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Biological Insect Control Using Metarhizium Anisopliae: Morphological, Molecular, and Ecological Aspects

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Biological Insect Control Using Metarhizium Anisopliae: Morphological, Molecular, and Ecological Aspects Ciência Rural, SantaBiological Maria, insectv.44, n.4, control p.645-651, using Metarhizium abr, 2014 anisopliae: morphological, molecular, and ecological aspects. 645 ISSN 0103-8478 Biological insect control using Metarhizium anisopliae: morphological, molecular, and ecological aspects Controle biológico de insetos utilizando Metarhizium anisopliae: aspectos morfológicos, moleculares e ecológicos Patricia Vieira TiagoI* Neiva Tinti de OliveiraI Elza Áurea de Luna Alves LimaI ABSTRACT INTRODUCTION Microbial control of insects is based on the rational use of pathogens to maintain environmentally balanced pest Biological control consists of the population levels, and Metarhizium anisopliae has been the introduction of benefi cial predatory or parasitic most studied and most utilized fungal species for that purpose. species into cultivation systems where they were The natural genetic variability of entomopathogenic fungi is previously absent or present only at low population considered one of the principal advantages of microbial insect control. The inter- and intraspecifi c variability and the genetic levels. This technique is designed to negatively diversity and population structures of Metarhizium and other affect specifi c target species that could otherwise entomopathogenic fungi have been examined using ITS-RFLP, become pests or infectious agents (GLIESSMAN, ISSR, and ISSP molecular markers. The persistence of M. anisopliae in the soil and its possible effects on the structures of 2001). Susceptibility to pests is a general refl ection resident microbial communities must be considered when selecting of plant health, which can be negatively infl uenced isolates for biological insect control. by poor soil fertility management (NICHOLLS & Key words: biological control, Metarhizium anisopliae, molecular ALTIERI, 2007). One of the objectives of biological markers. control is to assure that the benefi cial organism to be introduced can complete its lifecycle at the site, RESUMO and then reproduce with suffi cient effi ciency to become a permanent resident of the agrosystem. O controle microbiano consiste na utilização racional de patógenos, visando à manutenção da população de Frequently, however, the niche conditions available insetos em equilíbrio no ambiente. Metarhizium anisopliae é a to the benefi cial introduced organism do not espécie mais estudada e utilizada no controle biológico de insetos. fully satisfy its long-term needs, requiring its A variabilidade genética dos fungos entomopatogênicos pode ser considerada uma das principais vantagens no controle microbiano reintroduction (GLIESSMAN, 2001). Changes in de insetos e pode ser detectada por meio de marcadores production practices and the use of agricultural moleculares, como ITS-RFLP, ISSR e ISSP. Esses marcadores additives are often necessary for biological control são usados para a caracterização inter e intraespecífi ca de Metarhizium e outros fungos entomopatogênicos e poderão to be successful. Integrated Pest Management (IPM) auxiliar na compreensão da diversidade genética e da estrutura is an alternative to unilateral intervention strategies das populações destes fungos. A persistência de M. anisopliae no using agrochemicals, with a wider focus on the solo e seu possível efeito na estrutura da comunidade microbiana ecology of the insect pests as well as the crop plants, deste solo são características importantes e pouco estudadas, que devem ser consideradas no processo de seleção de isolados para o based on the use of complementary tactics and the controle biológico de insetos. adoption of cultivation techniques that favor plant diversity. Pest control in this type of approach is Palavras-chave: controle biológico, Metarhizium anisopliae, marcadores moleculares. initially based on natural agents such as pathogens, IDepartamento de Micologia, Centro de Ciências Biológicas (CCB), Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves s/n, 50670-420, Recife, PE, Brasil. E-mail: [email protected] *Autor para correspondência. Received 01.29.13 Approved 10.10.13 Returned by the author 01.27.14 CR-2013-0122.R2 Ciência Rural, v.44, n.4, abr, 2014. 