Suppression of root rot of mung bean (Vigna radiata L.) by Streptomyces sp. is associated with induction of peroxidase and polyphenol oxidase

Five strains of Streptomyces sp. were evaluated in vitro for their ability of inhibiting the mycelial growth of Macrophomina phaseolina, the causal agent of root rot of mung bean (Vigna radiata L.). Among the Streptomyces sp. strains tested, PDK showed the maximum in vitro inhibition of mycelial growth of M. phaseolina and recorded an inhibition zone of 21?mm. The strains CBE, MDU, SA and ANR recorded inhibition zones of 18, 16, 13 and 11?mm, respectively. These Streptomyces sp. strains were tested for their growth-promoting efficiency on mung bean seedlings. Among them, CBE and PDK recorded the maximum increase in shoot length, root length and seedling vigour compared with control, followed by MDU. Three Streptomyces sp. strains (CBE, MDU and PDK) that showed higher levels of inhibition of growth of M. phaseolina in dual culture assay and plant growth-promoting activity were tested for their biocontrol activity against root rot under greenhouse and field conditions. Seed treatment or soil application with powder formulation of Streptomyces sp. strains CBE, MDU and PDK was effective in controlling root rot disease; but, combined application through seed and soil increased the efficacy in both the greenhouse and field trials. Among the treatments, seed treatment plus soil application with powder formulation of Streptomyces sp. strain CBE proved to be most effective, which reduced the root rot incidence from 26.8% (with non-bacterised seeds) to 4.0% in Trial I and from 32.0 to 4.9% in Trial II. The above treatment recorded the highest yield in both the field trials, and the yield increase was 78 and 74% over control in Trial I and Trial II, respectively. Isozyme analysis of the Streptomyces sp.-treated plants indicates that seed treatment plus soil application strongly induce the activities of peroxidase (PO-1 and PO-2) and polyphenol oxidase (PPO-2 and PPO-3) in mung bean. Among the three strains tested, Streptomyces sp. strain MDU- treated plants showed higher levels of activities of PO and PPO. Based on the above findings, it can be concluded that both the direct inhibition of pathogen and induced resistance might be involved in the control of root rot of mung bean by Streptomyces sp.     

Incidence of charcoal rot of sorghum and soil populations of Macrophomina phaseolina associated with sorghum and native vegetation in Somalia

Studies were conducted in the Bay Region of Somalia to determine the incidence of charcoal rot in sorghum incited by Macrophomina phaseolina and the soil population of M. phaseolina in sorghum fields and areas of native vegetation. Charcoal rot was detected in 34 of 40 sorghum fields. Incidence (percent sorghum hills with diseased plants) in the four regional districts was 21, 70, 20 and 35% (mean of 15 hills/field and 10 fields/district) for Baidoa, Burhakaba, Dinsoore and Quansadhere, respectively. Soil collected from the 40 sorghum fields and from 40 native vegetation sites (10 in each district) all contained M. phaseolina. Mean soil populations for sorghum fields and native vegetation sites were 25.2 and 2.5 microsclerotia g^–1 soil, respectively. Soil populations of M. phaseolina in sorghum fields were significantly (p=0.05) different from populations in native vegetation sites and ranged from 7 to 107 and 0.1 to 14.0 microsclerotia g^–1 soil, respectively. Incidence of charcoal rot in sorghum was positively correlated with soil populations of M. phaseolina (r=0.53, p= 0.01). Correlation between ratoon failure and disease incidence was also highly significant (r=0.48, p=0.01). Correlation between ratoon failure and soil population of M. phaseolina was not significant (r=0.17, p=0.10). Only one out of 30 seed lots of sorghum grown in the Bay Region contained seed infected with M. phaseolina.     

