treatment of the minitubers with an extract of OMB inoculated with S-66 and S-67, intermediate with that inoculated with S-64 and lowest with S-63. The pathogenic isolates were identified by gray aerial mycelia, melanin pigment productivity, the type of spore chain morphology and carbon utilization asS. scabies strains S-63, S-64 and S-68, andS. acidiscabies strains S-66 and S-67. Strains S-63 and S-64 produced 0.65 and 1.60 mg ThxA per L of OMB, respectively, strains S-66 and S-67 producing similar amounts,viz. 2.36 and 2.10 mg/L, respectively. The optimal temperature for production (by both species) was 28 °C. Production of ThxA byS. scabies strain S-64 andS. acidiscabies strain S-66 was suppressed at least 50-fold at 0.5 and 0.3 % of glucose, respectively. Fructose enhanced the production by both species.     

Designing a crop rotation strategy to manage Streptomyces scabies causing potato scab in north India

Common scab of potato caused by various species in the genus Streptomyces has assumed serious proportions in certain potato-growing regions of North India. Although it does not have significant effect on tuber yield, it downgrades quality and reduces marketability. Being soil-borne, this disease is often difficult to manage. Therefore, the present studies were conducted to evaluate different potato-based crop rotations prevalent in north India as a part of a strategy to manage Streptomyces scabies population in scab sick fields. Our results showed that S. scabies population in soil can be effectively reduced either by keeping the field fallow after potato or by including mungbean or sunhemp as green manure, in a single year cropping sequence over a period of four years. Three crop rotations, viz. fallow–rice–potato, mungbean–rice–potato and sunhemp–rice–potato, showed maximum decrease in the population of S. scabies in soil resulting in reduced scab severity. The mungbean- and sunhemp-based crop rotations also enhanced rhizospheric soil microflora especially fluorescent pseudomonads and Trichoderma spp.     

Genetic diversity of Streptomyces spp. causing common scab of potato in eastern Canada

Common scab is an important disease of potato caused by Streptomyces scabies and other closely related species. In this study, the genetic diversity of Streptomyces spp. causing common scab of potato in eastern Canada was for the first time investigated. Forty-one Streptomyces spp. isolates were retrieved from necrotic lesions of potato tubers harvested from different regions of the Canadian provinces New-Brunswick, Nova Scotia and Prince-Edward-Island. Most isolates were closely related to known pathogenic S. scabies strains on the basis of partial 16S ribosomal (r) RNA and rpoB gene sequence analyses. Two isolates were identified as pathogenic species of Streptomyces acidiscabies. To our knowledge, this species has never been previously isolated in these areas. Genome fingerprinting studies using repetitive elements (rep) polymerase chain reactions (PCR) revealed 10 distinct genetic groups in eastern Canada. The geographical distribution of the genetic groups was region-dependant. Pathogenicity- and virulence-related genes (txtA, txtC, and tomA) were PCR-amplified from each isolate, and nucleotide sequence analysis of partial gene fragments revealed slight polymorphisms in both txtA and txtC genes. No genetic variation was noted in the partial tomA gene sequences.     

Major Streptomyces species associated with fissure scab of potato in South Africa including description of Streptomyces solaniscabiei sp. nov

