Biochemical and nanotechnological approaches to combat phytoparasitic nematodes.

The foundation of most food production systems underpinning global food security is the careful management of soil resources. Embedded in the concept of soil health is the impact of diverse soil-borne pests and pathogens, and phytoparasitic nematodes represent a particular challenge. Root-knot nematodes and cyst nematodes are severe threats to agriculture, accounting for annual yield losses of US$157 billion. The control of soil-borne phytoparasitic nematodes conventionally relies on the use of chemical nematicides, which can have adverse effects on the environment and human health due to their persistence in soil, plants, and water. Nematode-resistant plants offer a promising alternative, but genetic resistance is species-dependent, limited to a few crops, and breeding and deploying resistant cultivars often takes years. Novel approaches for the control of phytoparasitic nematodes are therefore required, those that specifically target these parasites in the ground whilst minimizing the impact on the environment, agricultural ecosystems, and human health. In addition to the development of next-generation, environmentally safer nematicides, promising biochemical strategies include the combination of RNA interference (RNAi) with nanomaterials that ensure the targeted delivery and controlled release of double-stranded RNA. Genome sequencing has identified more than 75 genes in root knot and cyst nematodes that have been targeted with RNAi so far. But despite encouraging results, the delivery of dsRNA to nematodes in the soil remains inefficient. In this review article, we describe the state-of-the-art RNAi approaches targeting phytoparasitic nematodes and consider the potential benefits of nanotechnology to improve dsRNA delivery.

Host-virus interaction between tobacco mild green mosaic virus strain U2 and tropical soda apple resulting in systemic hypersensitive necrosis and the host range, survival, spread, and molecular characterization of the virus.

BACKGROUND: Tobacco mild green mosaic virus strain U2 (TMGMV-U2) is a registered active ingredient in a bioherbicide to control tropical soda apple (TSA), Solanum viarum, an invasive weed. As required for registration, we developed empirical data on the host-virus interaction and the virus's host range, survival, spread, and genomic sequence. RESULTS: TMGMV-U2 killed TSA plants by causing systemic hypersensitive necrosis (SHN). It elicited local lesions in inoculated leaves which was followed by the plant's wilting and death. It moved from inoculated terminal leaves through the vasculature to roots and then to newly developed leaves. Phloem death was implicated in wilting and plant death. The SHN response was attenuated in plants grown at constant 32 °C. TMGMV-U2 titer in TSA was low compared to a systemically susceptible tobacco. The virus remained infective for up to 6 months in infected dead TSA tissues and in soil in which infected plants had grown. Susceptible tobacco and pepper plants grown in soil that previously had infected dead TSA or in soil amended with the virus remained asymptomatic and virus-free. A susceptible pepper crop grown in a field block following two consecutive crops of TMGMV-U2-infected susceptible tobacco grew disease-free and virus-free and without yield loss. Purified TMGMV-U2 was infective for 1 year when stored at -20 °C or 5 °C and for 1 month at room temperature. No virus spread was found in the field. Genomic analyses confirmed the registered isolate to be a U2 strain and free of satellite TMV. The TMGMV-U2-susceptible species preponderantly belonged to the Solanaceae. A few hosts that were killed belonged to this family. Several new hosts to TMGMV-U2 were found. These data enabled registration of TMGMV-U2. CONCLUSION: TMGMV-U2 can be used safely as a bioherbicide without risks to nontarget plants and the environment. © 2023 Society of Chemical Industry.

Use of plant viruses as bioherbicides: the first virus-based bioherbicide and future opportunities.

Until recently, only a few plant viruses had been studied for use as biological control agents for weeds, but none had been developed into a registered bioherbicide. This position changed in 2014, when the US Environmental Protection Agency granted an unrestricted Section 3 registration for tobacco mild green mosaic virus (TMGMV) strain U2 as a herbicide active ingredient for a commercial bioherbicide (SolviNix LC). It is approved for the control of tropical soda apple (TSA, Solanum viarum), an invasive 'noxious weed' in the United States. TSA is a problematic weed in cattle pastures and natural areas in Florida. The TMGMV-U2 product kills TSA consistently, completely, and within a few weeks after its application. It is part of the TSA integrated best management practice in Florida along with approved chemical herbicides and a classical biocontrol agent, Gratiana boliviana (Coleoptera: Chrysomelidae). TMGMV is nonpathogenic and nontoxic to humans, animals, and other fauna, environmentally safe, and as effective as chemical herbicides. Unlike the insect biocontrol agent, TMGMV kills and eliminates the weed from fields and helps recycle the dead biomass in the soil. Here the discovery, proof of concept, mode of action, risk analyses, application methods and tools, field testing, and development of the virus as the commercial product are reviewed. Also reviewed here are the data and scientific justifications advanced to answer the concerns raised about the use of the virus as a herbicide. The prospects for discovery and development of other plant-virus-based bioherbicides are discussed. © 2023 Society of Chemical Industry.

