Chemical,physical and biological control of carrot seedling diseases

In naturally infested soil containingPythium ultimum, P. acanthicum andPhytophthora megasperma, onlyP. ultimum was associated with root rot and damped-off seedlings. Damping-off was promoted by low soil temperatures and by flooding. Seedling stands were markedly reduced when seed was pre-incubated in soil at 12°C but not at 25°C or 35°C. Dusting carrot seed with metalaxyl significantly increased seedling stands in the field at rates from 1.5–6 g kg⁻¹ seed and in both flooded and unflooded, naturally infested soil at 3.15 g kg⁻¹. In greenhouse experiments using artifically infested soil,P. ultimum andP. paroecandrum caused damping-off of carrot seedlings andRhizoctonia solani reduced root and shoot weights.R. solani caused damping-off in nutrient-enriched soil.P. acanthicum andP. megasperma were not pathogenic to seedlings, although both fungi colonized roots. Soil populations of allPythium spp., particularlyP. ultimum, increased during growth of seedlings and population growth ofP. megasperma was promoted by periodic flooding. Infestation of soil withP. acanthicum did not reduce damping-off of carrot seedlings byP. ultimum orP. paroecandrum, but significantly increased root and shoot weights and decreased root colonization byR. solani P. acanthicum has potential as a biocontrol agent againstR. solani.     

Radiometric assessment of tillage and seed treatment effect on soybean root rot caused by Fusarium spp. in central Minnesota

Soybean root rot, caused primarily by Fusarium solani f. sp. phaseoli in a complex with F. oxysporum and Rhizoctonia solani, has become an increasing problem for soybeans, dry beans, and other rotation crops in central Minnesota due to soil conditions associated with reduced tillage. This study was conducted, in two field sites in central Minnesota located near Staples and Verndale, to develop methods for nondestructive assessment of root rot severity using plant radiometric properties. Soybean canopy reflectance was measured with a hand-held multi-spectral radiometer. Prior to the radiometer measurements, attempts were made to create differing root rot situations with moldboard or chisel tillage, and with or without a biological seed treatment. Root rot severity was estimated using a visual disease severity scale. Colony-forming units (CFU) were determined to estimate soil populations of pathogenic F. solani and F. oxysporum. Results from the Verndale site consistently showed significant treatment effects in the measured canopy radiometric parameters, and in the visual disease rating and yield (significant for seed treatment). Values of a simple ratio vegetation index from this site exhibited negative relationships with disease rating and F. oxysporum CFU, and a positive linear relationship with yield. Treatment effects were generally not significant at the Staples site because of low initial F. oxysporum populations. The results indicate that remote sensing is potentially a rapid, nondestructive means for assessment of root rot diseases in soybean.     

Effect of solarization of soil within plastic bags on root rot of gerbera (Gerbera jamesonii L.)

Solarization of soil, (potting mix = coarse sand:Eucalyptus marginata fines = 1∶1) infested with 3 fungi pathogenic to gerbera (Phytophthora cryptogea, Fusarium oxysporum andRhizoctonia solani), for 3 to 4 weeks within transparent polyethylene bags controlled root rot of gerbera. Solarization for 2 weeks however, was less effective. All plants grown in the infested potting mix which had been kept in shade for 2, 3 or 4 weeks were severely attacked. Solarization of soil within plastic bags for 4 weeks also increased availability of nutrients such as NH₄ ⁺-N, PO₄ ⁻ and K⁺ in comparison to bagged soil kept in the shade for the same period.     

Resident bacteria, nitric oxide emission and particle size modulate the effect of Brassica napus seed meal on disease incited by Rhizoctonia solani and Pythium spp

