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Fernando Fonseca Pereira De Paula - One of the best experts on this subject based on the ideXlab platform.
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hrnai mediated knock down of Sphenophorus levis v atpase e in transgenic sugarcane saccharum spp interspecific hybrid affects the insect growth and survival
Plant Cell Reports, 2021Co-Authors: Chakravarthi Mohan, Andrea Soarescosta, Fernando Fonseca Pereira De Paula, Priscila Yumi Tanaka Shibao, Danyelle Toyama, Marcos Antonio Sanches Vieira, Antonio Figueira, Danielle Camargo Scotton, Flavio HenriquesilvaAbstract:Transgenic sugarcane expressing V-ATPase subunit E dsRNA affects growth and survival of Sphenophorus levis. Plants being sessile organisms are constantly confronted with several biotic and abiotic stresses. Sugarcane (Saccharum spp) is a major tropical crop widely cultivated for its sugar and other by-products. In Brazil, sugarcane plantations account for significant production losses due to Sphenophorus levis (sugarcane weevil) infestations. With the existing control measures being less effective, there arises a necessity for advanced strategies. Our bioassay injection experiments with V-ATPase E dsRNA in S. levis larvae showed significant mortality and reduction in transcription levels. Furthermore, we down-regulated the V-ATPase E gene of S. levis in transgenic sugarcane using an RNAi approach. The resultant RNAi transgenic lines exhibited reduction in larval growth and survival, without compromising plant performance under controlled environment. Our results illustrate that RNAi-mediated down-regulation of key genes is a promising approach in imparting resistance to sugarcane weevil.
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pectinases from Sphenophorus levis vaurie 1978 coleoptera curculionidae putative accessory digestive enzymes
Journal of Insect Science, 2015Co-Authors: Danilo Elton Evangelista, Fernando Fonseca Pereira De Paula, Andre Rodrigues, Flavio HenriquesilvaAbstract:The cell wall in plants offers protection against invading organisms and is mainly composed of the polysaccharides pectin, cellulose, and hemicellulose, which can be degraded by plant cell wall degrading enzymes (PCWDEs). Such enzymes are often synthesized by free living microorganisms or endosymbionts that live in the gut of some animals, including certain phytophagous insects. Thus, the ability of an insect to degrade the cell wall was once thought to be related to endosymbiont enzyme activity. However, recent studies have revealed that some phytophagous insects are able to synthesize their own PCWDEs by endogenous genes, although questions regarding the origin of these genes remain unclear. This study describes two pectinases from the sugarcane weevil, Sphenophorus levis Vaurie, 1978 (Sl-pectinases), which is considered one of the most serious agricultural pests in Brazil. Two cDNA sequences identified in a cDNA library of the insect larvae coding for a pectin methylesterase (PME) and an endo-polygalacturonase (endo-PG)—denominated Sl-PME and Sl-endoPG, respectively—were isolated and characterized. The quantitative real-time reverse transcriptase polymerase chain reaction expression profile for both Sl-pectinases showed mRNA production mainly in the insect feeding stages and exclusively in midgut tissue of the larvae. This analysis, together Western blotting data, suggests that Sl-pectinases have a digestive role. Phylogenetic analyses indicate that Sl-PME and Sl-endoPG sequences are closely related to bacteria and fungi, respectively. Moreover, the partial genomic sequences of the pectinases were amplified from insect fat body DNA, which was certified to be free of endosymbiotic DNA. The analysis of genomic sequences revealed the existence of two small introns with 53 and 166 bp in Sl-endoPG, which is similar to the common pattern in fungal introns. In contrast, no intron was identified in the Sl-PME genomic sequence, as generally observed in bacteria. These data support the theory of horizontal gene transfer proposed for the origin of insect pectinases, reinforcing the acquisition of PME genes from bacteria and endo-PG genes from fungi.
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expression purification crystallization and preliminary x ray diffraction analysis of the pectin methylesterase from the sugar cane weevil Sphenophorus levis
Acta Crystallographica Section F-structural Biology and Crystallization Communications, 2014Co-Authors: Danilo Elton Evangelista, Fernando Fonseca Pereira De Paula, Flavio Henriquesilva, A S Godoy, Igor PolikarpovAbstract:Pectin methylesterase removes the methyl groups from the main chain of pectin, the major component of the middle lamella of the plant cell wall. The enzyme is involved in plant cell-wall development, is part of the enzymatic arsenal used by microorganisms to attack plants and also has a wide range of applications in the industrial sector. Therefore, there is a considerable interest in studies of the structure and function of this enzyme. In this work, the pectin methylesterase from Sphenophorus levis was produced in Pichia pastoris and purified. Crystals belonging to the monoclinic space group C2, with unit-cell parameters a = 122.181, b = 82.213, c = 41.176 A, β = 97.48°, were obtained by the sitting-drop vapour-diffusion method and an X-ray diffraction data set was collected to 2.1 A resolution. Structure refinement and model building are in progress.
