Combination of Pseudomonas aeruginosa and Pochonia chlamydosporia for Control of Root-Infecting Fungi in Tomato

A plant growth‐promoting rhizobacterium, Pseudomonas aeruginosa strain IE‐6, and a fungal antagonist, Pochonia chlamydosporia, were tested for their ability to inhibit mycelial growth of root‐infecting fungi under laboratory conditions including Macrophomina phaseolina, Fusarium oxysporum, F. solani and Rhizoctonia solani. Biocontrol effectiveness of the bacterium and the fungus alone or in combination was also determined for the control of root‐infecting fungi under field conditions. In a dual‐culture plate assay, the colonies of P. chlamydosporia and P. aeruginosa met each other and no further growth of either organism occurred. Against M. phaseolina, F. solani and R. solani, an ethyl acetate extract of the culture filtrates of P. aeruginosa inhibited fungal growth greater than the hexane extract, but against F. oxysporum the hexane extract caused greater inhibition of fungal growth. By contrast, against M. phaseolina, F. oxysporum and F. solani, the hexane extract of P. chlamydosporia was more effective in the inhibition of fungal growth than the ethyl acetate fraction. Ethyl acetate extracts of P. aeruginosa at 1.0 mg/ml not only inhibited the radial colony growth of R. solani but also lysed the fungal mycelium. P. aeruginosa produced siderophores and hydrogen cyanide under laboratory conditions. Field experiments conducted in 1997 and repeated in 1998 revealed that Pochonia chlamydosporia and P. aeruginosa significantly suppressed the root‐infecting fungi M. phaseolina, F. oxysporum, F. solani and R. solani and that the combination of the two caused greater inhibition of the fungal pathogens than either alone. Application of P. chlamydosporia and P. aeruginosa as a soil drench also resulted in enhanced growth of tomato plants.     

Cross-pathogenicity of Rhizoctonia solani strains on pasture legumes in pasture-crop rotations

In Australia, in the past, pasture legumes were rotated mainly with cereals, but increasingly these rotations now involve pasture legumes with a wider range of crops, including legumes. This increasing frequency of the leguminous host in the rotation system may be associated with increased root rots in legumes in the current pasture-crop rotations. The primary aim of this study was to see whether the pathogenicity on pasture legumes of strains of Rhizoctonia solani sourced from lupins and cereals (common crops in rotation with pastures) is associated with increased incidence of root rots in pasture legumes in the disease conducive sandy soils of the Mediterranean regions of southern Australia. The second aim was to determine sources of resistance among newly introduced pasture legumes to R. solani strains originating from rotational crops as this would reduce the impact of disease in the pasture phase. Fifteen pasture legume genotypes were assessed for their resistance/susceptibility to five different zymogram groups (ZG) of the root rot pathogen R. solani under glasshouse conditions. Of the R. solani groups tested, ZG1–5 and ZG1–4 (both known to be pathogenic on cereals and legumes) overall, caused the most severe root disease across the genotypes tested, significantly more than ZG6 (known to be pathogenic on legumes), in turn significantly >ZG4 (known to be pathogenic on legumes) which in turn was >ZG11 (known to be pathogenic on legumes including tropical species). Overall, Ornithopus sativus Brot. cvs Cadiz and Margurita, Trifolium michelianum Savi. cvs Paradana and Frontier and T. purpureum Loisel. cv. Electro showed a significant level of resistance to root rot caused by R. solani ZG11 (root disease scores ≤1.2 on a 1–3 scale where 3 = maximum disease severity) while O. sativus cvs Cadiz and Erica showed a significant level of resistance to root rot caused by R. solani ZG4 (scores ≤1.2). O. compressus L. cvs Charano and Frontier, O. sativus cv. Erica, and T. purpureum cv. Electro showed some useful resistance to root rot caused by R. solani ZG6 (scores ≤1.8). This is the first time that cvs Cadiz, Electro, Frontier, Margurita and Paradana have been recognised for their levels of resistance to root rot caused by R. solani ZG11; and similarly for cvs Cadiz and Erica against ZG4; and for cvs Charano, Erica, and Electro against ZG6. These genotypes with resistance may also serve as useful sources of resistance in pasture legume breeding programs and also could potentially be exploited directly into areas where other rotation crops are affected by these R. solani strains. None of the tested genotypes showed useful resistance to R. solani ZG1–4 (scores ≥2.0) or ZG1–5 (scores ≥2.5). This study demonstrates the relative potential of the various R. solani ZG strains, and particularly ZG1–4, ZG1–5, ZG4 and ZG6 to attack legume pastures and pose a significant threat to non-pasture crop species susceptible to these strains grown in rotation with these pasture legumes. Significantly, the cross-pathogenicity of these strains could result in the continuous build-up of inoculum of these strains that may seriously affect the productivity eventually of legumes in all rotations. In particular, when choosing pasture legumes as rotation crops, caution needs to be exercised so that the cultivars deployed are those with the best resistance to the R. solani ZGs most likely to be prevalent at the location.     

