Effect of Iranian strains of Pseudomonas spp. on the control of root-knot nematodes on Pistachios

The role of some Iranian strains of Pseudomonas spp. as biocontrol agents against Meloidogyne incognita and their ability to colonise pistachio roots was investigated. The results of in vitro experiments indicated that all tested bacteria produced significant suppression of M. incognita and showed that all strains were able to kill M. incognita juveniles with strain VUPf428 achieving about 99% mortality at 72 h. The results of in vivo treatments indicated that the best strains that could build high populations in soil infested with nematodes were VUPf5, VUPf52 and VUPf205. These isolates also caused highest reduction in galling and nematode multiplication in a greenhouse test although all strains native to Iran could colonise pistachio roots in pots. Some strains could produce secondary metabolites such as siderophores, proteases and volatile metabolites at high population levels.     

Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24

Pseudomonas fluorescens 2P24 is a biocontrol agent isolated from a wheat take-all decline soil in China. This strain produces several antifungal compounds, such as 2,4-diacetylphloroglucinol (2,4-DAPG), hydrogen cyanide and siderophore(s). Our recent work revealed that strain 2P24 employs a quorum-sensing system to regulate its biocontrol activity. In this study, we identified a quorum-sensing system consisting of PcoR and PcoI of the LuxR–LuxI family from strain 2P24. Deletion of pcoI from 2P24 abolishes the production of the quorum-sensing signals, but does not detectably affect the production of antifungal metabolites. However, the mutant is significantly defective in biofilm formation, colonization on wheat rhizosphere and biocontrol ability against wheat take-all, whilst complementation of pcoI restores the biocontrol activity to the wild-type level. Our data indicate that quorum sensing is involved in regulation of biocontrol activity in P. fluorescens 2P24.     

Gluconic acid: an antifungal agent produced by Pseudomonas species in biological control of take-all

Pseudomonas strain AN5 (Ps. str. AN5), a non-fluorescent Australian bacterial isolate, is an effective biological control (biocontrol) agent of the take-all disease of wheat caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Ps. str. AN5 controls Ggt by producing an antifungal compound which was purified by thin layer and column chromatography, and identified by NMR and mass spectroscopic analysis to be d-gluconic acid. Commercially bought pure gluconic acid strongly inhibited Ggt. Two different transposon mutants of Ps. str. AN5 which had lost take-all biocontrol did not produce d-gluconic acid. Gluconic acid production was restored, along with take-all biocontrol, when one of these transposon mutants was complemented with the corresponding open reading frame from wild-type genomic DNA. Gluconic acid was detected in the rhizosphere of wheat roots treated with the wild-type Ps. str. AN5, but not in untreated wheat or wheat treated with a transposon mutant strain which had lost biocontrol. The antifungal compounds phenazine-1-carboxylic acid and 2,4-diacetylphloroglucinol, produced by other Pseudomonads and previously shown to be effective in suppressing the take-all disease, were not detected in Ps. str. AN5 extracts. These results suggest that d-gluconic acid is the most significant antifungal agent produced by Ps. str. AN5 in biocontrol of take-all on wheat roots.     

Variability among strains of Trichoderma harzianum in their ability to reduce take-all and to produce pyrones

Antagonism tests on agar-plates and glasshouse screening indicated that three isolates of Trichoderma harzianum varied in their ability to antagonize the take-all fungus (Gaeumannomyces graminis var. tritici). Isolate 71 which was the most effective in suppressing take-all of wheat, produced two pyrones and other undetermined analogues. Isolates of T. koningii and T. hamatum shown to suppress take-all, produced a simple pyrone compound. Although T. harzianum isolates 70 and 73 did not produce any pyrones, they reduced the disease albeit to a much lesser extent than isolate 71; with isolate 73 showing distinct host growth promotion effects. It is proposed that the success of isolate 71 of T. harzianum was related to the pyrones it produces and that the ability of isolates 70 and 73 to reduce take-all may be related to mechanisms other than those involving antibiotics.     

