Bioherbicidal Activity of Albifimbria verrucaria (Formerly Myrothecium verrucaria) on Glyphosate-Resistant Conyza canadensis.

The effects of the bioherbicidal activity of the fungal phytopathogen, Albifimbria verrucaria (AV), formerly Myrothecium verrucaria, on glyphosate-resistant and -susceptible Conyza canadensis (horseweed) were examined in greenhouse and field studies. Spray applications of mycelial formulations of AV infected both glyphosate-resistant and -susceptible C. canadensis plants at various growth stages. Young plants in the rosette leaf stage of growth were controlled more efficaciously than were older plants that had bolted or that were in the inflorescence stage; nevertheless, severe injury and mortality also occurred in mature plants. The results indicate that this bioherbicidal fungus can infect and control C. canadensis, thereby demonstrating the potential of this fungus as a bioherbicidal agent against this troublesome weed, which has become resistant to various herbicides.

Effects of the Fungal Bioherbicide, Alternaria cassia on Peroxidase, Pectinolytic and Proteolytic Activities in Sicklepod Seedlings.

Certain plant pathogens have demonstrated potential for use as bioherbicides for weed control, and numerous studies have been published on this subject for several decades. One of the early examples of an important fungal bioherbicide is Alternaria cassiae, isolated from the weed sicklepod (Senna obtusifolia). To gain further insight into biochemical interactions of this fungus and its host weed, we examined the effects of this bioherbicide on various enzymes associated with plant defense. Young sicklepod seedlings were challenged with A. cassiae spore inoculum and enzyme activities associated with plant defense (peroxidase, proteolytic, and pectinolytic) were assayed periodically over a 96-h time course on plants grown in continuous darkness or continuous light. Peroxidase activity increased with time in untreated control seedlings in both light and dark, but the effect was greater in the light. In A. cassiae-treated plants, peroxidase was elevated above that in control tissue at all sample times resulting in a 1.5 -fold increase above control in light-grown tissue and a 2- to 3-fold increase in dark-grown tissue over 48-96 h. Differences in leucine aminopeptidase activity in control versus A. cassiae-treated tissues were not significant until 48-96 h, when activity was inhibited in fungus-treated tissues by about 32% in light-grown tissue and 27% in dark-grown tissue after 96 h. Proteolytic activity on benzoyl-arginine-p-nitroanilide was not significantly different in treated versus control tissue in either light or dark over the time course. Pectinase activity increased in treated tissues at all time points as early as 16 h after spore application in light- or dark-grown plants. The greatest increases were 1.5-fold above control levels in light-grown plants (40-64 h) and 2-fold in plants grown in darkness (72-96 h). Data suggests that peroxidase may be involved as defense mechanism of sicklepod when challenged by A. cassia and that this mechanism is operative in young seedlings under both light and dark growth conditions. Differential proteolytic activity responses on these two substrates suggests the presence of two different enzymes. Increased pectinase activity during pathogenesis suggests that A. cassiae-sicklepod interaction results in an infectivity mechanism to degrade pectic polymers important to sicklepod cell wall integrity. These studies provide important information on some biochemical interactions that may be useful for improvements to biological weed control programs utilizing plant pathogens. Such information may also be useful in genetic selection and manipulation of pathogens for weed control.

Taxonomic Evaluation of a Bioherbicidal Isolate of Albifimbria verrucaria, Formerly Myrothecium verrucaria.

The fungal genus Myrothecium was once polyphyletic but a recent reconsideration of the family Stachybotryaceae spilt it into several genera. The ex-neotype specimen of the species Myrothecium verrucaria is now recognized as Albifimbria verrucaria. The well-studied plant pathogen and candidate bioherbicide CABI-IMI 368023, previously identified as M. verrucaria, was analyzed morphologically and genetically and found to be most consistently aligned with the other representatives of A. verrucaria.

Survey of the genomic landscape surrounding the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant Amaranthus palmeri from geographically distant populations in the USA.

