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Interaction of the bioherbicide Myrothecium verrucariawith Technical-Grade Glyphosate on Glyphosate-Susceptible and -Resistant Palmer Amaranth

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American Journal of Plant Sciences 2018年第11期9卷 2306-2319页

作者： Robert E. Hoagland C. Douglas Boyette Robin H. Jordan Kenneth C. Stetina Crop Production Systems Research Unit USDA-ARS Stoneville MS USA Biological Control of Pests Research Unit USDA-ARS Stoneville MS USA

Previously we found that a strain of Myrothecium verrucaria (MV) exhibited bioherbicidal activity against several important weeds, and that some commercial formulations of glyphosate applied with MV resulted in synergistic interactions that improved weed control efficacy. We also found that MV had bioherbicidal activity against glyphosate-resistant Palmer amaranth. We have also reported that some commercial formulations are inhibitory to MV. Our objectives were to test the effect of unformulated glyphosate (high purity, technical-grade glyphosate) alone and in combination with MV for bioherbicidal activity on glyphosate-susceptible and -resistant Palmer amaranth biotypes under greenhouse conditions and to examine technical-grade glyphosate on the growth of this bioherbicide. High purity glyphosate (without adjuvants/surfactants) was not toxic to MV growth and sporulation at concentrations up to 2.0 mM when grown on agar supplemented with the herbicide. Both biotypes were injured by MV and MV plus glyphosate treatments as early as 19 h after application (3 h after a dew period of 16 h). These injury effects increased and were more evident through the 6-day time course, when after 120 h the MV plus glyphosate treatment had killed all glyphosate-susceptible and -resistant plants. The interaction of glyphosate plus MV was synergistic toward the control of Palmer amaranth. Data strongly suggest that the active ingredient is responsible for the synergy previously found when this bioherbicide was combined with some commercial formulations of glyphosate. Results demonstrated that MV can control both glyphosate-resistant and -susceptible Palmer amaranth seedlings and act synergistically with high-purity glyphosate to provide improved weed control.

关键词： Amaranthus palmeri Biocontrol Agent bioherbicide Biological Weed Control Glyphosate-Resistance Myrothecium verrucaria Palmer Amaranth Pigweed

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Extending the Shelf-Life of Myrothecium verrucaria, a bioherbicide

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American Journal of Plant Sciences 2017年第12期8卷 3272-3484页

作者： Robert E. Hoagland C. D. Boyette K. C. Stetina United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Crop Production Systems Research Unit Stoneville MS USA United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Biological Control of Pests Research Unit Stoneville MS USA

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°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.

关键词： bioherbicide Plant Pathogen Weed Control Lyophilization Freeze-Drying Mycelium Mycoherbicide Myrothecium verrucaria

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Biological Control of the Weed Sesbania exaltata Using a Microsclerotia Formulation of the bioherbicide Colletotrichum truncatum

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American Journal of Plant Sciences 2014年第18期5卷 2672-2685页

作者： Clyde D. Boyette Hamed K. Abbas Bobbie Johnson Robert E. Hoagland Mark A. Weaver United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Biological Control of Pests Research Unit Stoneville MS USA United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Crop Production Systems Research Unit Stoneville MS USA

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 (80%. Results suggest that seasonal environmental conditions are important in the efficacy of this C. truncatum-rice product formula

关键词： bioherbicide Biological Weed Control Colletotrichum truncatum Mycoherbicide Sesbania exaltata Microsclerotia Solid Growth Media Substrate

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Interaction of the bioherbicide Myrothecium verrucaria and Glyphosate for Kudzu Control

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American Journal of Plant Sciences 2014年第26期5卷 3943-3956页

作者： Clyde Douglas Boyette Robert E. Hoagland Mark A. Weaver Kenneth C. Stetina United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Biological Control of Pests Research Unit Stoneville MS USA United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Crop Production Systems Research Unit Stoneville MS USA