Biocontrol of Rhizoctonia crown and root rot of sugar beet by binucleate Rhizoctonia spp. and Laetisaria arvalis

Two isolates of Laetisaria arvalis and 10 of binucleate Rhizoctonia spp. (BNR) from the Ohio sugar beet production area, were tested in the greenhouse and field for biocontrol of Rhizoctonia crown and root rot of sugar beet, caused by Rhizoctonia solani anastomosis group 2, type 2. L. arvalis was ineffective in standard greenhouse tests, and the single isolate used in the field was generally ineffective. Seven of 10 BNR isolates effectively controlled crown and root rot in greenhouse tests. Delayed application of biocontrol agents to plants 5 – 10 wk old was generally more effective than applications made at planting. A BNR isolate significantly reduced % plant loss and disease ratings and increased yield in a 1985 field test as compared with the control infested with R. solani alone. Two BNR isolates were effective in a 1986 field test and increased yields c. 22% in comparison to a L. arvalis treatment, which did not differ from the R. solani-infested control. The Ohio binucleate Rhizoctonia isolates appear to have considerable potential as applied biocontrol agents and may play a role in the natural ecology of R. solani in the sugar beet production area of Ohio.     

Control of Mn status and infection rate by genotype of both host and pathogen in the wheat take-all interaction

Take-all is a world-wide root-rotting disease of cereals. The causal organism of take-all of wheat is the soil-borne fungus Gaeumannomyces graminis var tritici (Ggt). No resistance to take-all, worthy of inclusion in a plant breeding programme, has been discovered in wheat but the severity of take-all is increased in host plants whose tissues are deficient for manganese (Mn). Take-all of wheat will be decreased by all techniques which lift Mn concentrations in shoots and roots of Mn-deficient hosts to adequate levels. Wheat seedlings were grown in a Mn-deficient calcareous sand in small pots and inoculated with four field isolates of Ggt. Infection by three virulent isolates was increased under conditions which were Mn deficient for the wheat host but infection by a weakly virulent isolate, already low, was further decreased. Only the three virulent isolates caused visible oxidation of Mn in vitro. The sensitivity of Ggt isolates to manganous ions in vitro did not explain the extent of infection they caused on wheat hosts. In a similar experiment four Australian wheat genotypes were grown in the same Mn-deficient calcareous sand and inoculated with one virulent isolate of Ggt. Two genotypes were inefficient at taking up manganese and were very susceptible to take-all, one was very efficient at taking up manganese and was resistant to take-all, and the fourth genotype was intermediate for both characters. All genotypes were equally resistant under Mn-adequate conditions.     

Pseudomonas protegens CS1 from the lemon phyllosphere as a candidate for citrus canker biocontrol agent

• Citrus canker is a worldwide‐distributed disease caused by Xanthomonas citri subsp. citri. One of the most used strategies to control the disease is centred on copper‐based compounds that cause environmental problems. Therefore, it is of interest to develop new strategies to manage the disease. Previously, we reported the ability of the siderophore pyochelin, produced by the opportunistic human pathogen Pseudomonas aeruginosa, to inhibit in vitro several bacterial species, including X. citri subsp. citri. The action mechanism, addressed with the model bacterium Escherichia coli, was connected to the generation of reactive oxygen species (ROS). This work aimed to find a non‐pathogenic strain from the lemon phyllosphere that would produce pyochelin and therefore serve in canker biocontrol.
• An isolate that retained its capacity to colonise the lemon phyllosphere and inhibit X. citri subsp. citri was selected and characterised as Pseudomonas protegens CS1. From a liquid culture of this strain, the active compound was purified and identified as the pyochelin enantiomer, enantio‐pyochelin.
• Using the producing strain and the pure compound, both in vitro and in vivo, we determined that the action mechanism of X. citri subsp. citri inhibition also involved the generation of ROS. Finally, the potential application of P. protegens CS1 was evaluated by spraying the bacterium in a model that mimics the natural X. citri subsp. citri infection.
• The ability of P. protegens CS1 to reduce canker formation makes this strain an interesting candidate as a biocontrol agent.

     

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.     

