Extending the host range of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene

Spore formulations of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene (ATCC No. 20358) (CGA) were evaluated for control of three weed species: northern jointvetch (Aeschynomene virginica), Indian jointvetch (A. indica), and hemp sesbania (Sesbania exaltata) in greenhouse experiments. Mortality, dry weight reduction, and plant height reduction of A. virginica seedlings ranged from 98% to 100%, 15 days after inoculation with CGA in water, in an invert emulsion, or in Silwet L-77 surfactant. However, CGA in water caused no effects of these parameters on S. exaltata, and only slight effects on A. indica. A. indica and S. exaltata were also severely injured (mortality, dry weight and plant height reduction, 98–100%) by CGA in the invert emulsion or in Silwet L-77. The CGA in Silwet formulation incited severe disease development more rapidly than the invert emulsion or water formulations of CGA in all species. These results suggest that the host range of CGA can be expanded though formulation modification to enable this bioherbicide to control multiple weeds, thus making this product more economically acceptable.     

Adjuvants enhance the biological control potential of an isolate of Colletotrichum gloeosporioides for biological control of sicklepod (Senna obtusifolia)

In greenhouse experiments, unrefined corn oil, Silwet L-77, and an invert emulsion were tested as adjuvants with the mycoherbicidal fungus Colletotrichum gloeosporioides, a weakly virulent pathogen of sicklepod (Senna obtusifolia). A 1:1 (v/v) fungus/corn oil tank mixture containing 0.2% (v/v) Silwet L-77 surfactant reduced the dew period requirements for maximum weed infection and mortality from 16 to 8 h, and delayed the need for free moisture for greater than 48 h. This formulation also resulted in the ability of the pathogen to infect and kill weeds in larger (>5 leaf) growth stages. The invert emulsion resulted in similar effects upon these parameters. These results suggest that invert emulsions, unrefined corn oil and Silwet L-77 surfactant greatly improve the bioherbicidal potential of this pathogen for control of sicklepod, a serious weed pest in the southeastern US.     

Effect of row spacing on biological control of sicklepod (Senna obtusifolia) with Colletotrichum gloeosporioides

In field experiments conducted over 3 years, the mycoherbicidal fungus Colletotrichum gloeosporioides, formulated either in 20% (v/v) unrefined corn oil and 0.2% Silwet L-77 surfactant or with an invert emulsion, provided season-long control of Senna obtusifolia in narrow (51 cm) rows of soybean. However, in wide (102 cm) rows, one application of either formulation failed to provide season-long control of S. obtusifolia, and two applications were required to achieve season-long weed control. In narrow (51 cm) rows, one application of the fungus either in unrefined corn oil or an invert emulsion controlled S. obtusifolia an average of >90%, and a second application was not required for season long weed control. Soybean yields in wide-row plots treated with two applications of either the fungus/corn oil or fungus/invert emulsion, or with a single application of the fungal treatments in narrow-row soybean plots, were not significantly different from weed-free control plots, or from plots treated with the herbicide chlorimuron. These results suggest that row spacing can affect mycoherbicidal efficacy of this fungus for controlling S. obtusifolia.     

Bioherbicidal enhancement and host range expansion of a mycoherbicidal fungus via formulation approaches

Eastern black nightshade (Solanum ptycanthum; EBN) is a problematic weed partly due to its tolerance or resistance to certain herbicides. We examined the effects of an invert emulsion (IE) on the host range and weed control efficacy of the fungus Colletotrichum coccodes (NRRL strain 15547) for biocontrol of EBN. Greenhouse tests demonstrated that several other solanaceous weeds were also infected and killed, and field tests revealed >90% EBN control and dry weight reduction in plants treated with the fungus-IE formulation. These results demonstrate that this IE formulation can promote the efficacy of this bioherbicidal pathogen.     

