Evaluation of Curvularia lunata Strain B6 as a Potential Mycoherbicide to Control Barnydrgrass (Echinochloa crus-galli)

The virulent strain B6 of Curvularia lunata was screened out from 65 fungus strains isolated from the diseased leaves of barnyardgrass （Echinochloa crus-galli）. Greenhouse and field studies were conducted to evaluate the feasibility of the strain being exploited as a mycoherbicide for barnyardgrass control in paddy fields. The results of pathogenicity experiments showed that this strain was highly pathogenic to barnyardgrass at the 1- to 2.5-leaf stages. The fresh weight reduction increased with the increase of inoculated conidial concentrations and the prolongation of dew duration. Strain B6 provided excellent barnyardgrass control when it was applied at the concentration of 1×104-1×106 conidia mL-1 in paddy fields with water layer. This strain was very safe to rice and the most plant species except wheat, barley and corn. Findings of this study indicated that this strain could be a potential mycoherbicide for barnyardgrass control in paddy fields in the future.

Extending the Shelf-Life of <i>Myrothecium verrucaria</i>, a Bioherbicide

The shelf-life of a bioherbicide product is an important factor with regard to its commercial potential. The bioherbicidal efficacy of freshly fermented Myrothecium verrucaria (strain IMI 368023) (MV) mycelia formulations and MV mycelia preparations that had been freeze-dried and then stored at -20&deg;C for 8 years was compared. Two concentrations of each formulation (1.0x and 0.5x) were tested, utilizing bioassays on seedlings of the weed, hemp sesbania (Sesbania exaltata) under greenhouse conditions or in darkness utilizing hydroponically grown seedlings. Freeze drying of freshly prepared MV mycelium produced a light, brownish-colored powder. Efficacy tests of this reconstituted 8-year-old dried material showed that some bioherbicidal activity was lost during long-term storage, i.e., ~20% and ~60% seedling dry weight reduction at the 1.0x and 0.5x rate, respectively. Although plant mortality was greater in the fresh mycelial preparations treatments versus the freeze-dried and stored samples at all time points in the time-course, the stored material still caused >80% mortality, 15 days after treatment. Comparative disease progression ratings also showed a similar trend. Overall results show that freeze-drying MV is a useful method to reduce the bulk and cumbersomeness of storing heavy liquid fermentation product, while retaining bioherbicidal activity. These findings increase the utility of this bioherbicide and offer the potential to use the dried material in soil treatments or in a more concentrated form than attainable via the fermented product.

Biological Control of the Weed Sesbania exaltata Using a Microsclerotia Formulation of the Bioherbicide <i>Colletotrichum truncatum</i>

Colletotrichum truncatum, grown on rice grain (3 to 4 weeks, 22°C to 24°C) produced a fungus-infested rice mixture of microsclerotia and conidia (spores) in a ratio of ~9:1, respectively. Greenhouse tests of this formulation (0.4 to 50 mg finely-ground fungus-rice product) which applied pre-emergence to 5 cm2 of soil surface, caused 22% to 96% hemp sesbania plant mortality, after 14 days. Post-emergence treatment (fungus-rice aqueous formulation;2.4 × 105 microsclerotia ml-1, 30% unrefined corn oil and 0.2% Silwet L-77 surfactant) of weeds surviving the pre-emergence application, resulted in 93% mortality, after 14 days. Based on greenhouse results, field tests were undertaken: 1) pre-emergence treatment (fungus-rice formulation at 2.4 × 105 microsclerotia cm-2), 2) post-emergence (fungus-rice product in 30% unrefined corn oil, 0.2% Silwet) only treatment, applied 15 days after planting and 3) pre-emergence treatment followed by post-emergence treatment (fungus-rice product in 30% unrefined corn oil, 0.2% Silwet) applied 15 days after planting to surviving weeds. Control treatments were: 1) autoclaved rice product sans fungus, 2) unrefined corn oil (30% unrefined corn oil, 0.2% Silwet in water) and 3) untreated plants. Planting dates were: early season (April-May), early-mid season (June-July), late-mid season (July-August), and late season (September-October). Weed mortality was recorded at 15 days for the pre-plus post-treatment, and at 30 days after planting for the pre-emergence only and the post-treatment only. The early season, pre-emergence treatment caused 67% hemp sesbania mortality (3-yr average) within 15 days and the post-emergence treatment caused 91% mortality of the surviving weeds. In the late-mid-season, pre-emergence treatment caused minimal (<5%) mortality at 15 days, but mortality in the post-emergence treatment was >80%. Results suggest that seasonal environmental conditions are important in the efficacy of this C. truncatum-rice product formulation when applied pre- or post-emergence to this onerous weed.

Biological Control Potential of <i>Colletotrichum gloeosporioides</i>for Coffee Senna (<i>Cassia occidentalis</i>)

A fungal pathogen, Colletotrichum gloeosporioides was isolated from a greenhouse-grown seedling of coffee senna (Cassia occidentalis) and evaluated as a mycoherbicide for that weed. Host range tests revealed that coffee senna, wild senna (C. marilandica), and sicklepod (C. obtusifolia) were also affected by this pathogen, but 35 other crop and weed species, representing 8 botanical families were not affected. The fungus sporulated prolifically on solid and liquid media with maximum spore germination and growth occurring at 20°C - 30°C. Optimal environmental conditions included at least 12 h of free moisture (dew) at 20°C - 30°C. Spray mixtures containing approximately 1.0 × 105 or more conidia·ml–1 gave maximum control when coffee senna seedlings were sprayed until runoff occurred. Coffee senna seedlings that were in the cotyledon to first-leaf growth stage were most susceptible to this pathogen. Weed control efficacy studies under field conditions demonstrated that control of coffee senna was directly proportional to the inoculum concentration applied. Results of these tests suggest that this fungus has potential as a mycoherbicide to control coffee senna, a serious weed in the southeastern U.S.

Hot Water Treatment Enhances the Bioherbicidal Efficacy of a Fungus

Hemp sesbania (Sesbania exaltata) plants (>30 cm tall) sprayed with hot water (45&deg;C – 95&deg;C), followed by spray applications of fungal spores of Colletotrichum truncatum (CT) at 1.0 × 107 spores/ml-1 and 22&deg;C – 25&deg;C, suspended either in: 0.2% Silwet L-77 surfactant (SW);unrefined corn oil (CO)/distilled water (1:1, v:v);or 0.2% SW in CO were controlled by 80% - 95%, 12 days after treatment (DAT) under greenhouse conditions. These treatments also reduced dry weight accumulation of this weed. Plants treated with hot water without CT were also injured at temperatures ≥35&deg;C (5% mortality), and 60% mortality at 95&deg;C. Artificial dew treatments (25&deg;C, 12 h), imposed on plants after the treatment protocols above, had little or no effect on weed mortality or dry weight reduction compared to treated plants without dew. Under field conditions, 85% control of hemp sesbania was achieved 12 - 15 DAT when a pre-treatment with hot water (65&deg;C) was followed immediately with a CT application at the spore concentration as described above. Plants in field tests treated with CT without a hot water treatment were visually unaffected, with no mortality or plant biomass reductions recorded 15 DAT. These results suggest that use of hot water may be an important tool for improving the infectivity and bioherbicidal potential of some plant pathogens.