Streptomyces species are the causal agents of several scab diseases on potato tubers. A new type of scab symptom, caused by Streptomyces species, was observed in South Africa from 2010 onwards. The disease was initially thought to be caused by a single Streptomyces species, however, subsequent isolations from similar symptoms on other potato tubers revealed diversity of the Streptomyces isolates. The objective of this study was to characterise these isolates in order to determine what are the major species involved in the disease. This was done by sequencing and phylogenetic analyses of the 16S rDNA as well as five housekeeping genes, investigation of growth on different culture media, standard phenotypic tests and scanning electron microscopy of culture morphology. The presence of the pathogenicity island (PAI) present in plant pathogenic Streptomyces species was also investigated. The genomes of eight isolates, selected from the three main clades identified, were sequenced and annotated to further clarify species boundaries. Three isolates of each of the three main clades were also inoculated onto susceptible potato cultivars in order to establish the pathogenicity of the species. The results of the phylogenetic and genome analyses revealed that there are three main species involved, namely, Streptomyces werraensis, Streptomyces pseudogriseolus and a novel Streptomyces species that is described here as Streptomyces solaniscabiei sp. nov., with strain FS70^T (=?PPPPB BD 2226^T?=?LMG 32103^T) as the type strain. The glasshouse trial results showed that all three of the Streptomyces species are capable of producing fissure scab symptoms. None of the Streptomyces isolates from fissure scab contained the full PAI and the mechanism of disease initiation still needs to be determined. Genomic comparisons also indicated that S. gancidicus Suzuki 1957 (Approved Lists 1980) is a later heterotypic synonym of S. pseudogriseolus Okami and Umezawa 1955 (Approved Lists 1980).

     

Species' identification and microarray-based comparative genome analysis of Streptomyces species isolated from potato scab lesions in Norway

Streptomyces strains were isolated from scab lesions on potatoes collected from different parts of Norway. Twenty-eight plant-pathogenic strains, as tested on seedlings of radish and on potato, were identified on the basis of physiological and molecular criteria. Polymerase chain reaction (PCR) analysis, using species-specific primers, and sequencing of the 16S rRNA gene identified 14 nonmelanin-producing strains to S. turgidiscabies. Fourteen melanin-producing strains were detected with primers specific to S. scabies, but whole-genome microarray analysis, based on 12 766 probes designed for 8848 predicted open reading frames (ORFs) of S. scabies, showed that the 14 strains were different from S. scabies. They were subsequently identified to be S. europaeiscabiei based on the internal transcribed spacer (ITS) sequences of the rRNA genes. This is the first report of the occurrence of S. turgidiscabies and S. europaeiscabiei in Norway. The putative 762 genes exhibiting the highest sequence differences between strains of S. europaeiscabiei and S. scabies according to microarray analysis were concentrated in relatively few gene ontology (GO) categories, including 'symbiosis and mutualism through parasitism', 'cell death' and 'responses to biotic stimulus', whereas genes related to primary metabolism appeared to be more conserved. Microarray data and 16S rRNA gene phylogeny showed, consistently, that there were two genetically distinguishable groups of S. europaeiscabiei on the basis of differences in 131 genes. The results provide novel information about the genetic variability of S. europaeiscabiei and the gene-specific variability between the genomes of S. europaeiscabiei and S. scabies. The usefulness of a custom-designed, whole-genome oligonucleotide microarray in a survey of bacterial plant pathogens was demonstrated.     

Interactions and biocontrol of pathogenic Streptomyces strains co-occurring in potato scab lesions

Aims:  To test interactions between pathogenic strains of Streptomyces turgidiscabies , S. scabies and S. aureofaciens . To study biological control of S. turgidiscabies and S. scabies using the nonpathogenic Streptomyces strain (346) isolated from a scab lesion and a commercially available biocontrol agent ( S. griseoviridis strain K61; 'Mycostop').
Methods and Results:  Pathogenic strains of S. turgidiscabies and S. aureofaciens inhibited growth of S. scabies in vitro , whereas strain 346 and S. griseoviridis inhibited the pathogenic strains and were subsequently tested for control of scab in the greenhouse and field. Strains 346 and K61 suppressed development of common scab disease caused by S. turgidiscabies in the greenhouse. Strain 346 reduced incidence of S. turgidiscabies in scab lesions on potato tubers in the field.
Conclusions:  Streptomyces turgidiscabies shows antagonism against S. scabies that occurs in the same scab lesions and shares the ecological niche in the field. Biocontrol of S. turgidiscabies is possible with nonpathogenic Streptomyces strains but interactions may be complicated.
Significance and Impact of the Study:  Streptomyces turgidiscabies may have potential to displace S. scabies under the Scandinavian potato growing conditions. Biological control of the severe potato scab pathogen, S. turgidiscabies , is demonstrated for the first time. The results can be applied to enhance control of common scab.     

