• Food Science and Biotechnology
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• v.15 no.2
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• pp.232-237
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• 2006

Bacillus cereus group comprising B. cereus, B. thuringiensis, and B. mycoides was differentiated by polymerase chain reaction (PCR) and colony morphology. Prevalence of B. cereus group in rice and distribution of enterotoxin genes were determined as possible food poisoning agents. PCR using primers targeted for gyrB and cry genes could distinguish B. thuringiensis from B. cereus, and B. mycoides was differentiated by rhizoid morphological characteristics on nutrient agar. Among 136 rice and their processed products, prevalence of B. cereus group was 40%. B. cereus group consisted of 54 B. cereus, 11 B. thuringiensis, and 1 B. mycoides. Major isolates were B. cereus, with B. thuringiensis detected up to 10% among edible rice tested. Five enterotoxin genes, hbl, nhe, bceT, entFM, and cytK, were broadly distributed among B. cereus group, especially in B. cereus and B. thuringiensis. Prevalence of B. cereus group in rice and enterotoxin distribution suggest B. thuringiensis and B. cereus are toxigenic strain that should be controlled in rice and its products.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.361-367
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• 2010

Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

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

• Food Science and Biotechnology
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• v.15 no.4
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• pp.625-630
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• 2006

Polymerase chain reaction (PCR) method was developed for the specific detection of insect-resistant rice containing cry1Ac gene derived from Bacillus thuringiensis (Bt). Primers were designed from the 35S promoter, NOS terminator, cry1Ac gene, and sucrose phosphate synthase (SPS) for general screening of Bt rice. By sequencing the PCR products from the two putative kinds of Bt rice, we designed a specific primer from the junction region between the cry1Ac gene and the NOS terminator that had been inserted into Bt rice. The construct-specific primer was employed to amplify a 147 bp product in the two lines of Bt rice. No amplified products were observed from the other Bt crops with various Bt genes introduced. In qualitative PCR analysis, the limit of detection was 0.005 ng from genomic DNA of Bt rice. In addition, PCR analysis was performed on 64 kinds of rice presently available in the Korean market, and no Bt rice was detected. This method presented in this paper can be used as a highly sensitive and specific detection method of Bt rice.

• Journal of Applied Biological Chemistry
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• v.54 no.4
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• pp.296-302
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• 2011

Insect-resistant transgenic rice was developed by inserting the mCry1Ac1 a modified gene from the soil bacterium Bacillus thuringiensis (Bt). For biosafety assessment, we studied the effects on survival of cantor Daphnia magna, a commonly used as a model organism in ecotoxicological studies. D. magna fed on Bt rice and its near non-genetically modified (GM) counterparts (Nakdong) grown in the same environment (100% ground rice suspension). The Bt rice was comfirmed to have the insertion of T-DNA and protein expression by the polymerase chain reaction and ELISA analysis. Feeding study showed similar cumulative immobility and abnormal response of D. magna between Bt rice and non-GM counterparts. 48 h-$EC_{50}$ values of Bt rice and non-GM rice showed 4,429 and 2,889 mg/L respectively. The rice no observed effect concentration (NOEC) values for D. magna was suggested 1,000 mg/L. We conclude that the tested Bt-rice and Nakdong similar cumulative immobility for D. magna the widely used model organism. We found out that there is strong possibility that the growth of Bt rice didn't affect to non-target insects.

• Korean journal of applied entomology
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• v.48 no.1
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• pp.101-108
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• 2009

For biological control of larva of Cnaphalocrocis medinalis and Naranga aenescens do considerable damage to crops by folding and scraping the leaf tissue of rice, a large number of Bacillus thuringiensis isolates have been obtained from soil samples in Korea and the pesticidal activity was assayed against two insect pest species described above. Among 53 Bt isolates tested in bioassay, 18 and 13 isolates showed over 90% mortality against C. medinalis and N. aenescens, respectively. Some isolates (11 isolates including CAB141) presented dual activity against C. medinalis and N. aenescens. These isolates showed over 96% control effect in pest control in laboratory against larvae of C. medinalis. Also, it was investigated that pupation, pupal length, and adult emergence of larvae exposed to Bt suspension decreased. Especially, the pupal length of C. medinalis after being fed corn seedling leaves treated Bt suspension for 10 days, were much smaller than that of control.