Kudzu is an exotic invasive weed in the southeastern U.S. that is difficult to control with current commercial herbicides. Some success for its control has been achieved using a bioherbicidal agent, Myrothecium verrucaria (MV). Spore and mycelial formulations of MV were tested alone and in combination with glyphosate for control of kudzu (Pueraria lobata) under greenhouse and field conditions in naturally-infested areas. In greenhouse and field experiments, kudzu control increased as the concentration of spores or mycelia increased. Glyphosate alone provided 10%, 35%, 50% and 60% control in field experiments at 0.25, 0.50, 0.75 and 1.0X rates, respectively and MV alone spores provided 15%, 50%, 65% and 85% control at 0.25, 0.50, 0.75 and 1.0X rates, respectively. However, when MV spores were combined with glyphosate, significantly higher control occurred than that caused by either component alone. Similar levels of control were observed for MV mycelial formulations applied alone or with glyphosate at equivalent concentrations of the fungus. The rate of disease progression was more rapid and severe at all fungal spore or mycelial formulations and herbicide rates when these propagules were applied in combination with glyphosate. In field tests, 24 h after application, only 20% of kudzu plants were severely damaged by MV alone (0.25X), whereas 80% were severely diseased when MV spores and glyphosate were mixed and applied at 0.25X rates each. A similar trend occurred with the MV mycelial formulation applied at these rates. Synergist interactions on kudzu control were observed, especially when lower levels of MV (spores or mycelia) and glyphosate were combined and applied to kudzu in the greenhouse or in the field. These results suggest that it may be possible to incorporate glyphosate to improve the bioherbicidal control potential and reduce herbicide and inoculum requirements of M. verrucaria spores or mycelium for controlling kudzu.

关键词： bioherbicide Glyphosate Kudzu Myrothecium verrucaria Pueraria lobata

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Interactions and Effects on Cysteine Synthase Activity of Aminooxyacetate and Boc-Aminooxyacetate on the bioherbicides Colletotrichum truncatumand Alternaria cassiaand Their Weed Hosts

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American Journal of Plant Sciences 2021年第5期12卷 759-770页

作者： Robert E. Hoagland Kangetsu Hirase C. Douglas Boyette Crop Production Systems Research Unit USDA-ARS Stoneville USA Functional Chemicals Laboratory Mitsui Chemicals Chiba Japan Biological Control of Pests Research Unit USDA-ARS Stoneville USA

Aminooxyacetate (AOA) is a pyridoxal phosphate antagonist that inhibits various plant enzymes (including transaminases) which require pyridoxal phosphate as a cofactor and it exhibits phytotoxic and herbicidal properties. We examined AOA and its analog, N-t-butoxycarbonyl-AOA (Boc-AOA) for phytotoxicity, interactions with weed pathogens (bioherbicides), and effects on an important pyridoxal requiring enzyme, cysteine synthase (CS, E.C. 4.***.99.8). Studies were performed on two weeds, i.e., hemp sesbania [Sesbania exaltata (Raf.) Rybd. Ex A.W. Hill] and sicklepod (Senna obtusifolia), and two pathogens, (Colletotrichum truncatum and Alternaria cassiae), that are bioherbicidal agents against hemp sesbania and sicklepod, respectively. Pathogenicity tests, and assays for extractable, and in vitro CS activities were utilized. Phytotoxicity bioassays indicated that the bulky t-butoxycarbonyl moiety substitution on the AOA molecule did not substantially hinder expression of biological activity of Boc-AOA in these tests. Generally, spray application of the compounds to young dark-grown seedlings caused little growth effects, but root-feeding of the chemicals reduced growth

关键词： Aminooxyacetate bioherbicide Cysteine Synthase Pyridoxal Phosphate Antagonist Sicklepod Senna obtusifolia Hemp Sesbania Sesbania exaltata Transaminase

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Isolation, Identification, and Herbicidal Activity of Metabolites Produced by Pseudomonas aeruginosa CB-4

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Journal of Integrative Agriculture 2014年第8期13卷 1719-1726页

作者： YANG Juan CAO Hong-zhe WANG Wei ZHANG Li-hui DONG Jin-gao Mycotoxin and Molecular Plant Pathology Laboratory Agricultural University of Hebei

CB-4, a bacterial strain with highly effective herbicidal activity, was isolated from infected corn leaves. Through morphology, physiological and biochemical tests, and 16 S ribosomal DNA gene sequencing methods, CB-4 was identified as Pseudomonas aeruginosa. We conducted activity-evaluation experiments in the laboratory to assess the herbicidal potential of metabolites produced by strain CB-4. Crude extracts of strain CB-4 have high inhibition activity on Digitaria sanguinalis. In general, the root and shoot growth parameters of D. sanguinalis were significantly reduced by metabolites of strain CB-4. The IC50 of the culture filtrate extracts for the radicula and coleoptile of D. sanguinalis were 0.299 and 0.210 mg mL-1, respectively. Component 2 of the herbicidal activity of the crude toxin from strain CB-4 was successfully purified for the first time by using high-speed counter current chromatography with a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water（4：5：4：5, v/v） and high-performance liquid chromatography. We concluded that the metabolites of strain CB-4 have the potential to be developed as a microbe-based herbicide.