Streptomyces maghwi, a new species producing roflamycoin

A new Streptomyces species is described for which the name S. maghwi is proposed. The organism is characterized by a pink mass of aerial mycelium, spiral spore chains, spores with smooth surfaces and a nonchromogenic vegetative mycelium. S. maghwi produces roflamycoin (Schlegel et al. 1981) formerly known as flavomycoin (Schlegel et al. 1971). The type strain of S. maghwi is deposited with CBS, Netherland.     

Responses of potato cultivars to the common scab pathogens, Streptomyces scabies and S. turgidiscabies

A glasshouse experiment was conducted to study the responses to Streptomyces scabies and S. turgidiscabies in potato cultivars Bellona, Matilda and Sabina (Solanum tuberosum). Potatoes were grown in a peat‐sand mixture inoculated with one of the two strains of either S. scabies or S. turgidiscabies. Logit models were used to analyse the data on disease incidence and severity, whereas the data on emergence and yield were tested by analysis of variance. S. turgidiscabies, a recently described potato pathogen in Finland, possessed a high ability to cause superficial, raised and pitted lesions on all three cultivars tested. Symptoms induced by S. turgidiscabies were similar to those of S. scabies, regardless of the cultivar, which suggests that the two causal organisms of common scab cannot be distinguished based on symptoms. Infection by S. turgidiscabies and S. scabies delayed emergence, had the tendency to decrease the yield, and increased the proportion of small tubers in the yield, regardless of the potato cultivar. Differences in the levels of resistance to common scab were evident between potato cultivars, since cvs. Matilda and Bellona showed higher disease incidence and more severe scab symptoms than cv. Sabina.     

Neosartorya takakii, a new species from soil in Brazil

Among the isolates from soil of grassland in Roraima State, Brazil, a new species ofNeosartorya, N. takakii is described and illustrated.Neosartorya takakii differs from the other known species of the genus in having lenticular ascospores with two distinct equatorial crests and with roughly circularly arranged projections on the convex walls. A comparative SEM view of ascospores from six relatedNeosartorya species is also provided.     

Collagen hydrolysis by a new Streptomyces species

A soil streptomycete hydrolysed collagen extracted from bovine Achilles tendon, calf skin, carp swim bladder and rat tail tendon. Enzyme activity was highest with rat tail tendon collagen as substrate. The taxonomy of the streptomycete was studied according to internationally approved methods. Comparative studies revealed that it resembled Streptomyces humidus , S. rochei and S. distatochromogenes in some respects, but differed in many cultural and physiological characteristics. Based on these differences the organism was given a new identity as Streptomyces species A.     

TxtH is a key component of the thaxtomin biosynthetic machinery in the potato common scab pathogen Streptomyces scabies

Streptomyces scabies causes potato common scab disease, which reduces the quality and market value of affected tubers. The predominant pathogenicity determinant produced by S. scabies is the thaxtomin A phytotoxin, which is essential for common scab disease development. Production of thaxtomin A involves the nonribosomal peptide synthetases (NRPSs) TxtA and TxtB, both of which contain an adenylation (A-) domain for selecting and activating the appropriate amino acid during thaxtomin biosynthesis. The genome of S. scabies 87.22 contains three small MbtH-like protein (MLP)-coding genes, one of which (txtH) is present in the thaxtomin biosynthesis gene cluster. MLP family members are typically required for the proper folding of NRPS A-domains and/or stimulating their activities. This study investigated the importance of TxtH during thaxtomin biosynthesis in S. scabies. Biochemical studies showed that TxtH is required for promoting the soluble expression of both the TxtA and TxtB A-domains in Escherichia coli, and amino acid residues essential for this activity were identified. Deletion of txtH in S. scabies significantly reduced thaxtomin A production, and deletion of one of the two additional MLP homologues in S. scabies completely abolished production. Engineered expression of all three S. scabies MLPs could restore thaxtomin A production in a triple MLP-deficient strain, while engineered expression of MLPs from other Streptomyces spp. could not. Furthermore, the constructed MLP mutants were reduced in virulence compared to wild-type S. scabies. The results of our study confirm that TxtH plays a key role in thaxtomin A biosynthesis and plant pathogenicity in S. scabies.     