646 Tiago et al. parasites and predators, with the use of agrotoxins structures or blastospores and appressoria are produced being contemplated only as a last resort. However, as by M. anisopliae through mycelial differentiation. biological pest control methods do not demonstrate Blastospores can function in certain cases as immediate results in agro-industrial systems with reproductive units and are produced in submerged large-scale production and commercialization goals cultures (JACKSON & JARONSKI, 2009) and in (as agrotoxins), commercial groups tend to avoid the hemolymph of insect hosts (ALVES, 1998). The the costs and labor related to their development appressoria, formed at the extremity of the hyphae, and perfection. Nonetheless, growing energy costs, may be involved in fungus pathogenicity and have environmental degradation, and infl ation all reinforce the function of initiating epicuticular and procuticle the argument that immediate fi nancial gains should penetration of the insect tegument (ALVES, 1998). not be the principal motivating force in agricultural The production of microsclerotia by isolates of M. production (ALTIERI, 2002). In spite of the strong anisopliae has been observed after cultivation in liquid economic pressure on agricultural production, many media with different concentrations of carbon and farmers are making the transition to practices that are carbon-nitrogen (JACKSON & JARONSKI, 2009). more environmentally friendly and have the potential The fungal-host relationship occurs to contribute to long-term agricultural sustainability through the adhesion and germination of conidia with biological control being one of th e principal tools on the surface of the insect, followed by hyphae in this conversion process (GLIESSMAN, 2001). penetration through the cuticle. The process of host Microbial control is an aspect of biological colonization initiates after penetration, with the insect control and consists of the rational use of penetrating hyphae becoming thicker and ramify pathogens to maintain pest balances in agricultural within the tegument and the hemocoel of the insect, environments, with increases in the numbers of forming blastospores. The hyphae continue to grow other natural enemies often being observed in and invade various internal organs after the death of fi elds where microbial control has been used. the host and will subsequently emerge from the insect Successful programs of microbial control using body and produce conidia that disseminate and infect entomopathogenic fungi to combat arthropod pests in other individuals (ALVES, 1998). soils and aquatic environments have been developed, Molecular studies of the processes of principally utilizing the genera Metarhizium, host infection have shown them to be complex and Beauveria, Sporothrix, Lecanicillium, Nomuraea, multifactorial. The adhesion and penetration steps Hirsutella, Aschersonia, Isaria, Paecilomyces, and have been most closely examined and appear to be Enthomophthora (ALVES & LOPES, 2008). Species decisive to infection. The participation of an adhesin within the genus Metarhizium are pathogenic fungi coded by the gene Mad1 in the adhesion of conidia having broad ranges of insect hosts. M. anisopliae to the cuticle of Manduca sexta Linnaeus larva was was found to be a species complex composed of nine demonstrated using mutants in which this gene was species based on multilocus phylogeny (BISCHOFF deleted, with these mutants demonstrating signifi cant et al., 2009). The objective of this study was to analyze some morphological, molecular and ecological decreases in conidial germination, suppression of aspects of M. anisopliae. the formation of blastospores, and reduced virulence (WANG & St. LEGER, 2007a). COSENTINO- Metarhizium anisopliae GOMES et al. (2013) described that the inhibition of Metarhizium anisopliae, a anamorphic phosphatase activity in the conidia of M. anisopliae fungus which belong to the phylum Ascomycota, reduced adhesion to the integument of Dysdercus is the most intensively studied species of the genus peruvianus (Hemiptera: Pyrrhocoridae) and Metarhizium, considering that the teleomorph (indirectly) its infection. Cordyceps brittlebankisoides [= Metacordyceps The participation of perilipin (proteins brittlebankisoides (Liu, Liang, Whalley, Yao & Liu) that surround lipidic droplets in the cell interior) Sung, Sung, Hywel-Jones & Spatafora] was isolated in appressoria differentiation in M. anisopliae has from insect larva (Coleoptera: Scarabaeidae) and also been reported. The deactivation of the Mpl1 identifi ed as M. anisopliae var. majus [= M. majus gene in some strains generates defi ciencies in the (Johnston) Bischoff, Rehner & Humber] (LIU et al., infection process due to the formation of appressoria 2001). The reproductive structures of M. anisopliae with lower concentrations of lipidic droplets and (the anamorph, the most commonly encountered form) resultantly lower levels of osmotic pressure - resulting comprise conidiophores and conidia. Leveduriform in diffi culties in terms of hyphal penetration (WANG Ciência Rural, v.44, n.4, abr, 2014. Biological insect control using Metarhizium anisopliae: morphological, molecular, and ecological aspects. 647 & St. LEGER, 2007b). Defective appressoria were regions composed of 18S, 5.8S and 28S genes that are also observed after the deletion of the mapka1
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