THE CONTROL OF DISEASES OF LETTUCE BY THE USE OF ANTAGONISTIC ORGANISMS I. THE CONTROL OF BOTRYTIS CINEREA PERS.

The antagonism of a number of bacteria, actinomycetes and fungi to Botrytis cinerea in pure culture on lettuce extract agar has been investigated. A considerable number were antagonistic at 25° C., a few showed antagonism at 15° C. and a limited number grew and were active at 5° C.
Certain of these organisms were able to control rot of detached lettuce leaves when inoculated before or simultaneously with B. cinerea. The latter is unable to penetrate areas of dead tissue colonized by selected antagonists.
Control of rot was also obtained when leaves of growing plants were similarly inoculated.
Substantial control of disease occurred when young potted plants in frames were sprayed with suspensions of selected antagonists in 1.0 % glucose solution.
The practicability of this method of control is discussed.     

THE CONTROL OF FUSARIUM BLIGHT IN OAT SEEDLINGS WITH ANTAGONISTIC SPECIES OF CHAETOMIUM

Forty-seven isolates representing twenty-seven species of Chaetomium were tested for their ability to control seedling blight of oat seedlings caused by Fusarium spp., chiefly F. nivale (Fr.) Ces. In pot tests the majority of the isolates were ineffective, but certain isolates of Chaetomium cochliodes Pall, and C. globosum Kunze gave a large measure of control and were as effective as an organic mercury seed dressing. Control of disease was also obtained when living material was attached to the surface of oat seed and when infested seed was soaked in cell-free filtrates from cultures of isolates which reduced disease in vivo.
Substantial control of disease was obtained when infested seed was sown in soil to which cultures of C. cochliodes had been added 10 months previously. This antagonist was able to grow through unsterile soil in the presence of certain added substrates.
The results of pot experiments were confirmed in field experiments.
Isolates of Chaetomium which controlled disease in pot and field experiments were not strikingly antagonistic to Fusariirm nivale in pure culture on agar media.     

Biological Control of Basal Stem Rot of Chrysanthemum By Antagonists*

The basal stem rot of chrysanthemum caused by Rhizoctonia solani is an important chrysanthemum disease in Taiwan, Control of the disease using the antagonists of R.solani, including members of the genera Aspergillus, Gliocladium, Paecilomyces, Penicillhium, Tricboderma and Bacillus, was studied. The antagonists were grown in solid media or culture broths for preparations of rhe inoculums which were u.sed as soil amendments or coating materials for applications on the disease control. Sawdusts, rice husks and sawdust composts were used as supporting material to prepare the solid media. By planting of the chrysanthemums, the solid media growth with antagonists were put in the nursery sand beds, or the chrysanthemum cutting seedlings were coated with antagonists of culture broths; whilst the sand beds were inoculated with R. solani. The results indicated that all antagonists could protect the chrysanthemum from the infection of R. solani, and the effects of the solid media were higher than the culture broths. When the chrysanthemum seedlings were treated with the extracts of the antagonists, the disease could also be depressed. The different control effects on Rhizactonia disease of chrysanthemums resulted from the species of the antagonists and their delivery systems to be used.     

Timelapse scanning reveals spatial variation in tomato (Solanum lycopersicum L.) root elongation rates during partial waterlogging

Aims

Effects of soil amendments with crop residues on suppression of damping-off of sugar beet were examined by growing the seedlings in pasteurized, Rhizoctonia solani (AG2-2 IIIB)-infested soil at different temperatures. Dried residues of five dasheen or taro (Colocasia esculenta (L.) Schott) cultivars were compared with those of peanut (Arachys hypogaea L.) and Brassica rapa Olsson for their effects on disease suppression.

Methods and Results

When the seedlings were grown at 17/12 °C (day/night), all residues equally suppressed the disease when amended into the soil just before sowing. At 22/17 or 32/27 °C, damping-off developed in non-treated soil within 10 days, and differential suppressive effects by the residues became apparent by 21 days. When non-pasteurized, non-treated soil was infested with the pathogen, seedling survival was markedly better than in the same but pasteurized, infested soil. Yet, the effect was not different within the entire temperature ranges. Growth of both R. solani and the seedlings peaked near 25 °C and leveled off at higher temperatures.