利用纤维二糖差向异构酶制备乳果糖的研究进展

乳果糖是由D-半乳糖和D-果糖两个基团通过β-1,4糖苷键连接而成的还原型二糖;乳果糖口服液具有治疗慢性便秘和肝性脑病的功效,在100多个国家作为常见非处方药(OTC)使用,需求量十分巨大;乳果糖还可以作为益生元改善人体肠道菌群关系。乳果糖的生产依赖化学法,其催化剂对人体有害,下游分离难度大。近年来,纤维二糖差向异构酶被发现能够高效催化乳糖制备乳果糖,该技术绿色环保、步骤简单,具有很强的产业化前景。本文结合自身研究经历对纤维二糖差向异构酶的研发情况进行总结,并综述了乳果糖酶法制备技术的现状。     

Isolation screening and characterisation of local beneficial rhizobacteria based upon their ability to suppress the growth of Fusarium oxysporum f. sp. radicis-lycopersici and tomato foot and root rot

Local beneficial rhizobacteria were selected based upon their ability to control the fungus Fusarium oxysporum f. sp. radicis-lycopersici which causes crown and root rot of tomato. Seven out of 384 strains prevailed in multiple and dual cultures and were identified as Pseudomonas chlororaphis (one strain), Bacillus cereus (one strain), Serratia marcescens (three strains) and Serratia rubidaea (two strains), by sequencing the 16S rRNA or the 16S and 23S rRNA inter-spacer region. The seven selected rhizobacteria were tested for their biocontrol and growth-promoting effects in planta, and their antifungal properties in vitro. All strains significantly reduced disease severity under controlled conditions, in a gnotobiotic system and in pots. Moreover, one P. chlororaphis and one S. marcescens strain significantly decreased disease severity to the level of the healthy control under natural conditions in pots experiments. The inhibitory activity of bacterial liquid cultures' metabolites on the fungus was demonstrated for all strains in vitro, using filter paper, thin layer chromatography and microtiter bioassays. Genes encoding phenazines were tentatively detected by PCR in the P. chlororaphis strain and chitinase-encoding genes were detected in one S. rubidaea and all three S. marcescens strains. Production of phenazine-1-carboxamide and hydrogen cyanide was evidenced for the P. chlororaphis strain while protease activity and production of siderophore-like compounds was confirmed in all bacterial strains. Possible use of these strains as biological control agents and their impact on natural biocontrol of pathogens in soils is discussed.     

Relationship of in vitro and in planta screening: improving the selection process for biological control agents against Fusarium root rot in row crops

Fusarium root rot in row crops is typically managed by cultural practices and fungicide seed treatments. Biological control using microbial agents is another option but needs further development for improved disease management. Screening to identify biocontrol agents are crucial. However, relationships among the steps and how to improve the screening process are unresolved questions. Strains of Burkholderia (4), Bacillus (5) and Trichoderma (26) were studied in vitro against six Fusarium pathogens. All the bacteria and five selected Trichoderma strains were tested in planta in the greenhouse against diseases of Fusarium graminearum and Fusarium oxysporum. Burkholderia ambifaria C628, Bacillus simplex R180, and all Trichoderma isolates showed high reduction in disease levels in corn, soybean and wheat, ranging from 16 to 63%. Responses of the biocontrol agents during in vitro and in planta screening did not always correlate. In vitro and in planta tests should be considered independently in selecting biocontrol candidates.     

Plant Growth‐Promoting Bacteria from Solarized Soil with the Ability to Protect Melon Against Root Rot and Vine Decline Caused by Monosporascus cannonballus