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Expression, purification, crystallization and preliminary X-ray diffraction analysis of the pectin methylesterase from the sugar cane weevil Sphenophorus levis
Chester, 2014Co-Authors: Evangelista, Danilo Elton, Fernando Fonseca Pereira De Paula, A S Godoy, Silva, Flavio Henrique, Polikarpov IgorAbstract:Pectin methylesterase removes the methyl groups from the main chain of pectin, the major component of the middle lamella of the plant cell wall. The enzyme is involved in plant cell-wall development, is part of the enzymatic arsenal used by microorganisms to attack plants and also has a wide range of applications in the industrial sector. Therefore, there is a considerable interest in studies of the structure and function of this enzyme. In this work, the pectin methylesterase from Sphenophorus levis was produced in Pichia pastoris and purified. Crystals belonging to the monoclinic space group C2, with unit-cell parameters a = 122.181, b = 82.213, c = 41.176 Å, β= 97.48° , were obtained by the sittingdrop vapour-diffusion method and an X-ray diffraction data set was collected to 2.1 Å resolution. Structure refinement and model building are in progress.FAPESPCNP
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biologia molecular aplicada a identificacao de alvos para o controle do bicudo da cana de acucar Sphenophorus levis
2012Co-Authors: Fernando Fonseca Pereira De PaulaAbstract:O bicudo da cana-de-acucar, Sphenophorus levis, e um inseto que na fase larval se alimenta do rizoma da cana-de-acucar cavando galerias que causam danos e levam a morte das plantas. Os metodos convencionais de controle disponiveis nao tem sido eficientes contra esse inseto. O objetivo desse estudo foi gerar conhecimento sobre a biologia desse inseto em nivel molecular utilizando uma abordagem transcriptomica para a identificacao de possiveis genes alvo, visando futuros estudos para desenvolvimento de plantas transgenicas resistentes ao ataque deste inseto. Apos o processamento e montagem das sequencias utilizando o programa dCAS, 3804 sequencias foram agrupadas em 201 contigs e 1363 singlets, os quais foram manualmente anotados. Foram identificados diversos genes de degradacao de parede celular, incluindo pectinases e celulases. Pela primeira vez um contig contendo uma sequencia que codifica uma invertase foi identificado em um coleoptero. As provaveis enzimas digestivas totalizam 19,3% e os genes desconhecidos representam 28,8% do total de sequencias expressas. Considerando a predominância de catepsinas L dentre as enzimas digestivas identificadas no transcriptoma (54,3%) e a sua importância para a hidrolise de proteinas nos processos digestivos, um clone de cDNA codificando uma catepsina L, denominado Sl-CathL, foi escolhido para a expressao recombinante em celulas de Pichia pastoris, caracterizacao e inibicao in vitro utilizando a cistatina da cana-de-acucar CaneCPI-4 (Ki = 0,196 nM). Os ensaios de imunolocalizacao evidenciaram a producao da Sl-CathL no epitelio do intestino medio e a secrecao de vesiculas, contendo a enzima, no lumen do intestino. Larvas do inseto S. levis tambem apresentaram sensibilidade para disparar a maquinaria de RNA de interferencia induzida por injecoes de dsRNA na cavidade corporea e o silenciamento genico especifico foi confirmado por qRT-PCR. As larvas que receberam injecoes com dsRNA da V-ATPase E morreram dentro de tres semanas, enquanto as larvas que tiveram a serpina 1 silenciada exibiram atraso no desenvolvimento, aprisionamento na fase pupal, ou morreram como adultos farados. Desse modo, concluimos neste trabalho iniciado a partir da analise do transcriptoma, que a inibicao da principal enzima digestiva pela Cane-CPI-4 e o silenciamento genico via RNAi sao alternativas promissoras para o estabelecimento de plantas resistentes ao inseto e podem ser aplicadas no desenvolvimento de plantas de cana-de-acucar transgenicas com o objetivo de aumentar sua resistencia ao Sphenophorus levis.%%%%The sugarcane weevil, Sphenophorus levis, is an insect that feeds on the rhizome of sugarcane in its larval stage, boring channels that cause damage and death to the plant. Conventional methods of insect control have not been efficient. The aim of the present study was to generate knowledge on the biology of this insect on the molecular level using a transcriptomic approach to determine potential targets genes for the engineering of insect-resistant plants. After sequence processing and assembly…
Flavio Henriquesilva - One of the best experts on this subject based on the ideXlab platform.