Conclusions

These results suggest that damping-off was suppressed by antagonistic soil microorganisms, and individual residues elevated their effects differently. Under cool conditions, the antagonists dominated the pathogen to suppress the disease. Under warmer conditions, pathogenesis overcame antagonism depending on the residue, resulting in differential effects of disease suppression.     

STUDIES IN THE ANTAGONISM BETWEEN BLASTOMYCES LUTEUS AND SPECIES OF VERTICILLIUM

Blastomyces luteus has been shown to be antagonistic in culture to both Verticillium albo-atrum and V. dahliae over a wide range of temperature and irrespective of the pH value of the medium. When B. luteus is grown on Dox's and potato-dextrose solutions it produces an exudate which, added to agar media, has an inhibiting effect upon the growth of these two species of Verticillium. The effectiveness of this inhibiting material is slightly reduced by boiling for 15 min.
When 'spent' Dox's liquid medium, which has supported the growth of B. luteus , is injected into tomato and antirrhinum seedlings inoculated with Verticillium no control of the disease is effected, but when B. luteus mycelium is added to soil infected with Verticillium , marked reduction in the incidence of disease results when the antagonistic organism and the pathogens have co-existed in the soil a sufficient length of time for the exudate of the former to be effective upon the development of the latter.     

STUDIES ON THE HOST RANGE, PROPERTIES AND MODE OF TRANSMISSION OF BEET RINGSPOT VIRUS

An apparently undescribed mechanically transmissible virus has been named beet ringspot virus (BRV). It occurs naturally in Scotland in sugar-beet, turnip, swede, potato and many kinds of weed plants. BRV is readily distinguished from raspberry ringspot virus by the symptoms produced in Chenopodium amaranticolor , French bean, tobacco and Petunia hybrida plants. BRV lost infectivity when heated for 10 min. at 63°C. but not at 60°C.: at 20°C. its longevity in vitro was about 3 weeks. BRV was precipitated by ammonium sulphate, ethanol and acetone.
Protection experiments with tobacco plants, and serological tests, gave no evidence that BRV was related to tobacco ringspot, raspberry ringspot, potato bouquet or tobacco rattle viruses, but showed that viruses isolated from different host plants and from different localities were strains of BRV.
BRV is soil-borne: in glasshouse experiments sugar-beet, beetroot, potato, turnip, swede, French bean, Fragaria vesca , oat and wheat plants often became systemically infected when grown in soil from the site of a disease outbreak, but the virus was restricted to the roots of many infected plants. When sugar-beet seedlings were grown in virus-containing soil, BRV was first detected in their roots, where its concentration increased, before progressively increasing amounts of virus were found in the shoots.
Soils from five localities were found to contain BRV. BRV has been found only where the soil is light in texture, and often in fields where raspberry ringspot virus occurs.     

THE BIOLOGY OF THE RICE WEEVIL, CALANDRA ORYZAE (L.)

The length of the developmental instars of Calandra oryzae has been estimated at relative humidities of 50, 60, 70 and 80% at 21° C., at 70% at 18, 25 and 30° C., and at 80% at 18° C., by the dissection daily of samples of infested wheat-grains. The results are expressed as a median obtained by the method of probit analysis. Comparison of this method, with estimates of median and mean obtained by orthodox arithmetical methods from similar work on Rhizopertha , show that the probit method gives good estimates.
About 90% of the eggs laid are fertile. Normally only one adult will develop in a grain, all other individuals being destroyed by cannibalism. The sex ratio is unity. It was not possible to cross C. oryzae and C. granaria.
The daily oviposition rate of C. oryzae at 17, 21 and 25° C. increases with relative humidity. There is a critical point at about 60% r.h . below which egg laying declines rapidly, and mortality is high. At 100% r.h . the oviposition rates per female per day are approximately 1.3, 2.5 and 3.4 at 17, 21 and 25° C. respectively.
In experimental conditions most eggs per grain are obtained by giving isolated females one grain each, but more eggs are laid by females given more than one grain. Daily egg output is reduced by grouping females or including males. In culture, depths of grain up to 7 cm. do not discourage egg laying.     

