Laboratory Investigation of Cauliflower-Fungus-Insect Interactions for Cabbage Maggot Control.

The cabbage maggot (also known as cabbage root fly [CRF]; Delia radicum L.) is a serious pest of brassicas. The pest's soil-dwelling larvae are especially damaging to young brassica transplants. In light of toxic soil insecticide phase-out novel biocontrol management solutions are sought for. Our research is focused on the development of a biological control strategy involving cauliflower plantlet inoculation with insect pathogenic fungi. This article presents the results of a laboratory investigation of cauliflower × microbe × CRF interactions. Seven isolates of fungi (entomopathogenic and rhizosphere-competent fungi and soil saprotrophs) were tested for their pathogenicity to CRF and their effects on cauliflower plantlets. The laboratory experiments were performed in sterilized substrate. Several strains significantly increased CRF mortality, some at par with a commercial bioinsecticide based on B. bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae). All strains colonized the rhizoplane, however to varying extent. Some isolates were also reisolated from within healthy plant tissues and thus identified as endophytes. The method of applying conidia had a significant effect on survival and weight of seedlings and rhizoplane and endophytic colonization rates. Two Metarhizium brunneum Petsch (Hypocreales: Clavicipitaceae) isolates exhibited plant growth promotion effects when ungerminated seeds were coated with conidia. The ecological implications of plant × microbe × pest interactions and options for improving the effectiveness of a fungal-based biological CRF management strategy are discussed.

Comparison of cauliflower-insect-fungus interactions and pesticides for cabbage root fly control.

Cabbage root fly (Delia radicum L.) control represents a major challenge in brassica production, therefore different management strategies for its control were tested in conventionally managed open field cauliflower production. Strategies included treatments with low-risk methods such as nitrogen lime, the insecticide spinosad and the Beauveria bassiana ATCC 74040-based biopesticide Naturalis. Their effects were compared with treatments based on nonformulated fungal species Metarhizium brunneum, B. bassiana, Clonostachys solani, Trichoderma atroviride, T. koningiopsis, and T. gamsii and commercial insecticides λ-cyhalothrin and thiamethoxam. Spinosad and thiamethoxam were pipetted to individual plants before transplanting; λ-cyhalothrin was sprayed after transplanting; nitrogen lime was applied at first hoeing. Nonformulated fungi were delivered onto cauliflower plantlets' roots as a single pretransplantation inoculation. The cabbage root fly population dynamics exhibited a strong spatiotemporal variation. The lowest number of cabbage root fly pupae recovered from cauliflower roots in the field experiments was recorded in plants treated with spinosad (significant reduction), followed by Naturalis and one of the tested M. brunneum strains (nonsignificant reduction). Significantly more pupae were counted in the nitrogen lime treatment. The field experiments showed that a single drench of cauliflower plantlets with spinosad offered consistent and enduring cabbage root fly control. Naturalis and nonformulated fungal isolates did not decrease cabbage root fly pressure significantly, apparently due to lack of statistical power. The implications of the substantial intra- and inter-annual pest pressure variation and the benefits of using single plant treatments are discussed, and recommendations for improvement of rhizosphere-competence utilizing biological control strategies provided.

Evolutionary patchwork of an insecticidal toxin shared between plant-associated pseudomonads and the insect pathogens Photorhabdus and Xenorhabdus.

BACKGROUND: Root-colonizing fluorescent pseudomonads are known for their excellent abilities to protect plants against soil-borne fungal pathogens. Some of these bacteria produce an insecticidal toxin (Fit) suggesting that they may exploit insect hosts as a secondary niche. However, the ecological relevance of insect toxicity and the mechanisms driving the evolution of toxin production remain puzzling. RESULTS: Screening a large collection of plant-associated pseudomonads for insecticidal activity and presence of the Fit toxin revealed that Fit is highly indicative of insecticidal activity and predicts that Pseudomonas protegens and P. chlororaphis are exclusive Fit producers. A comparative evolutionary analysis of Fit toxin-producing Pseudomonas including the insect-pathogenic bacteria Photorhabdus and Xenorhadus, which produce the Fit related Mcf toxin, showed that fit genes are part of a dynamic genomic region with substantial presence/absence polymorphism and local variation in GC base composition. The patchy distribution and phylogenetic incongruence of fit genes indicate that the Fit cluster evolved via horizontal transfer, followed by functional integration of vertically transmitted genes, generating a unique Pseudomonas-specific insect toxin cluster. CONCLUSIONS: Our findings suggest that multiple independent evolutionary events led to formation of at least three versions of the Mcf/Fit toxin highlighting the dynamic nature of insect toxin evolution.

Evaluation of insect associated and plant growth promoting fungi in the control of cabbage root flies.

