Purification and Characterization of a Paralytic Polypeptide from Larvae of Myrmeleon bore

Matsuda, Kazuhiko; Suzuki, Hiroyuki; Nakanishi, Fernanda Akemi; Shio, K.; Komai, Koichiro; Nishimura, Kenji

doi:10.1006/bbrc.1995.2448

Cited by 26 publications

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“…In general, it takes several minutes for antlions to paralyze prey, but the time varies with the size of prey and the point of the antlion bite. As for factors inducing paralysis, we have isolated ALMBtoxin, which has a molecular mass of 170 kDa, from the regurgitant of antlions (Matsuda et al, 1995). However, we have also found that E. aerogenes and B. cereus produce insecticidal proteins which can kill insects rapidly by injection (Yoshida et al, 2001;Nishiwaki et al, 2004), although injection with washed insecticidal bacteria (see Section 2 for preparation) did not induce rapid paralysis.…”

Section: Discussionmentioning

confidence: 95%

“…Similar to other fluid-feeding carnivorous insects, antlions inject a regurgitant into their prey through their mandibles for extra-oral digestion before sucking out the prey's fluid (Cohen, 1995). From the regurgitant, a single polypeptide with a molecular weight of 170 kDa, antlion Myrmeleon bore toxin (ALMB-toxin), has been isolated as one of the insecticidal factors (Matsuda et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

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Insecticidal bacteria isolated from predatory larvae of the antlion species Myrmeleon bore (Neuroptera: Myrmeleontidae)

Nishiwaki

Ito

Shimomura

et al. 2007

Journal of Invertebrate Pathology

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Insecticidal bacteria isolated from predatory larvae of the antlion species Myrmeleon bore (Neuroptera: Myrmeleontidae)

Nishiwaki

Ito

Shimomura

et al. 2007

Journal of Invertebrate Pathology

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“…Because ant lions can kill prey larger than themselves, it was postulated that they used toxins. From the regurgitated fluid of Myrmeleon bore larvae, we have purified an insecticidal protein with a molecular mass of 170 kDa (19) and found that it was produced in the larval region from the thorax to the abdomen (33). In addition to the toxin produced by ant lions, insecticidal proteins were found to be produced by bacterial pathogens isolated from M. bore larvae (21,34).…”

mentioning

confidence: 99%

Cloning, Functional Characterization, and Mode of Action of a Novel Insecticidal Pore-Forming Toxin, Sphaericolysin, Produced by Bacillus sphaericus

Nishiwaki

Nakashima

Ishida

et al. 2007

Appl Environ Microbiol

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An insecticidal protein produced by Bacillus sphaericus A3-2 was purified to elucidate its structure and mode of action. The active principle purified from the culture broth of A3-2 was a protein with a molecular mass of 53 kDa that rapidly intoxicated German cockroaches (Blattela germanica) at a dose of about 100 ng when injected. The insecticidal protein sphaericolysin possessed the undecapeptide motif of cholesterol-dependent cytolysins and had a unique N-terminal sequence. The recombinant protein expressed in Escherichia coli was equally as potent as the native protein. Sphaericolysin-induced hemolysis resulted from the protein's poreforming action. This activity as well as the insecticidal activity was markedly reduced by a Y159A mutation. Also, coapplication of sphaericolysin with cholesterol abolished the insecticidal action, suggesting that cholesterol binding plays an important role in insecticidal activity. Sphaericolysin-lysed neurons dissociated from the thoracic ganglia of the German cockroaches. In addition, sphaericolysin's activity in ganglia was suppressed by the Y159A mutation. The sphaericolysin-induced damage to the cockroach ganglia was greater than the damage to the ganglia of common cutworms (Spodoptera litura), which accounts, at least in part, for the higher sensitivity to sphaericolysin displayed by the cockroaches than that displayed by cutworms.Using entomopathogens as biopesticides can reduce the use of synthetic pesticides. Entomopathogens have been isolated from soils and the carcasses of insects, although the natural resources of the pathogens are not limited to these. The larvae of Myrmeleontidae insects, referred to as ant lions, suck out the body fluids of their prey. Because ant lions can kill prey larger than themselves, it was postulated that they used toxins. From the regurgitated fluid of Myrmeleon bore larvae, we have purified an insecticidal protein with a molecular mass of 170 kDa (19) and found that it was produced in the larval region from the thorax to the abdomen (33). In addition to the toxin produced by ant lions, insecticidal proteins were found to be produced by bacterial pathogens isolated from M. bore larvae (21, 34). Thus, we have further isolated bacteria from the ant lion's crop, which serves as a reservoir of regurgitated fluid, to evaluate their toxicity to insects. It was found that some bacterial species obtained from the crop exhibit insecticidal actions against Spodoptera litura cutworms when injected (22). Of these, the A3-2 isolate was closely related to Bacillus sphaericus subgroup IIA on the basis of 16S rRNA gene sequencing and DNA-DNA hybridization tests. This Bacillus species is currently used as a biopesticide with mosquitocidal action involving protein toxins, notably, binary toxin, Mtx1, and Mtx2 (2, 6). However, it is not clear whether A3-2 uses such mosquitocidal toxins to kill insects other than mosquitoes.The aim of this study is to determine an insecticidal factor produced by A3-2 and elucidate its mode of action. Here we report that th...

