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Pathogenicity of Isolates of Serratia Marcescens towards Larvae of the Scarab Phyllophaga Blanchardi (Coleoptera).

Pineda-Castellanos ML, Rodríguez-Segura Z, Villalobos FJ, Hernández L, Lina L, Nuñez-Valdez ME - Pathogens (2015)

Bottom Line: Oral and injection bioassays using healthy Phyllophaga blanchardi larvae fed with the S. marcescens isolates showed different degrees of antifeeding effect and mortality.S. marcescens (Sm81) cell-free culture supernatant caused significant antifeeding effect and mortality to P. blanchardi larvae by oral bioassay and also mortality by injection bioassay.This insecticidal protein could have applications in agricultural biotechnology.

View Article: PubMed Central - PubMed

Affiliation: Centro de Investigación en Dinámica Celular, Instituto de Ciencias Básicas y Aplicadas, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, CP 62209, Cuernavaca, Morelos, Mexico. mlpc@uaem.mx.

ABSTRACT
Serratia marcescens is a Gram negative bacterium (Enterobacteriaceae) often associated with infection of insects. In order to find pathogenic bacteria with the potential to control scarab larvae, several bacterial strains were isolated from the hemocoel of diseased Phyllophaga spp (Coleoptera:Scarabaeidae) larvae collected from cornfields in Mexico. Five isolates were identified as Serratia marcescens by 16S rRNA gene sequencing and biochemical tests. Oral and injection bioassays using healthy Phyllophaga blanchardi larvae fed with the S. marcescens isolates showed different degrees of antifeeding effect and mortality. No insecticidal activity was observed for Spodoptera frugiperda larvae (Lepidoptera: Noctuidae) by oral inoculation. S. marcescens (Sm81) cell-free culture supernatant caused significant antifeeding effect and mortality to P. blanchardi larvae by oral bioassay and also mortality by injection bioassay. Heat treated culture broths lost the ability to cause disease symptoms, suggesting the involvement of proteins in the toxic activity. A protein of 50.2 kDa was purified from the cell-free broth and showed insecticidal activity by injection bioassay towards P. blanchardi. Analysis of the insecticidal protein by tandem- mass spectrometry (LC-MS/MS) showed similarity to a Serralysin-like protein from S. marcescens spp. This insecticidal protein could have applications in agricultural biotechnology.

No MeSH data available.


Related in: MedlinePlus

Skim milk plate assay to evaluate protease activity by S. marcescens strains and cell-free culture supernatants. Panel A. Protease activity of bacteria. Panel B. Protease activity of cell-free culture supernatants.
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pathogens-04-00210-f005: Skim milk plate assay to evaluate protease activity by S. marcescens strains and cell-free culture supernatants. Panel A. Protease activity of bacteria. Panel B. Protease activity of cell-free culture supernatants.

Mentions: It has been reported that a 61 kDa protease from S. marcescens HR3 culture broth has insecticidal activity towards the grassland locust Myrneleotettix palpalis Zub [9]. A characteristic of the genus Serratia is the production of extracellular proteases [6], hence, it was hypothesized that the Sm81 protease was the causative agent for the insecticidal activity observed for the cell-free culture supernatants. The S. marcescens isolates and cell-free supernatants were screened for extracellular protease activity on agar-milk plates as reported [10]. Milk clearing zones were produced by the five isolates as expected (Figure 5) demonstrating proteolytic activity. It was observed that the halo diameter sizes of one colony grown for 24 h were about 10–12 mm (Table 1).


Pathogenicity of Isolates of Serratia Marcescens towards Larvae of the Scarab Phyllophaga Blanchardi (Coleoptera).

Pineda-Castellanos ML, Rodríguez-Segura Z, Villalobos FJ, Hernández L, Lina L, Nuñez-Valdez ME - Pathogens (2015)

Skim milk plate assay to evaluate protease activity by S. marcescens strains and cell-free culture supernatants. Panel A. Protease activity of bacteria. Panel B. Protease activity of cell-free culture supernatants.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493471&req=5

pathogens-04-00210-f005: Skim milk plate assay to evaluate protease activity by S. marcescens strains and cell-free culture supernatants. Panel A. Protease activity of bacteria. Panel B. Protease activity of cell-free culture supernatants.
Mentions: It has been reported that a 61 kDa protease from S. marcescens HR3 culture broth has insecticidal activity towards the grassland locust Myrneleotettix palpalis Zub [9]. A characteristic of the genus Serratia is the production of extracellular proteases [6], hence, it was hypothesized that the Sm81 protease was the causative agent for the insecticidal activity observed for the cell-free culture supernatants. The S. marcescens isolates and cell-free supernatants were screened for extracellular protease activity on agar-milk plates as reported [10]. Milk clearing zones were produced by the five isolates as expected (Figure 5) demonstrating proteolytic activity. It was observed that the halo diameter sizes of one colony grown for 24 h were about 10–12 mm (Table 1).

Bottom Line: Oral and injection bioassays using healthy Phyllophaga blanchardi larvae fed with the S. marcescens isolates showed different degrees of antifeeding effect and mortality.S. marcescens (Sm81) cell-free culture supernatant caused significant antifeeding effect and mortality to P. blanchardi larvae by oral bioassay and also mortality by injection bioassay.This insecticidal protein could have applications in agricultural biotechnology.

View Article: PubMed Central - PubMed

Affiliation: Centro de Investigación en Dinámica Celular, Instituto de Ciencias Básicas y Aplicadas, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, CP 62209, Cuernavaca, Morelos, Mexico. mlpc@uaem.mx.

ABSTRACT
Serratia marcescens is a Gram negative bacterium (Enterobacteriaceae) often associated with infection of insects. In order to find pathogenic bacteria with the potential to control scarab larvae, several bacterial strains were isolated from the hemocoel of diseased Phyllophaga spp (Coleoptera:Scarabaeidae) larvae collected from cornfields in Mexico. Five isolates were identified as Serratia marcescens by 16S rRNA gene sequencing and biochemical tests. Oral and injection bioassays using healthy Phyllophaga blanchardi larvae fed with the S. marcescens isolates showed different degrees of antifeeding effect and mortality. No insecticidal activity was observed for Spodoptera frugiperda larvae (Lepidoptera: Noctuidae) by oral inoculation. S. marcescens (Sm81) cell-free culture supernatant caused significant antifeeding effect and mortality to P. blanchardi larvae by oral bioassay and also mortality by injection bioassay. Heat treated culture broths lost the ability to cause disease symptoms, suggesting the involvement of proteins in the toxic activity. A protein of 50.2 kDa was purified from the cell-free broth and showed insecticidal activity by injection bioassay towards P. blanchardi. Analysis of the insecticidal protein by tandem- mass spectrometry (LC-MS/MS) showed similarity to a Serralysin-like protein from S. marcescens spp. This insecticidal protein could have applications in agricultural biotechnology.

No MeSH data available.


Related in: MedlinePlus