Multifarious plant growth promoting characteristics of chickpea rhizosphere associated Bacilli help to suppress soil-borne pathogens

Wilt and root rot are the major constraints in chickpea production and very difficult to manage through agrochemicals. Hence, for an ecofriendly and biological management, 240 strains of Bacillus and Bacillus derived genera were isolated from chickpea rhizosphere, further narrowed down to 14 strains on the basis of in vitro production of indole acetic acid, siderophore, phosphate solubilization, hydrolytic enzymes and were evaluated for antagonism against chickpea pathogens (Fusarium oxysporum f. sp. ciceri race 1, F. solani and Macrophomina phaseolina). The strains were identified on the basis of physiological characters and 16S RNA gene sequencing. The genotypic comparisons of strains were determined by BOX-polymerase chain reaction profiles and amplified rDNA restriction analysis. These isolates were evaluated in greenhouse assay in which B. subtilis (B-CM191, B-CV235, B-CL-122) proved to be effective in reducing wilt incidence and significant enhancement in growth (root and shoot length) and dry matter of chickpea plants. PCR amplification of bacillomycin (bmyB) and β-glucanase genes suggests that amplified genes from the Bacillus could have a role to further define the diversity, ecology, and biocontrol activities in the suppression of soil-borne pathogens.     

Control of charcoal root rot in <Emphasis Type="Italic">Pinus radiata</Emphasis> nurseries with antagonistic bacteria

The objective of this study was to determine the potential of antagonistic bacteria to control charcoal root rot of coniferous seedlings caused by Macrophomina phaseolina (Tassi) Goid. in forest nurseries. Bacterial isolates were collected from nurseries located between Region Metropolitana and the VIII Region of Chile. Antagonists were initially evaluated in in vitro assays based on the ability to inhibit mycelial growth of M. phaseolina, and subsequently in two trials in a Pinus radiata nursery with a natural infestation of the pathogen. For nursery trials, the isolates were selected according to in vitro and field trial pathogen controls. The bacteria were applied as seed treatments and via water irrigation. The trials were conduced in a completely randomized block design. Among 568 bacterial isolates tested in vitro, 19.8% displayed some capacity to inhibit the mycelial growth of M. phaseolina, with inhibition between 1.7% and 67.6%. In the first nursery trial, Bacillus amyloliquefaciens VII 015, Bacillus pumilus IX 030, Bacillus stearothermophilus TM 008 and other two Bacillus sp. (VI 009 and IX 049) strains, significantly reduced the total, pre- and post-emergency mortality of seedlings, but no isolate reduced the incidence of M. phaseolina in seedlings. In the second trial, Bacillus sp. IX 049, VI 099, B. subtilis (IX 007) and a non-identified isolate V 005, decreased the incidence of charcoal root rot. It is concluded that the best of these bacterial antagonists have the potential to control M. phaseolina in P. radiata nurseries.     

Nitric Oxide Production by Necrotrophic Pathogen Macrophomina phaseolina and the Host Plant in Charcoal Rot Disease of Jute: Complexity of the Interplay between Necrotroph–Host Plant Interactions

M. phaseolina, a global devastating necrotrophic fungal pathogen causes charcoal rot disease in more than 500 host plants. With the aim of understanding the plant-necrotrophic pathogen interaction associated with charcoal rot disease of jute, biochemical approach was attempted to study cellular nitric oxide production under diseased condition. This is the first report on M. phaseolina infection in Corchorus capsularis (jute) plants which resulted in elevated nitric oxide, reactive nitrogen species and S nitrosothiols production in infected tissues. Time dependent nitric oxide production was also assessed with 4-Amino-5-Methylamino-2′,7′-Difluorofluorescein Diacetate using single leaf experiment both in presence of M. phaseolina and xylanases obtained from fungal secretome. Cellular redox status and redox active enzymes were also assessed during plant fungal interaction. Interestingly, M. phaseolina was found to produce nitric oxide which was detected in vitro inside the mycelium and in the surrounding medium. Addition of mammalian nitric oxide synthase inhibitor could block the nitric oxide production in M. phaseolina. Bioinformatics analysis revealed nitric oxide synthase like sequence with conserved amino acid sequences in M. phaseolina genome sequence. In conclusion, the production of nitric oxide and reactive nitrogen species may have important physiological significance in necrotrophic host pathogen interaction.     

