• BMB Reports
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• v.38 no.1
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• pp.41-48
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• 2005

In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AM1-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM1 and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages ($LD_{50}$) and the lethal time(s) ($LT_{50}$) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.2
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• pp.183-197
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• 2007

Field trial was conducted at the Research Farm of the Katsina State Agricultural and Rural Development Authority during the farming season of 2004 with a view to evaluate/determine the efficacy of some selected plant-derived biopesticides against the insect pests of cowpea as well as their effect on yield. The variety of the cowpea used was IT86D-719 and the plant derived biopesticides evaluated during the trial were chilli pepper, garlic, ginger, neem, sweetsop and tobacco. The experimental field was ploughed, harrowed and thereafter ridged before the commencement of the 2004 planting season. A total of twenty one (21) experimental plots were demarcated and arranged into seven treatment plots. The size of each plot was $5m{\times}4m$ while interspaces between adjacent plot and blocks were 1m and 2m, respectively. Results of the experiments showed that all the plant-derived biopesticide treatments were significantly (p>0.05) better than control treatment. The order of effectiveness of the treatments was tobacco (80-90%), sweetsop (75-85%), garlic (70-80), neem (72-78%), chilli pepper (60-70%) and ginger (30-50%). Furthermore, yield result corresponded positively with the effectiveness of the treatments. Results of the present finding suggest the use of tobacco, sweetsop and garlic as promising biopesticides in the control of cowpea insect pests.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1057-1062
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• 2004

The entomopathogenic bacterium Bacillus thuringiensis is the most widely used biopesticide. Insecticidal proteins, coded by genes located in plasmids, form typical parasporal, crystalline inclusions during sporulation. We isolated a Bacillus thuringiensis strain having insecticidal activity against larvae of the house fly (M. domestica) from the soils at a pig farm in Korea, and named it Bacillus thuringiensis SM. The culture filtrate from Bacillus thuringiensis SM showed strong lethality (83.3%) against M. domestica larvae. The parasporal crystal is enclosed within the spores' outermost envelope, as determined by transmission electron microscopy, and exhibited a bipyramidal form. The crystal proteins of strain SM consisted of five proteins with molecular weights of approximately ~130, ~80, ~68, ~42, and ~27 kDa on a 10% SDS-PAGE (major band, a size characteristic of Cry protein). Examination of antibiotic resistance revealed that the strain SM showed multiple resistant. The strain SM had at least three different plasmids with sizes of 6.6, 9.3, and 54 kb. Polymerase chain reactions (PCRs) revealed the presence of cry1, cry4A2, and cry11A1 genes in the strain SM. The cry1 gene profile of the strain SM appeared in the three respective products of 487 bp [cry1A(c)], 414 bp [cry1D], and 238 bp [cry1A(b)]. However, the strain SM has not shown the cry4A2 md cry11A1 genes. In in vivo toxicity assays, the strain SM showed high toxicity on fly larvae (M. domestic) [with $LC_{50}$ of 4.2 mg/ml, $LC_{90}$ of 8.2 mg/ml].

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.50-56
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• 2003

A freshwater bloom-forming cyanobacterium, Microcystis aeruginosa, and local soil isolate Scytonema sp. strain BT 23 were demonstrated to contain biotoxic secondary metabolites with pesticidal and mosquito larvicidal activities. A purified toxic constituent from M aeruginosa showed an absorption maximum at 230 nm and its toxicity symptoms, Rf value on TLC, and retention time observed ill an HPLC analysis were similar to those of the hepatotoxic heptapeptide microcystin-LR. The bioactive constituent of the Scytonema sp. was less polar in nature and exhibited two peaks at 240 and 285 m. When applied to two cruciffrous pests, Pieris brassicae and Plutella flostella, the crude extracts and toxic principles from the two cyanobacteria showed significant antifeedant activity in a no-choice bioassay, and at higher concenuations exhibited contact toxicity to the insect larvae. The purified toxin from M. aeruginosa was found to be more effective and produced 97.5 and $92.8\%$ larval mortality in the two pests, fo11owing 2 h of toxin treatment at a concentration of $25{\mu}g$ Per leaf disc (2.5 cm dia.). Meanwhile, similar treatment with the purified toxin from Sytonema sp. stain BT 23 only produced 73 and $78\%$ mortality in the two pests. The cyanobacterial constituents also showed significant activity against Culex and Anopheles larvae. The M. aeruginosa toxin ($20{\mu}g\;ml^-1$) caused 98.2 and $88.1\%$ mortality in the Culex and Anopheles larvae, respectively, while the purified toxin from the Sytonema sp. was less toxic and only produced a 96.3 and $91.2\%$ mortality, respectively, at a much higher concentration ($40{\mu}g\;ml^-1$). Accordingly, the current results point to certain hitherto unknown biological properties of cyanobacterial biotoxins.

• Weed & Turfgrass Science
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• v.6 no.1
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• pp.1-10
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• 2017

Weed management under organic farming systems is very problematic since organic agriculture does not allow synthetic herbicides. Bioherbicide is needed to develop for weed management in organic agriculture systems. This review covers current status and perspectives of bioherbicide development for effective nonchemical weed management in organic farming systems. Bioherbicides are products of natural origin derived from living organisms, and more specifically bacteria, fungus and plants including natural metabolites for weed control. Bioherbicides derived from microorganisms or natural molecules are currently available on the pesticide markets. Devine, Lockdown (Collego), BioMal, Camperico, Organo-Sol and Opportune were derived from bacteria, Woad Warrior, Smolder, Mygogen, Chontrol Paste, Starritor and Phoma derived from fungus, and Katoun (pelargonic acid) and Beloukha were derived from plants. Corn gluten meal products and plant essential oils products are also available for nonselective weed control in organic agriculture. Organic weed management methods may be more feasible in small scale farming and high-value crops, and bioherbicides may be applied with other weed control practices in organic farming systems.