Impact of biocontrol agents in combination with Prosopis juliflora (Swartz) DC. in controlling the root-infecting fungi of leguminous crops

Biocontrol agents, viz., Rhizobium meliloti, Pseudomonas aeruginosa, Aspergillus niger and Trichoderma harzianum, are used as seed dressing and soil is amended with Prsosopis juliflora (Swartz) DC. plant parts like stem, leaves and flower at 1% w/w for the control of root-rot fungi. All antagonists suppressed the infection of root-rot fungi viz., Fusarium spp., Rhizoctonia solani and Macrophomina phaseolina whereas the infection of R. solani and M. phaseolina was controlled when cowpea (Vigna unguiculata L.) and mungbean (Vigna radiata L.) seeds were treated with P. aeruginosa and T. harzianum and the soil was amended with P. juliflora leaves’ powder at 1% w/w. However, germination of both the crops was observed in all treatments. Growth parameters like shoot and root length, shoot and root weight, and leaf area significantly increased in all the treatments as compared to the control parameters. P. aeruginosa and T. harzianum in combination with soil amendment with P. juliflora plant parts at 1% w/w were the most effective for the control of root-rot fungi of leguminous plants.     

Induction of resistance in Phaseolus vulgaris L. cv. Lima against some soil-borne fungal pathogens by pre-inoculation with tobacco necrosis virus (TNV) reacting hypersensitivity

Abstract

Tobacco necrosis virus (TNV) was tested to induce systemic acquired resistance (SAR) in Phaseolus vulgaris cv. Lima against three important soil-borne fungal pathogens viz: Rhizoctonia solani, Macrophomina phaseolina and Fusarium oxysporum. Application of TNV as a local infection of seven-day old primary leaves of Phaseolus vulgaris cv. Lima resulted in reduction of the mean disease rating of root-rot and damping-off caused by the tested fungal pathogens. The pre-inoculated plants with TNV showed a significant enhancement in their content of photosynthetic pigments (chlorophyll a, b and carotenoids) compared to those inoculated with fungal pathogens only. The percentage of cell membrane stability and ion leakage of viral-treated plants were significantly increased confirming the healthy cytological status of the treated plants. Results demonstrated that inoculation of the primary leaves of beans with TNV before infection with the fungal pathogens leads to changes in protein patterns and showed differences compared with control and caused the appearance of at least one new protein band compared with only fungal-infected plants. Also, an increase in peroxidase activity emerged in the thickness of the isozymic pattern in addition to the synthesis of new bands which was observed as a result of TNV application before infection with the three fungal pathogens. Induction of the synthesis of a new protein and increasing peroxidase activity in the inoculated plants enhanced the defense system against the target pathogen. The results greatly supported the successful application of TNV in the induction of systemic acquired resistance in P. vulgaris cv. Lima against the fungal pathogens.     

Biocontrol of fungal root rot diseases of crop plants by the use of rhizobia and bradyrhizobia

Twenty-oneRhizobium andBradyrhizobium strains were testedin vitro against the mycelial growth of three pathogenic fungi on solid and liquid media. All tested rhizobia and bradyrhizobia significantly suppressed the growth of the three soil-borne root-infecting fungi (Fusarium solani, Macrophominia phasolina andRhizoctonia solani) either in the absence or presence of iron. This indicates that the siderophore played a minor role in the biocontrol potential ofRhizobium andBradyrhizobium against pathogenic fungi. Pot experiments revealed that the numbers of propagules causing disease after 4 weeks of planting varied with species and host plant. The three most activeRhizobium andBradyrhizobium strains (R. leguminosarum bv.phaseoli TAL 182,B. japonicum TAL 377 andBradyrhizobium sp. (lupin) WPBS 3211 D) tested under greenhouse conditions for their ability to protect one leguminous (soybean) and two non-leguminous (sunflower and okra) seedlings from root rot caused byFusarium solani, Macrophominia phaseolina andRhizoctonia solani provided significant suppression of disease severity compared with nonbacterized control in both leguminous and non-leguminous seedlings.Bradyrhizobium sp. (lupin) WPBS 3211 D provided the lowest degree of resistance against all the tested pathogens with all host plants. *** DIRECT SUPPORT *** A00EN058 00013     

