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Studies on Mycosis of Metarhizium (Nomuraea) Rileyi on Spodoptera Frugiperda Infesting Maize in Andhra Pradesh, India M

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Studies on Mycosis of Metarhizium (Nomuraea) Rileyi on Spodoptera Frugiperda Infesting Maize in Andhra Pradesh, India M Visalakshi et al. Egyptian Journal of Biological Pest Control (2020) 30:135 Egyptian Journal of https://doi.org/10.1186/s41938-020-00335-9 Biological Pest Control RESEARCH Open Access Studies on mycosis of Metarhizium (Nomuraea) rileyi on Spodoptera frugiperda infesting maize in Andhra Pradesh, India M. Visalakshi1* , P. Kishore Varma1, V. Chandra Sekhar1, M. Bharathalaxmi1, B. L. Manisha2 and S. Upendhar3 Abstract Background: Mycosis on the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), infecting maize was observed in research farm of Regional Agricultural Research Station, Anakapalli from October 2019 to February 2020. Main body: High relative humidity (94.87%), low temperature (24.11 °C), and high rainfall (376.1 mm) received during the month of September 2019 predisposed the larval instars for fungal infection and subsequent high relative humidity and low temperatures sustained the infection till February 2020. An entomopathogenic fungus (EPF) was isolated from the infected larval instars as per standard protocol on Sabouraud’s maltose yeast extract agar and characterized based on morphological and molecular analysis. The fungus was identified as Metarhizium (Nomuraea) rileyi based on ITS sequence homology and the strain was designated as AKP-Nr-1. The pathogenicity of M. rileyi AKP-Nr-1 on S. frugiperda was visualized, using a light and electron microscopy at the host-pathogen interface. Microscopic studies revealed that all the body parts of larval instars were completely overgrown by white mycelial threads of M. rileyi, except the head capsule, thoracic shield, setae, and crotchets. The cadavers of larval instars of S. frugiperda turnedgreenonsporulationand mummified with progress in infection. In vitro pathogenicity tests revealed the potential of AKP-Nr-1 strain of M. rileyi in management of S. frugiperda. Short conclusion: The results indicated the potential of M. rileyi AKP-Nr-1 as biocontrol agent for management of the fall armyworm. This AKP-Nr-1 strain of M. rileyi needs further evaluation under field conditions to evaluate its efficacy against S. frugiperda and its effects on other hosts. Keywords: Metarhizium rileyi, Spodoptera frugiperda, Maize, Efficacy, Microscopy Background maize farmers and poses a major threat to food security and Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) agricultural trade in Africa (Igyuve et al. 2018). (Lepidoptera: Noctuidae) is a serious polyphagous pest of Various control approaches like cultural methods and voracious nature with a wide host range and causes signifi- use of chemical pesticides and natural plant products are cant losses to agricultural crops. Damage due to the pest in vogue for the management of the FAW in maize. larval instars was reported on foliage, leaf whorls, tassel, and However, overuse of chemical insecticides had witnessed cobs based on the stage of infection (Goergen et al. 2016). deleterious effects on soil health and the environment as FAWattackonmaizehasthepotential to cause yield losses well as beneficial flora and fauna like earthworms, natural in a range from 8.3 to 20.6 million tons per annum, in the predators, and parasites (FAO 2018). Use of biocontrol absence of any control measure. This is a nightmare to agents in sustainable management of insect pests is “oflate” gaining importance in view of environmental safety. Fungal * Correspondence: [email protected] bioagents infecting insects can provide an effective alterna- 1Department of Entomology, Regional Agricultural Research Station, tive for ecofriendly management of insect pests. Visakhapatnam District, Anakapalle, Andhra Pradesh 531001, India Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Visalakshi et al. Egyptian Journal of Biological Pest Control (2020) 30:135 Page 2 of 10 Mycosis of entomopathogenic fungus (EPF) on various 48″ N, 83° 01′ 12″ E), Visakhapatnam, Andhra Pradesh, insect pests was reported worldwide under favourable en- India, during 2019–2020 crop seasons. The maize hy- vironmental conditions for fungal growth and multiplica- brid, Syngenta S6668, was sown in the first fortnight of tion (Vimala et al. 2003; Meyling and Eilenberg 2007; August 