Interactions of two selected field isolates of Colletotrichum gloeosporioides f. sp. aeschynomene on Aeschynomene virginica

The fungal plant pathogen, Colletotrichum gloeosporioides f. sp. aeschynomene (CGA), has been used successfully since 1982 to control northern jointvetch (NJV), Aeschynomene virginica, in rice and soybeans in the USA as COLLEGO^®, a bioherbicidal formulation of a single isolate of this pathogen. The interactions and fitness of different field isolates of this fungus and effective bioherbicide have not been fully examined. We examined CGA population structure and inter-isolate competition using molecular markers in field and greenhouse experiments. NJV plants were first inoculated with one isolate or a mixture of two isolates, Cla-5a and 3-1-3, followed by a challenge inoculation containing either one or both isolates, 4 or 5 days later. Plants first inoculated with Cla-5a alone, or in mixture with 3-1-3, had Cla-5a comprising approximately 80% of the population after 21 days regardless of challenge inoculation. In contrast, when the primary treatment was 3-1-3 alone, wounding, or benzodiathiazole, 3-1-3 detection only approached 50% of the population following challenge inoculation. Primary inoculation with Cla-5a significantly reduced the number of lesions caused by 3-1-3 challenge, whereas primary inoculation with 3-1-3 did not significantly affect the number of lesions caused by Cla-5a challenge. Size of lesions caused by the two isolates following challenge inoculations and dispersal of the isolates from lesions to healthy plants were not affected by the primary inoculation. These results suggest a specific interaction by Cla-5a with the host defense response creating a more favorable environment for its own colonization of NJV to the exclusion of 3-1-3.     

CgOpt1, a putative oligopeptide transporter from Colletotrichum gloeosporioides that is involved in responses to auxin and pathogeniCity

Background  

The fungus Colletotrichum gloeosporioides f. sp. aeschynomene produces high levels of indole-3-acetic acid (IAA) in axenic cultures and during plant infection. We generated a suppression subtractive hybridization library enriched for IAA-induced genes and identified a clone, which was highly expressed in IAA-containing medium.     

Indole-3-Acetic Acid Biosynthesis in Colletotrichum gloeosporioides f. sp. aeschynomene

We characterized the biosynthesis of indole-3-acetic acid by the mycoherbicide Colletotrichum gloeosporioides f. sp. aeschynomene. Auxin production was tryptophan dependent. Compounds from the indole-3-acetamide and indole-3-pyruvic acid pathways were detected in culture filtrates. Feeding experiments and in vitro assay confirmed the presence of both pathways. Indole-3-acetamide was the major pathway utilized by the fungus to produce indole-3-acetic acid in culture.     

Induction of infection in Sesbania exaltata by Colletotrichum gloeosporioides f. sp. aeschynomene formulated in an invert emulsion

World Journal of Microbiology and Biotechnology - In greenhouse experiments, an invert emulsion (MSG 8.25) was tested with spores of the mycoherbicidal fungus Colletotrichum gloeosporioides f. sp....     

Biological control of alligatorweed (Alternanthera philoxeroides) with a Fusarium sp.

Alligatorweed (Alternanthera philoxeroidesG.) has become a serious weed in different crops in China. A fungal pathogen was found in Chongqing and Sichuan Provinces and was identified as a species in the Fusarium genus. The fungus produced macroconidia and chlamydospores abundantly on potato sucrose agar (PSA) plates. The bestconidial production and germination and colonygrowth of Fusarium sp. were at 23–31°C and pH 6.7–7.0. Light period and flooding did not affect fungal growth and conidium formation. The herbicides, glyphosate and paraquat, inhibited the fungal development in vitro. The fungus did not affect seed germination and seedling growth of paddy rice, wheat, maize, oilseed rape and broad bean inlaboratory or greenhouse trials. Inoculum density and wetness duration influenced the efficiency of Fusarium sp. to control alligatorweed; a concentration of 1.0 × 10⁵ spores^–1 ml and 12 h of high humidity duration after inoculation produced goodinfections on the weed at 23°C in the laboratory. When the fungus was applied to alligatorweed grown in greenhouse and in the field, good biocontrol efficiency was obtained: the plants started to wilt after four to five (greenhouse) or six days (field), and were killed 9–10 (greenhouse) or 13–14 (field) days after spraying the fungal inoculum. This was similar to the control efficiency resulting from glyphosate treatment. Therefore, this Fusarium sp. appeared to be a good candidatefor further studies and a promising biocontrol agent to manage alligatorweed in some terrestrial and aquatic crops.     

In Planta Production of Indole-3-Acetic Acid by Colletotrichum gloeosporioides f. sp. aeschynomene

The plant pathogenic fungus Colletotrichum gloeosporioides f. sp. aeschynomene utilizes external tryptophan to produce indole-3-acetic acid (IAA) through the intermediate indole-3-acetamide (IAM). We studied the effects of tryptophan, IAA, and IAM on IAA biosynthesis in fungal axenic cultures and on in planta IAA production by the fungus. IAA biosynthesis was strictly dependent on external tryptophan and was enhanced by tryptophan and IAM. The fungus produced IAM and IAA in planta during the biotrophic and necrotrophic phases of infection. The amounts of IAA produced per fungal biomass were highest during the biotrophic phase. IAA production by this plant pathogen might be important during early stages of plant colonization.     

