The legume family (Fabaceae) is the third largest family of flowering plants worldwide and often damaged by stink bugs, particularly Riptortus clavatus Thunberg (Heteroptera: Alydidae). This insect selectively feeds and oviposits on these plants, resulting in >50% of damage to soybeans harvested in autumn. The most common control methods for this insect are periodic application of chemical pesticides, pheromone traps, and bait crops, but still they encounters residual problem, insect resistance or low control efficacy. The current management system needs alternative control methods, which possibly combined with the present methods. Some entomopathogenic fungi have high virulence against the nymphs and adults of R. clavatus in a favorable conditions, reaching ∼80% mortality with mycosis in 3∼5 days. The genera of Beauveria and Metarhizium are possible control agents for effective and safe management of the serious pest. Fungal infection in R. clavatus was observed using an egfp-expressing fungal transformant. Thermotolerant entomopathogenic fungal conidia can be produced in cereal substrates, which enables conidia to be stable for long times. Our considerations need to be given to the combinations with other control methods, such as chemical pesticides or pheromone trap.

Antimicrobial peptides (AMPs) can be produced in mealworms. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEMBbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbs-cecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae) causes serious damage to Leguminosae. Herein an entomopathogenic fungal virulence assay system against bean bugs was established to construct a fungal database which can be used in integrated pest management (IPM). First to obtain as many bean bugs as possible at the same stage, host plant-preference and developmental synchronization of bean bugs were investigated. In the preference assay, five pairs of adults were infested in a plastic cage, where a pot of green bean, pea or cowpea was previously placed. The highest fecundity and the fastest development of bean bug was observed in the green bean cage. Secondly, in the synchronization experiment, eggs were collected from the cage of adults in 1, 3, 5 and 7 days after oviposition and transferred to a fresh cage with green beans. From the every 4 days of survey, similar stages of bean bugs were found in the cages with the oviposition for 1 and 3 days, rather than the longer times of oviposition. A fungal bioassay against bean bugs was conducted using the bean bugs from the above insect rearing system. Ten Beauveria bassiana isolates were cultured on quarter-strength Sabouraud dextrose agar (¼SDA) for 7 days at 25°C. Ten 4th instar of nymphs were placed on a cultured plate for 1 hour and tranferred to a fresh moisturized plate with grains of green bean. ERL836 isolate treatment showed the highest virulence and fungal mycosis was observed on the bean bugs. In conclusion, these results can be useful to establish an entomopathogenic fungal database for IPM.

Antimicrobial peptides (AMPs) can be produced in mealworms, currently being used as animal feeds, by the infection of genetically engineered-entomopathogenic fungi. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEM-Bbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbscecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Entomopathogenic fungi formulated as wettable powders and suspension concentrates have been sprayed to crop pests for pest management. However, the use of fungal granules to control paddy field pests has not been fully explored. Herein, several Beauveria bassiana isolates (ERL 1170, 1578 and 836) were produced as granules using a millet-based solid culture. The granules were applied to the rice nursery 3 days before transplanting and their control efficacy against rice water weevils was determined in paddy fields. The solid cultures produced ~1×108conidiag-1ofmilletgrains10daysaftertheinoculation. The granules were applied to the soil in the rice nursery 3 days before the rice seedlings were transplanted in the paddy fields. Rice in plots with granules of ERL1578 had 17.3% leaf damage (74% control efficacy) 14 days post application, whereas rice plants in the non-treated control had 66.5% damage. Rice plants treated in the nursery with ERL1170 and ERL836 had 52~54% damage. In the rice plots previously treated with ERL1578 the smallest numbers of larvae and adults were observed 38 days post application. In laboratory conditions, ERL1578-treated larvae were tuned pink and covered with mycelial mass. Applications of millet-based B. bassiana granules on rice nursery soil can be an effective and efficient biological control strategy for the management of rice water weevils. This method is relatively inexpensive and requires less labor compared to practices involving the spraying of fungi directly on rice in paddy fields.

Brown planthopper, Nilaparvata lugens is giving enormous damage to rice production. In this work, virulence of four Beauveria bassiana isolates against brown planthoppers was investigated by applying fungal granules on the water of pots, and we further examined the growth of hypha on the rice plants from the water surcae to explain the insecticidal mode of action. We used Beauveria bassiana (Bb) ERL 836, 1170, 1575 and 1578 isolates, which produced ~2×108 conidia/g of millet grain in a solid culture. Rice seeding was grown in breeding boxes at 28±2℃ for 5 days. Mycotized millet grains were treated on the water in a box at 1 g/box and the rice seeding was infested with 18~25 brown planthopper adults per box. A chemical pesticide and water treatment served as controls. Among the treatments, Bb ERL 836-treated plants had the lowest damage, rather than the other fungal treatments in laboratory assays. Hyphal growth on the stem of rice plants was observed in Petridish conditions under a fluorescent microscopy. This work suggests a possible control of brown planthoppers using entomopathogenic fungi.