Amendment of orchard soil with low-glucosinolate Brassica napus (rape) seed meal (RSM) suppresses infection of apple roots by Rhizoctonia solani but increases incidence of Pythium spp. infection. Following incorporation of Brassica sp. seed meals, soils were monitored for changes in populations of selected saprophytic and plant pathogenic microorganisms. When conducted in pasteurized soil, which possessed high numbers of Bacillus spp. and lower than detectable numbers of Streptomyces spp., RSM amendment did not provide control of R. solani. Populations of streptomycetes in RSM-amended soil increased to stable levels >20-fold higher than in non-amended soil. Disease suppressiveness was restored to pasteurized RSM-amended soil by adding any of several Streptomyces strains. Maximal rates of nitrification in orchard soil, determined by nitric oxide emission, were observed within two weeks following RSM amendment and inhibition of nitrification via application of nitrapyrin abolished the capacity of RSM to suppress R. solani infection of apple roots when seedlings were planted one day after soil amendment. Apple seedling mortality and Pythium spp. root infection were highest for seedlings planted immediately following incorporation of B. napus cv. Athena RSM, particularly when meal was added in a flake rather than powder form. Lower infection frequencies were observed for seedlings planted four weeks after RSM incorporation, even for soil in which densities of culturable Pythium spp. had not declined. Our results demonstrate that suppression of Rhizoctonia root rot in response to RSM amendment requires the activity of the resident soil microbiota and that initial disease control is associated with the generation of nitric oxide through the process of nitrification.     

Biological control of root rot fungus <Emphasis Type="Italic">Macrophomina phaseolina</Emphasis> and growth enhancement of <Emphasis Type="Italic">Pinus roxburghii</Emphasis> (Sarg.) by rhizosphere competent <Emphasis Type="Italic">Bacillus subtilis</Emphasis> BN1

Bacterial isolates having antifungal and good plant growth-promoting attributes were isolated from chir-pine (Pinus roxburghii) rhizosphere. An isolate, Bacillus subtilis BN1 exhibited strong antagonistic activity against Macrophomina phaseolina, and other phytopathogens including Fusarium oxysporum and Rhizoctonia solani. It was characterized and selected for the present studies. BN1 resulted in vacuolation, hyphal squeezing, swelling, abnormal branching and lysis of mycelia. The cell-free culture filtrate of BN1 inhibited the growth of M. phaseolina. Pot trial study resulted in statistically significant increase in seedling biomass besides reduction in root rot symptoms in chir-pine seedlings. BN1 treatment resulted in 43.6% and 93.54% increases in root and shoot dry weights respectively, as compared to control. Also, 80–85% seed viability was recorded in treatments receiving BN1 either alone or in the presence of M. phaseolina, compared to 54.5% with M. phaseolina. Bioinoculant formulation study suggested that maximum viability of bacteria was in a sawdust-based carrier. B. subtilis BN1 produced lytic enzymes, chitinase and β-1,3-glucanase, which are known to cause hyphal degradation and digestion of the cell wall component of M. phaseolina. In the presence of M. phaseolina, population of B1 was 1.5 × 10⁴ c.f.u. g⁻¹ root after one month, which increased to 4.5 × 10⁴ c.f.u. g⁻¹ root in three months. Positive root colonization capability of B. subtilis BN1 proved it as a potent biocontrol agent.     

Rhizotron studies on Zea mays L. to evaluate biocontrol activity of Bacillus subtilis

The effect of Bacillus as a biocontrol agent against some root-rot fungi was tested using maize (Zea mays L.) in rhizotrons placed in a growth chamber with relative humidity 60% with a 12 h photoperiod and day and night temperatures of 24 and 18°C respectively. Rhizoctonia solani Kühn caused pre-emergence damping-off in the untreated maize seeds showing weak and soft roots as observed through the perspex rhizotrons. Image analysis was used to quantify the effects of Bacillus treatment on seedlings infected with Pythium sp. Bacillus B77 and B81 were most effective in the control of the pathogen, R. solani which achieved a biocontrol activity of 24 and 35% respectively with regard to shoot dry biomass while B81 achieved 48% biocontrol with reference to root dry biomass. There was no effect on the root area. For root dry biomass, B81, B69, B11 and B77 showed higher biocontrol activity in comparison to the control. Pythium sp. caused pre- and post emergence damping- off in the untreated seeds. Root rot of the maize seedlings caused by Pythium sp. was slightly controlled by Bacillus B69 and B81 which achieved biocontrol activity of 18 and 11% respectively. For the biocontrol of Fusarium solani, Bacillus B77, B69, B81 achieved biocontrol activity of 50, 48 and 33% respectively with reference to root dry biomass.     