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“…Although some toxins are apparently insect derived (43,67), compounds produced by bacterial isolates cultured from the Japanese ant lion, Myrmeleon bore, have been implicated as contributing to prey paralyzation and/or death and include GroEL from Enterobacter aerogenes and sphingomyelinase C from Bacillus cereus (47,66).…”

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confidence: 99%

Culture-Independent Characterization of the Microbiota of the Ant LionMyrmeleon mobilis(Neuroptera: Myrmeleontidae)

Dunn

Stabb

2005

Appl Environ Microbiol

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Ant lions are insect larvae that feed on the liquefied internal components of insect prey. Prey capture is assisted by the injection of toxins that are reportedly derived from both the insect and bacterial symbionts. These larvae display interesting gut physiology where the midgut is not connected to the hindgut, preventing elimination of solid waste until adulthood. The presence of a discontinuous gut and the potential involvement of bacteria in prey paralyzation suggest an interesting microbial role in ant lion biology; however, the ant lion microbiota has not been described in detail. We therefore performed culture-independent 16S rRNA gene sequence analysis of the bacteria associated with tissues of an ant lion, Myrmeleon mobilis. All 222 sequences were identified as Proteobacteria and could be subdivided into two main groups, the ␣-Proteobacteria with similarity to Wolbachia spp. (75 clones) and the ␥-Proteobacteria with similarity to the family Enterobacteriaceae (144 clones). The Enterobacteriaceae-like 16S rRNA gene sequences were most commonly isolated from gut tissue, and Wolbachia-like sequences were predominant in the head and body tissue. Fluorescence in situ hybridization analyses supported the localization of enterics to gut tissue and Wolbachia to nongut tissue. The diversity of sequences isolated from freshly caught, laboratory-fed, and laboratory-starved ant lions were qualitatively similar, although the libraries from each treatment were significantly different (P ‫؍‬ 0.05). These results represent the first culture-independent analysis of the microbiota associated with a discontinuous insect gut and suggest that the ant lion microbial community is relatively simple, which may be a reflection of the diet and gut physiology of these insects.

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Purification and Characterization of a Paralytic Polypeptide from Larvae of Myrmeleon bore

Cited by 26 publications

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Insecticidal bacteria isolated from predatory larvae of the antlion species Myrmeleon bore (Neuroptera: Myrmeleontidae)

Insecticidal bacteria isolated from predatory larvae of the antlion species Myrmeleon bore (Neuroptera: Myrmeleontidae)

Cloning, Functional Characterization, and Mode of Action of a Novel Insecticidal Pore-Forming Toxin, Sphaericolysin, Produced by Bacillus sphaericus

Culture-Independent Characterization of the Microbiota of the Ant LionMyrmeleon mobilis(Neuroptera: Myrmeleontidae)

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