• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2003.10a
• /
• pp.114-115
• /
• 2003

A Bacillus thuringiensis isolate, Bt 2385-1, which showed toxicity to lepidopteran, was isolated from Korean soil sample and characterized. PCR-RFLP showed that this isolate contains two novel cryl-type crystal protein genes. In this study, we designed cryl-type specific primer set (ATG1-F and N400-R) to clone the toxic domain of the all cryl-type genes. The two novel rlyl-type toxin genes in addition to crylJal gene were cloned and sequenced. (omitted)

• Korean journal of applied entomology
• /
• v.31 no.3
• /
• pp.247-255
• /
• 1992

A laboratory bioassay that incorporates Bacillus thuringiensis (Bt) purified crystal protein toxins into an artificial diet has identified three toxins, CryIA(b), CryIA(c), and CryIIA, to by effective against the yellow stemborer, Scirpophaga incertulas(Walker). Research is aimed at engineering rice that incorporates genes of one of or more of these toxins so as to mimic the insecticidal action of the insect to Bt. The paper discusses potential strategies for slowing the rate of adaptation that include the use of multiple Bt toxins, promoters that express the toxins only in specific plant tissues at specific times, and mixing transgenic and non-transgenic plants.

• Proceedings of the Korean Society of Sericultural Science Conference
• /
• 2003.04a
• /
• pp.72-72
• /
• 2003

A new Bacillus thuringiensis strain (Kl), having high toxicities to Plutella xylostella and Spodoptera exigua was isolated from Korean soil sample. It was determined to belong to subsp. kurstaki (H3a3b3c) and produced bipyramidal inclusion. PCR-RFLP analysis showed that this isolate contains three novel cryl-type crystal protein genes in addition to crylAa and crylE genes. (omitted)

• International Journal of Industrial Entomology and Biomaterials
• /
• v.11 no.1
• /
• pp.57-61
• /
• 2005

A strain of Bacillus thuringiensis that showed signifi­cantly high toxicity to Plutella xylostella was isolated from a dust sample collected from Chinese tobacco warehouse and characterized. The isolate named B. thuringiensis LY-99 was determined to belong to subsp. alesti (H3a3c) by an H antisera agglutination test and produced bipyramidal inclusions. Plasmid and crystal protein patterns of the LY-99 were different from those of the reference strain, subsp. alesti. PCR analysis with specific primers revealed that this isolate contained abundant cry genes including crylAa, crylAc, crylB, crylD, crylE, crylF and cry2 genes, which was absolutely different from cry gene profile of the subsp. alesti. In addition, insecticidal activity of the LY-99 against P. xylostella larvae was about 44 times higher than that of the subsp. alesti.

• The Korean Journal of Food And Nutrition
• /
• v.9 no.4
• /
• pp.459-465
• /
• 1996

Bacillus thuringiensis serovar. darmstadiensis produced bipyramidal endo-toxin. The toxin protein was purified by Renografin-76 step gradient centrifugation and investigated by electron microscope. Analysis of total plasmid DNA patterns showed that four different size of plasmids existed in wild type B. thuringiensis serovar. darmstadiensis. Total plasmids DNA was isolated and transformed into pst I site of pBR322 cloning vector. Ten clones containing crystal toxin gene were forst screened colony hybridization by using PUYBT 9044 probe ontained B. thuringiensis kurskaki HD 1 toxin gene. Cloned-DNA was digested with EcoR1 and HindIII and transformed to pIBI30 sequencing vector. Finally, 2.6kb and 3.6kb size fragments contatined toxin-gene were cloned with restriction analysis.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.8 no.1
• /
• pp.89-93
• /
• 2004