THE USE OF TETRA- AND PENTA-CHLORONITRO-BENZENES IN THE CONTROL OF BOTRYTIS DISEASE AND RHIZOCTONIA ATTACK OF LETTUCE

Botrytis disease in the 'spring' lettuce crop was controlled by six talc dusts containing s, 10 or 20 % of tetra- or penta-chloronitrobenzene. All the dusts increased equally the final stand in the frames, but the number of seedlings with stem lesions decreased with increasing concentrations of both compounds. At similar concentrations the tetra- was more effective than the penta-compound. The percentage survival in the field was increased by the dust treatments. The 20% dust of the penta- was only as effective as the 5% dust of the tetra-compound.
The dusts affected the leaves, stunted the seedlings and delayed hearting. At similar concentrations the tetra- was more harmful than the penta-compound. The 5 % dust of the tetra- was less damaging than the 20% dust of the penta-compound.
The spread of Rhizoctonia solani was reduced by incorporating the dusts with soil. Again, the 5% tetra- dust was as effective as the penta- dust at 20%. At 12·5 g. of dust per 20 kg. of soil, none of the dusts retarded the rate of emergence of lettuce seedlings but the highest concentration of the tetra- series reduced the number emerging.
Foliage applications of two dusts, 5% tetra- and 20% penta- dusts, controlled the spread of R. solani.     

Biological control of bacterial spot of tomato caused by Xanthomonas campestris pv. vesicatoria by Rahnella aquatilis

Xanthomonas campestris pv. vesicatoria strain 2 was isolated from infected tomato seedlings grown in open field in Egypt. This strain produced irregular yellow-necrotic areas on tomato leaves and spotting of the stem. In an attempt to control this disease biologically, four experiments were conducted and tomato seedlings were pretreated, before the pathogen, with either of two antagonistic strains of Rahnella aquatilis through leaves, roots, soil or seeds. In all experiments, seedlings pretreated with R. aquatilis showed reduced susceptibility toward X. c. pv. vesicatoria. They also contained reduced protein concentration and showed reduced number of protein bands in SDS-PAGE analysis as well as increased fresh and dry weight relative to control seedlings inoculated with the pathogen only. This indicates that R. aquatilis reduced the deleterious effect and the stress exerted by X. c. pv. vesicatoria on tomato seedlings. Foliar application of R. aquatilis was the most effective method in disease reduction which could be attributed to the direct effect of the antagonistic bacteria on the pathogen. The highest amounts of fresh and dry weight ere obtained from seed treatment, which might suggest that bacterial seed inoculation provides earlier protection than could be achieved with foliar, soil or root treatment.     

WHITE ROT (SCLEROTIUM CEPIVORUM) OF ONIONS IN WORCESTERSHIRE WITH SPECIAL REFERENCE TO CONTROL BY SEED TREATMENT WITH CALOMEL

Observations in Worcestershire have shown that heavy losses of salad onions due to white rot may occur on sites which have not been known to carry a previous susceptible crop. Reasons for this are discussed. During five seasons a good control of the disease has been obtained by treating the seed with pure calomel applied with a resin-alcohol sticker. The treatment also gave some control of damping off of seedlings due to Corticium solani. Broadcasting 4 % calomel dust at 2½-3 cwt. per acre was less satisfactory in controlling white rot. In one season promising results were obtained with seed treatment of bulb onions.     

EXPERIMENTS ON THE CONTROL OF BOTRYTIS DISEASE OF LETTUCE

Experiments were made during 1955-57 on the fungicidal control of Botrytis attack on lettuce, cv. May Queen, matured in frames from November to April, and cv. Trocadero Improved, grown as seedlings in frames from November to March and then matured in the field from March to June. The frame lettuce were treated during their whole period of growth and the field lettuce in the seedling stage only.
On both types, 0.33% thiram sprays applied on up to five occasions gave the best control of disease: 5% tecnazene dust was effective on the frame crop, in 2 out of 3 years when applied to the plants and also when incorporated in the top soil at the rate of 15 g. per sq.yd. Tecnazene applications did not give a good control of the disease on the field crop. Sprays with 0.25% captan were almost as effective as the thiram treatments on the field lettuce but not on the frame crop.     