This study was undertaken to isolate indigenous plant growth‐promoting (PGP) bacteria from solarized soil effective in the biocontrol of Monosporascus cannonballus, the cause of root rot and vine decline of melon, which is one of the most destructive soilborne diseases of this crop worldwide. The screening strategy resulted in the selection of two interesting PGP bacteria as biocontrol candidates against M. cannonballus belonging to the same microbial community. The two bacterial species, identified according to phenotypic, physiological tests and analysis of the 16S rDNA sequence as Bacillus subtilis/amyloliquefaciens (BsCR) and Pseudomonas putida (PpF4), showed PGP traits and in vitro antagonistic activity towards M. cannonballus. Antagonism by BsCR was characterized by a consistent inhibition of the pathogen in vitro growth; PpF4 strongly inhibited the development of perithecia of the pathogen. Under greenhouse conditions, the selected bacteria were tested for their biocontrol activity in the pathosystem melon‐M. cannonballus. BsCR alone and in combination with PpF4 determined a consistent decrease in the disease symptoms. BsCR and the combination of the bacterial strains significantly increased root biomass in both inoculated and un‐inoculated plant. Upon seed treatment with BsCR, the accumulation and isoenzyme induction of peroxidase in roots as biochemical marker for induction of resistance were found, thus indicating that BsCR may reduce the disease severity also by the activation of the plant defence responses. The study highlights the synergistic biocontrol potential of B. subtilis BsCR and P. putida PpF4 in the integrated management of root rot and vine decline of melon caused by M. cannonballus.     

Evaluation of endophytic actinobacteria as antagonists of Gaeumannomyces graminis var. tritici in wheat

Endophytic actinobacteria isolated from healthy cereal plants were assessed for their ability to control fungal root pathogens of cereal crops both in vitro and in planta. Thirty eight strains belonging to the genera Streptomyces, Microbispora, Micromonospora, and Nocardioidies were assayed for their ability to produce antifungal compounds in vitro against Gaeumannomyces graminis var. tritici (Ggt), the causal agent of take-all disease in wheat, Rhizoctonia solani and Pythium spp. Spores of these strains were applied as coatings to wheat seed, with five replicates (25 plants), and assayed for the control of take-all disease in planta in steamed soil. The biocontrol activity of the 17 most active actinobacterial strains was tested further in a field soil naturally infested with take-all and Rhizoctonia. Sixty-four percent of this group of microorganisms exhibited antifungal activity in vitro, which is not unexpected as actinobacteria are recognized as prolific producers of bioactive secondary metabolites. Seventeen of the actinobacteria displayed statistically significant activity in planta against Ggt in the steamed soil bioassay. The active endophytes included a number of Streptomyces, as well as Microbispora and Nocardioides spp. and were also able to control the development of disease symptoms in treated plants exposed to Ggt and Rhizoctonia in the field soil. The results of this study indicate that endophytic actinobacteria may provide an advantage as biological control agents for use in the field, where others have failed, due to their ability to colonize the internal tissues of the host plant.     

Genomic-assisted characterisation of Pseudomonas sp. strain Pf4, a potential biocontrol agent in hydroponics

In an attempt to select potential biocontrol agents against Pythium spp. and Rhizoctonia spp. root pathogens for use in soilless systems, 12 promising bacteria were selected for further investigations. Sequence analysis of the 16S rRNA gene revealed that three strains belonged to the genus Enterobacter, whereas nine strains belonged to the genus Pseudomonas. In in vitro assays, one strain of Pseudomonas sp., Pf4, closely related to Pseudomonas protegens (formerly Pseudomonas fluorescens), showed noteworthy antagonistic activity against two strains of Pythium aphanidermatum and two strains of Rhizoctonia solani AG 1-IB, with average inhibition of mycelial growth >80%. Strain Pf4 was used for in vivo treatments on lamb’s lettuce against R. solani root rot in small-scale hydroponics. Pf4-treated and untreated plants were daily monitored for symptom development and after two weeks of infection, a significant protective effect of Pf4 against root rot was recorded. The survival and population density of Pf4 on roots were also checked, demonstrating a density above the threshold value of 10⁵?CFU?g^?1 of root required for disease suppression. Known loci for the synthesis of antifungal metabolites, detected using PCR, and draft-genome sequencing of Pf4 demonstrated that Pseudomonas sp. Pf4 has the potential to produce an arsenal of secondary metabolites (plt, phl, ofa and fit-rzx gene clusters) very similar to that of the well-known biocontrol P. protegens strain Pf-5.     