Biological control of anthracnose (Colletotrichum gloeosporioides) in yam by Streptomyces sp.MJM5763

Aim: To find a suitable biocontrol agent for yam anthracnose caused by Colletotrichum gloeosporioides. Methods and Results: An actinobacterial strain, MJM5763, showing strong antifungal activity, multiple biocontrol and plant growth‐promoting traits was isolated from a yam cultivation field in Yeoju, South Korea. Based on morphological and physiological characteristics and analysis of the 16S rDNA sequence, strain MJM5763 was identified as a novel strain of Streptomyces and was designated as Streptomyces sp. MJM5763. Treatment with MJM5763 and the crude culture filtrate extract (CCFE) was effective in suppressing anthracnose in detached yam leaves in vitro and reduced incidence and severity of anthracnose in yam plants under greenhouse conditions. The CCFE treatment was the most effective of all the treatments and reduced the anthracnose severity by 85–88% and the incidence by 79–81%, 90 days after inoculation with the pathogen. CCFE treatment was also effective under field conditions and showed a reduction of 86 and 75% of anthracnose severity and incidence, respectively. Conclusion: Streptomyces sp. strain MJM5763 was effective in biocontrolling anthracnose in yam caused by C. gloeosporioides. Significance and Impact of the Study: Streptomyces sp. MJM5763 is a potential alternative to chemical fungicides for reducing yield losses to anthracnose in yam.     

Soybean and sunflower oils increase the infectivity of Colletotrichum gloeosporioides f. sp. aeschynomene to Northern Jointvetch

Soybean and sunflower oils increased the level of infection of northern jointvetch, Aeschynomene virginica, plants by Colletotrichum gloeosporioides f. sp. aeschynomene. Inoculation of seedlings with spore suspensions containing 10% (v:v) soybean oil or 10% sunflower oil resulted in more disease than when inoculated with suspensions of spores in water alone. The lengths of the dew periods required to establish equivalent levels of disease by spore suspensions containing 10% soybean or 10% sunflower oil were approximately 4–8 h less compared to aqueous suspensions. Incubation of spores in 10% soybean oil followed by removal and resuspension in water did not affect the infectivity of spores when compared to spores incubated in aqueous suspensions. Spore germination and appressoria formation were unaffected by either of the oils tested in in vitro assays; however, in in vivo assays, 10% soybean oil and 10% sunflower oil increased spore germination in comparison to spores that were suspended in water.     

<Emphasis Type="Italic">Sesbania exaltata</Emphasis> biocontrol with <Emphasis Type="Italic">Colletotrichum truncatum</Emphasis> microsclerotia formulated in ‘Pesta’ granules

The weed Sesbania exaltata (Raf.) Rydb. ex A.W. Hill (hemp sesbania) was effectively controlled in soybean [Glycine max (L.) Merr.] field test plots over a 2-year testing period (1995–1996) with microsclerotia of the bioherbicidal fungus, Colletotrichum truncatum, formulated in wheat gluten-kaolin granules called ‘Pesta’. Weed control averaged 84% and 88%, respectively, in plots treated pre-plant incorporated (PPI) at ‘Pesta’ rates of 168 or 336 kg ha⁻¹, and 71% and 78%, respectively, in plots treated pre-emergence (PE) at ‘Pesta’ rates of 168 or 336 kg ha⁻¹ over the testing period. Post-emergence (POE) control averaged 30% and 50%, respectively, for the 168 and 336 kg ha⁻¹ treatments, and was significantly less effective than either PE or PPI treatments. Although pathogenesis and mortality occurred in hemp sesbania tissues, satisfactory weed control was not achieved in plots treated at rates of 17 or 84 kg ha⁻¹ with any of the application methods. Soybean yields were significantly greater in test plots treated PPI or PE, as compared to yields from test plots treated either POE, with inert ‘Pesta’ granules, or from untreated controls. Microsclerotia formulated in ‘Pesta’ granules exhibited excellent shelf-life, retaining high viability after storage for 10 years at 4 °C. These results suggest that microsclerotia of C. truncatum formulated in ‘Pesta’ granules offers an effective method for controlling this important weed and preserving the activity of this bioherbicide.     

