Antimycin A-producing nonphytopathogenic Streptomyces turgidiscabies from potato

Aim:  To detect if substances with mammalian cell toxicity are produced by Streptomyces turgidiscabies and Streptomyces scabiei isolated from potato scab lesions. Methods and Results:  In vitro cultures of phytopathogenic and nonphytopathogenic strains of S. scabiei and S. turgidiscabies, isolated from scab lesions of potato tubers originating from nine different cultivars from Finland and Sweden, were tested for toxicity using the rapid spermatozoan motility inhibition assay, previously shown useful in the detection of many different Streptomyces toxins and antimicrobial compounds. Purified toxins were used as reference. Three nonphytopathogenic strains of S. turgidiscabies were found to produce antimycin A when cultured on solid medium. Conclusions:  Boar sperm-motility-inhibiting substances are produced by strains of S. turgidiscabies and S. scabiei. The most powerful inhibitory substance, produced by three nonphytopathogenic S. turgidiscabies strains, was identified as antimycin A. The phytotoxic compounds thaxtomin A and concanamycin A did not inhibit sperm motility even at high doses. Significance and Impact of the Study:  The presence of antimycin A-producing Streptomyces strains, nonpathogenic to potato, was unexpected but important, considering the high mammalian toxicity of this cytochrome bc-blocking antibiotic.     

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.     

植物毒素thaxtomins生物合成及其分子调控研究进展

马铃薯疮痂病(potato scab)是世界范围内广泛存在的土传细菌性病害,难以防治。植物毒素thaxtomins由疮痂病链霉菌(Streptomyces scabies)次级代谢产生,是马铃薯疮痂病的主要致病原因,对马铃薯等作物产业造成严重危害。鉴于疮痂病链霉菌在农业上的重要作用,其中thaxtomins生物合成过程和分子调控得到越来越多的关注,并取得了较好的进展。本文综述了thaxtomins的结构特征、生物合成与异源表达,并重点介绍了疮痂病链霉菌中thaxtomins生物合成的分子调控机制等方面的研究进展,有利于深入认知疮痂病链霉菌次级代谢调控网络,为未来开发新型马铃薯疮痂病的防治策略提供理论指导。     

Phenolic acid content in potato peel determines natural infection of common scab caused by <Emphasis Type="Italic">Streptomyces</Emphasis> spp.

Common scab of potato caused by the actinomycete Streptomyces scabies is a common pathogen in almost all the potato growing areas of the world. Twenty cultivars of potato were screened in naturally scab infested farmers fields at two locations Tikari and Bachhawan, Varanasi, in two successive crop seasons (2006–2007 and 2007–2008). Among the cultivars, five cultivars were recorded to be least susceptible and the others ranged from medium susceptible to very highly susceptible. Most of the cultivars showed a stable resistance reaction in both the years. Qualitative as well as quantitative estimation of phenolic acids present in peels of the potato cultivars showed their possible role in protection of the potato cultivars against common scab. All the red skinned potato cultivars that were least susceptible to common scab infection were usually found to be rich in phenolic acid contents in their peels. This showed a positive correlation between cultivar resistance to common scab and phenolic acid content in the peel.     

Decreased severity of potato common scab after foliar sprays of 3,5 -dichlorophenoxyacetic acid, a possible antipathogenic agent

Glasshouse tests on the potato cultivars Majestic and Maris Bard measured the effects of single early foliar sprays of ring-substituted phenoxyacetic acids (0.9 × 10–³ M) on the incidence of potato common scab, caused by soil-borne Streptomyces scabies. The most effective compound was 3,5-dichlorophenoxyacetic acid (3,5-D), which decreased scab by about 90%; its action was preventative, early sprays being more effective than late However, it slightly decreased yield, and increased the number of tubers per plant and the proportion of deformed tubers. The trichloro- and tetrachloro- acids containing the 3,5-dichloro- group also decreased scab, though to lesser extents; the most active of these was the 2,3,4,5-tetrachloro- acid, which had the same side effects as 3,5-D. The herbicide 2,4-D almost halved scab incidence, but seriously decreased yield. Other acids tested had little or no effect on scab. In tests against S. scabies in culture, 3,5-D was no more toxic than other disubstituted acids which were inactive, or weakly active, against the disease in vivo. 3,5-D may be an ‘antipathogenic agent’, preventing scab development by altering tuber metabolism.     