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hrnai mediated knock down of Sphenophorus levis v atpase e in transgenic sugarcane saccharum spp interspecific hybrid affects the insect growth and survival
Plant Cell Reports, 2021Co-Authors: Chakravarthi Mohan, Andrea Soarescosta, Fernando Fonseca Pereira De Paula, Priscila Yumi Tanaka Shibao, Danyelle Toyama, Marcos Antonio Sanches Vieira, Antonio Figueira, Danielle Camargo Scotton, Flavio HenriquesilvaAbstract:Transgenic sugarcane expressing V-ATPase subunit E dsRNA affects growth and survival of Sphenophorus levis. Plants being sessile organisms are constantly confronted with several biotic and abiotic stresses. Sugarcane (Saccharum spp) is a major tropical crop widely cultivated for its sugar and other by-products. In Brazil, sugarcane plantations account for significant production losses due to Sphenophorus levis (sugarcane weevil) infestations. With the existing control measures being less effective, there arises a necessity for advanced strategies. Our bioassay injection experiments with V-ATPase E dsRNA in S. levis larvae showed significant mortality and reduction in transcription levels. Furthermore, we down-regulated the V-ATPase E gene of S. levis in transgenic sugarcane using an RNAi approach. The resultant RNAi transgenic lines exhibited reduction in larval growth and survival, without compromising plant performance under controlled environment. Our results illustrate that RNAi-mediated down-regulation of key genes is a promising approach in imparting resistance to sugarcane weevil.
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transgenic sugarcane overexpressing canecpi 1 negatively affects the growth and development of the sugarcane weevil Sphenophorus levis
Plant Cell Reports, 2017Co-Authors: Vanessa Karine Schneider, Andrea Soarescosta, M Chakravarthi, Carolina Werner Ribeiro, S M Chabregas, M C Falco, Flavio HenriquesilvaAbstract:Transgenic sugarcane expressing CaneCPI-1 exhibits resistance to Sphenophorus levis larvae. Transgenic plants have widely been used to improve resistance against insect attack. Sugarcane is an economically important crop; however, great losses are caused by insect attack. Sphenophorus levis is a sugarcane weevil that digs tunnels in the stem base, leading to the destruction of the crop. This insect is controlled inefficiently by chemical insecticides. Transgenic plants expressing peptidase inhibitors represent an important strategy for impairing insect growth and development. Knowledge of the major peptidase group present in the insect gut is critical when choosing the most effective inhibitor. S. levis larvae use cysteine peptidases as their major digestive enzymes, primarily cathepsin L-like activity. In this study, we developed transgenic sugarcane plants that overexpress sugarcane cysteine peptidase inhibitor 1 (CaneCPI-1) and assessed their potential through feeding bioassays with S. levis larvae. Cystatin overexpression in the transgenic plants was evaluated using semi-quantitative RT-PCR, RT-qPCR, and immunoblot assays. A 50% reduction of the average weight was observed in larvae that fed on transgenic plants in comparison to larvae that fed on non-transgenic plants. In addition, transgenic sugarcane exhibited less damage caused by larval attack than the controls. Our results suggest that the overexpression of CaneCPI-1 in sugarcane is a promising strategy for improving resistance against this insect.
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pectinases from Sphenophorus levis vaurie 1978 coleoptera curculionidae putative accessory digestive enzymes
Journal of Insect Science, 2015Co-Authors: Danilo Elton Evangelista, Fernando Fonseca Pereira De Paula, Andre Rodrigues, Flavio HenriquesilvaAbstract:The cell wall in plants offers protection against invading organisms and is mainly composed of the polysaccharides pectin, cellulose, and hemicellulose, which can be degraded by plant cell wall degrading enzymes (PCWDEs). Such enzymes are often synthesized by free living microorganisms or endosymbionts that live in the gut of some animals, including certain phytophagous insects. Thus, the ability of an insect to degrade the cell wall was once thought to be related to endosymbiont enzyme activity. However, recent studies have revealed that some phytophagous insects are able to synthesize their own PCWDEs by endogenous genes, although questions regarding the origin of these genes remain unclear. This study describes two pectinases from the sugarcane weevil, Sphenophorus levis Vaurie, 1978 (Sl-pectinases), which is considered one of the most serious agricultural pests in Brazil. Two cDNA sequences identified in a cDNA library of the insect larvae coding for a pectin methylesterase (PME) and an endo-polygalacturonase (endo-PG)—denominated Sl-PME and Sl-endoPG, respectively—were isolated and characterized. The quantitative real-time reverse transcriptase polymerase chain reaction expression profile for both Sl-pectinases showed mRNA production mainly in the insect feeding stages and exclusively in midgut tissue of the larvae. This analysis, together Western blotting data, suggests that Sl-pectinases have a digestive role. Phylogenetic analyses indicate that Sl-PME and Sl-endoPG sequences are closely related to bacteria and fungi, respectively. Moreover, the partial genomic sequences of the pectinases were amplified from insect fat body DNA, which was certified to be free of endosymbiotic DNA. The analysis of genomic sequences revealed the existence of two small introns with 53 and 166 bp in Sl-endoPG, which is similar to the common pattern in fungal introns. In contrast, no intron was identified in the Sl-PME genomic sequence, as generally observed in bacteria. These data support the theory of horizontal gene transfer proposed for the origin of insect pectinases, reinforcing the acquisition of PME genes from bacteria and endo-PG genes from fungi.