Bioherbicidal activity from washed spores of <Emphasis Type="Italic">Myrothecium verrucaria</Emphasis>

The fungal plant pathogen, Myrothecium verrucaria, is highly virulent to several important weed species and has potential utility as a bioherbicide. However the production of macrocyclic trichothecene mycotoxins by this fungus presents significant safety concerns. It was discovered that trichothecenes are removed from M. verrucaria spores by repeated washes with water. These washed spores retained bioherbicidal efficacy against kudzu when tested in field trials and on sicklepod when tested under greenhouse conditions. Changes in the growth medium combined with washing spores with water resulted in greater than 95% reduction in roridin A and verrucarin A. Washing spores reduced trichothecene concentrations in spore preparations with no significant effect on plant biomass reduction, thus demonstrating the possibility of M. verrucaria formulations with improved safety to researchers, producers and applicators.     

Biological and genetic characterisation of Phoma macrostoma isolates with bioherbicidal activity

The fungus Phoma macrostoma Mont. isolate 94-44B was registered as a bioherbicide for control of broadleaved weeds in Canada and the USA in 2011 and 2012, respectively. To obtain the registrations, the fungus had to be characterised both biologically and genetically. The objectives of this study were to demonstrate that bioherbicidal activity was associated with specific genetic markers and to determine whether bioherbicidal activity was a general trait of the species or only selected isolates. A collection of 64 isolates of P. macrostoma was established. A greenhouse bioassay and bioherbicidal-specific primers were used to determine bioherbicidal activity of all isolates. Only isolates originating from Canada thistle demonstrated the ability to reduce dandelion seedlings and display the 853 bp amplicon for the bioherbicidal-specific primer. Bioherbicidal isolates were consistently differentiated from all other isolates with two main genotypic groupings (I and II) arising from internal transcribed spacer (ITS) and amplified fragment length polymorphisms (AFLP) sequence analyses. Using AFLP, two biotypes of bioherbicidal isolates were also differentiated by the presence or absence of an AFLP marker at a single polymorphic locus. The genetic divergence among the bioherbicidal and nonbioherbicidal isolates of P. macrostoma was only 2.21% which was lower than that reported for other related Phoma sp. Other than the bioherbicidal trait, there was no apparent affiliation of the genetics with known varietal types, host or geographic origin. ITS sequence analysis and AFLP fingerprinting may be used as tools to detect bioherbicidal isolates of P. macrostoma.     

In vitro,greenhouse and field assessments of cassava lines for resistance to anthracnose disease caused by Colletotrichum gloeosporioides f.sp. manihotis

Fifty-three cassava lines were selected from breeding populations at the International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria and screened in vitro for resistance to cassava anthracnose disease (CAD). The in vitro inoculation of stem cuttings with the fungus Colletotrichum gloeosporioides f.sp. manihotis showed significant differences (p ± 0.05) in acervuli production and in the sensitivity of the cassava lines to the fungal infection after 7 days of incubation at 25 °C. Cassava lines 88/01084, 91/00595, 91/00475, 91/00344, 91/00684, 91/00313, 91/00422, and 91/00344 were highly resistant, with necrotic lesion sizes less than 7 mm. In contrast pedigree lines 88/02549, 89/0008, 91/00390 and 91/00402 were highly susceptible with the largest necrotic lesion size being greater than 20 mm. Ten cassava lines from the in vitro screening that showed varying levels of resistance to CAD were selected, based on their flowering abilities for diallel hydridization trials, and were further screened in greenhouse and field trials for CAD resistance. The greenhouse and field screening showed significant varietal differences (p ± 0.05) in sensitivity to the fungus. In all cases, the progeny lines showed correlated levels of resistance irrespective of the type of screening or assessments. Correlation analysis of the in vitro, greenhouse and field assessments showed that there was a good correspondence among all three methods of evaluating for CAD. This revised version was published online in June 2006 with corrections to the Cover Date.     