Conserving and Enhancing Biological Control of Nematodes

Conservation biological control is the modification of the environment or existing practices to protect and enhance antagonistic organisms to reduce damage from pests. This approach to biological control has received insufficient attention compared with inundative applications of microbial antagonists to control nematodes. This review provides examples of how production practices can enhance or diminish biological control of plant-parasitic nematodes and other soilborne pests. Antagonists of nematodes can be enhanced by providing supplementary food sources such as occurs when organic amendments are applied to soil. However, some organic amendments (e.g., manures and plants containing allelopathic compounds) can also be detrimental to nematode antagonists. Plant species and genotype can strongly influence the outcome of biological control. For instance, the susceptibility of the plant to the nematode can determine the effectiveness of control; good hosts will require greater levels of suppression than poor hosts. Plant genotype can also influence the degree of rhizosphere colonization and antibiotic production by antagonists, as well the expression of induced resistance by plants. Production practices such as crop rotation, fallow periods, tillage, and pesticide applications can directly disrupt populations of antagonistic organisms. These practices can also indirectly affect antagonists by reducing their primary nematode host. One of the challenges of conservation biological control is that practices intended to protect or enhance suppression of nematodes may not be effective in all field sites because they are dependent on indigenous antagonists. Ultimately, indicators will need to be identified, such as the presence of particular antagonists, which can guide decisions on where it is practical to use conservation biological control. Antagonists can also be applied to field sites in conjunction with conservation practices to improve the consistency, efficacy, and duration of biological control. In future research, greater use should be made of bioassays that measure nematode suppression because changes in abundance of particular antagonists may not affect biological control of plant parasites.     

CONTROL OF THE INTERNODAL INTERCALARY MERISTEM OF CYPERUS ALTERNIFOLIUS

In the growing culm of C. alternifolius, surgical removal of parts indicated that the stimulus for the prolonged activity of the internodal intercalary meristem (IM) came from the matured leaves and upper internode and that buds were not involved in maintaining internodal growth. Decapitation of the culm resulted in cessation of internodal extension. Various growth regulators were applied to the decapitated internode, and both the total extension and growth rates were analyzed statistically. Gibberellin A₃ (GA) and benzyladenine (BA) substituted for the excised parts in their effect on internodal extension. Indoleacetic acid (IAA) had little effect. (2-chloroethyl) trimethylammonium chloride (CCC) inhibited internodal growth, and its effects were reversed by GA. IAA was antagonistic to BA but not to GA. BA and GA were somewhat antagonistic. The quantitative effects of growth regulators on epidermal and ground parenchyma cell length and number of interstomatal cells were examined. Extension induced by GA was due to both cell division and cell elongation in the IM. Cells were longer, and fewer stomates differentiated than in the control. In internodes induced to extend by GA + BA cell division, cell length, and stomate differentiation were similar to the control. The results indicate that prolonged internodal IM activity is maintained by cytokinins and gibberellins coming from the matured upper portions of the culm. Changes in the levels of these regulators during growth presumably result in the histological gradient in the internode.     

Sclerotium cepivorum Berk. in onion (Allium cepa L.) crops: isolation and characterization of bacteria antagonistic to the fungus in Queensland

Bacteria antagonistic to the fungal pathogen Sclerotium cepivorum Berk., which causes onion white rot disease, were isolated and characterized. Eighty percent of the bacterial antagonists isolated from soil samples and sclerotia of S. cepivorum recovered from soil were identified as belonging to the genus Bacillus. The other antagonists were coryneforms and Pseudomonas spp. In contrast, 90% of the antagonists isolated from onion roots were mostly Pseudomonas spp. and the remainder were Erwinia spp. The significance of these antagonistic bacteria is discussed in relation to the control of white rot disease of onion.     

Cold tolerance in tomato. II. Early seedling growth of Lycopersicon spp.

