ABSTRACT
BACKGROUND: Piercing/sucking insect pests in the order Hemiptera causes substantial crop losses by removing photoassimilates and transmitting viruses to their host plants. Cloning and heterologous expression of plantderived insect resistance genes is a promising approach to control aphids and other sap-sucking insect pests. While expression from the constitutive 35S promoter provides broad protection, the phloem-specific rolC promoter provides better defense against sap sucking insects. The selection of plant-derived insect resistance genes for expression in crop species will minimize bio-safety concerns. RESULTS: Pinellia ternata leaf agglutinin gene (pta), encodes an insecticidal lectin, was isolated and cloned under the 35S and rolC promoters in the pGA482 plant transformation vector for Agrobacterium-mediated tobacco transformation. Integration and expression of the transgene was validated by Southern blotting and qRT-PCR, respectively. Insect bioassays data of transgenic tobacco plants showed that expression of pta under rolC promoter caused 100% aphid mortality and reduced aphid fecundity up to 70% in transgenic tobacco line LRP9. These results highlight the better effectivity of pta under rolC promoter to control phloem feeders, aphids. CONCLUSIONS: These findings suggested the potential of PTA against aphids and other sap sucking insect pests. Evaluation of gene in tobacco under two different promoters; 35S constitutive promoter and rolC phloemspecific promoter could be successfully use for other crop plants particularly in cotton. Development of transgenic cotton plants using plant-derived insecticidal, PTA, would be key step towards commercialization of environmentally safe insect-resistant crops.
Subject(s)
Aphids/pathogenicity , Pest Control, Biological , Pinellia/chemistry , Plant Viruses , Tobacco , Blotting, Southern , Polymerase Chain Reaction , Promoter Regions, Genetic , Plants, Genetically Modified , Plant Leaves/chemistry , Transgenes , Disease Resistance , Crop ProtectionABSTRACT
Se encontraron cuatro especies de hongos Entomophthorales, Pandora neoaphidis, Zoophthora radicans, Entomophthora planchoniana (Entomophthorales: Entomophthoraceae) y Neozygites fresenii (Neozygitales: Neozygitaceae) infectando a Aphis craccivora, Therioaphis trifolii, Acyrthosiphon pisum y a especies no identificadas pertenecientes al género Acyrthosiphon en cultivos de alfalfa (Medicago sativa L.), en la Argentina. Los muestreos fueron realizados en cinco sitios (Ceres, Rafaela, Sarmiento, Monte Vera y Bernardo de Irigoyen) de la provincia de Santa Fe. Zoophthora radicans fue el patógeno más importante registrado principalmente en Acyrthosiphon sp. Zoophthora radicans fue exitosamente aislado y mantenido en cultivos puros. Este estudio documenta por primera vez en la Argentina la presencia de hongos Entomophthorales infectando áfidos en alfalfa
Four species of entomophthoroid fungi, Pandora neoaphidis (Entomophthorales: Entomophthoraceae), Zoophthora radicans (Entomophthorales: Entomophthoraceae), Entomophthora planchoniana (Entomophthorales: Entomophthoraceae) and Neozygites fresenii (Neozygitales: Neozygitaceae) were found to infect Aphis craccivora, Therioaphis trifolii, and Acyrthosiphon pisum and unidentified species of Acyrthosiphon on lucerne in Argentina. Samples were collected from five sites (Ceres, Rafaela, Sarmiento, Monte Vera and Bernardo de Irigoyen) in the province of Santa Fe. In this study, Zoophthora radicans was the most important pathogen and was recorded mainly on Acyrthosiphon sp. Zoophthora radicans was successfully isolated and maintained in pure cultures. This study is the first report of entomophthoroid fungi infecting lucerne (Medicago sativa L.) aphids in Argentina
Subject(s)
Aphids/pathogenicity , Argentina/epidemiology , Medicago sativa/toxicity , Entomophthorales/isolation & purification , Entomophthorales/classificationABSTRACT
Pandora neoaphidis overwintering had been investigated by monitoring its prevalence in Myzus persicae populations in open fields. Cabbage plants in field plots were weekly taken after mycosis initiation, to count and examine the living and dead aphids infected by P. neoaphidis. Based on the field data, infection levels (I) varied with field temperature (T), relative humidity (RH) and aphid count (numbers of living aphids per plant, N) over days (D), fitting well to the modified logistic equation I=0.91/[1+exp(8.5+(2.0H T H RH-20.2NI0)D)] (r²=0.897), where H T indicated daily hours of low temperature (<4°C), H RH daily hours of high air humidity (>90% RH) and I0 primary infection level. The model demonstrated the abiotic and biotic factors influencing P. neoaphidis mycosis development in winter, and also verifies the fungal overwintering by infecting available host aphids without a resting stage. Ultimately, P. neoaphidis mycosis reduced 81.4% of aphid populations, presenting great potential for biocontrol.
