Electrophoretic karyotype of Metarhizium anisopliae

Shimizu, Susumu; Arai, Youko; Matsumoto, Tsuguo

doi:10.1016/0022-2011(92)90094-k

Cited by 20 publications

(19 citation statements)

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“…Should that be the case, and in light of the fact that plant pathogenicity genes are located on dispensable chromosomes or portions of chromosomes, such a phenomenon could not only have signi¢cant evolutionary implications for the origin of pathogenicity genes, but it will also increase chances to identify and isolate such genes in the future. The underlying mechanism will also help to explain why there are high levels of chromosomelength and number di¡erences between di¡erent strains of M. anisopliae [4,25] and as well as in other entomopathogenic fungi [26^28].…”

Section: Discussionmentioning

confidence: 99%

Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae

Wang

2002

FEMS Microbiology Letters

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In an initial attempt to characterise single-spore isolates from Metarhizium anisopliae wild-type strain V275, three genetically identical spontaneous pr1A-and pr1B-deficient mutants were detected. These mutants were identified by Nested-PCR amplification and verified by Southern hybridisation. Sequencing and comparison of the highly conserved sequences of 28S rDNA domains 9^11 confirmed that the mutant isolates were Metarhizium anisopliae, but each had both morphological and genetic differences from the wild-type parent. RAPD analyses indicated that the overall genetic similarity between wild-type strain and the mutant was less than 70%. Enzyme assays proved that the mutant isolates had significantly lower Pr1 and elastase activities. Bioassays showed reduced lethal activity of the mutants (20%) on Tenebrio molitor, but lethal activities were similar to those of the wild-type parent on the larvae of Galleria mellonella. This paper shows for the first time that a stable mutant strain lacking the most important pr1 genes is still able to infect its respective hosts. Our observations suggest that it is not uncommon for Metarhizium anisopliae to lose its virulence genes during maintenance on artificial medium. ß

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Section: Discussionmentioning

confidence: 99%

Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae

Wang

2002

FEMS Microbiology Letters

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“…1). Three bands in Group III were only resolved using B electrophoresis conditions (Shimizu et al, 1992) (data not shown). These parameters were useful to show that some single bands showed in Group III with condition A were doublets.…”

Section: Resultsmentioning

confidence: 97%

“…The plugs were inserted into the gel wells and sealed with the same agarose used to prepare the gel. The electrophoresis conditions (pulse intervals and durations) were: A) 50 min, 45min and 37 min, during 73h, 18h and 73h, respectively, with a voltage of 46V; B) 90 and 60 min during 72h each one with a voltage of 40V (Shimizu et al, 1992). During the run, the temperature was kept at 12 o C. Gels were stained in ethidium bromide (0.5 mg/mL) for 20 min, destained in distilled water for 20 min and photographed under ultraviolet transillumination using Ilford 50 film.…”

Section: Pulsed Field Gel Electrophoresis Conditionsmentioning

confidence: 99%

“…The results derived from RAPD analysis (Fungaro et al, 1996) showed for the first time that the genetic variability among insect isolates was much lower than the genetic variability observed among soil isolates, suggesting that this fungus was developed a certain degree of host specificity. An effective technique designated pulsed-field gel electrophoresis (PFGE), was used to separate chromosome-sized DNA molecules of M. anisopliae (Shimizu et al, 1992 andValadaresInglis andPeberdy, 1998). However, so far only a few isolates have been tested by this technique and most of them derived from a single species host.…”

Section: Introductionmentioning

confidence: 99%

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Pulsed field gel electrophoresis reveals chromosome length and number differences in Brazilian strains of Metarhizium Anisopliae

Kava-Cordeiro

Queiroz

Pizzirani-Kleiner

et al. 2005

Braz. arch. biol. technol.

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“…Due to the reduced number of bands (3-8) generated by PCR amplification using several combinations of selective AFLP primers, which was attributed to the small fungal genome size (Beauveria bassiana: ∼40 Mbp, Metarhizium anisopliae: ∼30 Mbp) [18,19], the analysis was based on a double PCR amplification approach using the preselective combination of EA/MC primers. This strategy produced clear and informative fingerprint patterns for all DNA samples analyzed ( Fig.…”

Section: Modified Aflp Analysesmentioning

confidence: 99%

Diversidad genética de una colección de hongos entomopatógenos de Ecuador utilizando un enfoque AFLP modificado

Arahana

Bastidas

Torres

et al. 2013

Av. Cienc. Ing. (Quito)

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The genetic diversity of 41 Ecuadorian entomopathogenic fungal strains plus one isolate from the USA, from a collection maintained by INIAP (Instituto Nacional Autónomo de Investigaciones Agropecuarias) was determined using a modified AFLP (Amplified Fragment Length Polymorphism) approach. We found genetic similarity indexes lower than 0.50 and 121 polymorphic bands. The AMOVA analysis revealed that between-group and within-group variation contributed in similar amounts (59% and 41%, respectively) to the whole genetic variation detected. The dendrogram built using Jaccard's genetic coefficient shows twelve groups, where seven of them contain isolates clustered by genus. From these seven groups, five of them contain isolates clustered by the host from which they were recovered. The Bootstrap values show twelve reliable phylogenetic relations with values higher than 70% of confidence. The Principal Components Analysis (PCA) produced six clusters; four of them contain isolates associated by genus. The results suggest the existence of a considerable genetic diversity within the INIAP's entopathogenic fungi collection, and a clustering tendency related to the host from which they were isolated. We did not find common genomic regions among the most virulent entomopathogenic fungi strains. The high genetic diversity found within this collection represents a potential source of genotypes with potent bioinsecticide activity.Keywords. Biocontrol, Genetic Diversity, AFLP, Entomopathogenic fungi, Beauveria, Metarhizium, Verticillium ResumenSe determinó la diversidad genética de 41 cepas ecuatorianas de hongos entomopatógenos y un aislamiento de los EEUU, de una colección mantenida por el INIAP (Instituto Nacional Autónomo de Investigaciones Agropecuarias) utilizando un protocolo modificado de AFLP (Amplified Fragment Length Polymorphism). Se encontraron índices de similitud genéticos inferiores a 0.50 y 121 bandas polimórficas. El análisis AMOVA reveló que la variación entre grupos y dentro de grupos contribuían en cantidades similares (59% y 41% respectivamente) a la variación genética total detectada. El dendograma construido a partir del coeficiente genético de Jaccard muestra doce grupos, de los cuales siete contienen aislamientos agrupados por género. De estos siete grupos, cinco contienen aislamientos agrupados en base al huésped de donde se recolectaron. Los valores de Bootstrap muestran doce relaciones filogenéticas confiables con valores de confianza mayores al 70%. El análisis de componentes principales (PCA) produjo seis grupos; cuatro de ellos contienen aislamientos asociados por género. Estos resultados sugieren la existencia de una diversidad genética considerable dentro de la colección de hongos entomopatógenos del INIAP, y una tendencia de agrupamiento relacionada con el huésped de donde fueron aislados. No se encontraron regiones genómicas comunes dentro de las cepas más virulentas de hongos entomopatógenos. La elevada diversidad genética dentro de esta colección representa una fuente potencial de g...

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Electrophoretic karyotype of Metarhizium anisopliae

Cited by 20 publications

References 10 publications

Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae

Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae

Pulsed field gel electrophoresis reveals chromosome length and number differences in Brazilian strains of Metarhizium Anisopliae

Diversidad genética de una colección de hongos entomopatógenos de Ecuador utilizando un enfoque AFLP modificado

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