Biological Control (Jul 2024)
Gamma irradiation induced thermotolerance in Metarhizium spp. enhancing biocontrol of Nilapavarta lugens (Stål)
Abstract
The Brown Planthopper (BPH), Nilaparvata lugens (Stål), is a devastating pest to rice crops in Southeast Asia, primarily due to its resistance to insecticides. As a result, the fungus Metarhizium spp. is used as an alternative control for BPH. However, their efficiency is limited due to delayed infectivity and sensitivity to environmental variables. In this study, two isolates of Metarhizium spp. MLUBN038 and MNMHN031, were induced for thermotolerance and biocontrol potential against Nilapavarta lugens (Stål). A multilocus phylogeny of four protein-coding genes including Beta-tubulin, rpb1, rpb2, and tef has identified the two isolates to be Metarhizium koreanum. Seventy-nine potential thermotolerant mutants were screened from the two isolates. These mutant isolates demonstrated significant tolerance to vegetative heat at 39 °C and enhanced resistance to wet heat stress at 45 °C after exposure for 60 h, outperforming the wild-type (WT) isolates. The mutant isolates also showed increased mycelial growth, conidial germination, and spore production compared to the WT isolates. Two mutant isolates, ML038_Mt 71 and MN031_Mt 46, exhibited the highest efficacy against BPH nymphs (4th-5th instars), resulting in a maximum mortality of 88.82 % and 83.55 %, respectively, compared to the WT (54.07 % and 47.54 %) after 14 days of inoculation in the greenhouse. These two highly virulent mutant isolates presented the lowest LT50 values of 2.88 and 3.72 days at a concentration of 1 × 109 conidia/mL. RAPD analysis revealed genetic diversity between the mutant and WT isolates, indicating distinct diversity profiles and supporting the successful generation of mutant variants. These findings suggest that the use of gamma irradiation and heat stress to induce mutations is crucial for the successful production of thermotolerant mutant isolates. Our results suggest that ML031_Mt 71 and MN031_Mt 46 have potential as effective biocontrol agents.