Suppression of Charcoal Rot of Chickpea by Fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> Under Saline Stress Condition

The ability of fluorescent Pseudomonas strain EKi, in production of biocontrol and plant growth promotory (PGP) metabolites under saline stress was evaluated. Strain EKi could tolerate NaCl up to 1,550 mM and showed biocontrol of Macrophomina phaseolina (76.19%) in the presence of up to 400 mM NaCl. Strain EKi was able to produce IAA, siderophore and pyocyanin with gradual reduction of up to 76.31, 45.46, and 48.99%, respectively, as NaCl concentration increased from 0 to 500 mM. Reduced growth rate resulted in delayed induction of IAA, siderophore and pyocyanin by the PGPR. Thin layer chromatography of chloroform extract from non-stressed and salt stressed EKi, and inhibition of M. phaseolina by purified pyocyanin clearly indicated its role in biocontrol. In vitro and in vivo results showed the growth promotion and charcoal rot disease suppression of chickpea by strain EKi under both non-stressed and saline stress. There was 76.75 and 65.25% reduction of disease incidence in non-saline and saline conditions, respectively, in vitro conditions. In presence of M. phaseolina strain EKi brought about 67.65 and 58.45% reduction of disease incidence in non-saline and saline soil, respectively.     

Evaluation of seed dressing and soil application formulations of Trichoderma species for integrated management of dry root rot of chickpea

The efficacy of the newly developed seed dressing and soil application formulations of Trichoderma viride, T. virens and T. harzianum were evaluated individually and in combinations under pot and field experiments for the management of dry root rot (Rhizoctonia bataticola) of chickpea (Cicer arientinum). In pot experiments, T. harzianum based seed dressing formulation, Pusa 5SD, and soil application formulations, Pusa Biogranule 5 (PBG 5) and Pusa Biopellet 10G (PBP 10G), were found to be effective in reducing dry root rot incidence in chickpea and increasing the seed germination, shoot and root lengths of the crop. Under field experiments, a combination of soil application of T. harzianum based PBP 10G and seed treatment with Pusa 5SD+carboxin was found to be the best by providing the highest seed germination, shoot and root lengths and grain yield and the lowest dry root rot incidence in chickpea.     

Biocontrol of chickpea root rot using endophytic actinobacteria

Eleven actinobacterial strains were isolated from different plants, lentil (Lens esculentus), chickpea (Cicer arietinum L.), pea (Pisum sativum), faba bean (Vicia faba) and wheat (Triticum vulgare) from Paskerville, South Australia. Isolates were characterized and identified morphologically as well as using 16S ribosomal RNA gene sequencing. Of the actinobacteria tested, 72% produced siderophores, 33% were positive for cyanogens production, and 11% showed phosphate solubility. All isolates had antimicrobial activity against Phytophthora medicaginis, Pythium irregulare and Botrytis cinerea. In a greenhouse experiment, actinobacteria with the highest biocontrol capabilities were tested for their ability to control Phytophthora root rot on chickpea. Both Streptomyces sp. BSA25 and WRA1 successfully suppressed Phytophthora root rot when coinoculated with either Mesorhizobium ciceri WSM1666 or Kaiuroo 3. Streptomyces sp. BSA25 with either rhizobial strain enhanced vegetative growth of root (7–11 fold) and shoot dry weights (2–3 fold) compared to infected control, whereas Streptomyces sp. WRA1 increased root and shoot dry weights by 8- and 4-fold, respectively when inoculated with M. ciceri WSM1666. We suggest that careful selection of actinobacteria should be considered when coinoculated with beneficial microorganisms as plant symbionts.     