Beauveria bassiana isolates have been used in integrated pest management, but little consideration has been given to the studies on fungal gene expressions and their functions. In this work, to determine the functions of genes, B. bassiana ERL1170 was transformed by restriction enzyme-mediated integration method, where pABeG with bar gene was used as a transformation vector. Among seven hundred of transformants, morphologically different ERL1170-pABeG-#160 transformant, particularly dysfunctional in conidiogenesis. The transformant had yellow hyphal growth on fourth strength Sabouraud dextrose agar (SDA/4) and produced very small amount of conidia (<1.0×105 conidia/cm2 agar) in 7 days, whereas wild type had white mycelial growth and significantly greater conidia (3.6×106 conidia/cm2 agar). Additionally under microscopic observation, hyphae of #160 seemed like indian club, compared to the straight forms of wild type hyphae. The next work is figure out possible genes contributing the conidiogenesis of B. bassiana.

Entomopathogenic fungus, Beauveria bassiana can be used in integrated crop management and pharmaceutical applications. Recently some efforts have been given to the fungus to improve its biological performances, but low fungal transformation efficiency is one of the limitations. In this work, B. bassiana ERL1170 isolate was used for fungal transformation by restriction enzyme-mediated integration, where pBARKS1-Bbs-cecropinA was linearied using HindIII. The fungal transformation comprised two steps, preparation of competent blastospores and integration of the plasmid into the cells. To prepare competent blastospores, 2-d cultured blastospores were individually treated with 0.1, 0.2, 0.4 and 0.8 M lithium acetate (LiAc). Secondly in the integration step, concentration of LiAc and calcium chloride (CaCl2), and time period of heat shock were investigated as follows: LiAc, 1, 2 and 4 M; CaCl2, 50, 100 and 200 mM; and heat shock at 42℃, 20, 40 and 60 min. Consequently, combination of 0.2 M LiAc in preparing competent blastospores and 2 M LiAc and 200 mM CaCl2 in the second step showed the highest transformation efficiency. This work would be helpful in the fungal transformation of B. bassiana.

To identify subspecies and stocks of minke whale meats purchased from Korean markets during 2005-2007, we first obtained their complete sequences of mitochondrial DNA cytochrome b and control region sequences, and compared these sequences to the corresponding sequences of the common minke whale (Balaenoptera acutorostrata), obtained from GenBank. From analyses with partial cytochrome b sequences (383 bp) and non-coding, partial control region sequences (463 bp), Korean mink whale meats are identified as products from the North Pacific minke whale (B. a. scammoni). In addition, the sequences of the partial control region from these meats showed G at site no. 298 and G or A at site no. 463, and the meats appeared to originate from the J stock within this subspecies. Thus, because the J stock has been protected since 1986, implementation of strict regulation measures to reduce their accidental fisheries by catch seems urgent. In addition, B. a. scammoni is distinct from B. a. acutorostrata, with an average Jukes-Cantor distance of 2.21% in the complete control region sequence analysis (935 bp) and 1.31% in the complete cytochrome b gene sequence analysis; the current results support the current subspecies classification, although further sequencing analyses with nuclear genes are necessary.

The objective of this study was to develop a reliable and reproducible descriptive analysis procedure for Korean style sweet pumpkin gruel (Hobakjuk). The sensory attributes of the sweet pumpkin gruel were developed and defined, the sample preparation method was standardized， and the sensory evaluation procedure for a sample was established. Seven types of sweet pumpkin gruel (five ready-to-eat type vs. two ready-to-heat type) were selected to be analyzed. Panel training and descriptive analysis were carried out with these 7 samples. A total of 12 sensory attributes (2 aroma/odor, 5 taste/flavor, 4 texture/mouthfeel, and 1 aftertaste attributes) were developed to describe the sensory characteristics of the sweet pumpkin gruel. The definition and reference standards for each sensory attribute were determined to clearly understand each attribute. In the main experiment, trained panelists evaluated the sensory characteristics of the 7 gruel samples based on a fifteen-point intensity scale using the developed attributes. The results were statistically analyzed by analysis of variance, principal component analysis, and cluster analysis. The results showed that the 7 sweet pumpkin gruel samples significantly diffieredin their intensities of all attributes except for sweet pumpkin aroma and viscosity. The ready-to-eat style samples were distinctly characterized by their sweet pumpkin aroma and flavor, whereas the ready-to-heat style samples were markedly characterized by their low intensity of gelatinized starch and pumpkin flavor retention.