2019 in 6 blocks of 27 m2 each. Maize cultivation Ingle 2014; and Patil and Abhilash 2014). Prevalence, dis- was carried out as per the regular cultural practices and tribution, and mycosis of entomo-fungal pathogens were data on FAW incidence was recorded on first appear- found influenced by weather parameters like temperature, ance of the pest. rainfall, and humidity (Maurya et al. 2013). Some of these fungi were isolated in pure form, evaluated and identified Correlation of weather parameters with mycosis on fall as potential biocontrol agents for pest management owing armyworm in maize to their specificity and ease of multiplication. Observations on FAW incidence in maize were recorded Among various EPF, Metarhizium (Nomuraea) rileyi at fortnightly intervals from September 2019 to February (Farlow) Samson was reported to infect several lepidop- 2020. Data on weather parameters, viz., temperature, teran pests and found highly effective against noctuid relative humidity, and rainfall, were recorded during this defoliators like Spodoptera litura Fabricius, Helicoverpa period and correlated with M. rileyi natural infection. armigera (Hubner), Anticarsia gemmatalis (Hubner), The data were analysed statistically (Panse and Sukatme and Trichoplusia ni Hubner (Keller and Zimmermann 1985). The data was subjected to stepdown regression by 1989) for their ecofriendly management. M. rileyi is a Efroymson (1960) was followed for generation a prediction dimorphic, ubiquitous fungus with yeast-like hyphal bodies system, using XLSTAT software version 2016.03.30882. and true mycelial filaments and named initially as Botrytis rileyi (Farlow) and later as Spicaria rileyi (Farlow) Charles. Isolation of entomopathogenic fungus (EPF) The fungus was re-described and placed in the genus, Larval instars of S. frugiperda infected with EPF were Nomuraea by Kish et al. (1974). Based on molecular collected from the maize field in the research farm of analysis, using RAPD, internal transcribed spacer (ITS) se- Regional Agricultural Research Station (RARS), Anakapalli, quence analysis, amplified length polymorphism (AFLP), during October 2019 and the pathogen was isolated on and telomeric finger printing methods, Boucias et al. Sabouraud’s maltose yeast extract agar (SMYA) by standard (2000) stated that N. rileyi isolates were more closely re- protocol of Vimala et al. (2002). The dead larvae collected lated to Metarhizium anisopliae and M. flavoviride than to from the field were surface sterilized by immersing in 4% N. atypicola and N. anemonoides. More recently, N. rileyi sodium hypochlorite solution for 1 min, followed by rinsing has been changed to M. rileyi based on its morphological in 3 changes of sterile distilled water. The surface sterilized and molecular characterization by Kepler et al. (2014). diseased specimens were cut in a sterile watch glass and a M. rileyi produces conidia on conidiophores, which small portion of the infected tissue was transferred to a are airborne and the infective propagules that will initiate sterile culture plate containing Sabouraud’s maltose agar the pathogenesis. The conidia adhere to the host surfaces, media fortified with 2% yeast extract. The plates were incu- germinate and produce germ tube that penetrates the host bated at 26 + 1 °C for 8 days and the colonies formed were cuticle, and colonizes the haemocoel of the insect leading further purified by subculture on SMYA medium. to death of the insect (Srisukchayakul et al. 2005). Previous studies revealed a marked variability among the Pathogenicity test isolates of various EPF, collected from different geograph- Pathogenicity of the isolated fungus was carried out as ical locations and their efficacy against insect pests (Vimala per the method of Dutta et al. (2014) with slight modifi- et al. 2003). Therefore, it is important to exploit native EPF cation. Laboratory reared 3rd instar larvae of S. frugi- and to test their potential against target pests for developing perda (N = 30 in 3 replications) were inoculated by a myco-insecticide for specific geographic locations. spraying with conidial suspension of M. rileyi containing Hence, the present study was taken up to characterize 2×108spores/ml. The inoculated larvae were kept in the EPF, naturally infecting S. frugiperda on maize and plastic beakers covered with muslin cloth. Thoroughly to visualize host colonization through light and electron washed maize leaf bits were provided
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