Temperature response,pathogenicity, seed infection and mutant evaluation against Macrophomina phaseolina causing charcoal rot disease of sesame

Charcoal rot caused by Macrophomina phaseolina is a serious disease of sesame in Pakistan. M. phaseolina sesame isolate was subjected to growth rate test at 10, 15, 20, 25, 30, 35 and 40°C. The optimum temperature for fungal growth and microsclerotia production was found to be 30–35°C. Gray to black, radial fungal colonies with intermediate mycelial growth and jet black oval to round microsclerotia were observed at this optimum range. M. phaseolina was found to be pathogenic against all the 18 tested plant species and this pathogenicity proved its necrophytic behavior. Seed infection efficiency of M. phaseolina was 100% with significant reduction in seed index. For two consecutive years 21 mutants/varieties were screened in the field for their reactions to charcoal rot disease. During 2007 three mutants NS11704S1, NS11304S2 and NS26004 were ranked as resistant while others were moderately resistant to highly susceptible. During 2008 all mutants showed a susceptible to highly susceptible reaction with variable disease reactions. All over screening results revealed that four mutants viz, NS13P1, NS163-1, NS270P1 and NS26004 showed about 50% stand with consistent performance during both years under optimum disease conditions and can be used to manage the disease following the disease management strategies, however in the future improvement for high seed yield along with resistance is a prerequisite for sustainable high production.     

Effect of Indole-3-Acetic Acid (IAA) Produced by Pseudomonas aeruginosa in Suppression of Charcoal Rot Disease of Chickpea

The production of indole-3-acetic acid (IAA), by rhizobacteria, has been associated with plant growth promotion, especially root initiation and elongation. Isolate TO3 selected from 103 fluorescent pseudomonads, identified as Pseudomonas aeruginosa, showed maximum production of IAA. Isolate TO3 having biocontrol activity against Macrophomina phaseolina also showed production of siderophore and HCN was used to screen the role of bacterial IAA in reducing the level of charcoal rot disease occurrence in chickpea. Four IAA defective stable mutants of isolate TO3 having biocontrol activity against M. phaseolina were developed through 5-bromouracil mutagenesis. Mutant TO₅₂ showed 76.47% reduction in production of IAA. Standard IAA was used in similar concentration as present in cell-free culture supernatant of wild isolate TO3 and its mutant TO₅₂. The in vitro and in vivo study showed that IAA-defective mutant TO₅₂ caused reduced biocontrol and plant growth promotory activity than wild isolate TO3. Standard IAA showed comparable biocontrol activity to the culture supernatant. To some extent better biocontrol and growth promotory activity in supernatant than standard IAA indicates the synergistic role of siderophore and HCN. The study clearly reports the role of bacterial IAA in suppression of charcoal rot disease of chickpea.     

Occurrence of Charcoal Rot Caused by Macrophomina phaseolina,an Emerging Disease of Adzuki Bean in China

Adzuki bean (Vigna angularis) is an important legume crop in China. Soil‐borne charcoal rot caused by Macrophomina phaseolina (Tassi) Goid is an important and devastating disease of many crops including legumes. During late August and early September, 2014, symptoms similar to charcoal rot were observed on adzuki bean plants in Yulin City of Shanxi Province, and Fangshan County of Beijing, China. This study was conducted to determine the causal agent of the emerging disease on adzuki bean. Four fungal isolates were obtained and identified as M. phaseolina based on morphological and molecular characteristics, including species‐specific primer detection and sequences of internal transcribed spacer (ITS) of nuclear ribosomal DNA. The resulting sequences showed 99% identity with more than 60 M. phaseolina strains from diverse hosts. The virulence on adzuki bean was verified using pathogenicity tests, producing symptoms similar to those observed in the fields. To our knowledge, this is the first report of M. phaseolina causing charcoal rot on adzuki bean.     

Evaluation of bacteria isolated from rice rhizosphere for biological control of charcoal rot of sorghum caused by <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis> (Tassi) Goid.