BACKGROUND: Glyphosate resistance in Amaranthus palmeri, one of the most prevalent herbicide-resistant weeds in the USA, is attributable to amplification and increased expression of the gene encoding the target site of glyphosate, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). The EPSPS gene and the surrounding 287 kilobases (kb) of amplified sequence are unique to glyphosate-resistant plants and termed the EPSPS cassette. It has only been sequenced in one A. palmeri population from Mississippi. This research compares EPSPS cassettes in seven resistant and five sensitive populations from geographically distant locations within the USA, including Mississippi, Arizona, Kansas, Maryland, Delaware and Georgia. RESULTS: Polymerase chain reaction (PCR) products from 40 primer pairs specific to the cassette were similar in size and sequence in resistant populations. Several primer pairs failed to generate PCR products in sensitive populations. Regions of the cassette sequenced in the resistant populations were found to be nearly identical to those from Mississippi. Gene expression analysis showed that both EPSPS and another gene in the cassette, a reverse transcriptase, were elevated in all resistant populations tested relative to the sensitive populations. CONCLUSION: EPSPS cassettes from distant resistant populations were nearly homologous. Considering the complexity of the cassette, and the degree of similarity among some cassette sequences, the results are consistent with the hypothesis that glyphosate resistance probably evolved once and then rapidly spread across the USA. © 2017 Society of Chemical Industry.

Bioherbicidal activity from washed spores of Myrothecium verrucaria.

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.

Immunohistochemical investigation of the necrotrophic phase of the fungus Colletotrichum gloeosporioides in the biocontrol of hemp sesbania (Sesbania exaltata; Papilionaceae).

PREMISE OF THE STUDY: Fungal plant pathogens exert much of their effect on plant cells through alterations in the host cell walls. However, obtaining biochemical proof for this change is difficult because of the relatively small number of cells that are affected by the pathogen relative to the bulk of host tissue. In this study, we examined the differences in host wall composition between infected and uninfected areas of seedlings of the weed hemp sesbania (Sesbania exaltata) that were treated with the biocontrol agent Colletotrichum gloeosporioides. • METHODS: To determine the changes in cell wall composition, we used semi-thin sections and a battery of antibody probes that recognize components of the cell wall and immunogold-silver cytochemistry to visualize the probes. • KEY RESULTS: A loss of specific plant cell wall polysaccharides in the region surrounding the primary fungal infection and the creation of a defensive layer by the plant to limit the fungal invasion were the two most obvious changes noted in this study. At the invasion site, there was significant loss of rhamnogalacturon-1 (RGI) and esterified and de-esterified homogalacturonan (HG)-reactive epitopes from the cell walls. In contrast, boundary tissue between the vascular tissue and the fungal lesion reacted more strongly with antibodies that recognize arabinogalactan proteins (AGPs) and xyloglucans than in unaffected areas. • CONCLUSIONS: These data strongly indicate a role of pectinases in the invasion of the biocontrol agent and the importance of extensins, AGPs, and xyloglucans as defense by the host.

Influence of simulated Quinclorac drift on the accumulation and movement of herbicide in tomato (Lycopersicon esculentum) plants.

Quinclorac (3,7-dichloro-8-quinolinecarboxylic acid) is a herbicide commonly used in rice, and its drift has been suspected of causing injury to off-target tomato fields throughout Arkansas. Studies were conducted to evaluate the effects of single and multiple simulated quinclorac drift applications on tomato plant growth and development. Residues extracted from tomato plants treated with 0.42 g of ai ha(-1) were below the detection limit of liquid chromatography-double mass spectrometry (LC-MS/MS) analysis. Quinclorac residue levels and half-lives in tomato tissue increased as the application rate and number of applications increased. From 3 to 72 h after (14)C-quinclorac treatment of plants, most of the absorbed (14)C was retained in the treated leaf, and translocations of (14)C out of the treated leaf of vegetative and flowering tomato plant tissues were similar. Of the (14)C that translocated out of the treated leaf, the greatest movement was acropetally. The flower cluster contained 1% of the total absorbed (14)C, which suggests the potential for quinclorac translocation into tomato fruit. More extensive research will be required to understand the impact that quinclorac may have on tomato production in the area.