Hypocotyl and root growth elongation of etiolated seedlings was measured non-destructively for the wild tomato accessions LA 460 ( Lycopersicon chilense Dun.), PI 126435, PI 127831 and PI 127832 ( L. peruvianum Mill.) and controls PI 120256 and T3 ( L. esculentum Mill.) on slant boards at 10, 15 and 20°C. Both hypocotyl and root elongation over time were fitted by a logistic growth function with three parameters estimated for each seedling by non-linear least squares regression. Analysis of variance of these equation parameters indicated linear decreases of both hypocotyl and root growth rate parameters with temperature. All four wild accessions maintained greater hypocotyl growth rate parameters at 10°C than the fast-germinating cultivated accession PI 120256, but not significantly greater than T3. Hypocotyl growth rates of the wild accessions were less inhibited at 10°C relative to 20°C than were either cultivated accession. These results suggest that these wild accessions have greater chilling tolerance than cultivated controls for early seedling growth, and may have potential use for genetically improving emergence times for tomatoes sown in cold soil.     

THE PRODUCTION OF ANTIBIOTICS IN SOIL: II. PRODUCTION OF GRISEOFULVIN BY PENICILLIUM NIGRICANS

A study has been made of the conditions affecting production of griseofulvin by Penicilliiim nigricans in two types of soil, an acid, sandy podsol from Wareham Heath and a garden soil. The characteristic morphogenetic response of many fungi to low concentrations of griseofulvin was made the basis of a highly specific bioassay.
The essential prerequisites for production of griseofulvin in either soil were sterilization and enrichment with organic matter; no griseofulvin could be detected in autoclaved soil which had not been supplemented or in normal soil even when organically enriched. Garden soil was a better medium for growth of P. nigricans and production of griseofulvin than Wareham soil although this soil could be improved in this respect by liming.
The yield of griseofulvin was decreased in soil re-infected by other soil organisms, particularly by some which were known to produce antifungal antibiotics, e.g. Penicilliunr expansum, P. frequentons and two strains of Trichoderma viride. The antagonism shown to Penicilliunz nigricuns was not entirely a matter of antibiotic activity, as some fungi believed not to produce antifungal substances had an antagonistic effect. These were mostly fungi with a characteristically rapid growth rate, e.g. Mucor rarnmannianus and one strain of Trichoderma riride. In some cases Penicillium nigricans was itself antagonistic to other fungi irrespective of their ability to produce antibiotics or of their fast-growing habit.
The results were compared with those obtained from a previous study of the soil conditions affecting the production of gliotoxin by Trichoderma viride. A higher level of nutrient was required for the production of griseofulvin, and the effect of antagonism by other soil micro-organisms was more important than in the production of gliotoxin by T. viride in the soil.     

Survival ofFrankia strains introduced into soil

Factors affecting the establishment of Alnus/Frankia symbioses were studied partly by following the survival ofFrankia strains exposed to different soil conditions, and partly by investigating the effect of pH on nodulation. TwoFrankia strains were used, both of the Sp⁻ type (sporangia not formed in nodules). One of the strains sporulated heavily, while the other formed mainly hyphae. The strains originated fromAlnus incana root nodules growing in soils of pH 3.5 and 5.0. The optimum pH for their growth in pure culture was found to be 6.7 and 6.2, respectively. The strains were introduced into twoFrankia-free soils, peat and fine sand. Their survival, measured as the persistance of nodulation capacity using the plant infection technique, was followed for 14 months. The survival curves of the strains were similar despite the morphological differences between the strains in pure culture. The nodulation capacities declined over time both at 14 and 22°C. Survival was better in soils limed to a pH above 6 than in soils at their original pH (peat 2.9, fine sand 4.2). The effect of pH on nodule formation in Alnus seedlings by theFrankia strains was studied in liquid culture. The number of nodules increased linearly within the pH range studied (3.5–5.8). No nodules were formed at pH 3.5.     