Anti-inflammatory effects of ethanolic extract of Angelica gigas (EAG 50, 150, and 500 mg/kg) were investigated in thermal burn-paw edema model. Immersion of rat hind legs into 48.5℃ water for 50 scincreased the leg volume by 0.46-0.55 ml. EAG attenuated not only the paw volume in a dose-dependent manner, but also blood lymphocyte counts and interleukin-6 level increased by burns, without influence on the level of tumor-necrosis factor-α. Moreover, EAG reduced alkaline phosphatase level, a parameter of hepatic injury, which was increased by thermal burns. The results indicate that EAG attenuates inflammatory aymptoms induced by thermal burn and EAG or its conmarin derivatives could be a promising anti-inflammatory drug candidate in inflammation diseases.

Inhibitory effect of colchicine on growth and gravitropic responses in Arabidopsis root was explored to find whether there was an involvement of ethylene production. It has been known that cytoskeleton components are implicated in sedimentation of statoliths to respond to gravitropism and growth. The root growth was inhibited by 25% and 40% over control for 8 hr treatment of colchicine at a concentration of 10-5 M and 10-7 M, respectively. The roots treated with colchicine at the concentration of 10-7 M showed the same pattern as control in 3 hr, however, gravitropic response was decreased in the next 5 hr. The colchicine treatment at the concentration of 10-5 M inhibited the gravitropic response resulting in 60° of curvature. In order to better understand the role of colchicine, the production of ethylene was measured with and without the treatment of colchicine. Colchicine increased the ethylene production by 20% when compared to control via the activation of ACC oxidase and ACC synthase activity. These results suggest that the inhibition of the growth and gravitropic responses of Arabidopsis roots by the treatment of colchicine could be attributed to the rearrangement of microtubule, and increase of ethylene production.

Adenostemma madurense DC. (Asteraceae, Adenostemma L.), which is known to be distributed in Japan, Taiwan, Thailand and Nepal was recently found on Jeju Island in Korea. It is morphologically allied to A. madurense but distinguished from A. lavenia (L.) Kuntze by having such characters as broadly ovate or ovate to oblong leaves, 15-21 ㎝ long × 7-12 ㎝ wide and smooth achenes with slightly muricate at the upper. Furthermore, A. madurense is found in dry mountain slopes in evergreen forests while Adenostemma lavenia occures in the wetland and side of pond. This taxon was named ‘San-mul-meo-wi’ in Korean based on its habitat. We provide a description, illustrations, photographs, and the key to the related species in Korea.

A new spray chrysanthemum cultivar, 'Yellow PangPang' was released by National Horticultural Research Institute (NHRI), Rural Development Administration (RDA), in 2005. The cross between 'Restone' and 'Focus' was made in 2001. After evaluation for the characteristics including shading culture in summer and retarding culture in spring and consecutive selection from 2002 to 2005, final selection was made. The natural flowering time of "Yellow PangPang" is late October, but year-round flowering is possible by shading and illumination. It has pompon type flowers with yellow petals and greenish yellow flower center. It shows very strong stem hardness, especially flower neck, and excellent flower setting. The diameter of flower is 3.9 cm. The number of flowers per stem is 11.3 and the days to flower under the short day treatment is about 54 in autumn season. It has long vase life of 21 days in autumn.

A new spray chrysanthemum cultivar, 'Innocence' was released by National Horticultural Research Institute (NHRI), Rural Development Administration (RDA), in 2005. It was selected from the progenies of open pollination of 'Sei-Alps' in 2002. After evaluation for the characteristics including shading culture in summer and retarding culture in spring between 2002 and 2005, final selection was made. The natural flowering time of "Innocence" is late October, but year-round flowering is possible by shading and illumination. It has round and wide petal, single type flowers with pure white petals and yellowish green flower center. The flowers are large and volumed. The plant shows vigorous growth. The diameter of flower is 7.4 cm. The number of flowers per stem is 9.3 and number of petals per flower is 27.7. The days to flower under short day treatment is about 57 and its vase life is 19 days in autumn season.

A new spray chrysanthemum cultivar, 'Pink Pride' was released by National Horticultural Research Institute (NHRI), Rural Development Administration (RDA), in 2005. The cross between 'Grand Pink' and 'YeonJa' was made in 2001. The characteristics were investigated from 2002 to 2005 for the evaluation and selection of this variety, including shading culture in summer and retarding culture in spring. The natural flowering time of "Pink Pride" is late October, but year-round flowering is possible by shading and illumination. It has pollenless, single type flowers with pink petals and green flower center. The growth of plant is very vigorous and resistant to white rust (Puccinia horiana Henn). The diameter of flower is 6.6 cm. The number of flowers per stem is 11.7 and number of petals per flower is 27. The days to flower under short day treatment is about 56 and its vase life is 19 days in autumn season.