Biological control of take-all in wheat by endophytic Bacillus subtilis E1R-j and potential mode of action

The bacterial strain E1R-j, isolated as an endophyte from wheat roots, exhibited high antifungal activity to Gaeumannomyces graminis var. tritici (Ggt). Strain E1R-j was identified as Bacillus subtilis based on morphological, physiological and biochemical methods as well as on 16S rDNA analysis. This strain inhibited mycelium growth in vitro of numerous plant pathogenic fungi, especially of Ggt, Coniothyrium diplodiella, Phomopsis sp. and Sclerotinia sclerotiorum. In greenhouse experiments, soil drenches with cell densities of 10⁶, 10⁹ and 10¹² CFU ml⁻¹ E1R-j reduced significantly take-all disease, caused by Ggt, in wheat seedling by 62.6%, 68.6% and 70.7%, respectively, compared to the inoculated control, 4 weeks after sowing. Growth parameters such as lengths and fresh weights of roots and shoots of Ggt-inoculated control plants were significantly lower compared to Ggt-inoculated and E1R-j treated plants. Field experiments in the season 2006/2007, heights of wheat plants in the Ggt inoculated plots were significantly reduced compared to the non inoculated treatments. Yield parameters such as kernels per head and thousand kernel weight (TKW) in inoculated control plants were lower compared to the other treatments. In the experimental year 2007/2008, independent treatments with the bacterial strain E1R-j and the fungicide Triadimefon reduced take-all disease in wheat roots by 55.3% and 61.9%, compared to the inoculated control plants. In this season plant height in inoculated control was significantly lower and also the yield parameters seeds per head and especially TKW were drastically reduced compared to the other treatments. E1R-j treatment alleviated the detrimental effects of take-all on grain yield parameters to a similar extent as Triadimefon application. SEM studies revealed that in the presence of E1R-j, hyphae of Ggt showed leakage, appeared ruptured, swollen and shriveled. Following root drench, strain E1R-j was able to colonize endophytically roots and leaves of wheat seedlings. While the population of the bacterial strain in wheat roots steadily increased from the second to the fourth leaf stage, in the leaf tissue the population of the strain rapidly declined. TEM studies also showed that cells of E1R-j were present in roots of wheat seedlings and effectively retarded infection and colonization of Ggt in root tissue; suppression of Ggt by E1R-j was accompanied by disintegration of hyphal cytoplasm. In addition, in the presence of E1R-j cells in Ggt-infected root tissue morphological defense reactions were triggered such as formation of wall appositions and papillae. The results presented indicate that the endophytic strain E1R-j of B. subtilis meets demands required for biocontrol of take-all.     

Clonostachys rosea BAFC3874 as a Sclerotinia sclerotiorum antagonist: mechanisms involved and potential as a biocontrol agent

Aims: To establish the modes of action of the antagonistic fungal strain Clonostachys rosea BAFC3874 isolated from suppressive soils against Sclerotinia sclerotiorum and to determine its potential as a biocontrol agent. Methods and Results: The antagonistic activity of C. rosea BAFC3874 was determined in vitro by dual cultures. The strain effectively antagonized S. sclerotiorum in pot‐grown lettuce and soybean plants. Antifungal activity assays of C. rosea BAFC3874 grown in culture established that the strain produced antifungal compounds against S. sclerotiorum associated with secondary metabolism. High mycelial growth inhibition coincided with sclerotia production inhibition. The C. rosea strain produced a microheterogeneous mixture of peptides belonging to the peptaibiotic family. Moreover, mycoparasitism activity was observed in the dual culture. Conclusions: Clonostachys rosea strain BAFC3874 was proved to be an effective antagonist against the aggressive soil‐borne pathogen S. sclerotiorum in greenhouse experiments. The main mechanisms involve peptaibiotic metabolite production and mycoparasitism activity. Significance and Impact of the Study: Clonostachys rosea BAFC3874 may be a good fungal biological control agent against S. sclerotiorum. In addition, we were also able to isolate and identify peptaibols, an unusual family of compounds in this genus of fungi.     