A total of 360 bacteria, isolated from the rhizospheres of a system of rice intensification (SRI) fields, were characterized for the production of siderophore, fluorescence, indole acetic acid (IAA), hydrocyanic acid (HCN) and solubilization of phosphorus. Of them, seven most promising isolates (SRI-156, -158, -178, -211, -229, -305 and -360) were screened for their antagonistic potential against Macrophomina phaseolina (causes charcoal rot in sorghum) by dual culture assay, blotter paper assay and in greenhouse. All the seven isolates inhibited M. phaseolina in dual culture assay, whereas six isolates solubilized phosphorous (except SRI-360), all seven produced siderophore, four produced fluorescence (except SRI-178, -229 and -305), six produced IAA (except SRI-305) and five produced HCN (except SRI-158 and -305). In the blotter paper assay, no charcoal rot infection was observed in SRI-156-treated sorghum roots, indicating complete inhibition of the pathogen, while the roots treated with the other isolates showed 49–76% lesser charcoal rot infection compared to the control. In the antifungal activity test (in green house on sorghum), all the isolates increased shoot dry mass by 15–23% and root dry mass by 15–20% (except SRI-158 and -360), over the control. In order to confirm the plant growth-promoting (PGP) traits of the isolates, the green house experiment was repeated but, in the absence of M. phaseolina. The results further confirmed the PGP traits of the isolates as evidenced by increases in shoot and root dry mass, 22–100% and 5–20%, respectively, over the control. The sequences of 16S rDNA gene of the isolates SRI-156, -158, -178, -211, -229, -305 and -360 were matched with Pseudomonas plecoglossicida, Brevibacterium antiquum, Bacillus altitudinis, Enterobacter ludwigii, E. ludwigii, Acinetobacter tandoii and P. monteilii, respectively in BLAST analysis. This study indicates that the selected bacterial isolates have the potential for PGP and control of charcoal rot disease in sorghum.     

Plant growth-promoting traits of Streptomyces with biocontrol potential isolated from herbal vermicompost

Three strains of Streptomyces (CAI-21, CAI-26 and MMA-32) were earlier reported by us as having potential for biocontrol of charcoal rot of sorghum, caused by Macrophomina phaseolina (Tassi) Goid., and plant growth promotion (PGP) of the plant. In the present investigation, the three Streptomyces were characterised for their physiological traits (tolerance of salinity, temperature, pH and resistance to antibiotics) and further evaluated in the field for their PGP of rice, grown by a system of rice intensification methods. All three Streptomyces were able to grow in NaCl concentrations of up to 12% (except MMA-32), at pH values between 5 and 13 and temperatures between 20 and 40°C. They were highly resistant to ampicillin and trimethoprim (>800 ppm), sensitive to chloramphenicol, kanamycin and nalidixic acid (50–100 ppm) and highly sensitive to streptomycin and tetracycline (5–25 ppm). When evaluated for their PGP activity on seedlings of rice, % germination and shoot and root lengths were significantly enhanced over the control. In the field, the Streptomyces strains significantly enhanced the panicle length, filled grain numbers and weight, panicle weight, 1000 seed weight, tiller numbers, total dry matter, root length (39–65%), root volume (13–30%), root dry weight (16–24%), grain yield (9–11%) and stover yield (11–22%) over the control. In the rhizosphere soil (0–15 cm from root) at harvest, the population of actinomycetes was significantly enhanced as was microbial biomass carbon (27–83%) and nitrogen (24–43%), dehydrogenase activity (34–152%), available P (13–34%) and N (30–53%) and % organic carbon (26–28%). This study further confirms that the selected Streptomyces have PGP properties.     