Calcium and temperature effects on seedling exudation and root rot infection of common bean on an acid sandy soil

Soil born fungi such as Phytium ultimum, Fusarium ssp., and Rhizoctonia solani (Kühn) severely restrict stand establishment of common bean (Phaseolus vulgaris L.) on acid soils of the Tropics. Calcium application is known to alleviate fungal infection in many legumes but the causes are still unclear. To investigate environmental factors and physiological mechanisms involved, growth chamber experiments were conducted with an acid sandy soil from Mexico. Treatments were soil liming at a rate of 0.67 g Ca(OH)₂ kg^-1, gypsum application at 0.49 g CaSO₄ 2H₂O kg^-1 soil placed around the seed, and an untreated control. Beans were grown under three temperature regimes with constant night and one constant day vs. two sinusoidal day temperatures. To examine patterns of seed and seedling exudation at regular intervals leachates of germinating seeds were collected on filter paper soaked with equilibrium solutions from soils of the three treatments. The severity of root rot in the control treatment was highest when plants were stressed by temperature extremes. At a sinusoidal day temperature peaking at 40°C soil liming and gypsum application to the seed increased the number of healthy seedlings similarly by over 60%. However, only liming which effectively eliminated growth constraints by low pH and high aluminum concentrations led to an increase in hypocotyl elongation by 22% and in total root length by 8%. Both calcium amendments increased the calcium and potassium contents in the hypocotyl tissue. From seeds exposed to the equilibrium solution of unlimed soil with pH 3.7, 1 mM Ca, and 0.6 mM Al considerable amounts of amino acids and carbohydrates were leached. In contrast, exposure to the equilibrium solution from limed soil with pH 4.3, 3 mM Ca, and negligible concentrations of Al led to a net uptake of amino acids and decreased leaching of carbohydrates. Exposure to the equilibrium solution of the gypsum treatment with pH 3.6, 20 mM Ca, and 1.2 mM Al resulted in a somewhat smaller net uptake of amino acids compared to liming. During germination pH around the seeds steeply increased in the untreated control but significantly less with both amendments. The results indicate that pH and the Ca/Al ratio in the soil solution around bean seeds determine their pattern of exudation and solute uptake. For bean germination and early growth on acid soils locally placed application of small amounts of gypsum as seed pelleting seems as effective as soil liming in reducing the incidence of root rot. The results indicate that this may be accomplished by decreasing the amount of leachates available for fungal development.     

Fungicidal control of damping-off and seedling root rot in Brassica species caused by Rhizoctonia solani in the growth chamber*

Eight fungicides (benodanil, carboxin, cyproconazole, fenpropimorph, fur-mecyclox, iprodione, pencycuron and tolclofos-methyl) were evaluated, under growth chamber conditions, as seed treatments against pre-emergence damping-off and post-emergence seedling root rot in six Brassica species. Five cultivars of B. rapa, four cultivars of B. juncea, four cultivars of B. napus and one cultivar/ strain from each of B. carinata, B. nigra and B. oleracea were grown in soilless mix infested with an isolate of Rhizoctonia solani AG-2-1. B. nigra and B. juncea were considerably less susceptible to R. solani than the four other species. Cyproconazole at 0.05-0.1 g a.i./kg seed and the other fungicides at 2–4 g a.i./ kg seed provided almost complete control of pre-emergence damping-off in most Brassica species and their cultivars. Their efficacy varied against the post-emergence seedling root rot. Furmecyclox, iprodione, tolclofos-methyl and pencycuron consistently gave good control of seedling root rot in all Brassica species and their cultivars. Benodanil and fenpropimorph provided moderate control, and carboxin and cyproconazole gave poor control against root rot. Efficacy of carboxin, cyproconazole, benodanil and fenopropimorph against seedling root rot varied significantly (P ≤ 0.05) among cultivars within a Brassica species.     

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.     