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a novel β fructofuranosidase in coleoptera characterization of a β fructofuranosidase from the sugarcane weevil Sphenophorus levis
Insect Biochemistry and Molecular Biology, 2014Co-Authors: Rafael Pedezzi, Walter R Terra, Fernando P. P. Fonseca, Celio Dias Santos, Luciano Takeshi Kishi, Flavio HenriquesilvaAbstract:β-fructofuranosidases or invertases (EC 3.2.1.26) catalyze the hydrolysis of sucrose into fructose and glucose. β-fructofuranosidases have been widely described in microorganisms, but were not known in the animal kingdom until very recently. There are studies reporting lepidopteran β-fructofuranosidases, but no β-fructofuranosidase gene sequence or encoding transcript has previously been identified in beetles. Considering the scarcity of functional studies on insect β-fructofuranosidases and their apparent non-occurrence among coleopterans, the aim of the present study was to investigate the occurrence and characterize a β-fructofuranosidase transcript identified in a cDNA library from the sugarcane weevil, Sphenophorus levis (Curculionidae). To validate that the β-fructofuranosidase sequence (herein denominated Sl-β-fruct) is indeed encoded by the S. levis genome, PCRs were performed using genomic DNA extracted from the larval fat body as well as DNA from the midgut with microbial content. Amplification of Sl-β-fruct gene using larval fat body DNA indicated its presence in the insect's genomic DNA. The Sl-β-fruct gene was cloned in Pichia pastoris to produce the recombinant enzyme (rSl-β-fruct). Molecular weight of the recombinant protein was about 64 kDa, indicating possible glycosylation, since the theoretical weight was 54.8 kDa. The substrate specificity test revealed that rSl-β-fruct hydrolyzes sucrose and raffinose, but not melibiose or maltose, thereby confirming invertase activity. The pH curve revealed greatest activity at pH 5.0, demonstrating rSl-β-fruct to be an acidic β-fructofuranosidase. Quantitative PCR (qRT-PCR) analyses indicated that the production of mRNA only occurs in the midgut and reaches the greatest expression level in 30-day-old larvae, which is the expected pattern for digestive enzymes. Chromatography of glycosidases from S. levis midguts showed two enzymes acting as β-fructofuranosidase, indicating the presence of a Sl-β-fruct isoform or a β-fructofuranosidase from insect intestinal microbiota. Moreover, it was found that α-glucosidases do not act on sucrose hydrolysis. Phylogenetic analyses indicated this enzyme to be similar to enzymes found in other coleopteran and lepidopteran β-fructofuranosidases, but also closely similar to bacterial enzymes, suggesting potential horizontal gene transfer. Despite this, the enzyme seems to be restricted to different groups of bacteria, which suggests distinct origin events. The present study expands the concept of the occurrence of β-fructofuranosidase in insects. Despite the few descriptions of this gene in the animal kingdom, it is possible to state that β-fructofuranosidase is crucial to the establishment of some insects throughout their evolutionary history, especially members of the Lepidoptera and Coleoptera clades.
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expression purification crystallization and preliminary x ray diffraction analysis of the pectin methylesterase from the sugar cane weevil Sphenophorus levis
Acta Crystallographica Section F-structural Biology and Crystallization Communications, 2014Co-Authors: Danilo Elton Evangelista, Fernando Fonseca Pereira De Paula, Flavio Henriquesilva, A S Godoy, Igor PolikarpovAbstract:Pectin methylesterase removes the methyl groups from the main chain of pectin, the major component of the middle lamella of the plant cell wall. The enzyme is involved in plant cell-wall development, is part of the enzymatic arsenal used by microorganisms to attack plants and also has a wide range of applications in the industrial sector. Therefore, there is a considerable interest in studies of the structure and function of this enzyme. In this work, the pectin methylesterase from Sphenophorus levis was produced in Pichia pastoris and purified. Crystals belonging to the monoclinic space group C2, with unit-cell parameters a = 122.181, b = 82.213, c = 41.176 A, β = 97.48°, were obtained by the sitting-drop vapour-diffusion method and an X-ray diffraction data set was collected to 2.1 A resolution. Structure refinement and model building are in progress.
Luis Garrigos Leite - One of the best experts on this subject based on the ideXlab platform.
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Attraction of the sugarcane billbug, Sphenophorus levis, to vinasse and its volatile composition
Chemoecology, 2020Co-Authors: Lizandra F. Martins, Mateus Tonelli, José Mauricio S. Bento, César J. Bueno, Luis Garrigos LeiteAbstract:The expansion of sugarcane plantations in Brazil and the discarding of vinasse into the sugarcane field have been speculated to contribute to the growing population of the billbug Sphenophorus levis . This beetle attacks the root system and forms galleries in rhizomes, causing damage or even the death of host plants. It has been suspected that vinasse, a residue from ethanol production, can release volatiles that are attractive to the insect; however, no study has addressed this hypothesis so far. The aim of this study was to investigate the attractiveness of vinasse to S. levis adults and identify the volatile compounds released by this substance using gas chromatography–mass spectrometry (GC–MS). We found that vinasse was more attractive to S. levis than sugarcane stems, molasses and wastewater under laboratory conditions, but not than cane stems at field conditions. Our GC–MS analysis revealed the presence of primary alcohols, terpenes and organic carboxylic acids in vinasse. When a mixture of the commercial synthetic compounds identified in the chemical analysis was tested in the laboratory, a strong attraction of the insects to the mixture was observed. Our results help to explain how vinasse can contribute to the infestations of S. levis in sugarcane fields and shed new light on the development of strategies to control this pest using chemical attractants.