Characterization of a <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> strain from tomato rhizosphere and its use for integrated management of tomato damping-off

A highly antagonistic Pseudomonas fluorescens strain was isolated from tomato rhizosphere and characterized for its in vitro and in vivo biocontrol potential against Pythium aphanidermatum. The identified Pseudomonas fluorescens strain (PfT-8) was capable of producing high levels of chitinase, β-1,3-glucanase, cellulase, fungitoxic metabolites and siderophores. Seven different carrier formulations including a talc-based powder, lignite-based powder, peat-based powder, lignite + fly ash-based powder, wettable powder, bentonite-paste and polyethylene glycol (PEG) paste were prepared utilizing PfT-8. Shelf life was evaluated for up to 6 months of storage at ambient room temperature (28 °C). Biocontrol efficacy of formulations was studied under greenhouse and field conditions. The formulations were stable up to at least 2 months of storage at ambient room temperature. Among the formulations, peat, lignite, lignite+fly-ash and bentonite paste based formulations maintained higher propagule number than others and also showed greater biocontrol potential. However, propagule number gradually decreased with time. Several organic amendments including farm yard manure (FYM), leaf-compost, poultry manure, press mud, vermi-compost and neem cake were incorporated into soil to study their influence on P. fluorescens colonization in the rhizosphere and on potential disease control. Soil incorporation of organic amendments and specifically poultry manure and FYM, significantly reduced damping-off incidence and also augmented the rhizosphere population of the marked␣P.␣fluorescens strain that was resistant to streptomycin and rifampicin. An integrated␣approach of damping-off management employing dual inoculation of PfT-8 in seed and soil coupled with soil application of organic amendments including poultry manure or␣FYM was evaluated under field conditions. Under these conditions, damping-off incidence substantially reduced by up to 90% and further the healthy plant stand, plant biomass and plant rhizosphere population of P. fluorescens increased significantly.     

Effects of Temperature on Germination of Peronospora parasitica Conidia and Infection of Brassica oleracea

The effect of temperature on germination of a South African isolate of Peronospora parasitica , and infection of Brassica oleracea was studied. The optimum condition for germination was 20° C at 100% relative humidity. The percentage germination obtained was 80–98% and 70–80% between 15 and 25° C at 100% relative humidity, after a 12 and 6h incubation period, respectively. Optimum temperature for germ tube growth was also 20° C. The temperature range for maximum infection of seedlings of a highly susceptible cabbage cultivar and subsequent disease development in vitro was 15–25° C and 90–100% infection was achieved after 48 h of incubation. At<15°C and 26–30° C infection percentage was decreased to 40–50% and 35–40%, respectively. No disease incidence was recorded at temperatures above 35° C. A scanning electron microscope study of the infection process showed that penetration of cotyledons by germ tubes was mostly via stomata and occasionally directly through the cuticle. Results are discussed in relation to the need for future studies of P. parasitica in South Africa.     

Influence of soil factors,fertilizers and manures on pathogenicity ofRhizoctonia solani onVigna species

Rhizoctonia solani caused maximum mortality of mung bean seedlings at 20°C, and the disease incidence decreased with increase of temperature; 30° was optimum for mycelial growth of the fungusin vitro. The fungus grew best in nutrient broth of pH 5.5 but infected mung bean and pea seedlings more severely in neutral and alkaline river sand than in the sand adjusted to acidic reaction. The disease incidence was higher in adequately moist sandy loam and less in soil under moisture stress. Incidence of cowpea seedling rot was higher in heavy-textured loam and silt loam soils than in light-textured sandy- and loamy sand. Addition of montmorillonite and kaolinite in the sandy soil increased the disease incidence, but these clays reduced fungus growth in culture. More seedling rot occurred in the sandy soil fertilized with urea, potassium nitrate, monocalcium phosphate, or potassium dihydrogen phosphate while soil application of ammonium nitrate, potassium chloride, or potassium sulphate decreased the disease. In tests with combined soil application of N (as urea), P (as monocalcium phosphate) and K (as potassium chloride), disease incidence was more in all combinations having P. Among the six micronutrients tested, only boron reduced the disease incidence significantly both in presence and absence of NPK fertilizers. Farm-yard manure and biogas sludge aggravated seedling rot but their water extracts decreased it. Humic acid, extracted from farm-yard manure, increased the disease incidence but was inhibitory to fungus growth in culture. Green manure also resulted in more disease.