Effect of the entomopathogenic fungus Lecanicillium lecanii in combination with the botanical insecticide Bino2® on larvae of the oak leaf roller (Tortrix viridana L.)

Document Type : Scientific article

Authors

1 Assistant Professor, Plant Protection Research Department, Agricultural and Natural Resources Research and Education Center, Kurdistan Province, Agricultural Research, Education and Extension Organization, Sanandaj, I. R. Iran

2 Associate Professor, Department of Plant Protection, Faculty of Agriculture, Urmia University, Urmia, I. R. Iran

3 Postdoctoral Researcher in Entomology, Department of Plant Protection, Faculty of Agriculture, Urmia University, Urmia, I. R. Iran

Abstract

Background and Objective: Tortrix viridana L., the European oak leaf roller, is one of the most destructive pests of oak forests, causing reduced photosynthesis, suppressed growth, premature defoliation, and ultimately tree decline by feeding on buds and young leaves. Owing to the limitations associated with chemical insecticides, biological control agents and botanical insecticides have gained increasing attention as sustainable components of integrated pest management (IPM). The entomopathogenic fungus Lecanicillium lecanii infects insects through direct penetration of the cuticle, proliferates within the hemocoel, and causes host mortality while also suppressing feeding and reproduction. Likewise, Beno2, a botanical insecticide derived from the root extract of Sophora flavescens, contains alkaloids such as matrine that exhibit insecticidal and antifeedant activities with lower environmental risks than conventional insecticides. Therefore, combining these two control agents may enhance pest suppression through synergistic interactions while reducing reliance on chemical pesticides.
Material and Methods: This laboratory study was conducted to evaluate the compatibility of L. lecanii isolate 229 with the botanical insecticide Beno2 and to investigate their lethal and sublethal effects against larvae of the oak leaf roller (T. viridana). Egg masses were collected from the Pirdanan forests of Piranshahr, and third- and fourth-instar larvae were reared under controlled laboratory conditions. Fungal bioassays were performed using conidial suspensions ranging from 10⁴ to 10⁸ conidia mL⁻¹, and larval mortality was recorded nine days after treatment. Similarly, Beno2 bioassays were conducted using a range of concentrations based on the recommended field dose, with mortality assessed 24 and 48 h after application. Fungal compatibility with Beno2 was evaluated by measuring conidial germination on media containing half, the recommended, and twice the recommended dose of the insecticide. Sublethal effects were assessed using LC₂₅ of each agent individually and a combined treatment of LC₁₂.₅ + LC₁₂.₅. Life-table parameters, including survival, developmental duration, adult longevity, fecundity, intrinsic rate of increase (r), net reproductive rate (R₀), mean generation time (T), and finite rate of increase (λ), were estimated using the age-stage, two-sex life table approach.
Results: Bioassay results demonstrated that larval mortality was concentration-dependent for both control agents. The LC₅₀ values of L. lecanii isolate 229 were estimated at 2.01 × 10⁵ and 7.78 × 10⁵ conidia mL⁻¹ for third- and fourth-instar larvae, respectively. The 48-h LC₅₀ values of Beno2 were 275.50 and 308.09 μL L⁻¹ for the respective larval instars. Compatibility assays showed that half of the recommended Beno2 dose had no significant inhibitory effect on conidial germination, with germination exceeding 92%, whereas the recommended dose reduced germination and twice the recommended dose completely inhibited it. Sublethal treatments, including fungal LC₂₅, Beno2 LC₂₅, and the combined LC₁₂.₅ + LC₁₂.₅ treatment, significantly affected the biological performance of the pest. All treatments prolonged the immature developmental period and shortened adult longevity. Reproductive performance and population growth parameters were also significantly reduced. The greatest reductions in the intrinsic rate of increase (r) and net reproductive rate (R₀) were observed in the combined treatment, indicating a synergistic interaction between L. lecanii and Beno2 in suppressing pest survival and population growth. Moreover, the significant increase in mean generation time (T) suggests a slower rate of population development in subsequent generations.
Conclusion: Overall, the results demonstrated that both L. lecanii and the botanical insecticide Beno2 are effective against the oak leaf roller (T. viridana), and their combined application at appropriate concentrations can enhance efficacy while reducing the need for higher application rates. Because the sublethal combination treatment substantially suppressed population growth and reproductive potential, this strategy represents a promising environmentally friendly option for the integrated management of oak leaf roller populations. Further field studies are recommended to evaluate the long-term effectiveness and stability of this approach under natural forest conditions.

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Forest Research and Development
Volume 12, Issue 2
September 2026
Pages 277-297

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