Biocontrol activity of salt tolerant Streptomyces isolates against phytopathogens causing root rot of sugar beet

Biological control of fungi causing root rot on sugar beet by native Streptomyces isolates (C and S2) was evaluated in this study. The dry weight and colony forming unit (CFU) of S2 and C increased when 300 mM NaCl was added to medium. The in vitro antagonism assays showed that both isolates had inhibitory effect against Rhizoctonia solani AG-2, Fusarium solani and Phytophthora drechsleri. In dual culture, Streptomyces isolate C inhibited mycelial growth of R. solani, F. solani and P. drechsleri 45%, 53% and 26%, respectively. NaCl treatment of medium increased biocontrol activity of soluble and volatile compounds of isolate C and S2. After salt treatment, growth inhibition of R. solani, F. solani and P. drechsleri by isolate C increased up to 59%, 70% and 79%, respectively. To elucidate the mode of antagonism, protease, chitinase, beta glucanase, cellulase, lipase and α-amylase activity and siderophore and salicylic acid (SA) production were evaluated. Both isolates showed protease, chitinase and α-amylase activity. Also, biosynthesis of siderophore was detectable for both isolates. Production of siderophore and activity of protease and α-amylase increased after adding salt for both isolates. In contrast, chitinase activity decreased significantly. Production of SA, beta glucanase and lipase by isolate S2 and biosynthesis of cellulase by isolate C were observed in presence and absence of NaCl. Soil treatment with Streptomyces isolate C inhibited root rot of sugar beet caused by P. drechsleri, R. solani and F. solani. Results of this study showed that these two Streptomyces isolates had potential to be utilized as biocontrol agent against fungal diseases especially in saline soils.     

Mechanisms of biocontrol of soil-borne plant pathogens by Rhizobacteria

Bacterial antagonism, responsible for biological control, may operate by antiobiosis, competition or parasitism. Parasitism relies on lytic enzymes for the degradation of cell walls of pathogenic fungi. Serratia marcescens was found to be an efficient biocontrol agent of Sclerotium rolfsii and Rhizoctonia solani under greenhouse conditions. Populations of 10⁵ or 10⁶ colony forming units g^-1 soil were the most effective. Drench and drip application of S. marcescens suspension were more effective in controlling S. rolfsii than spraying, mixing in soil or seed coating. The highest population density of the bacteria in the rhizosphere was found on the proximal portion of the root, decreasing significantly until the tips, where it increased again. The isolated Serratia, found to possess chitinolytic activity, was able to release N-acetyl D-glucosamine from cell walls of S. rolfsii. The gene coding for chitinase was cloned into Escherichia coli and the enzyme was uniquely excreted from the bacterium into its growth medium. When S. rolfsii was sprayed by partially purified chitinase produced by the cloned gene, rapid and extensive bursting of the hyphal tips was observed. This chitinase preparation was effective in reducing disease incidence caused by S. rolfsii in beans and R. solani in cotton, under greenhouse conditions. A similar effect was obtained when a viable E. coli cell, containing the plasmid with the chitinase gene (pLCHIA), was applied. It appears that genetic engineering of the lytic enzymes, such as chitinase which play an important role in plant disease control, may improve the efficacy of biocontrol agents.     

Efficiency of different application methods of biocontrol agents and biocides in control of Fusarium root rot on some citrus rootstocks

Abstract

The efficiency of two biocontrol agents (Trichoderma harzianum NB and Bacillus subtilis NB) and two commercial biocides (Plant Guard and Rhizo-N) in controlling Fusarium root rot disease on some citrus rootstocks was evaluated under artificially infested soil in green house.

Fusrium root rot on citrus rootstocks seedlings i.e. sour orange (SO), volkamer lime (VL), rangpur lime (RP) and cleopatra mandarin (CL) was successfully controlled by dipping the root system of such seedlings in water suspensions of each biological treatment i.e. Trichoderma harzianum (spore suspension 5×10⁶ spore/ml), Bacillus subtilis (cell suspension 8×10⁷ cell/ml), Plant Guard (3 g/l) and Rhizo-N (4 g/l), then transplanted into artificially infested soil with Fusarium solani and drenched with enough water suspension of such biological treatments. Plant Guard (3 g/l) and Rhizo-N (4 g/l) were highly effective treatments in decreasing infection and severity of the disease, Fusarium density in rhizosphere soil and colonization of Fusarium solani in the roots of all tested seedlings.

Meanwhile, root dipping or soil drenching with the same treatments individually gave the least effect in reducing root rot incidenceon all tested rootstocks compared with application of the two methods together.

It should be noted that using biocontrol agents and commercial biocides could be successfully used in controlling root rot pathogens on citrus in commercial greenhouses or under field conditions before transplanting in new reclaimed lands in the desert.     