Subject(s)
Abiotic Factors , Aphids/pathogenicity , Environmental Monitoring , Fungi , Hibernation , Models, Anatomic , Mycoses , Vegetables , Brassica , Environmental Microbiology , Food Samples , Methods , Prevalence , MethodsABSTRACT
El pulgón del ciprés Cinara cupressi, se considera uno de los insectos invasores más dañinos del mundo, ha provocado una gran mortalidad en especies Cupresáceas nativas y exóticas en varios países. En Chile, fue detectado el año 2003 y para el año 2008 esta plaga ya se encontraba distribuida en todo el país, afectando especies cupresáceas forestales exóticas como Cupressus macrocarpa y especies nativas, tales como, Austrocedrus chilensis (Ciprés de la Cordillera) y Fitzroya cupressoides (Alerce), que están categorizadas en el estado de conservación vulnerable y en peligro respectivamente. El área forestal de Chile ha potenciado y privilegiado el uso de controladores biológicos como parte del manejo integrado de plagas. Es por esto que se ha utilizado el parasitoide Pauesia juniperorum. Sin embargo, se han detectado bajos niveles de parasitismo. Se propone el uso de hongos entomopatógenos como alternativa y complemento al control biológico. Se prospectaron hongos entomopatógenos en la Región ecológica mediterránea per-húmeda y en la Región oceánica con influencia mediterránea. Como resultado se han identificado seis géneros de hongos descritos como patógenos de insectos. Sin embargo, bajo condiciones de laboratorio sólo cepas del género Verticillium fueron virulentas, causando sobre el 80 por ciento de mortalidad acumulada a los 7 días. No hubo diferencias significativas entre las cepas de Verticillium y un producto químico (pirimicarb) aplicado como control, aunque el hongo fue más lento. Las cepas más virulentas fueron identificadas como Verticillium lecanii, Ve 1 y Ve 2, con un TL50 de: 3.2 y 3.1 días y un DL50 : 1.24 y 1.37 conidias mL-1 respectivamente, sugiriendo el uso de estos hongos para controlar C. cupressi en Chile.
The cypress aphid, Cinara cupressi, is considered one of the most important invasive species causing high mortality in exotic and native species of Cupressaceae in several countries in the world. In Chile it was detected in 2003 and in 2008 was distributed throughout the country affecting the exotic forest species Cupressus macrocarpa and the native forest species Austrocedrus chilensis (Ciprés de la Cordillera) and Fitzroya cupressoides (Alerce), both classified as vulnerable and endangered species respectively. Efforts to their management have focused on biological control by using the parasitoid Pauesia juniperorum but until now it has not reached satisfactory control. We propose using entomopathogenic fungi, as alternative and complementary biocontrol. Entomopathogenic fungi were prospected in the ecological region Mediterranean per-humid and in the ecological region Oceanic with mediterranean influence in both colonies of C. cupressi. There were identified six genera of fungi described as insect pathogens. However, in laboratory assays only Verticillium strains were virulent, causing about 80 percent cumulative mortality at seven days. There were not significant differences among strains of Verticillium and chemical (pirimicarb) applied as control, although fungi were slower. The most virulent strains were two Verticillium lecanii, Ve 1 and Ve 2, with an LT50 of: 3.2 and 3.1 days and LD50 : 1.24 and 1.37 conidia mL-1 respectively, suggesting the use of these fungi to control C. cupressi in Chile.
Subject(s)
Aphids/pathogenicity , Cupressus/microbiology , Fusarium , Fungi/isolation & purification , Fungi/classification , Fungi/growth & development , Fungi/pathogenicity , Paecilomyces , Pest Control, Biological , Verticillium , Chile , TreesABSTRACT
El pulgón del ciprés (Cinara cupressi), es una plaga exótica distribuida en todo el territorio continental chileno que ha provocado daños importantes tanto enespecies forestales introducidas como nativas. Se han estudiado algunos aspectos de la biología de estepulgón, mediante control biológico y productos químicos, los cuales no presentan actualmente resultadossatisfactorios. Durante el tiempo que esta plaga se ha establecido en Chile, no se ha realizado un estudio sistemático de hongos entomopatógenos asociados aeste pulgón, aunque se cuenta con antecedentes de un control promisorio en pulgones con estos agentes enotros países. Con el objetivo inicial de aislar en una primera fase del presente estudio hongos patogénicospotenciales para el control biológico de C. cupressi, iniciamos su búsqueda en dos regiones ecológicas del sur de Chile (Región ecológica Mediterránea Per-Húmeda y Región ecológica Oceánica con influencia Mediterránea), seleccionándose en cada una de ellas 6sitios de muestreo donde en cada uno se recolectaron 10 ramas con colonias de pulgones que fueron mantenidas en una cámara bioclimática (20 +/- 2 °C, 16:8 h) por 7 días, para estimular el desarrollo fúngico. Se aislaron integrantes de lo géneros Verticillium, Paecilomyces(ambos con mayor frecuencia de presencia), Fusarium y un entomophthoral en estudio.
The cypress aphid (Cinara cupressi) is an exotic plague distributed throughout the Chilean continental territory which has caused significant damage both inintroduced forest species as well as in native ones. Some characteristics as to the biology of this aphid have beenstudied by means of biological control and chemicals yet they have not revealed any satisfactory results up tonow. During the time of occurrence of this plague in Chile systematic study on entomopathogenic fungi associated to this aphid has failed to be accomplishedalthough information about a promissory control in aphids with these agents has been reported in other countries. In order to carry out an isolation of potential pathogenic fungi for the biological control of C.cupressiin the first phase of this present research, we began to look for them in two ecological regions from southernChile (Ecological Mediterranean Per-humid Region and Ecological Oceanic Region having Mediterraneaninfluence). Six sampling sites were selected in each of them while 10 branches infected with aphid colonies were kept in a bioclimatic chamber (20+2ºC, 16:8 h) for 7 days to stimulate fungal growth. Fungi of generaVerticillium, Paecilomyces (both showing the highest occurrence frequency), Fusarium and an entomophtoral under study were isolated.