Efficacy assessment of antifungal metabolites from Chaetomium globosum No.05, a new biocontrol agent,against Setosphaeria turcica

Northern corn leaf blight (NCLB), an important and potentially destructive corn foliar disease, is caused by Setosphaeria turcica. The intent of this study was to evaluate antifungal metabolites from Chaetomium globosum (Cg) strain No.05 to suppress NCLB in maize. This strain significantly suppressed mycelial growth of numerous phytopathogenic fungi especially S. turcica on potato dextrose agar medium. The secondary metabolites of the strain inhibited mycelial growth and conidial germination of S. turcica. When co-inoculated at three droplets (5 μL/droplet) of conidial suspension (5 × 10⁴ conidia/mL) on each 8-cm-long detached leaf, 20% culture filtrates completely suppressed disease incidence of northern corn leaf blight. The application of the culture filtrates at 2 h post-inoculation (hpi) of S. turcica in greenhouse studies showed a 81.9% inhibition of NCLB on the seedlings, while culture filtrates applied before pathogen inoculation showed even higher rates of disease reduction. The application of the culture filtrates had no observed effects on the treated maize leaves or seedlings. Two active compounds, isolated from the extracts, were identified as chaetoglobosin A and chaetoglobosin C based on the spectroscopic analysis. Both in vitro and in planta bioassay experiments showed that chaetoglobosin A displayed potent biocontrol efficiency against S. turcica. To the best of our knowledge, this is the first report of the evaluation of the inhibitory effects of C. globosum and chaetoglobosin A against S. turcica both in vitro and on detached maize leaves.     

Potential of endophytic Streptomyces spp. for biocontrol of Fusarium root rot disease and growth promotion of tomato seedlings

Sixteen endophytic actinobacteria isolated from roots of native plants were evaluated for their antagonistic potential against soil-borne phytopathogenic fungi. Among them, three strong antagonistic isolates were selected and characterised for in vitro plant-growth-promoting and biocontrol traits, including production of hydrogen cyanide, indole-3-acetic acid and siderophores, chitinase and β-1,3-glucanase activities, and inorganic phosphate solubilisation. In all trials, the strain Streptomyces sp. SNL2 revealed promising features. The selected actinobacteria were investigated for the biocontrol of Fusarium oxysporum f. sp. radicis lycopersici and for growth promotion of tomato (Solanum lycopersicum L. cv. Aïcha) seedlings in autoclaved and non-autoclaved soils. All seed-bacterisation treatments significantly reduced the root rot incidence compared to a positive control (with infested soil), and the isolate SNL2 exhibiting the highest protective activity. It reduced the disease incidence from 88.5% to 13.2%, whereas chemical seed treatment with Thiram® provided 14.6% disease incidence. Furthermore, isolate SNL2 resulted in significant increases in the dry weight, shoot and root length of seedlings. 16S rDNA sequence analysis showed that isolate SNL2 was related to Streptomyces asterosporus NRRL B-24328^T (99.52% of similarity). Its interesting biocontrol potential and growth enhancement of tomato seedlings open up attractive uses of the strain SNL2 in crop improvement.     

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.     

Enhancement of the biocontrol agent Candida oleophila (strain O) survival and control efficiency under extreme conditions of water activity and relative humidity

The objective of this work is to evaluate the ability of some additive substances in protecting the biocontrol agent Candida oleophila (strain O) against the adverse effects of environmental factors, such as water activity (a_w, 0.93 and 0.98) and relative humidity (75% and 98%). The protection obtained with various protectant substances, skimmed milk (SM), peptone, maltose, sucrose, sorbitol, lactose and polyethylene glycol was assayed under in vitro and in vivo conditions. The yeast cells with the highest level of protecting agents (1%) had higher viability than those with low protectant levels (0.1% and 0.5%). SM, sucrose and sorbitol improved significantly the C. oleophila survival on apple fruit surface by 80.8%, 42.26% and 37.27% and gave a significant protection (from 96% to 100%) against Penicillium expansum under dried conditions. The highest strain O density and efficacy was obtained with SM. Under experimental conditions reflecting practical conditions, SM applied in combination with the strain O resulted in improved biocontrol efficacy by 74.65%. Therefore, SM could be used as material substrate with the best sugar protectants during the formulation process of this antagonistic yeast for eventual pre-harvest application.