Soil Type Dependent Rhizosphere Competence and Biocontrol of Two Bacterial Inoculant Strains and Their Effects on the Rhizosphere Microbial Community of Field-Grown Lettuce

Rhizosphere competence of bacterial inoculants is assumed to be important for successful biocontrol. Knowledge of factors influencing rhizosphere competence under field conditions is largely lacking. The present study is aimed to unravel the effects of soil types on the rhizosphere competence and biocontrol activity of the two inoculant strains Pseudomonas jessenii RU47 and Serratia plymuthica 3Re4-18 in field-grown lettuce in soils inoculated with Rhizoctonia solani AG1-IB or not. Two independent experiments were carried out in 2011 on an experimental plot system with three soil types sharing the same cropping history and weather conditions for more than 10 years. Rifampicin resistant mutants of the inoculants were used to evaluate their colonization in the rhizosphere of lettuce. The rhizosphere bacterial community structure was analyzed by denaturing gradient gel electrophoresis of 16S rRNA gene fragments amplified from total community DNA to get insights into the effects of the inoculants and R. solani on the indigenous rhizosphere bacterial communities. Both inoculants showed a good colonization ability of the rhizosphere of lettuce with more than 10⁶ colony forming units per g root dry mass two weeks after planting. An effect of the soil type on rhizosphere competence was observed for 3Re4-18 but not for RU47. In both experiments a comparable rhizosphere competence was observed and in the presence of the inoculants disease symptoms were either significantly reduced, or at least a non-significant trend was shown. Disease severity was highest in diluvial sand followed by alluvial loam and loess loam suggesting that the soil types differed in their conduciveness for bottom rot disease. Compared to effect of the soil type of the rhizosphere bacterial communities, the effects of the pathogen and the inoculants were less pronounced. The soil types had a surprisingly low influence on rhizosphere competence and biocontrol activity while they significantly affected the bottom rot disease severity.     

Bacterial antagonists of Sunflower (Helianthus annuus L.) fungal pathogens

Bacteria isolated on nutrient agar and King's medium B from sunflower leaves, crown and roots inhibited in vitro growth of the leaf spot and wilt pathogens Alternaria helianthi, and Sclerotium rolfsii, respectively, and also the root rot pathogensRhizoctonia solani and Macrophomina phaseolina. Antagonistic bacteria from leaves were mainly actinomycetes and pigmented Gram-positive bacteria, while those from roots and crowns were identified asPseudomonas fluorescens-putida, P. maltophilia, P. cepacia, Flavobacterium odoratum andBacillus sp. In soil bioassays, when used as seed inoculum in the presence ofS. rolfsii, P. cepacia strain N24 increased significantly the percentage of seedling emergence. Bacterial strains which exhibited broad spectrum in vitro antagonistic activity were tested for colonisation of sunflower roots, when used as a seed inoculum. Good colonisers (10⁴ to 10⁶ bacteria/g root) were consistent in their ability to reduce disease and fungal wilt. A seedling having a primary root length < 5 cm with fewer lateral roots, necrosed cotyledons or crown and a wilted shoot indicated its diseased status. On an average, only 30% of seedlings were diseased when treated with the antagonistic strains, in the presence of the pathogen, while 60% of the seedlings were diseased in the presence of the pathogen alone. In microplots treated with strain N24, only 1 to 3% of the seedlings were wilted, while 14% of the seedlings were wilted in the presence of the pathogen alone. The results obtained show that bacterial antagonists of sclerotial fungi can be used as seed inocula to improve plant growth through disease suppression     

Development of Pusa 5SD for seed dressing and Pusa Biopellet 10G for soil application formulations of Trichoderma harzianum and their evaluation for integrated management of dry root rot of mungbean (Vigna radiata)