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eficiencia de nematoides entomopatogenicos e inseticidas quimicos contra Sphenophorus levis e leucothyreus sp em cana de acucar
Tropical agricultural research, 2012Co-Authors: Luis Garrigos Leite, Paulo Sergio Machado Botelho, Fernando Martins Tavares, Antonio Batista Filho, Ricardo Antonio Polanczyk, Fabio Silber SchmidtAbstract:The sugarcane billbug, Sphenophorus levis (Coleoptera: Dryophthoridae), is an important pest of sugarcane crops in the Sao Paulo State, Brazil. Their larvae damage plants rhizome, causing losses of up to 30 t ha -1 year -1 of sugarcane. In order to evaluate the effectiveness of entomopathogenic nematodes and chemical insecticides against the sugarcane billbug and the scarab beetle Leucothyreus sp., three field trials were carried out in Brazil. For the first one, 10 treatments were considered: 1) thiametoxan (Actara 250WG) 500 g ha -1 of commercial product (c.p.); 2) thiametoxan 1.0 kg ha -1 c.p.; 3) thiametoxan 1.5 kg ha -1 c.p.; 4) Steinernema brazilense 10 8 infective juveniles (IJ) ha -1 + thiametoxan 250 g ha -1 c.p.; 5) S. brazilense (10 8 IJ ha -1 ) + thiametoxan 500 g ha -1 c.p.; 6) Heterorhabditis indica (10 8 IJ ha -1 ) + thiametoxan 250 g ha -1 c.p; 7) H. indica (10 8 IJ ha -1 ) + thiametoxan 500 g ha -1 c.p.; 8) S. brazilense (10 8 IJ ha -1 ); 9) H. indica (10 8 IJ ha -1 ); and 10) Control. The second one consisted of five treatments: 1) thiametoxan (Actara 250WG) 800 g ha -1 c.p.; 2) fipronil (Regente 800WG) 250 g ha -1 c.p.; 3) S. brazilense (10 8 IJ ha -1 ); 4) S. brazilense (10 8 IJ ha -1 ) + thiametoxan 200 g ha -1 c.p.; and 5) Control. In the third one, all treatments already mentioned for the second trial plus the mixture S. brazilense (10 8 IJ ha -1 ) + fipronil (Regente 800WG) 62.5 g ha -1 c.p. were considered. All treatments with the insecticides and nematodes provided higher sugarcane yield, when compared to their respective controls, which were most damaged by S. levis , but without statistical significance. S. brazilense was more efficient on controlling this pest, if compared to H. indica , but without statistical significance. S. brazilense provided a 50% control of Leucothyreus sp. larvae, meanwhile the best treatments against this scarab were the mixtures of this nematode with fipronil (78%) and thiametoxan (83%).
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Efficiency of entomopathogenic nematodes and chemical insecticides against Sphenophorus levis and Leucothyreus sp. on sugarcane
'FapUNIFESP (SciELO)', 2012Co-Authors: Luis Garrigos Leite, Paulo Sergio Machado Botelho, Fernando Martins Tavares, Antonio Batista Filho, Ricardo Antonio Polanczyk, Fabio Silber SchmidtAbstract:The sugarcane billbug, Sphenophorus levis (Coleoptera: Dryophthoridae), is an important pest of sugarcane crops in the São Paulo State, Brazil. Their larvae damage plants rhizome, causing losses of up to 30 t ha-1 year-1 of sugarcane. In order to evaluate the effectiveness of entomopathogenic nematodes and chemical insecticides against the sugarcane billbug and the scarab beetle Leucothyreus sp., three field trials were carried out in Brazil. For the first one, 10 treatments were considered: 1) thiametoxan (Actara 250WG) 500 g ha-1 of commercial product (c.p.); 2) thiametoxan 1.0 kg ha-1 c.p.; 3) thiametoxan 1.5 kg ha-1 c.p.; 4) Steinernema brazilense 108 infective juveniles (IJ) ha-1 + thiametoxan 250 g ha-1 c.p.; 5) S. brazilense (108 IJ ha-1) + thiametoxan 500 g ha-1 c.p.; 6) Heterorhabditis indica (108 IJ ha-1) + thiametoxan 250 g ha-1 c.p; 7) H. indica (108 IJ ha-1) + thiametoxan 500 g ha-1 c.p.; 8) S. brazilense (108 IJ ha-1); 9) H. indica (108 IJ ha-1); and 10) Control. The second one consisted of five treatments: 1) thiametoxan (Actara 250WG) 800 g ha-1 c.p.; 2) fipronil (Regente 800WG) 250 g ha-1 c.p.; 3) S. brazilense (108 IJ ha-1); 4) S. brazilense (108 IJ ha-1) + thiametoxan 200 g ha-1 c.p.; and 5) Control. In the third one, all treatments already mentioned for the second trial plus the mixture S. brazilense (108 IJ ha-1) + fipronil (Regente 800WG) 62.5 g ha-1 c.p. were considered. All treatments with the insecticides and nematodes provided higher sugarcane yield, when compared to their respective controls, which were most damaged by S. levis, but without statistical significance. S. brazilense was more efficient on controlling this pest, if compared to H. indica, but without statistical significance. S. brazilense provided a 50% control of Leucothyreus sp. larvae, meanwhile the best treatments against this scarab were the mixtures of this nematode with fipronil (78%) and thiametoxan (83%)
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virulencia de nematoides entomopatogenicos nematoda rhabditida a Sphenophorus levis coleoptera curculionidae