Chemical control of potato common scab disease under field conditions

The aim of this study was to evaluate certain fungicides against Streptomyces scabies (Thaxter), the main causal agent of common scab disease in potato and other crops, in vitro and in vivo. Fourteen isolates of S. scabies were isolated from naturally infected potato tubers showing common scab symptoms. All isolates were pathogenic to potato tubers and produced typical symptoms of common scab. Isolate (No. 11) caused highest disease index (DSI) followed by isolates 10, 8 and 5 (61.13%). Seven fungicides (Rizolex, Capitan, Moncut, Ridomil, Maxim, Topsin, and Oxyplus) were screened in vitro for their toxicity against the pathogen isolate (No. 11). Results showed that four of them (Rizolex, Capitan, Moncut, and Ridomil) exhibited inhibition zone ranging from 5.33 to 26.33?mm. Capitan, Ridomil, and Rizolex were able to reduce DSI under field condition but they varied in their effects. Capitan was the best fungicide which aids in the reduction of disease (33.8%) followed by Ridomil (31.5%) while Rizolex (21.2%) was the lowest one.     

Bacillus altitudinis strain AMCC 101304: a novel potential biocontrol agent for potato common scab

ABSTRACT

Potato common scab, caused by Streptomyces spp., is one of the leading causes of heavy commercial losses in the potato industry and is thus one of the most serious plant diseases worldwide. This study identified and assessed potential biocontrol agents against potato common scab. In total, 110 isolates were obtained through antagonistic tests; among which, Bacillus sp. strain AMCC 101304 was found to be most effective at inhibiting the potato common scab pathogen, Streptomyces scabies. Bacillus sp. strain AMCC 101304 was finally identified as Bacillus altitudinis by morphological observation, physiological and biochemical experimentation, as well as 16S rRNA sequencing and phylogenetic analysis. Pot experiments were conducted twice (in spring and autumn) to verify the biocontrol effect of B. altitudinis AMCC 101304 against potato common scab. In spring, the control efficiency reached 76.34%. In autumn, the disease incidence was reduced from 100% to 34.19% (one treatment with strain AMCC 101304) and 38.42% (two treatments with strain AMCC 101304), and the control efficiency reached 82.50% (one application) and 78.43% (two applications). The present study demonstrated the potential of an isolate, identified as B. altitudinis AMCC 101304, as an effective biocontrol agent for future use in the field.     

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.     

What does it take to be a plant pathogen: genomic insights from <Emphasis Type="Italic">Streptomyces</Emphasis> species

Plant pathogenicity is rare in the genus Streptomyces, with only a dozen or so species possessing this trait out of the more than 900 species described. Nevertheless, such species have had a significant impact on agricultural economies throughout the world due to their ability to cause important crop diseases such as potato common scab, which is characterized by lesions that form on the potato tuber surface. All pathogenic species that cause common scab produce a family of phytotoxins called the thaxtomins, which function as cellulose synthesis inhibitors. In addition, the nec1 and tomA genes are conserved in several pathogenic streptomycetes, the former of which is predicted to function in the suppression of plant defense responses. Streptomyces scabies is the oldest plant pathogen described and has a world-wide distribution, whereas species such as S. turgidiscabies and S. acidiscabies are believed to be newly emergent pathogens found in more limited geographical locations. The genome sequence of S. scabies 87-22 was recently completed, and comparative genomic analyses with other sequenced microbial pathogens have revealed the presence of additional genes that may play a role in plant pathogenicity, an idea that is supported by functional analysis of one such putative virulence locus. In addition, the availability of multiple genome sequences for both pathogenic and nonpathogenic streptomycetes has provided an opportunity for comparative genomic analyses to identify the Streptomyces pathogenome. Such genomic analyses will contribute to the fundamental understanding of the mechanisms and evolution of plant pathogenicity and plant-microbe biology within this genus.     