Morphological and biochemical responses of selected germplasm of tobacco to soil inoculation with Pythium aphanidermatum

The effect of inoculation with Pythium aphanidermatum was studied on total phenol (TP), salicylic acid (SA), chlorophylls and carotenoid contents of leaves and plant growth characteristics of five tobacco cultivars, namely RK-10 P3, RK-12 P3, RK-13 P4, RK-18 P8 and RK-26 P3, to assess cultivar response at biochemical and morphological levels. Root rot measured at 0–5 scale was 2.66 on cv. RK-10 P3, followed by 2. 33 on cv. RK-18 P8, 1.33 on cv. RK-26 P3 and 1.0 on cv. RK-13 P4. The cv. RK-12 P3 did not develop measurable root rot. The rhizosphere population of root rot fungus increased over time, being highest on the cv. RK-10 P3 (P ≤ 0.001), followed by cvs. RK-18 P8, RK-26 P3, RK-13 P4 and RK-12 P3. Inoculation with the fungus resulted in 5% (cv. RK-10 P3), 10.3% (cv. RK-18 P8, P ≤ 0.05), 10.9% (cv. RK-26 P3, P ≤ 0.05), 16.4% (cv. RK-13 P4, P ≤ 0.01) and 41.5% (cv. RK-12 P3, P ≤ 0.001) increase in the TP content of leaves. SA concentration in tobacco leaves increased marginally (0.8%–3%) in cvs. RK-10 P3, RK-18 P8 and RK-26 P3, but considerably (16%–17%, P ≤ 0.01) in cv. RK-13 and RK-12 P3 in comparison to uninoculated plants. Total chlorophyll content of leaves in response to inoculation with P. aphanidermatum decreased by 27% and 23% in tobacco cvs. RK-10 P3 and RK-18 P8 (P ≤ 0.001) and 17.6 (P ≤ 0.01) and 10.6% (P ≤ 0.05) in cv. RK-26 P3 and RK-13 P4, respectively. Reduction in chlorophylls a and b was 20% and 15% in cv. RK-10 P3 and 20% and 11% in cv. RK-18 P8. Total carotenoid contents of tobacco leaves decreased significantly in cvs. RK-10 P3 and RK-18 P8 (P ≤ 0.05). Significant and greater decrease in plant growth variables was recorded in the cultivars in which increase in TP and SA was lower and decrease in chlorophyll and carotenoids was greater. This study has revealed that greater synthesis of TP and SA may provide resistance in tobacco plants against P. aphanidermatum. The cv. RK-12 P3, in which greatest increase in the SA (17%) and TP (41.5%) was recorded, did not exhibit a significant decrease in plant growth variables and leaf pigments (P ≤ 0.05).     

Transmission of Fusarium solani var. coeruleum and F. sulphureum from seed potatoes to progeny tubers in the field

In experiments in three years, seed tubers were inoculated before planting with either Fusarium solani var. coeruleum or F. sulphureum to initiate a rot, or were contaminated by dipping in soil slurries containing spore suspensions of one or other of the pathogens. Transmission to progeny tubers was tested by uniformly wounding and incubating tubers and by dilution plating of soil samples. In two years, transmission of F. sulphureum was greater from highly contaminated than from rotting seed and was greater on cv. Pentland Crown than on cvs Desiree and Maris Piper. F. solani var. coeruleum appeared to be transmitted most readily from rotting seed and Maris Piper was the cultivar most extensively contaminated. In experiments with different harvest dates, transmission of both fungi from highly contaminated seed could be detected by late June or mid-July. More progeny tuber wounds rotted in F. sulphureum than in F. solani var. coeruleum plots and in one year, F. sulphureum caused more rots on cv. Record than on cv. Maris Piper. These differences between the pathogens may be related to their differing abilities to sporulate underground on the surface of seed tubers and on stem bases.     