Ophiobolin E and 8-epi-ophiobolin J produced by Drechslera gigantea, a potential mycoherbicide of weedy grasses.

Drechslera gigantea, a fungal pathogen isolated from large crabgrass (Digitaria sanguinalis) and proposed as a potential mycoherbicide of grass weeds, produces phytotoxic metabolites in liquid and solid cultures. Ophiobolin A and three minor ophiobolins i.e., 6-epi-ophiobolin A, 3-anhydro-6-epi-ophiobolin A and ophiobolin I were obtained from the liquid culture broths. Interestingly and unexpectedly, ophiobolins also appeared in cultures of this fungus and they were isolated together with the known ophiobolins B and J, and designed as ophiobolin E and 8-epi-ophiobolin J. They were characterized using essentially spectroscopic methods. It is noteworthy that D. gigantea produces such a plethora of bioactive organic substances. Some structure-activity relationship results are also discussed in this report.

Herbicidal potential of ophiobolins produced by Drechslera gigantea.

Drechslera gigantea, a potential mycoherbicide of grass weeds, was isolated in Florida from naturally infected large crabgrass (Digitaria sanguinalis); it produces phytotoxic metabolites in liquid culture. The main metabolite was identified by spectroscopic methods and optical properties as ophiobolin A (1), a well-known phytotoxic sesterterpene produced by several phytopathogenic fungi of important crops and already extensively studied for its interesting biological activities. The other three minor metabolites proved to be related to ophiobolin A and were identified using the same techniques as 6-epi-ophiobolin A and 3-anhydro-6-epi-ophiobolin A (2 and 3) and ophiobolin I (4). Assayed on punctured detached leaves of several grass and dicotyledon weeds, ophiobolin A proved to be on average more phytotoxic as compared to the other related compounds. Some structural features appear to be important for the phytoxicity, such as the hydroxy group at C-3, the stereochemistry at C-6, and the aldehyde group at C-7. Furthermore, grass weeds usually proved to be more sensitive to the phytotoxins than dicotyledons, on which ophiobolin A caused the appearance of large necrosis even at the lowest concentration assayed. This is the first report about the production of ophiobolins from D. gigantea and of the proposed use as potential natural herbicides against grass weeds.

Evaluation of Agar and Grain Media for Mass Production of Conidia of Dactylaria higginsii.

Growth and sporulation of Dactylaria higginsii were quantified on complex agar media containing biological materials (group 1) and chemically defined agar media (group 2), as well as on grains, and the inoculum produced on these various substrates was tested for virulence on Cyperus rotundus. The fungus grew well between 25 and 30°C on potato dextrose agar (PDA), with 27°C being the optimum temperature. Generally, conidial production was highly variable and lower on complex agar media than on chemically defined media. Addition of purple nutsedge leaves to PDA did not increase colony growth or conidial production when compared with una-mended PDA. Conidial production was lowest on brown rice compared with white rice or white rice with nutsedge leaves. Peak production on grain media occurred from day 12 in test 1 (2.4 × 106 spores/g of grain) and on day 16 in test 2 (2.5 × 106 spores/g of grain). Germination rate of conidia produced on white rice was 50% compared with the near 100% germination of conidia produced on PDA or on white rice amended with potato dextrose broth (PDB). Conidia produced on white rice or PDA, when tested fresh or after two washings, were less virulent on C. rotundus than conidia from white rice amended with PDB. After four washings, conidia from all three media produced the same level of disease severity. White rice supplemented with PDB and PDA in trays were suitable for mass production of conidia of D. higginsii.