Genetic background affects pathogenicity island function and pathogen emergence in Streptomyces

With few exceptions, thaxtomin A (ThxA), a nitrated diketopiperazine, is the pathogenicity determinant for plant‐pathogenic Streptomyces species. In Streptomyces scabiei (syn. S. scabies), the ThxA biosynthetic cluster is located within a 177‐kb mobile pathogenicity island (PAI), called the toxicogenic region (TR). In S. turgidiscabies, the ThxA biosynthetic cluster is located within a 674‐kb pathogenicity island (PAIst). The emergence of new plant pathogens occurs in this genus, but not frequently. This raises the question of whether the mobilization of these pathogenicity regions, through mating, is widespread and whether TR and PAIst can confer plant pathogenicity. We showed that ThxA biosynthetic clusters on TR and PAIst were transferred into strains from five non‐pathogenic Streptomyces species through mating with S. scabiei and S. turgidiscabies. However, not all of the transconjugants produced ThxA and exhibited the virulence phenotype, indicating that the genetic background of the recipient strains affects the functionality of the ThxA biosynthetic cluster and therefore would be expected to affect the emergence of novel pathogenic Streptomyces species. Thxs have been patented as natural herbicides, but have yet to be commercialized. Our results also demonstrated the potential of the heterologous production of ThxA as a natural and biodegradable herbicide in non‐pathogenic Streptomyces species.     

Isolation and characterization of an antibiotic produced by the scab disease-suppressive Streptomyces diastatochromogenes strain PonSSII

An antibiotic produced by the scab disease-suppressive Streptomyces diastatochromogenes strain PonSSII has been isolated and partially characterized. The antibiotic is produced throughout culture growth, with maximum amounts accumulating in the broth when the culture is in the early stationary phase of growth. The activity declines within about 30 h after the culture enters stationary phase. Purification techniques included chromatography on Amberlite XAD-2, DEAE Sephadex and SP Sephadex in addition to C18 HPLC with an average yield of 75%. This antibiotic only inhibits pathogenic strains of S. scabies that cause scab disease on potato and other tuberous vegetables and does not affect S. griseus, S. venezuelae, Actinomyces bovis, Nocardia asteroides, Clostridium perfringens, Bacillus subtilis, Staphylococcus aureus, S. epidermidis, Enterococcus faecalis, Micrococcus luteus, Serratia marcescens and Escherichia coli. The antibiotic has a molecular weight of 500 or less, and is stable for weeks at acidic pH but is very labile at alkaline pH conditions. Received 18 February 1997/ Accepted in revised form 11 August 1997     

Effect of carbohydrates on the production of thaxtomin A by <Emphasis Type="Italic">Streptomyces acidiscabies</Emphasis>

Several Streptomyces species cause plant diseases, including S. scabies, S. acidiscabies and S. turgidiscabies, which produce common scab of potato and similar diseases of root crops. These species produce thaxtomins, dipeptide phytotoxins that are responsible for disease symptoms. Thaxtomins are produced in vivo on diseased potato tissue and in vitro in oat-based culture media, but the regulation of thaxtomin biosynthesis is not understood. S. acidiscabies was grown in a variety of media to assess the impact of medium components on thaxtomin A (ThxA) production. ThxA biosynthesis was not correlated with bacterial biomass, nor was it stimulated by α-solanine or α-chaconine, the two most prevalent potato glycoalkaloids. ThxA production was stimulated by oat bran broth, even after exhaustive extraction, suggesting that specific carbohydrates may influence ThxA biosynthesis. Oat bran contains high levels of xylans and glucans, and both of these carbohydrates, as well as xylans from wheat and tamarind, stimulated ThxA production, but not to the same extent as oat bran. Starches and simple sugars did not induce ThxA production. The data indicate that complex carbohydrates may act as environmental signals to plant pathogenic Streptomyces, allowing production of thaxtomin and enabling bacteria to colonize its host.     