Use of Rhizobia in the Control of Root Rot Diseases of Sunflower, Okra, Soybean and Mungbean

Biocontrol potential of Rhizobium and Bradyrbizobium against soilborne root infecting fungi was tested. In vitro tests Rhizobium meliloti inhibited growth of Macrophomina phaseolina, Rhizoctonia solani and Fusarium solani while Bradyrhizobium japonicum inhibited M. phaseolina and R. solani producing zones of inhibition. In field R. meliloti, R. leguminosarum and B. japonicum used either as seed dressing or as soil drench reduced infection of M. phaseolina, R. solani and Fusarium spp., in both leguminous (soybean, mungbean) and non-leguminous (sunflower and okra) plants.     

Evaluation of Pseudomonas aeruginosa Z5 for biocontrol of cotton seedling disease caused by Fusarium oxysporum

Plant growth-promoting bacteria-mediated biocontrol of plant pathogens is renowned to enhance the growth of the plants using different direct or indirect mechanisms. The goal of the present investigation was the evaluation of Pseudomonas aeruginosa Z5 isolated from cotton grown in Pakistani soils for the suppression of Fusarium oxysporum associated with cotton seedling disease. In dual culturing techniques, four bacterial strains inhibited fungal pathogens, i.e. F. oxysporum, Fusarium moniliforme, Fusarium solani and Rhizoctonia solani, significantly with percent inhibition ranging from 25% to 91.5%. P. aeruginosa Z5 showed maximum suppression of all the tested pathogens. Net-house experiments showed that the application of P. aeruginosa Z5 both separately and in combination with Bacillus fusiformis S10 significantly reduced the disease incidence by suppressing F. oxysporum (the causal agent of cotton seedling disease) up to 64–65% and improved the percent germination as compared to the infected control plants. The production of antibiotics, proteases and siderophores may be the contributing factors for its antagonistic properties. Highest bacterial population (8.9 CFU/g root) observed on roots of cotton plants inoculated with P. aeruginosa Z5 showed its good colonisation aptitudes even in the presence of high inoculation of soil with F. oxysporum. Confocal laser scanning microscopy supported the root colonisation of cotton plants with fluorescently labelled P. aeruginosa Z5. Because of innate fungicidal potential, growth promoting P. aeruginosa Z5 can be used as a bioinoculant and an antagonist to suppress the growth of cotton root-associated fungal pathogen.     

Host Effect on Cross Protection Between Two Strains of Tobacco Mosaic Virus

The expression of cross protection between two strains of tobacco mosaic virus (TMV-C and TMV-P) differed in Arabidopsis thaliana cv. Columbia and Nicotiana tabacum cvs Samsun and Xanthi. Protection in A. thaliana cv. Columbia was expressed as a prevention of systemic movement of the challenge strain, regardless of the protecting strain of TMV, Protection in N. tabacum cvs Samsun and Xanthi was expressed as an inhibition of an early event in the infection process. The results presented indicate that the host may influence the mechanism by which cross protection is expressed between the same virus strains.     

Identification of some Bacteria from Paddy Antagonistic to Several Rice Fungal Pathogens

The effect of 23 bacterial strains from ricefields in the tropics on rice seed germination and on radicle and hypocotyl development of four rice cultivars was determined. There was a varietal difference in response to seed bacterization with the different bacterial strains. Germination of cv. IR58 increased from 78 to 93 %, that of cv. IR64, from 89 to 97 %. Less effects on germination of cvs IR42 and IR36 were observed. All strains inhibited the mycelial growth of Rhizoctonia solani in vitro. The three strains, identified as Bacillus subtilis, inhibited the mycelial growth of eight fungal pathogens whereas the other strains were pathogen-specific. Seed bacterization with these bacterial strains provided a sheath blight protection of 4. 5 to 73 % in the glasshouse trial. These 23 bacterial strains were identified by phenotypic tests using the API systems, morphological and biochemical features, and by comparison of electrophoretic patterns after sodium dodecyl sulphate polyacrylamide gel electrophoresis. Bacterial strains were identified (number of strains in brackets) as: Bacillus subtilis (3), Bacillus laterosporus (1), Bacillus pumilus (1), Pseudomonas aeruginosa (7), Pseudomonas belonging to section 1 (5), Erwina herbicola-like (1), and Serratia marcescens (1). The features of the other four strains were similar to Serratia except for the DNAase and lipase activities.     