Bragantia, 2011Co-Authors: F H C Giometti, Luis Garrigos Leite, Fernando Martins Tavares, Fabio Silber Schmit, Antonio Batista Filho, Raphaela DellacquaAbstract:The sugarcane billbug, Sphenophorus levis (Coleoptera: Curculionidae) is an important underground pest of the sugarcane crop in Sao Paulo state, causing losses up to 30 tons of cane/ha/year. This study aimed to evaluate the virulence of 17 strains of entomopathogenic nematodes on the genus Steinernema and Heterorhabditis against adults of S. levis, and the effect of the four most virulent ones in three dosage each one. The research was done in laboratory conditions, performing four tests related to the virulence study and one related to the dosage study. S. brazilense IBCB n6, Heterorhabditis sp. IBCB n10, Heter- orhabditis sp. IBCB n24 and Heterorhabditis sp. IBCB n44 were the most virulent ones, differing significantly from the respec - tive control with 45%, 40%, 20% and 31% mortality of insects using the concentration of 240 infective juveniles (IJ)/insect. For the dosage test, the strains IBCB n10, IBCB n24 and IBCB n44 of Heterorhabditis noteworthy concerning the virulence for adults of the sugarcane billbug, differing significantly from the control at the higher dosage (1200 IJ/insect) with insect mortality levels of 60%, 65,7% e 74,3%, respectively.
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Virulência de nematóides entomopatogênicos (Nematoda: Rhabditida) a Sphenophorus levis (Coleoptera: Curculionidae)
Instituto Agronômico de Campinas, 2011Co-Authors: Fernando Henrique Carvalho Giometti, Luis Garrigos Leite, Fernando Martins Tavares, Fabio Silber Schmit, Antonio Batista Filho, Raphaela Dell'acquaAbstract:O bicudo da cana-de-açúcar, Sphenophorus levis (Coleoptera: Curculionidae), é uma importante praga de solo dos canaviais no Estado de São Paulo, ocasionando prejuízos de até 30 t de cana/ha/ano. Visando ao controle biológico desta praga, esse estudo teve por objetivo avaliar a virulência de 17 isolados de nematóides entomopatogênicos do gênero Steinernema e Heterorhabditis contra adultos de S. levis, e o efeito dos quatro nematóides mais virulentos em três dosagens cada um. A pesquisa foi realizada em condições de laboratório, sendo realizados quatro ensaios para o estudo de virulência. Os nematóides selecionados como mais virulentos em cada teste foram S. brazilense IBCB n6, Heterorhabditis sp. IBCB n10, Heterorhabditis sp. IBCB n24 e Heterorhabditis sp. IBCB n44, sendo os únicos que diferenciaram significativamente das respectivas testemunhas com 45%, 40%, 20% e 31% de mortalidade do inseto na concentração de 240 juvenis infectivos (JI)/inseto. No teste de dose, os isolados de Heterorhabditis identificados como IBCB n10, IBCB n24 e IBCB n44 destacaram-se quanto à virulência para os adultos do bicudo da cana-de-açúcar, diferindo significativamente da testemunha na maior dose avaliada (1200 JI/inseto) com níveis de mortalidade do inseto de 60%, 65,7% e 74,3% respectivamente
Douglas S. Richmond - One of the best experts on this subject based on the ideXlab platform.
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characterizing billbug Sphenophorus spp seasonal biology using dna barcodes and a simple morphometric analysis
Insects, 2021Co-Authors: Marian M Rodriguezsoto, Douglas S. Richmond, Ricardo A. Ramirez, Xi Xiong, Laramy S EndersAbstract:Billbugs (Sphenophorus spp.) are a complex of grass-feeding weevil species that reduce the aesthetic and functional qualities of turfgrass. Effective billbug monitoring and management programs rely on a clear understanding of their seasonal biology. However, our limited understanding of regional variation in the species compositions and seasonal biology of billbugs, stemming primarily from our inability to identify the damaging larval stage to species level, has hindered efforts to articulate efficient IPM strategies to growers. We used a combination of DNA barcoding methods and morphometric measures to begin filling critical gaps in our understanding of the seasonal biology of the billbug species complex across a broad geographic range. First, we developed a DNA barcoding reference library using cytochrome oxidase subunit 1 (COI) sequences from morphologically identified adult billbugs collected across Indiana, Missouri, Utah and Arizona. Next, we used our reference library for comparison and identification of unknown larval specimens collected across the growing season in Utah and Indiana. Finally, we combined our DNA barcoding approach with larval head capsule diameter, a proxy for developmental instar, to develop larval phenology charts. Adult COI sequences varied among billbug species, but variation was not influenced by geography, indicating that this locus alone was useful for resolving larval species identity. Overlaid with head capsule diameter data from specimens collected across the growing season, a better visualization of billbug species composition and seasonal biology emerged. This approach will provide researchers with the tools necessary to fill critical gaps in our understanding of billbug biology and facilitate the development of turfgrass pest management programs.
