Pineda-Castellanos Mónica L, Rodríguez-Segura Zitlhally, Villalobos Francisco J, Hernández Luciano, Lina Laura, Nuñez-Valdez M Eugenia
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.
Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, 62209, Cuernavaca, Morelos, Mexico.
Pathogens. 2015 May 13;4(2):210-28. doi: 10.3390/pathogens4020210.
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.
粘质沙雷氏菌是一种革兰氏阴性菌(肠杆菌科),常与昆虫感染有关。为了寻找具有控制金龟子幼虫潜力的病原菌,从墨西哥玉米田采集的患病叶甲属(鞘翅目:鳃金龟科)幼虫的血腔中分离出了几种细菌菌株。通过16S rRNA基因测序和生化试验,鉴定出5株分离菌为粘质沙雷氏菌。用粘质沙雷氏菌分离株喂养健康的布兰查德叶甲幼虫进行口服和注射生物测定,结果显示出不同程度的拒食作用和死亡率。通过口服接种,未观察到草地贪夜蛾幼虫(鳞翅目:夜蛾科)有杀虫活性。粘质沙雷氏菌(Sm81)无细胞培养上清液通过口服生物测定对布兰查德叶甲幼虫有显著的拒食作用和死亡率,通过注射生物测定也有致死率。热处理的培养液失去了引起疾病症状的能力,这表明蛋白质参与了毒性活动。从无细胞培养液中纯化出一种50.2 kDa的蛋白质,通过注射生物测定对布兰查德叶甲显示出杀虫活性。通过串联质谱(LC-MS/MS)对杀虫蛋白进行分析,结果显示其与粘质沙雷氏菌属的一种类丝氨酸蛋白酶相似。这种杀虫蛋白可能在农业生物技术中有应用。