Influence of Host Plant Genotype, Presence of a Pathogen, and Coinoculation with Pseudomonas fluorescens Strains on the Rhizosphere Expression of Hydrogen Cyanide- and 2,4-Diacetylphloroglucinol Biosynthetic Genes in P. fluorescens Biocontrol Strain CHA0

The production of hydrogen cyanide (HCN) and 2,4-diacetylphloroglucinol (DAPG) is a major factor in the control of soil-borne diseases by Pseudomonas fluorescens CHA0. We investigated the impact of different biotic factors on the expression of HCN–in comparison to DAPG biosynthetic genes in the rhizosphere. To this end, the influence of plant cultivar, pathogen infection, and coinoculation with other biocontrol strains on the expression of hcnA-lacZ and phlA-lacZ fusion in strain CHA0 was monitored on the roots of bean. Interestingly, all the tested factors influenced the expression of the two biocontrol traits in a similar way. For both genes, we observed a several-fold higher expression in the rhizosphere of cv. Derakhshan compared with cvs. Goli and Naz, although bacterial rhizosphere colonization levels were similar on all cultivars tested. Root infection by Rhizoctonia solani stimulated total phlA and hcnA gene expression in the bean rhizosphere. Coinoculation of strain CHA0 with DAPG-producing P. fluorescens biocontrol strains Pf-68 and Pf-100 did neither result in a substantial alteration of hcnA nor of phlA expression in CHA0 on bean roots. To our best knowledge, this is the first study investigating the impact of biotic factors on HCN production by a bacterial biocontrol strain in the rhizosphere.     

Plant growth-promotion (PGP) activities and molecular characterization of rhizobacterial strains isolated from soybean (Glycine max L. Merril) plants against charcoal rot pathogen, Macrophomina phaseolina

Charcoal rot disease, caused by the fungus Macrophomina phaseolina, leads to significant yield losses of soybean crops. One strategy to control charcoal rot is the use of antagonistic, root-colonizing bacteria. Rhizobacteria A₅F and FPT₇21 and Pseudomonas sp. strain GRP₃ were characterized for their plant growth-promotion activities against the pathogen. Rhizobacterium FPT₇21 exhibited higher antagonistic activity against the pathogen on dual plate assay compared to strain A₅F and GRP₃. FPT₇21 and GRP₃ gave decreased disease intensity in terms of average number of pathogen-infested plants. Lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL), and peroxidase (POD) activities were estimated in extracts of plants grown from seeds that were treated with rhizobacteria, and inoculated with spore suspension of M. phaseolina. The activity of these enzymes after challenge with the test pathogen increased. Strains FPT₇21 and GRP₃ exhibited maximum increases in LOX, PAL and POD activity (U mg⁻¹ fresh leaf wt) compared to strain A₅F.     

Growth enhancement of Sesamum indicum L. by rhizosphere-competent Azotobacter chroococcum AZO2 and its antagonistic activity against Macrophomina phaseolina

Indigenous strains isolated from rhizosphere may contain highly competent genotypes to enhance the plant growth and often perform better than the introduced isolates. The present study deals with the characterisation of plant growth-promoting (PGP) attributes and antagonistic activity of Azotobacter chroococcum AZO2 against Macrophomina phaseolina causing charcoal rot disease and their effect on the growth of sesame (Sesamum indicum L.). Eight strains of Azotobacter were isolated from sesame rhizosphere on nitrogen-free medium, which exhibited significant PGP parameters such as phosphate solubilisation, indole acetic acid and siderophore production. The strain A. chroococcum AZO2 (EU274299) was characterised by 16S rDNA gene sequencing. Amplification of 781 bp nif H gene confirms nitrogenase activity of all the strains. A. chroococcum AZO2 exhibited strong antagonistic activities against M. phaseolina causing 81% colony growth inhibition and resulted in hyphal perforations, empty cell (halo) formation, hyphal twisting, shrinking and lysis of fungal mycelia along with degeneration of sclerotia. A. chroococcum AZO2 produced chitinase that caused degradation and digestion of the cell wall component of M. phaseolina. Different vegetative and reproductive parameters of sesame were found to be enhanced significantly upon application of A. chroococcum AZO2 + half doses of chemical fertilisers. A. chroococcum AZO2 was also found to be an effective root coloniser, plant growth promoter and potential antagonistic bacterium. It can be concluded that A. chroococcum AZO2 strain bears the characteristics of technological applications for inoculant preparation and growth enhancement of sesame besides being utilised as a better PGP bacterium as well as an effective agent for biocontrol of M. phaseolina.     