• Korean Journal of Medicinal Crop Science
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• v.28 no.1
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• pp.38-46
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• 2020

Background: Although Sancho (Zanthoxylum schinifolium Siebold & Zucc) oil has traditionally been used for its antibiotics properties, there is currently a lack of scientific evidence regarding its biological activities. In this study, we investigated the antimicrobial and antioxidant activities of Sancho oil against food-hazardous microorganisms, phytopathogens, and dermatophytes. Methods and Results: We investiated the antimicrobial activity of Sancho oil against 11 food-hazardous microorganisms, nine phytopathogens, and six dermatophytes. The Sancho oil was found to show the strongest antibacterial activity against Shigella flexneri and Listeria spp. Sancho oil also showed high antifungal activity against plant pathogens, particularly Fusarium oxysporum, and showed antimicrobial activity against dermatophytes such as Trichophyton rubrum, Microsporum canis and Candida albicans. The antioxidant activity of Sancho oil was measured using the DPPH method, and was found to be stronger than that of unrefined oil. Moreover, this activity increased with increasing oil concentration. Conclusions: We found that Sancho oil showed differing antimicrobial activities against food-hazardous microorganisms, dermatophytes, and plant pathogens. The antimicrobial activity spectrum of Sancho oil was not broad and varied among microbial strains. On the basis of our findings, we consider that Sancho oil could be used an antibacterial material for food-borne S. flexneri and Listeria spp., a biopesticide for Fusarium spp., and a treatment for dermatophytes such as T. rubrum.

• Korean journal of applied entomology
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• v.56 no.3
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• pp.289-294
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• 2017

The Oriental fruit fly, Bactrocera dorsalis, has been designated as a main quarantine insect pest. Sensitive monitoring and diagnosis against B. dorsalis are required for early quarantine treatment. This study formulated a wax type dispenser containing methyl eugenol and biopesticide to attract and kill this insect. It also developed diagnostic PCR primers against five major quarantine fruit flies of B. dorsalis, B. cucurbitae, B. tryoni, B. latifrons, and Ceratitis capitata. The lure and diagnostic primers were evaluated in a field located in Kota Kinabalu, Malaysia.

• The Plant Pathology Journal
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• v.28 no.1
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• pp.68-74
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• 2012

Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.547-559
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• 2007

Bacillus thuringiensis (Bt) was first described by Berliner [10] when he isolated a Bacillus species from the Mediterranean flour moth, Anagasta kuehniella, and named it after the province Thuringia in Germany where the infected moth was found. Although this was the first description under the name B. thuringiensis, it was not the first isolation. In 1901, a Japanese biologist, Ishiwata Shigetane, discovered a previously undescribed bacterium as the causative agent of a disease afflicting silkworms. Bt was originally considered a risk for silkworm rearing but it has become the heart of microbial insect control. The earliest commercial production began in France in 1938, under the name Sporeine [72]. A resurgence of interest in Bt has been attributed to Edward Steinhaus [105], who obtained a culture in 1942 and attracted attention to the potential of Bt through his subsequent studies. In 1956, T. Angus [3] demonstrated that the crystalline protein inclusions formed in the course of sporulation were responsible for the insecticidal action of Bt. By the early 1980's, Gonzalez et al. [48] revealed that the genes coding for crystal proteins were localized on transmissible plasmids, using a plasmid curing technique, and Schnepf and Whiteley [103] first cloned and characterized the genes coding for crystal proteins that had toxicity to larvae of the tobacco hornworm, from plasmid DNA of Bt subsp. kurstaki HD-1. This first cloning was followed quickly by the cloning of many other cry genes and eventually led to the development of Bt transgenic plants. In the 1980s, several scientists successively demonstrated that plants can be genetically engineered, and finally, Bt cotton reached the market in 1996 [104].

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.69-79
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• 2012

In an ongoing survey of the bioactive potential of microorganisms from Ladakh, India, the culture medium of a bacterial strain of a new Pseudomonas sp., strain ICTB-745, isolated from an alkaline soil sample collected from Leh, Ladakh, India, was found to contain metabolites that exhibited broad-spectrum antimicrobial and biosurfactant activities. Bioactivity-guided purification resulted in the isolation of four bioactive compounds. Their chemical structures were elucidated by $^1H$ and $^{13}C$ NMR, 2D-NMR (HMBC, HSQC, $^1H$,$^1H$-COSY, and DEPT-135), FT-IR, and mass spectroscopic methods, and were identified as 1-hydroxyphenazine, phenazine-1-carboxylic acid (PCA), rhamnolipid-1 (RL-1), and rhamnolipid-2 (RL-2). These metabolites exhibited various biological activities like antimicrobial and efficient cytotoxic potencies against different human tumor cell lines such as HeLa, HepG2, A549, and MDA MB 231. RL-1 and RL-2 exhibited a dose-dependent antifeedant activity against Spodoptera litura, producing about 82.06% and 73.66% antifeedant activity, whereas PCA showed a moderate antifeedant activity (63.67%) at 60 ${\mu}g/cm^2$ area of castor leaf. Furthermore, PCA, RL-1, and RL-2 exhibited about 65%, 52%, and 47% mortality, respectively, against Rhyzopertha dominica at 20 ${\mu}g/ml$. This is the first report of rhamnolipids as antifeedant metabolites against Spodoptera litura and as insecticidal metabolites against Rhyzopertha dominica. The metabolites from Pseudomonas sp. strain ICTB-745 have interesting potential for use as a biopesticide in pest control programs.