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.     

Glasshouse tests of chemicals for control of potato common scab

Quintozene is the only chemical used successfully in practice to control soil-borne Streptomyces scabies, the cause of potato common scab. However it may be carcinogenic, and the aim of this work was to find a substitute for it. About 100 chemicals, most of which had known fungitoxic action or were related to such chemicals, were tested in the glasshouse by growing potato plants in soil with which the chemicals had been mixed, usually at 50 ppm. Many of the chemicals failed to decrease the incidence of scab, or decreased the yield of tubers, or both. The most effective chemical was captafol, which was as effective as quintozene and, unlike quintozene, did not decrease yield. Captan and folpet, which are very closely related to captafol, were ineffective.     

Effect of Pathogen Isolate, Potato Cultivar, and Antagonist Strain on Potato Scab Severity and Biological Control

Antibiotic-producing Streptomyces spp. have shown potential in biocontrol of potato scab caused by Streptomyces scabies. However, results have been inconsistent among field trials. Pathogen isolate, antagonist strain and potato cultivar were investigated as potential sources of variation in the efficacy of potato scab biocontrol. Biocontrol success varied significantly among pathogen isolates and was not correlated with in vitro sensitivity to antibiotic inhibition. Antagonists also varied in their effectiveness as biocontrol agents, and the relative effectiveness of different antagonists varied among growing seasons. Finally, biocontrol varied among potato cultivars in the field. The diverse origins of significant variation in potato scab biocontrol suggest that consistent control in the field is likely to be difficult to achieve.     

Effects of foliar sprays of daminozide on the incidence of potato common scab

Single foliar sprays of the growth retardant daminozide (1.5–12 g/l) approximately halved the incidence of common scab, caused by soil-borne Streptomyces scabies, on potted potato plants in the glasshouse. Two analogues of daminozide (N-dimethylaminomaleamic and N-(dimethylamino)-methylsuccinamic acids) also decreased scab, but others were inactive. Of 22 other unrelated growth regulators and translocated chemicals tested as foliar sprays, only gibberellic acid (0.1 g/l) decreased scab incidence, but many of the tubers were distorted. Chlormequat chloride and chlorphonium chloride, as root treatments, were inactive. In other experiments with daminozide, scab incidence was decreased after application to soil. In tests with two plants per pot, spraying one of each pair decreased its rate of stem extension, but did not affect the other, indicating that little or no daminozide passed into the soil from the roots of the sprayed plant. The decrease in scab brought about by foliar sprays was not altered by varying their timing during the period before symptom development (1 to 5 wk after potting). In agar plate tests, daminozide was only weakly toxic to S. scabies. It is concluded that daminozide probably decreased scab by altering the physiology of the plants, so that scab symptoms did not develop.     

Development of a Genotyping Method for Potato Scab Pathogens Based on Multiplex PCR

Scab disease significantly damages potato and other root crops. Streptomyces scabiei, S. acidiscabiei, and S. turgidiscabiei are the best-known causal agents of this disease. We have developed a novel genotyping method for these potato scab pathogens using multiplex PCR, whose benefits include rapid and easy detection of multiple species. We designed a species-specific primer set (6 primers, 3 pairs) for the 16S rRNA genes and 16S–23S ITS regions of these potato scab pathogens. The specificity of the primer set was confirmed by testing 18 strains containing potato scab pathogens, other Streptomyces species, and strains of other genera. The application of the developed method to potato field soil and potato tissue samples resulted in the clear detection and identification of pathogens. Since this method is applicable to a large number of environmental samples, it is expected to be useful for a high-throughput analysis of soil and plant tissues of scab disease.     

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.