关键词： Pseudomonas aeruginosa bioherbicide biocontrol HSCCC HPLC

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Isolation and identification of Serratia marcescens Ha1 and herbicidal activity of Ha1 ‘pesta’ granular formulation

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Journal of Integrative Agriculture 2015年第7期14卷 1348-1355页

作者： YANG Juan WANG Wei YANG Peng TAO Bu YANG Zheng ZHANG Li-hui DONG Jin-gao Mycotoxin and Molecular Plant Pathology Laboratory Agricultural University of Hebei

A total of 479 bacterial strains were isolated from brine （Bohai, Qinhuangdao City, Hebei Province, China）. Bioassay results indicated that 4 strains named Hal, Hal7, Ha38, and Ha384 had herbicidal activity. And strain Hal had the highest effective herbicidal activity. As a result, this study aims to JdentJfy strain Hal, characterize its physiological and biological activities, evaluate the herbicidal activity of its metabolites, and develop a ＇pesta＇ formulation and assess its effectiveness on Digitaria sanguinalis. Hal was identified as Serratia marcescens based on 16S rDNA sequencing. This strain has a flagellum, a diameter of 0.5 to 0.8 IJm, and a length of 0.9 to 2.0 IJm. The indole test shows positive results, and the catalase enzyme exhibits strong positive reactions. Results further showed that the inhibitory concentration （IC50） of the crude extracts to D. sanguinalis radicula and coleoptile were 3.332 and 2.828 mg mL-1, respectively. Both the suppression of D. sanguinalis and the cell viability of the Hal formulation in ＇pesta＇ were higher when stored at 4℃ than at （25＋2）℃. These results indi- cated that S. marcescens Hal can potentially be used as a biocontrol agent against D. sanguinalis.

关键词： Serratia marcescens bioherbicide biocontrol pesta

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The Allelopathic Effects of Sunflower and Wheat Root Exudates on Sinapis arvensis and Sinapis alba

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Phyton-International Journal of Experimental Botany 2019年第4期88卷 413-423页

作者： Bengu Turkyilmaz Unal Mustafa Bayram NigdeÖmer Halisdemir University Faculty of Arts and SciencesDepartment of Biotechnology51240NigdeTurkey NigdeÖmer Halisdemir University Faculty of Arts and SciencesDepartment of Biology51240NigdeTurkey

In this study,we aimed to investigate the allelopathic effects of sunflower and wheat root exudates on the common weeds such as wild mustard and white mustard in our *** root exudates which were obtained by soaking 8 weeks old sunflower and wheat seedlings(20 or 40 seedlings)in 100 mL of distilled water for 3 days were applied to the leaves of wild mustard and white *** order to compare the allelopathic effect,the recommended dose(1 ***^(-1))and twice the recommended dose(2 ***^(-1))of Gromstor(Tribenuron-methyl),a herbicide preferred by farmers for the chemical control of these weeds was also *** allelopathy was performed for wild mustard and white mustard seedlings by the measurement of different physiological and biochemical parameters,such as chlorophyll a,chlorophyll b,total chlorophyll,carotenoid,proline,total protein amounts and superoxide dismutase enzyme *** amounts of total chl and carotenoid in wild mustard leaves decreased in all treatment groups compared to *** highest decrease in total chl(50.93%)and carotenoid(46.69%)was oberved in the treatment of 40 wheat seedlings.100 mL^(-1) distilled *** the white mustard leaves,the amount of total chl in all treatment groups except the treatment group of Gromstor 2 ***^(-1) and carotenoid in all treatment groups increased compared to the *** highest increases again were observed in 40 wheat seedlings.100 mL^(-1) distilled water *** proline amounts in wild mustard and white mustard increased in all treatment *** highest increase was observed for the treatment of 20 wheat seedlings.100 mL^(-1) distilled water in wild mustard(459.69%)and 40 sunflower seedlings.100 mL^(-1) distilled water in white mustard plant(104.70%).In superoxide dismutase enzyme activities,treatments decreased activity except treatment of 40 sunflower seedling root exudate in wild mustard,while increased activity outside commercial herbicide treatment in white *** r