Cell wall anionic polymers of Streptomyces sp. MB-8, the causative agent of potato scab

The cell wall of Streptomyces sp. MB-8 contains a major teichoic acid, viz., 1,3-poly(glycerol phosphate) substituted with N-acetyl-alpha-D-glucosamine (the degree of substitution is 60%), a minor teichoic acid, viz., non-substituted poly(glycerol phosphate), and a family of Kdn (3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid)-containing oligomers of the following general structure: [carbohydrate structure: see text]. The composition of the oligomers was established using MALDI-TOF mass spectroscopy. The present study provides the second example of the identification of Kdn as a component of cell wall polymers of streptomycetes, which are the causative agents of potato scab.     

Effect of Streptomyces melanosporofaciens strain EF-76 and of chitosan on common scab of potato

The efficiency of Streptomyces melanosporofaciens strain EF-76, a geldanamycin producer, and of chitosan, a polymer derived from chitin that elicits plant defense mechanisms, to protect potato tubers against common scab was evaluated under both controlled and field conditions (years 2000 and 2001). S. melanosporofaciens EF-76 reduced disease incidence in the greenhouse assay and in the 2001 field assay. EF-76 also reduced symptom severity on potato tubers grown under field conditions. Chitosan provided a protective effect against S. scabies,the causal agent of potato common scab during the 2000 field assay by reducing both disease incidence and symptom severity. Combination of S. melanosporofaciensEF-76 and chitosan ensures a level of protection that was at least equivalent to the protection conferred by one of the two products used alone. In some instances, an additive effect of protection was observed when both products were used in combination. Combination of S. melanosporofaciensEF-76 and of chitosan thus represents a promising method of biocontrol against common scab.     

Characterization of atypical Erwinia carotovora strains causing blackleg of potato in Brazil

AIMS: To determine the characteristics of bacteria associated with the blackleg disease of potato in Brazil and compare them with species and subspecies of pectolytic Erwinia. METHODS AND RESULTS: Biochemical and physiological characteristics of 16 strains from blackleg-infected potatoes in State of Rio Grande do Sul, Brazil, were determined and differentiated them from all the E. carotovora subspecies and E. chrysanthemi. Pathogenicity and maceration ability of the Brazilian strains were greater than those of E. carotovora subsp. atroseptica, the causal agent of potato blackleg in temperate zones. Analyses of serological reaction and fatty acid composition confirmed that the Brazilian strains differed from E. carotovora subsp. atroseptica, but the sequence of 16S rDNA gene and the 16S-23S intergenic spacer (IGS) region confirmed the Brazilian strains as pectolytic Erwinia. Restriction analysis of the IGS region differentiated the Brazilian strains from the subspecies of E. carotovora and from E. chrysanthemi. A unique SexAI restriction site in the IGS region was used as the basis for a primer to specifically amplify DNA from the Brazilian potato blackleg bacterium in PCR. CONCLUSIONS: The bacterium that causes the blackleg disease of potato in Brazil differs from E. carotovora subsp. atroseptica, the blackleg pathogen in temperate zones. It also differs from other subspecies of E. carotovora and from E. chrysanthemi and warrants status as a new subspecies, which would be appropriately named E. carotovora subsp. brasiliensis. SIGNIFICANCE AND IMPACT OF THE STUDY: The blackleg disease of potato is caused by a different strain of pectolytic Erwinia in Brazil than in temperate potato-growing regions. The Brazilian strain is more virulent than E. carotovora subsp. atroseptica, the usual causal agent of potato blackleg.