Multifunctional Exopolysaccharides from Pseudomonas aeruginosa PF23 Involved in Plant Growth Stimulation,Biocontrol and Stress Amelioration in Sunflower Under Saline Conditions

Isolate PF23 selected from among 110 fluorescent pseudomonads, identified as Pseudomonas aeruginosa, displayed salinity tolerance and exopolysaccharides (EPS) production up to 2,000 mM NaCl concentration. EPS-defective mutant PF23^EPS? of the isolate showed 86 % reduction in EPS production in comparison with wild strain. Defect in EPS production brought loss in salt tolerance capability. Purified EPS obtained from PF23 displayed multiple roles. At low concentration EPS functioned as biocontrol agent, at high concentration EPS behaved as osmoprotective or stress ameliorating metabolite and when introduced in saline soil, served as a plant growth promotor along with seed biopriming agent. Both in planta and in vivo studies were performed taking sunflower as a test crop and it was observed that PF23 showed plant growth promotion and significant biocontrol potential against dreadful phytopathogen Macrophomina phaseolina (under saline conditions). The mutant PF23^EPS? was ineffective under saline conditions both in growth enhancement as well as in disease suppression. The study reports a potent strain, Pseudomonas aeruginosa PF23, capable of enhancing production of sunflower crop in semiarid regions and minimizing the incidence of charcoal rot disease in sunflower.     

Isolation and identification of cultivated bacteria associated with soybeans and their biocontrol activity against <Emphasis Type="Italic">Phytophthora sojae</Emphasis>

One hundred bacteria, isolated from rhizospheric soil and rhizoplane of healthy soybean plants, were assayed for antifungal activity against six Phytophthora sojae isolates. Nine of the tested bacteria inhibited the hyphal growth of P. sojae in vitro. They were subsequently evaluated for their in vitro traits and identified using the 16S rRNA gene sequences. Four of them (Paenibacillus sp.,—S1; Streptomyces sp.,—S9, S10 and S11) were further selected on the basis of their strongest antagonistic activity in vitro against P. sojae race 4, the predominant race in most growing soybean areas in Canada, and tested for their beneficial effects on soybean plants in the greenhouse. Results showed that application of bacterial strain S11 as seed coating reduced the disease severity by 57.1% and increased the root and shoot weight by and 140 and 108% respectively, in comparison to the diseased control. Overall, a positive correlation was recorded between the in vitro and in planta effects of the selected bacteria. This is promising for further application as select environmentally safe biological control agents in the protection of soybean against root rot diseases.     

UVC-priming mediated modulation of forskolin biosynthesis key genes against Macrophomina root rot of Coleus forskohlii⿿A tissue culture based sustainable approach

Coleus forskohlii is susceptible to root rot disease that reduces yield of root specific metabolite forskolin. In this communication, we first time reported sustainable management of Macrophomina phaseolina root rot in C. forskohlii via Ultraviolet-C (UVC)-hormesis using tissue culture. Compact calli were developed using 2,4-dichlorophenoxy acetic acid (2.0 mg l^⿿1) and tolerance level of UVC exposure was optimized. Calli exhibited 88% viability, 8.4% relative growth in biomass and higher absorbance in 2,3,5-triphenyl tetrazolium chloride analysis subsequent to 10 min of UVC exposure. Pathogenicity of M. phaseolina was reduced in UVC-primed calli-regenerated plants compared with un-primed ones under controlled condition. Plant height, leaf number, shoot and root biomass significantly improved in UVC-primed calli-regenerated plants over un-primed ones when challenged by M. phaseolina in glasshouse. Significant elevation of economically important forskolin via up-regulation of key forskolin biosynthesis genes GGPP synthase, CfTPS2 and CfTPS3 highlighted the potentiality of UVC-priming to boost plant⿿s defence mechanism and protect from drastic economic loss.     