Evaluation of antifungal activity of food additives against soilborne phytopathogenic fungi

The efficacy of eight food additives as possible alternatives to synthetic fungicides for the control of soilborne pathogens, Fusarium oxysporum f. sp. melonis, Macrophomina phaseolina, Rhizoctonia solani, and Sclerotinia sclerotiorum was evaluated in this study. A preliminary selection of food additives was performed through in vitro tests. The ED₅₀, minimum inhibition concentration (MIC), and minimum fungicidal concentration (MFC) values showed that ammonium bicarbonate and potassium sorbate were more toxic to soilborne pathogens compared to other food additives with few exceptions and, therefore selected for further testing in soil. The inhibitory and fungistatic efficacy potassium sorbate were higher than that of ammonium bicarbonate in in vitro tests. Potassium sorbate completely inhibited F. oxysporum f. sp. melonis, M. phaseolina, and R. solani at 0.6% in soil tests. In contrast ammonium bicarbonate at 0.6% was inferior compared to potassium sorbate. Ammonium bicarbonate achieved to control all fungi at 2% that is the highest concentration used in this study. Potassium sorbate showed higher toxicity to all fungi compared to ammonium bicarbonate in soil tests. Both ammonium bicarbonate and potassium sorbate increased the pH of soil. The rate of pH increase was higher in ammonium bicarbonate.     

Plant defense approach of Bacillus subtilis (BERA 71) against Macrophomina phaseolina (Tassi) Goid in mung bean

This study was aimed to elucidate the mitigation mechanism of an endophytic bacterium, Bacillus subtilis (BERA 71) against Macrophomina phaseolina (Tassi) Goid disease in mung bean. M. phaseolina reduced the plant growth by inducing disease, hydrogen peroxide (H₂O₂) and lipid peroxidation. The inoculation of B. subtilis to diseased plants increased chlorophyll, ascorbic acids, and superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and glutathione reductase activities, and while inhibited H₂O₂ and lipid peroxidation for enhancing plant growth. In addition, B. subtilis association in plants mitigated the M. phaseolina infection due to increase of indole acetic acids and indole butyric acid, and also a decrease of abscisic acid. However, the nutrients (N, K, Ca, Mg, Zn, Cu, Mn and Fe) were increased, except Na in M. phaseolina diseased plants treated with B. subtilis. The result of this study suggests that B. subtilis interaction with plants can modulate the metabolism of pigments, hormones, antioxidants and nutrients against M. phaseolina to induce disease resistance in mung bean.     

Antagonistic potential of native strain Streptomyces aurantiogriseus VSMGT1014 against sheath blight of rice disease

A total of 132 actinomycetes was isolated from different rice rhizosphere soils of Tamil Nadu, India, among which 57 showed antagonistic activity towards Rhizoctonia solani, which is sheath blight (ShB) pathogen of rice and other fungal pathogens such as Macrophomina phaseolina, Fusarium oxysporum, Fusarium udum and Alternaria alternata with a variable zone of inhibition. Potential actinomycete strain VSMGT1014 was identified as Streptomyces aurantiogriseus VSMGT1014 based on the morphological, physiological, biochemical and 16S rRNA sequence analysis. The strain VSMGT1014 produced lytic enzymes, secondary metabolites, siderophore, volatile substance and indole acetic acid. Crude metabolites of VSMGT1014 showed activity against R. solani at 5 µg ml^?1; however, the prominent inhibition zone was observed from 40 to 100 µg ml^?1. Reduced lesion heights observed in culture, cells-free filtrate, crude metabolites and carbendazim on challenge with pathogen in the detached leaf assay. The high content screening test clearly indicated denucleation of R. solani at 5 µg ml^?1 treatment of crude metabolite and carbendazim respectively. The results conclude that strain VSMGT1014 was found to be a potential candidate for the control of ShB of rice as a bio fungicide.     