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Volatile and Contact Chemical Cues Associated with Host and Mate Recognition Behavior of Sphenophorus venatus and Sphenophorus parvulus (Coleoptera: Dryophthoridae)
Journal of Chemical Ecology, 2018Co-Authors: Alexandra G. Duffy, Matthew D. Ginzel, Gabriel P. Hughes, Douglas S. RichmondAbstract:Beetles in the genus Sphenophorus Schönherr, or billbugs, potentially utilize both volatile and non-volatile behavior-modifying chemical signals. These insects are widely distributed across North America, often occurring in multi-species assemblages in grasses. However, details about their host- and mate-finding behavior are poorly understood. This study tested the hypothesis that volatile organic compounds from host-plants and conspecifics direct the dispersal behavior of hunting billbug S. venatus Say. Further, we characterized the cuticular hydrocarbon profiles of two widespread pest species, S. venatus and bluegrass billbug S. parvulus Gyllenhaal, to assess the potential role of contact pheromones in mate-recognition. In Y-tube olfactometer bioassays, S. venatus males were attracted to a combination of conspecifics and Cynodon dactylon host-plant material, as well as C. dactylon plant material alone. S. venatus females were attracted to a combination of male conspecifics and host-plants but were also attracted to male conspecifics alone. Field evaluation of a putative male-produced aggregation pheromone, 2-methyl-4-octanol, identified from two congeners, S. levis Vaurie and S. incurrens Gyllenhaal, did not support the hypothesis that S. venatus and S. parvulus were also attracted to this compound. Gas chromatography-mass spectrometry analysis of S. venatus and S. parvulus whole-body cuticular extracts indicated a series of hydrocarbons with qualitative and quantitative interspecific variation in addition to intraspecific quantitative variation between males and females. This study provides the first evidence that S. venatus orients toward host- and insect-derived volatile organic compounds and substantiates the presence of species-specific cuticular hydrocarbons that could serve as contact pheromones for sympatric Sphenophorus species.
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billbug coleoptera dryophthoridae Sphenophorus spp seasonal biology and dna based life stage association in indiana turfgrass
Journal of Economic Entomology, 2018Co-Authors: Alexandra G. Duffy, Gareth S Powell, Jennifer M Zaspel, Douglas S. RichmondAbstract:Eleven species of billbugs (Coleoptera: Dryophthoridae: Sphenophorus spp. Schonherr) infest managed turfgrass in North America. However, the regional variation in species composition remains unresolved and the seasonal phenology of several species has not been well documented. The latter gap is largely due to the inability to identify the larval stage to species-a confounding problem with several sympatric insect species. We used field trapping (adults) and soil sampling (larvae and pupae) surveys along with a DNA-based life-stage association to characterize the biology of billbugs associated with turfgrass in the Midwestern United States. Pitfall trapping at four locations in Indiana revealed four billbug species: S. venatus Say, S. parvulus Gyllenhaal, S. minimus Hart, and S. inaequalis Say. Sphenophorus venatus was the most abundant species on warm-season turfgrass while S. parvulus was most abundant on cool-season turfgrass. Investigation of S. venatus seasonal biology revealed two overwintered life stages-larva and adult-which resulted in two overlapping cohorts and two larval generations. Degree-day models describing S. venatus activity were more accurate for first-generation adults and larvae than for overwintering life stages. Maximum-likelihood analyses provided the first molecular species identification of billbug larvae and direct evidence that S. venatus larvae are capable of overwintering above 40°N latitude. Findings clarify the utility of molecular markers (CO1, 18S, and ITS2) for describing billbug larval population dynamics and seasonal phenology in regions where several sympatric billbug species occur. These results support the development of sustainable management strategies based on billbug seasonal phenology in different regions of North America.
Ordaz González, Edith Lorena - One of the best experts on this subject based on the ideXlab platform.
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Identificación de Sphenophorus venatus vestitus, su distribución y asociación con Paspalum vaginatum Swartz en México.