Expression of a fusion protein between B. thuringiensis crystal protein, Cry1Ac1 and a scorpion insect toxin (AaIT, Androctonus australis Hector insect toxin) in acrystalliferous B. thuringiensis strain (Cry-B strain) was examined. The cry 1Ac1 gene was cloned in B. thuringiensis-E coli shuttle vector, pHT3101, under the control of the native cry 1Ac1 gene promoter (pProAc) and a gene encoding AaIT was inserted in XhoI site in the middle of the cry 1Ac1 gene (pProAc-ScoR). B. thuringiensis Cry-B strain carrying pProAc-ScoR (PyoAc-ScoR/CB) produced an inclusion body of irregular shape and the expressed fusion protein is approximately 65 kDa in size. Sporulated cells and spore-crystal mixtures of ProAc-ScoR/CB had insecticidal activity against Plutella xylostella larvae, showing $LT_50$ of ProAc-ScoR/CB (22.59 hrs) lower than that of ProAc/CB (30.06 hrs) at $1{\times}{10^7} {CEU/cm^2}$. These results suggest that the fusion protein including a B. thuringiensis crystal protein and an AaIT may be functionally expressed in B. thupingiensis. Moreover, we verified the additive toxicity of AaIT, which is a new feasible candidate for insect control.

• BMB Reports
• /
• v.40 no.5
• /
• pp.773-782
• /
• 2007

Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

• Korean journal of applied entomology
• /
• v.45 no.3 s.144
• /
• pp.357-362
• /
• 2006

Bacillus thuringiensis strains were isolated from the domestic soil and a strain was selected that had a new host range and high toxicity against agriculture insect pest. The 142 samples of soil were sampled from the mountains, paddy fields and patches, in Daejon, Chungnam, Chungbuk and Jeonbuk and used for the investigation. Sixteen B. t strains were isolated from 12 samples among collected samples. There were 11 strains that showed toxical activity on Plutella xylostella (Lepidoptera: Yponomeutidae), 7 steins on Spodoptera litura (Lepidoptera: Noctuidae), 5 strains on Arete coerulea (Lepidoptera: Noctuidae), 5 strains on Culex pipiens pallens (Diptera: Culicidae) among the 16 isolated B. t strains. But there were not any strains that showed activity against Hyphanria cunea (Lepidoptera: Arctiidae) and Sitophilus oryzae (Coleoptera: Rhynchophoridae). And also some of B. thuringiensis strains showed insecticidal activity with 2, 3 or 4 kinds of insects. But there were also 3 strains that did not show any activities to the 6 insects which were used in the experiment. When examined with a phase-contrast microscope, the insecticidal crystal protein produced from 16 selected strains had 13 bipyramidal and 3 spherical shapes. The insecticidal bioactivity of the S. litura showed 100% mortality when there were $1.3{\times}10^{7}\;(cfu/ml)$ of CAB109 isolates.

• Korean journal of applied entomology
• /
• v.47 no.1
• /
• pp.87-93
• /
• 2008

Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.

• Journal of Food Hygiene and Safety
• /
• v.38 no.3
• /
• pp.170-175
• /
• 2023

In this study, the effect of MnSO₄ on the insecticidal crystal (IC) produced by Bacillus thuringiensis for a rapid detection medium was analyzed. The strains used included one B. thuringiensis reference (KCTC 1511) and nine wild-type strains. The IC in B. thuringiensis was detected following the method published by the Ministry of Food and Drug Safety in Korea. In the nutrient agar to which 0.005% MnSO₄ was added, IC was observed on two of the three plates after 48 hours of incubation and on all three plates after 120 hours. In AK agar, IC was observed on one and two of the three plates after 48 and 96 hours of incubation, respectively. These results indicated that 0.005% MnSO₄ nutrient agar is more appropriate than AK agar for production of IC in B. thuringiensis. The effect of various MnSO₄ concentrations on IC production was studied after 24 hours of incubation. IC was produced on 1 of the 10 plates with 0.000% MnSO₄ nutrient agar, 2 of the 10 plates with 0.001% MnSO₄ nutrient agar, and 3 of the 10 plates with 0.002% MnSO₄ nutrient agar. IC was not observed for the other nutrient agars containing 0.003%-0.009% MnSO₄. These results indicated that nutrient agar with 0.002% MnSO₄ led to the most rapid production of IC by B. thuringiensis after 24 hours of incubation. However, the conditions for IC production by B. thuringiensis depended on the incubation conditions and strain activity. Therefore, further studies are needed to verify the effects of 0.002% MnSO₄ on the production of IC by various Bacillus thuringiensis strains.