Various seed dressing and soil application formulations were developed from Trichoderma viride, T. virens and T. harzianum to increase the shelf life of bio-formulations used to manage dry root rot (Rhizoctonia bataticola) of mungbean (Vigna radiata), a major yield limiting factor in mungbean production. The shelf life of the formulations developed in the present study was monitored by counting colony forming units (cfu) up to 25 months of storage at room temperature (26 ± 8 °C). A newly developed seed dressing formulation, Pusa 5SD based on peat powder (47.5%), Sabudana powder (Manihot esculenta) (47.5%) and carboxymethyl cellulose (5%) and a newly developed soil application formulation, Pusa Biopellet (PBP) based on sodium alginate, aluminium silicate, Sabudana powder and tap water (1:5:5:100 w/w/w/v) exhibited longer shelf life. Another formulation Pusa Biogranule (PBG) based on wheat and pulse brans varied in cfu counts during different periods of storage. Pusa 5SD could be used up to 25 months of storage while PBP 10G and PBG 5 could be used up to 15 months of storage (>10⁵ cfu). The efficacy of the formulations was evaluated in pot experiments against the disease. In these experiments, T. harzianum based PBP 10G and PBG 5 for soil application, and Pusa 5SD for seed treatment were found to be superior to others in reducing the dry root rot incidence, and increasing the seed germination and shoot and root lengths. However, a combination of soil application of PBP 10G (T. harzianum) and seed treatment with T. harzianum based Pusa 5SD + carboxin was found superior to the use of any of these formulations alone in reducing the dry root rot incidence (87.2%) and increasing the seed germination (43.0%), shoot length (40.3%), root length (37.0%) and grain yield (54.6%) of mungbean crop over those of untreated control under sick field conditions.     

关防风根腐病拮抗细菌筛选与鉴定

【背景】由木贼镰刀菌[Fusarium equiseti(Corda)Sacc.]引起的关防风根腐病,是近年来导致关防风产量及质量下降的主要土传病害之一。生物防治因其对环境安全及人畜无害等优势,成为目前植物病害防治的一种有效手段。【目的】挖掘关防风根际土壤中对木贼镰刀菌具有良好拮抗作用的生防菌株。【方法】采用稀释平板法分离根际土壤细菌;用滤纸片法和牛津杯法对拮抗细菌进行筛选和抑菌谱检测;用抗生素标记法标记拮抗细菌并测定其定殖能力;通过盆栽实验研究其对关防风根腐病的防效;通过形态学、生理生化特征及16S rRNA基因序列确定分类学地位。【结果】从健康关防风根际土壤中分离纯化了157株细菌,筛选获得对关防风根腐病菌具有显著抑菌作用的拮抗细菌SC-119,无菌滤液的抑菌率可达68.53%,而且兼具广谱抑菌能力和良好的定殖能力;盆栽实验表明,菌株SC-119对关防风根腐病的防治效果达到67.39%,其相对防效较接种哈茨木霉菌剂、枯草芽胞杆菌菌剂和代森锰锌分别提高了29.03%、32.26%和16.13%;对菌株SC-119进行分类学鉴定,确定其为萎缩芽胞杆菌(Bacillus atrophae...     

Interaction between zinc nutritional status of cereals and Rhizoctonia root rot severity

An inverse correlation between plant Zn concentration and the severity of Rhizoctonia root rot, described in an earlier paper, was examined in two experiments in a growth chamber. In the first experiment, wheat (Triticum aestivum cv Songlen) was planted in a Zn deficient soil with and without added Zn, and combined factorially with different inoculum densities of Rhizoctonia solani anastomosis group 8. When Zn was added, the percentage of seminal roots infected with R. solani was significantly lower compared to the treatments without added Zn, showing that low Zn potentiated the disease. A subsequent factorial experiment of four inoculum densities and six Zn levels, (0, 0.01, 0.04, 0.1, 0.4 and 6.0 mg Zn kg^–1 soil) was conducted to investigate the Zn effect in more detail. Disease severity was markedly decreased by the higher Zn applications; the disease score dropped sharply between treatments of Zn_0.04 and Zn_0.1, a difference which was reflected in the plant yield response to Zn. For both experiments the Zn concentrations in shoots were significantly different only among Zn treatments, not among the inoculum treatments. This indicated that inoculum density or disease severity did not reduce Zn concentration in the plant. Thus, disease did not exaggerate Zn deficiency, but rather, Zn sufficiency suppressed disease severity. A potentiating link between Zn nutrition and disease severity is thereby established, although this type of experiment did not indicate the mechanism of the Zn effect.     