Delia radicum L. or cabbage maggot is an important pest for Brassicaceous crops. There are currently no registered chemical control agents for its control in Slovenia. Fungal control agents for cabbage maggot were therefore sought among nine rhizosphere-compatible and plant growth-promoting, soil-adapted, and entomopathogenic species to cabbage maggots and were assayed in in vitro and soil laboratory bioassays. In the in vitro tests, the conidial suspensions were applied directly to cabbage maggot eggs. The soil tests mimicked pathways of natural exposure of various insect life stages to the fungal strains. Conidial concentrations used in soil tests were comparable to economic rates for in-furrow application. The following fungi were tested: Trichoderma atroviride P. Karst. (2 isolates), Trichoderma koningiopsis Samuels, C. Suárez & H.C. Evans (1), Trichoderma gamsii Samuels & Druzhin. (3), Beauveria brongniartii (Saccardo) Petch (1), Beauveria bassiana (Balsamo-Crivelli) Vuillemin (2), Metarhizium robertsii J.F. Bisch., Rehner & Humber (1), Metarhizium anisopliae (Metschn.) Sorokin (4), Purpureocillium lilacinum (Thom) Luangsa-ard, Houbraken, Hywel-Jones & Samson (2), and Clonostachys solani f. nigrovirens (J.F.H. Beyma) Schroers (2). Abbott's corrected mortality in the in vitro tests ranged from 0.0 +/- 18.9 to 47.6 +/- 9.0% and in the soil test from 2.4 +/- 13.0 to 68.2 +/- 21.5%. Seven isolates (B. bassiana [isolate 1174], C. solani [1828], M. anisopliae [1154 and 1868], T. atroviride [1872], T. koningiopsis [1874], and T. gamsii [1876]) caused significant cabbage maggot mortality in either in vitro or soil tests. The importance of fungal ecology as a criterion during the screening of potential biological control agents is discussed.

Direct plantlet inoculation with soil or insect-associated fungi may control cabbage root fly maggots.

A potential Delia radicum biological control strategy involving cauliflower plantlet inoculation with various fungi was investigated in a series of laboratory and glasshouse experiments. In addition to entomopathogenic fungi, fungi with a high rhizosphere competence and fungi with the ability to survive as saprotrophs in soil were tested. The following fungal species were evaluated in the experiments: Trichoderma atroviride, T. koningiopsis, T. gamsii, Beauveria bassiana, Metharhizium anisopliae, M. brunneum and Clonostachys solani. A commercial carbosulfan-based insecticide was used as a positive control. Additionally, two commercial products, one based on B. bassiana (Naturalis) and one on Bacillus thuringiensis (Delfin) were used as reference biocontrol agents. The aims were (i) to assess the pathogenicity of the selected fungal isolates to Delia radicum, (ii) to evaluate the fungal isolates' rhizosphere competence, with the emphasis on the persistence of the original inoculum on the growing roots, (iii) to assess possible endophytic plant tissue colonization, and (iv) to evaluate potential plant growth stimulating effects of the added inoculi. Significant pathogenicity of tested fungi against Delia radicum was confirmed in in vitro and glasshouse experiments. All tested fungi persisted on cauliflower rhizoplane. More importantly, the added fungi were found on thoroughly washed roots outside the original point of inoculation. This provided us with evidence that our tested fungi could be transferred via or grow with the elongating roots. In addition to colonizing the rhizoplane, some fungi were found inside the plant root or stem tissue, thus exhibiting endophytic characteristics. The importance of fungal ecology as a criterion in appropriate biological control agent selection is discussed.

Integrated Management of European Cherry Fruit Fly Rhagoletis cerasi (L.): Situation in Switzerland and Europe.

The European cherry fruit fly, Rhagoletis cerasi (L.) (Diptera: Tephritidae), is a highly destructive pest. The low tolerance for damaged fruit requires preventive insecticide treatments for a marketable crop. The phase-out of old insecticides threatens cherry production throughout the European Union (EU). Consequently, new management techniques and tools are needed. With the increasing number of dwarf tree orchards covered against rain to avoid fruit splitting, crop netting has become a viable, cost-effective method of cherry fruit fly control. Recently, a biocontrol method using the entomopathogenic fungus Beauveria bassiana has been developed for organic agriculture. However, for most situations, there is still a lack of efficient and environmentally sound insecticides to control this pest. This review summarizes the literature from over one hundred years of research on R. cerasi with focus on the biology and history of cherry fruit fly control as well as on antagonists and potential biocontrol organisms. We will present the situation of cherry fruit fly regulation in different European countries, give recommendations for cherry fruit fly control, show gaps in knowledge and identify future research opportunities.

Molecular analysis of a novel gene cluster encoding an insect toxin in plant-associated strains of Pseudomonas fluorescens.

Pseudomonas fluorescens CHA0 and the related strain Pf-5 are well-characterized representatives of rhizosphere bacteria that have the capacity to protect crop plants from fungal root diseases, mainly by releasing a variety of exoproducts that are toxic to plant pathogenic fungi. Here, we report that the two plant-beneficial pseudomonads also exhibit potent insecticidal activity. Anti-insect activity is linked to a novel genomic locus encoding a large protein toxin termed Fit (for P. fluorescensinsecticidal toxin) that is related to the insect toxin Mcf (Makes caterpillars floppy) of the entomopathogen Photorhabdus luminescens, a mutualist of insect-invading nematodes. When injected into the haemocoel, even low doses of P. fluorescens CHA0 or Pf-5 killed larvae of the tobacco hornworm Manduca sexta and the greater wax moth Galleria mellonella. In contrast, mutants of CHA0 or Pf-5 with deletions in the Fit toxin gene were significantly less virulent to the larvae. When expressed from an inducible promoter in a non-toxic Escherichia coli host, the Fit toxin gene was sufficient to render the bacterium toxic to both insect hosts. Our findings establish the Fit gene products of P. fluorescens CHA0 and Pf-5 as potent insect toxins that define previously unappreciated anti-insect properties of these plant-colonizing bacteria.