Assessment of potential antagonists for anthracnose (Colletotrichum gloeosporioides) disease of mango (Mangifera indica L.) in North Western Ethiopia (Pawe)

Mango (Mangifera indica L.) is considered as one of the most popular fruits among millions of people in the tropical area and increasingly in the developed countries. Anthracnose, caused by the fungus Colletotrichum gloeosporioides, is the most important pre- and post-harvest disease of mango. The objective of this research was to evaluate the prevalence of different promising antagonistic Trichoderma and Bacillus spp. on phyloplane of mango in Ethiopia and to evaluate their antagonistic potential against the pathogen. A total of 19 mango fields were surveyed and anthracnose affected all fields. Culture studies on potato dextrose agar for evaluation of antibiosis activity of Trichoderma spp. and Bacillus spp. revealed that they have inhibitory and lytic effect on C. gloeosporioides, which is an indication of their potential biocontrol agent for management of mango anthracnose as an alternative to chemical control. Significant differences (p?<?0.05) were observed among Bacillus isolates in causing lysis of pathogen mycelium, when inoculated on actively growing colony of C. gloeosporioides. Maximum reduction in growth rate of pathogen was observed with Bacillus spp. (B50), which restricted the growth to 2.7?mm compared to 8.3?mm in the control with 67.5% efficacies. There were similar effects (p?<?0.05) among Trichoderma spp. in formation of inhibition zones and lysis by varying degrees up to 59.7% efficacies in reducing linear growth of the pathogen in dual culture.     

Fungus gnats vector Fusarium oxysporum f.sp. radicislycopersici1

Fungus gnat adults transported Fusarium oxysporum f.sp. radicis-lycopersici from Petri dish culture and infected host plants to the roots and hypocotyls of healthy tomato and bean plants. The source of the fungus did not affect the ability of fungus gnats to transport the fungus to healthy hosts. The presence of fungus gnat larvae in media in which young tomato plants were grown did not increase the incidence of plant infection by the pathogen. Fungus gnat adults appear to aid in the dissemination of F. oxysporum f.sp. radicis-lycopersici.     

Feeding and maturation by soybean looper (Lepidoptera: Noctuidae) larvae on soybean affected by weed, fungus, and nematode pests

Feeding and maturation by the soybean looper, Pseudoplusia includens (Walker) (Lepidoptera: Noctuidae), were investigated in a 2-yr study on 'Davis' soybean, Glycine max (L.) Merr., grown alone and combined with the weed hemp sesbania, Sesbania exaltata (Raf.) Rybd. ex. A. W. Hill, the root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, and the charcoal rot fungus, Macrophomina phaseolina (Tassi) Goid. Of the three pests, hemp sesbania had the greatest effects on plant growth and insect feeding and maturation. When fed foliage from soybean stressed by hemp sesbania, soybean looper larvae remained longer in feeding stages, consumed more foliage, and showed altered weight gain compared with larvae fed control foliage. Results suggest that nutrient (s) critical for proper development of larvae may have been limited in weed-stressed soybean foliage. Less dramatic results were observed when larvae fed on foliage from soybean with roots colonized by the charcoal rot fungus. Such larvae consumed more foliage, weighed more, and showed a slight increase in larval feeding period, but only in 1 yr of the study. Colonization of soybean roots by the root-knot nematode had no consistent effects on either the soybean host or insect.     

Larval cannibalism in aphidophagous ladybirds: Influencing factors, benefits and costs

Spray retention on scentless chamomile and round-leaved mallow was characterized in relation to droplet size and travel speed. The effect of varying retention on biocontrol efficacy was studied using Colletotrichum truncatum and Colletotrichum gloeosporioides f. sp. malvae, the bioherbicide agents for the two weeds, respectively. In retention studies, a tracer dye solution was applied using a cabinet sprayer fitted with a fine, medium, or coarse nozzle at travel speeds of 0.5, 1.0, and 2.0 km/h that resulted in approximate application volumes of 500–2000 L/ha. Retention efficiency, a ratio between the volume retained on plants and actual volume applied, was calculated for each application. In general, finer sprays achieved higher retention efficiency on whole plants of both weeds. On an average, fine sprays resulted in 68% greater retention than coarse sprays on scentless chamomile and 22–59% on round-leaved mallow. Faster travel speeds increased spray retention only slightly. Applying the biocontrol agents at 500 L/ha with different droplet sizes showed different effects on weed control on scentless chamomile and round-leaved mallow. C. truncatum applied with fine droplets was more efficacious than treatments with coarser droplets on scentless chamomile. On round-leaved mallow, however, the efficacy of C. gloeosporioides f. sp. malvae did not appear to be substantially affected by the droplet size. This varying effect may be due to reduced retention on the vertical stems of round-leaved mallow, which is critical to biocontrol of this weed by C. gloeosporioides f.sp. malvae.