• The Korean Journal of Food And Nutrition
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• v.24 no.4
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• pp.657-663
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• 2011

$Bacillus$ $cereus$ is widely distributed on various foods and is known to cause clinical infections, food poisoning toxin induced diarrhea and vomiting. In this study, $B.$ $cereus$ group detected and analyzed rice, rice bran, and biofilm characterization of $B.$ $cereus$ confirmed. $B.$ $cereus$ was identified in approximately 34.6% of brown rice and 50.0% of rice bran. $B.$ $thuringiensis$ was detected in 3.9% of brown rice and 23% of rice bran, and $B.$ $mycoides$ was isolated from rice bran. The microtiter plate assay detected differences in biofilm-forming ability among $B.$ $cereus$ group isolates. Biofilm of $B.$ $cereus$ seemed to increase the MIC values of antimicrobial agent and antibiotic compounds compared with planktonic cells. Therefore, sufficient attention should be given to good manufacturing practice and good agriculture practice to avoid contamination of $B.$ $cereus$ group raw material including rice.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1223-1229
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• 2009

To construct a new recombinant strain of Bacillus velezensis that has antifungal and insecticidal activity via the expression of the insecticidal Bacillus thuringiensis crystal protein, a B. thuringiensis expression vector (pHT1K-1Ac) was generated that contained the B. thuringiensis cry1Ac gene under the control of its endogenous promoter in a minimal E. coli-B. thuringiensis shuttle vector (pHT1K). This vector was introduced into a B. velezensis isolate that showed high antifungal activities against several plant diseases, including rice blast (Magnaporthe grisea), rice sheath blight (Rhizotonia solani), tomato gray mold (Botrytis cinerea), tomato late blight (Phytophthora infestans), and wheat leaf rust (Puccinia recondita), by electroporation. The recombinant B. velezensis strain was confirmed by PCR using cry1Ac-specific primers. Additionally, the recombinant strain produced a protein approximately 130 kDa in size and parasporal inclusion bodies similar to B. thuringiensis. The in vivo antifungal activity assay demonstrated that the activity of the recombinant B. velezensis strain was maintained at the same level as that of wild-type B. velezensis. Furthermore, it exhibited high insecticidal activity against a lepidopteran pest, Plutella xylostella, although its activity was lower than that of a recombinant B. thuringiensis strain, whereas wild-type B. velezensis strain did not show any insecticidal activity. These results suggest that this recombinant B. velezensis strain can be used to control harmful insect pests and fungal diseases simultaneously in one crop.

• Korean journal of applied entomology
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• v.28 no.3
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• pp.167-174
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• 1989

A confined field assessment on the integrated control of vector mosquitoes (Anopheles sinen- sis and Culex tritaent'orhynchus) was conducted by timely treatment of Bacillus thuringiensis(H-14) formulation in the presence of larvivorous fish (Aplocheilus lalipes) in natural rice fields of $30,000M^2$ at Yongam, South Korea Cholla Province from July through October in 1988. In presence of larvivorous fish (Aplocheilus) at lower density of 0.6 fish per $M^2$ water surface, mosquito vector control rates ranged 55. 0~57. 6% from July through August, and when the mosquio density started to increase with over 10 larvae on the average per sample, B.t.(H-14) formulation treatment at the rate of 1. 0 Kg/ha made 100% vectors reduc¬tion in 24 hours, the control sustained above 98% until the test was terminated on October 11th. In the fish introduced rice paddies with Aphyocypris chinensis at the density of above 1. 5 fish per $M^2$, a satisfactory degree of vector control was obtained by 88. 2~96. 7% in 2 week period until September 21st. In the absence of larvivorous fishes, B.t. (H-14) treatment made 100% control in 24 hours, however, vector population rebound appeared in day 7, required to make additional treatment to suppress population down.

• Food Science and Biotechnology
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• v.15 no.3
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• pp.385-391
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• 2006

In Korea, different kinds of genetically modified (GM) crops are under development, including GM-rice expressing insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) modified to change a single amino acid. In this study, amino acid (aa) sequences of modified Cry proteins were compared to that of known allergens, and Cry proteins expressed in GM-rice were identified by using Cry protein specific polyclonal antibody. The antigen-antibody reactions were compared between GM and commercial rice to assess the allergic risk of Cry proteins. This analysis showed no known allergen to have more than 35% aa sequence homology with modified Cry proteins in Bt rice over an 80 aa window or to have more than 8 consecutive identical aa. Sera from allergic patients showed some IgE reactivity via immunoblotting and enzyme-linked immunosorbent assay (ELISA), although no differences were seen between GM and commercial rice. Based on these results we conclude that GM rice with modified Cry proteins has no differences in its protein composition or allergenicity relative to commercial rice.