• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.27-28
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• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• The Journal of Korean Society for Radiation Therapy
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• v.20 no.1
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• pp.45-53
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• 2008

Purpose: We are trying to analyze 130 patients' conditions by using our Helical Tomotherapy, which was installed in our center in Oct. 2007. We will be statistically approach this examination and analyze so that we will be able to figure out adaptive plans according to the change in place of the tumor, GTV (gross tumor volume), total amount of time it took, vector (${\upsilon}=\surd$x2+y2+z2) and the change in size of the tumor. Materials and Methods: Objectives were the patients who were medicated with Tomotherapy in our medical center since Oct. 2007 August 2008. The Average age of the patients were 53 years old (Minimum 25 years old, Maximum 83 years old). The parts of the body we operated were could be categorized as Head&neck (n=22), Chest (n=47), Abdomen (n=25), Pelvis (n=11), Bone (n=25). MVCT had acted on 2702 times, and also had acted on our adaptive plan toward patients who showed big difference in the size of tumor. Also, after equalizing our gained MVCT and kv-CT we checked up on the range of possible mistake, using x, y, z, roll and vector. We've also investigated on Set-up, MVCT, average time of operation and target volume. Results: Mean time on table was 22.8 minutes. Mean treatment time was 13.26 minutes. Mean correction (mm) was X=-0.7, Y=-1.4, Z=5.77, roll=0.29, vector=8.66 Head&neck patients had 2.96 mm less vector value in movement than patients of Chest, Abdomen, Bone. In increasing order, Head&neck, Bone, Abdomen, Chest, Pelvis showed the vector value in movement. Also, there were 27 patients for adaptive plan, 39 patients, who had long or multiple tumor. We could know that When medical treatment is one cure plan, it takes 32 minutes, and when medical treatment is two cure plan, it takes 40 minutes that one medical treatment takes 21 minutes, and the other medical treatment takes 19 minutes. Conclusion:With our basic tools, we could bring more accurate IMRT with MVCT. Also, through our daily image, we checked up on the change in tumor so that adaptive plan could work. It was made it possible to take the cure of long or multiple tumor, the cure in a nearby OAR, and the complicated cure that should make changes of gradient dose distribution.

• Korean Journal of Soil Science and Fertilizer
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• v.33 no.4
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• pp.221-233
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• 2000

Evaluation method of soil fertility by application of geographic information system (GIS) which includes landscape characteristics and soil map data was investigated from productivities of red pepper and tobacco grown on the fields with no fertilization. Total 131 fields experiments, 64 fields of red pepper and 67 fields of tobacco were conducted from 22 and 23 fields for red pepper and tobacco, respectively, located at Cheangweon and Eumseong counties in 1996, from 20 and 25 fields at Boeun and Goesan counties in 1997, and 22 and 19 fields at Jincheon and Chungju counties in 1998. All the experimental sites were selected on the basis of wide range of distribution in landscape and soil attributes. Dry weights and nutrients (N, P and K) uptakes by red pepper plant and tobacco leaves were considered as basic fertility of the soil (BFS). The BFS was estimated by twenty-five independent variables including 13 chemical properties and 12 GIS data. Twenty-five independent variables were classified by two groups, 15 quantitative variables and 10 qualitative variables, and were analyzed by multiple linear regression (MLR) of REG and GLM models of SAS. Dry weight of red pepper (DWRP) and dry weight of tobacco leaves (DWTL) every year showed high variations by five times in difference plots with minimum yield and maximum yield indicating the diverse soil fertility among the experimental fields. Evaluation for the BFS by the MLR including independent variables was better than that by simple regression showing gradual improvement by adding chemical properties, quantitative variables, and qualitative variables of the GIS. However the evaluation for the BFS by the MLR showed the better result for tobacco than red pepper. For example the variability in the DWTL by MLR was explained 34.2% by only chemical properties, 35.0% by adding quantitative variables, and 72.5% by adding both the quantitative and qualitative variables of the GIS compared with 21.7% by simple regression with $NO_3-N$ content in soil. Consequently, it is assumed that this approach by the MLR including both the quantitative and qualitative variables was available as an evaluation model of soil fertility for upland field.