Colegio de Postgraduados (COLPOS), 2014Co-Authors: Ordaz González, Edith LorenaAbstract:En el presente trabajo se realizó la identificación de cinco especies de Sphenophorus (Coleoptera: Dryophthoridae) asociadas a campos de golf en siete estados de México en varias plantas hospederas; Sphenophorus arizonensis Horn en el estado de Puebla en Paspalum vaginatum Swartz (nuevo registro estatal y de hospedero); Sphenophorus cicatristriatus Fahraeus se colectó en Puebla en Cynodon dactylon (L.) Pers.; Sphenophorus incurrens Gyllenhal fue colectado en Querétaro en Pennisetum clandestinum Hochst. (Nuevo registro estatal); Sphenophorus rectus (Say) se colectó en Veracruz en P. vaginatum (Nuevo hospedero); y Sphenophorus venatus vestitus Chittenden que fue colectado en Puebla, Veracruz, Guerrero, Baja California Sur y Nuevo León en C. dactylon y en Guanajuato en Lolium perenne L. (nuevo hospedero). Se realizó la comparación de sus caracteres morfológicos externos y de genitalia masculina para su identificación. Además de realizar un ensayo molecular usando el gen COI, para obtener la secuencia de ADN, de las especies del género. La información generada contribuye con las bases de la distribución y diversidad de las especies del género Sphenophorus en el país; con lo cual se busca a mediano plazo el mejoramiento de las técnicas de manejo de las especies de este género que atacan pastos en los campos de golf en México. _______________ ABSTRACT: In this study five species Sphenophorus (Coleoptera: Dryophthoridae) were found associated to golf courses in seven states of Mexico on several host plants; Sphenophorus arizonensis Horn in the state of Puebla in Paspalum vaginatum Swartz ( new state record and host ); Sphenophorus cicatristriatus Fahraeus was collected in Puebla in Cynodon dactylon (L. ) Pers; Sphenophorus incurrens Gyllenhal was collected in Querétaro in Pennisetum clandestinum Hochst. ( new state record) ; Sphenophorus rectus (Say) was collected in Veracruz in P. vaginatum (New host) and Sphenophorus venatus vestitus Chittenden was collected in Puebla , Veracruz, Guerrero, Baja California Sur , and Nuevo Leon in C. dactylon and Guanajuato in Lolium perenne L. (new host). External morphological characters and male genitalia were studied for identification. In addition, a molecular assay using the COI gene to obtain the DNA sequence of the genus was done. The information contributes to the knowledge of the distribution and diversity of species in the genus Sphenophorus in the country, which seeks to medium term, improved management of the species of this genus that attack grass in golf courses in Mexico
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Identificación de Sphenophorus venatus vestitus, su distribución y asociación con Paspalum vaginatum Swartz en México.
Colegio de Postgraduados (COLPOS), 2014Co-Authors: Ordaz González, Edith LorenaAbstract:En el presente trabajo se realizó la identificación de cinco especies de Sphenophorus (Coleoptera: Dryophthoridae) asociadas a campos de golf en siete estados de México en varias plantas hospederas; Sphenophorus arizonensis Horn en el estado de Puebla en Paspalum vaginatum Swartz (nuevo registro estatal y de hospedero); Sphenophorus cicatristriatus Fahraeus se colectó en Puebla en Cynodon dactylon (L.) Pers.; Sphenophorus incurrens Gyllenhal fue colectado en Querétaro en Pennisetum clandestinum Hochst. (Nuevo registro estatal); Sphenophorus rectus (Say) se colectó en Veracruz en P. vaginatum (Nuevo hospedero); y Sphenophorus venatus vestitus Chittenden que fue colectado en Puebla, Veracruz, Guerrero, Baja California Sur y Nuevo León en C. dactylon y en Guanajuato en Lolium perenne L. (nuevo hospedero). Se realizó la comparación de sus caracteres morfológicos externos y de genitalia masculina para su identificación. Además de realizar un ensayo molecular usando el gen COI, para obtener la secuencia de ADN, de las especies del género. La información generada contribuye con las bases de la distribución y diversidad de las especies del género Sphenophorus en el país; con lo cual se busca a mediano plazo el mejoramiento de las técnicas de manejo de las especies de este género que atacan pastos en los campos de golf en México. _______________ ABSTRACT: In this study five species Sphenophorus (Coleoptera: Dryophthoridae) were found associated to golf courses in seven states of Mexico on several host plants; Sphenophorus arizonensis Horn in the state of Puebla in Paspalum vaginatum Swartz ( new state record and host ); Sphenophorus cicatristriatus Fahraeus was collected in Puebla in Cynodon dactylon (L. ) Pers; Sphenophorus incurrens Gyllenhal was collected in Querétaro in Pennisetum clandestinum Hochst. ( new state record) ; Sphenophorus rectus (Say) was collected in Veracruz in P. vaginatum (New host) and Sphenophorus venatus vestitus Chittenden was collected in Puebla , Veracruz, Guerrero, Baja California Sur , and Nuevo Leon in C. dactylon and Guanajuato in Lolium perenne L. (new host). External morphological characters and male genitalia were studied for identification. In addition, a molecular assay using the COI gene to obtain the DNA sequence of the genus was done. The information contributes to the knowledge of the distribution and diversity of species in the genus Sphenophorus in the country, which seeks to medium term, improved management of the species of this genus that attack grass in golf courses in Mexico.Tesis (Maestría en Ciencias, especialista en Entomología y Acarología).- Colegio de Postgraduados, 2014.Consejo Nacional de Ciencia y Tecnología (CONACYT)