关键词： Allelopathy antioxidant enzymes bioherbicide photosynthetic pigments white mustard wild mustard

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Bioherbicidal Efficacy of a Myrothecium verrucaria-Sector on Several Plant Species

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American Journal of Plant Sciences 2016年第16期7卷 2376-2389页

作者： Robert E. Hoagland Clyde D. Boyette Kenneth C. Stetina Robin H. Jordan United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Crop Production Systems Research Unit Stoneville MS USA United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Biological Control of Pests Research Unit Stoneville MS USA

Comparative studies were conducted on mycelial preparations of the bioherbicide, Myrothecium verrucaria (MV) strain IMI 361690 and a recently discovered sector (MV-Sector BSH) of this fungus. The whitish sector was discovered, isolated, grown in pure culture on PDA and found to be a stable, non-spore producing mutant when cultured over several months under conditions that cause circadian sporulation during growth of its MV parent. Application of MV and MV-Sector BSH mycelial preparations to intact plants (hemp sesbania and sicklepod) and leaf discs (kudzu and glyphosate-resistant Palmer amaranth) showed that the sector efficacy was generally equal to, or slightly lower than MV. Bioassays of MV and this sector on seed germination and early growth of sicklepod and hemp sesbania seeds demonstrated that hemp sesbania seeds were slightly more sensitive to the fungus than sicklepod seeds and that the sector bioherbicidal activity was slightly less than that of MV. SDS-PAGE protein profiles of cellular extracts of MV and the sector and their respective culture supernatants showed several differences with respect to quantity and number of certain protein bands. Overall results showed that the isolate was a non-spore producing mutant with phytotoxicity to several weeds (including weeds tolerant or resistant to glyphosate), and that the phytotoxic effects were generally equivalent to those caused by MV treatment. Results of this first report of a non-sporulating MV mutant that suggest additional studies on protein analysis, and an extended weed host range under greenhouse and field conditions are needed in order to further evaluate its possible bioherbicidal potential.

关键词： bioherbicide Glyphosate-Resistance Palmer Amaranth Kudzu Hemp Sesbania Sicklepod Myrothecium verrucaria Fungal Sector

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Effects of Myrothecium verrucariaon Ultrastructural Integrity of Kudzu (Pueraria montana var. lobata) and Phytotoxin Implications

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American Journal of Plant Sciences 2012年第11期3卷 1513-1519页

作者： Robert E. Hoagland C. Douglas Boyette Kevin C. Vaughn Neal D. Teaster Ken Stetina United States Department of Agriculture-Agricultural Research Service (USDA-ARS) Crop Production Systems Research Unit Stoneville USA USDA-ARS Biological Control of Pests Research Unit Stoneville USA.

The fungus Myrothecium verrucaria (Alb. & Schwein.) (MV), originally isolated from diseased sicklepod (Senna obtusifolia L.), has bioherbicial activity against kudzu and several other weeds when applied with low concentrations of the surfactant Silwet L-77. To more fully understand the initial events of MV infection or disease progression, and to improve knowledge related to its mechanism of action, the effects of MV and its product (roridin A) on kudzu seedlings were examined at the ultrastructural level. Ultrastructural analysis of MV effects on kudzu seedlings revealed a rapid (~1 h after treatment) detachment of the protoplast from the cell wall and plasmodesmata appeared to be broken off and retained in the wall. These symptoms occurred well in advance of the appearance of any fungal growth structures. Some fungal growth was observed after severe tissue degeneration (24 to 48 h after treatment), but this occurred primarily at the extra-cellular location with respect to the kudzu tissues. Kudzu seedlings treated with roridin A, a trichothecene produced by the fungus, exhibited some symptoms similar to those induced by the fungus applied in spore formulations with surfactant. The overall results are the first to report the ultrastructural effects of this bioherbicide on plants and suggest that penetration of a phytotoxic substance(s) in the fungal formulation was facilitated by the surfactant, and that roridin A exerts phytotoxicity toward kudzu.

关键词： bioherbicide Biological Weed Control Kudzu Myrothecium verrucaria Ultrastructure Trichothecene

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