RSC3K of soybean cv. Kefeng No.1 confers resistance to soybean mosaic virus by interacting with the viral protein P3

Soybean mosaic virus (SMV) is one of the most devastating viral pathogens of soybean (Glycine max (L.) Merr). In total, 22 Chinese SMV strains (SC1–SC22) have been classified based on the responses of 10 soybean cultivars to these pathogens. However, although several SMV-resistance loci in soybean have been identified, no gene conferring SMV resistance in the resistant soybean cultivar (cv.) Kefeng No.1 has been cloned and verified. Here, using F₂-derived F₃ (F_2:3) and recombinant inbred line (RIL) populations from a cross between Kefeng No.1 and susceptible soybean cv. Nannong 1138-2, we localized the gene in Kefeng No.1 that mediated resistance to SMV-SC3 strain to a 90-kb interval on chromosome 2. To study the functions of candidate genes in this interval, we performed Bean pod mottle virus (BPMV)-induced gene silencing (VIGS). We identified a recombinant gene (which we named R_SC3K) harboring an internal deletion of a genomic DNA fragment partially flanking the LOC100526921 and LOC100812666 reference genes as the SMV-SC3 resistance gene. By shuffling genes between infectious SMV DNA clones based on the avirulent isolate SC3 and virulent isolate 1129, we determined that the viral protein P3 is the avirulence determinant mediating SMV-SC3 resistance on Kefeng No.1. P3 interacts with RNase proteins encoded by R_SC3K, LOC100526921, and LOC100812666. The recombinant R_SC3K conveys much higher anti-SMV activity than LOC100526921 and LOC100812666, although those two genes also encode proteins that inhibit SMV accumulation, as revealed by gene silencing in a susceptible cultivar and by overexpression in Nicotiana benthamiana. These findings demonstrate that R_SC3K mediates the resistance of Kefeng No.1 to SMV-SC3 and that SMV resistance of soybean is determined by the antiviral activity of RNase proteins.     

Induction of pathogenesis-related proteins during biocontrol of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> with <Emphasis Type="Italic">Pseudomonas aureofaciens</Emphasis> in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L. Merr.) plants

To investigate the biocontrol effectiveness of the antibiotic producing bacterium, Pseudomonas aureofaciens 63–28 against the phytopathogen Rhizoctonia solani AG-4 on Petri plates and in soybean roots, growth response and induction of PR-proteins were estimated after inoculation with P. aureofaciens 63–28 (P), with R. solani AG-4 (R), or with P. aureofaciens 63–28 + R. solani AG-4 (P + R). P. aureofaciens 63–28 showed strong antifungal activity against R. solani AG-4 pathogens in Petri plates. Treatment with P. aureofaciens 63–28 alone increased the emergence rate, shoot fresh weight, shoot dry weight and root fresh weight at 7 days after inoculation, when compared to R. solani AG-4; P + R treatment showed similar effects. Peroxidase (POD) and β-1,3-glucanase activity of P. aureofaciens 63–28 treated roots increased by 41.1 and 49.9%, respectively, compared to control roots. POD was 26% greater in P + R treated roots than R. solani treated roots. Two POD isozymes (59 and 27 kDa) were strongly induced in P + R treated roots. The apparent molecular weight of chitinase from treated roots, as determined through SDS-PAGE separation and comparison with standards, was about 29 kDa. Five β-1,3-glucanase isozymes (80, 70, 50, 46 and 19 kDa) were observed in all treatments. These results suggest that inoculation of soybean plants with P. aureofaciens 63–28 elevates plant growth inhibition by R. solani AG-4 and activates PR-proteins, potentially through induction of systemic resistance mechanisms.     

Evaluation of the ability of C. albicans to form biofilm in the presence of phage-resistant phenotypes of P. aeruginosa

Pseudomonas aeruginosa and Candida albicans are disparate microbial species, but both are known to be opportunistic pathogens frequently associated with nosocomial infections. The aim of this study was to provide a better understanding of the interactions between these microorganisms in dual-species biofilms. Several bacteriophage-resistant P. aeruginosa phenotypes have been isolated and were used in dual-species mixed-biofilm studies. Twenty-four and 48 h mixed-biofilms were formed using the isolated phenotypes of phage-resistant P. aeruginosa and these were compared with similar experiments using other P. aeruginosa strains with a defined lipopolysaccharide (LPS) deficiency based on chromosomal knockout of specific LPS biosynthetic genes. Overall, the results showed that the variants of phage-resistant P. aeruginosa and LPS mutants were both less effective in inhibiting the growth of C. albicans in mixed-biofilms compared to the wild-type strains of P. aeruginosa. Conversely, the proliferation of P. aeruginosa was not influenced by the presence of C. albicans. In conclusion, the ability of strains of P. aeruginosa to inhibit the formation of a biofilm of C. albicans appears to be correlated with the LPS chain lengths of phenotypes of P. aeruginosa, suggesting that LPS has a suppressive effect on the growth of C. albicans.