The role of cuticle and epidermal cell wall in resistance of rapeseed and mustard to Rhizoctonia solani

A study was conducted to determine whether the cuticles in two genera of the family Cruciferae are effective barriers to infection by Rhizoctonia solani, and whether differences in cuticle and epidermal cell wall thickness and morphology of epicuticular wax exist between resistant and susceptible cultivars. As Canola/rapeseed (Brassica napus) and mustard (Sinapis alba) plants develop from 1 to 3 weeks of age, they become increasingly resistant to R. solani AG2-1 seedling root rot. Seven-day-old seedlings of S. alba cultivars are invariably more resistant than B. napus cultivars. Brassica napus cultivars do not show an obvious cuticle layer at 1 week but at 3 weeks the presence of a cuticle is seen through autofluorescence with a concomitant increase in resistance to R. solani. Removal of the cuticle from 3-week-old hypocotyls by chloroform treatment results in a decrease in cuticular autofluorescence and a significant increase in disease severity in both resistant and susceptible cultivars. Three-week-old plants of S. alba have a much lower percent disease rating and a significantly (p=0.05) thicker cuticle layer than similar-age plants of B. napus. The results suggest that the cuticle plays an important role in the resistance of S. alba and older plants of B. napus to infection by R. solani.     

Bacterial Antagonist as Seed Treatment to Control Leaf Blight Disease of Bambara Groundnut (Vigna subterranea)

Summay Soil samples were taken from 48 fields in the southern part of Thailand in which either bambara groundnut (Vigna subterranea) or groundnut (Arachis hypogeae) had been planted. Bacillus spp. were isolated using soil dilution plates and heat treatment to screen for endospore-producing bacteria. Among 342 Bacillus spp. isolates tested, 168 isolates were not antagonistic to Bradyrhizobium sp. strain NC-92 using dual culture technique. Further testing found 16 isolates of Bacillus spp. had the ability to inhibit mycelial growth of Rhizoctonia solani, a causal agent of leaf blight of bambara groundnut. Among these isolates, Bacillus spp. isolate TRV 9-5-2 had the greatest activity in anti-microbial tests against R. solani. This isolate was later identified as B. firmus. A powder formulation of B. firmus was developed by mixing bacterial endospores, talcum, sodium carboxymethylcellulose (SCMC) and polyvinylpyrolidone (PVP). The formulations contained bacterial levels ranging from 10⁸ to 10¹⁰ c.f.u./g and the viability of bacteria in all formulations remained high after 1 year storage at room temperature (26–32 °C). All formulations showed satisfactory effectiveness in vitro in suppressing mycelial growth of R. solani using dual culture technique. The application of formulations as seed treatment showed that these formulations did not cause abnormality of seedling shape and had no effect on the germination of bambara groundnut seeds.     

一株抗茶炭疽菌和魔芋镰刀菌的淀粉酶产色链霉菌的分离、鉴定及生防潜能

【背景】放线菌是一类重要的生防菌,具有强大的代谢活性,能产生抗生素、酶、酶抑制剂和激素等天然产物抑制病原物生长。【目的】从茶树根际分离得到放线菌,研究候选放线菌对茶炭疽菌和魔芋镰刀菌的抑菌活性及其生防潜能。【方法】分别以茶炭疽病致病菌Colletotrichum camelliae和魔芋茎腐病致病菌Fusarium solani为指示菌,采用土壤稀释涂布法、平板对峙法和菌丝生长速率法,从茶树根际土壤中分离、筛选拮抗放线菌,并根据菌株的形态特征、生理生化特性和系统发育分析结果对其进行分类鉴定,并开展候选放线菌的产促生相关物质和分泌细胞壁水解酶能力的定性检测试验。【结果】共分离得到14株拮抗放线菌,菌株A-dyzsc04-2的拮抗效果最强,被鉴定为淀粉酶产色链霉菌（Streptomyces diastatochromogenes）。该菌株的活菌体对C. camelliae和F. solani的抑制率分别为66.71%±1.23%和71.59%±2.46%,其无菌发酵滤液对2种指示菌的抑菌率均大于90%;此外,菌株A-dyzsc04-2还具有产嗜铁素和葡聚糖酶以及溶解无机磷的能力。【结论】菌株A-dyzsc04-2是一株优良的生防菌,具有较高的开发利用价值,研究结果为菌株A-dyzsc04-2防治茶炭疽病和魔芋茎腐病提供了理论支撑。