Detection,identification and morphological characteristic of Macrophomina phaseolina: the charcoal rot disease pathogens isolated from infected plants in Northern Jordan

This is the first report about charcoal rot disease in Jordan. Twenty-five Macrophomina phaseolina isolates were collected from infected plants showing typical symptoms of charcoal rot disease. All of the 25 M. phaseolina isolates were pathogenic to cucumber plants under green house effect. The amplification of the isolated DNA from the 25 pathogenic fungal cultures using ITS specific primers (ITS 1?+?ITS 4) showed a single band of 580?bp. There was a significant variation of their mycelial linear growth rate on PDA medium. The 25 M. phaseolina isolates showed a wide heterogeneity in their mycelium colour, microsclerotia distribution, pycnidia formation and chlorate phenotypes. Based on the morphological characterisation, the 25 isolates were grouped into seven different groups as indicated in a dendrogram of their morphological variation. The overall means similarity matrix of the 25 M. phaseolina recovered isolates were 0.58. The means of similarity matrix of the 25 M. phaseolina was in between 0.83 and 0.14. The similarity coefficient between the 25 isolates varies between 0.27 and 1.0.     

Dry root rot (Rhizoctonia bataticola (Taub.) Butler): an emerging disease of chickpea – where do we stand?

Dry root rot caused by Rhizoctonia bataticola (Macrophomina phaseolina) of chickpea (Cicer arietinum L.) is gaining importance in the changed scenario of climate when growing crop is predisposed to high temperature and moisture stress. Being mainly a soil-inhabiting pathogen, many environmental and soil factors are responsible for the development of disease. No systematic research related to the biology, ecology and epidemiology of dry root rot in chickpea has been conducted so far. Research is needed to improve the identification and characterisation of variability within its epidemiological and pathological niches. Limited literature available on host plant resistance for dry root rot indicated lack of resistant sources for this disease. The present article discusses current status of the disease in the context of climate change and possible management options to alleviate the problem.     

seed rot and root rot complex of beans (Phaseolus vulgaris L.)

Fusarium solani (76%),Phyllosticta phaseolina (12%) andAspergillus flavus (10%) were isolated from bean(Phaseolus vulgaris) seeds showing lesions. Out of these the first two caused 30 per cent and 20 per cent seed rotting respectively after artificial seed inoculations. When inoculum was added into the soil before sowing the seed, 35 per cent and 10 per cent pre-emergence rot ocurred and there was 35 per cent and 15 per cent post-emergence blight when soil with healthy seedlings was inoculated withF. solani andP. phaseolina respectively.F. solani caused similar lesions on bean seeds when inoculated after an injury.F. oxysporum was isolated from the roots of dried bean plants showing necrosis.     

Endophytic actinomycetes from spontaneous plants of Algerian Sahara: indole-3-acetic acid production and tomato plants growth promoting activity

Twenty-seven endophytic actinomycete strains were isolated from five spontaneous plants well adapted to the poor sandy soil and arid climatic conditions of the Algerian Sahara. Morphological and chemotaxonomical analysis indicated that twenty-two isolates belonged to the Streptomyces genus and the remaining five were non-Streptomyces. All endophytic strains were screened for their ability to produce indole-3-acetic acid (IAA) in vitro on a chemically defined medium. Eighteen strains were able to produce IAA and the maximum production occurred with the Streptomyces sp. PT2 strain. The IAA produced was further extracted, partially purified and confirmed by thin layer chromatography (TLC) analysis. The 16S rDNA sequence analysis and phylogenetic studies indicated that strain PT2 was closely related to Streptomyces enissocaecilis NRRL B 16365^T, Streptomyces rochei NBRC 12908^T and Streptomyces plicatus NBRC 13071^T, with 99.52 % similarity. The production of IAA was affected by cultural conditions such as temperature, pH, incubation period and l-tryptophan concentration. The highest level of IAA production (127 μg/ml) was obtained by cultivating the Streptomyces sp. PT2 strain in yeast extract-tryptone broth supplemented with 5 mg l-tryptophan/ml at pH 7 and incubated on a rotary shaker (200 rpm) at 30 °C for 5 days. Twenty-four-hour treatment of tomato cv. Marmande seeds with the supernatant culture of Streptomyces sp. PT2 that contained the crude IAA showed the maximum effect in promoting seed germination and root elongation.