Immune-inducible antimicrobial peptides were produced using transgenic silkworms that expressed Rel family transcription factor, truncated BmRelish1 (BmRelish1t) genes under the control of the BmA3 promoter using the piggyBac vector. BmRelish1t gene contains all domains of Bmrelish: a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acids (AHAA) rich region except the Ankyrin repeat domain (ANK) and the death domain (DD). (1:1) Mixtures of the donor vector (pG-3xP3EGFP-BmA3BmRelish1t) and helper vector were micro-injected into 1,800 eggs of bivoltin silkworms, Baegokjam and EGFP-induced fluorescence was observed for 25 broods of transgenic silkworms under a florescence stereomicroscope. Analysis by real-time PCR indicated that transgenic silkworms expressing BmRelish1t recombinant proteins displayed higher mRNA expression levels of the Bombyx mori antimicrobial peptides such as lebocin, moricin, and nuecin than the normal silkworms. Moreover, transgenic silkworms expressing BmRelish1t showed antibacterial activity against Escherichia coli. We suggest that transgenic expression of BmRelish1t may find useful applications for the production of various antimicrobial peptides at the same time in transgenic silkworms.
Silkworm transgenesis is now a routine method leading to a satisfactory yield of transformed animals and the reliable expression of transgenes during multiple successive generations. However, the screening of G1 transgenic individuals from numerous progeny has proved to be difficult and time-consuming work. Previously, we characterized the promoter of heat shock protein 70 from Bombyx mori (bHsp70), which is ubiquitously expressed in all tissues and developmental stages. To investigate the utilization of the bHsp70 promoter to screen transgenic individuals, the EGFP marker gene was inserted into the piggyBac vector under the control of the bHsp70 promoter. Mixtures of the donor and helper vectors were micro-injected into 3,060 eggs of bivoltine silkworms (Keomokjam). EGFP fluorescence was observed in 17 broods of transgenic silkworms under a florescence stereomicroscope. Interestingly, this fluorescent marker protein was detected not only in parts of the embryo segments on the seventh day of the G1 embryonic developmental stage but it was also detected in a part of the body of G1 hatched larvae, in the middle silk gland of G1 fifth instar larvae, and in the wings of seven-day-old G1 pupae and G1 moths. Therefore, we suggest that the bHsp70 promoter can be used for the rapid and simple screening of transgenic silkworms.
본 연구는 환경스트레스 저항성이 증진된 페튜니아를 개발하기 위하여 NDPK2유전자 도입 형질전환 계통 NDPK2-7-1와 SOD2 유전자 도입 형질전환 계통 SOD2- 2-1-1-35간의 교잡에 의해 획득된 후대들의 비생물적 스트레스 저항성을 조사하기 위해 수행되었다. 비 생물적 스트레스 유발원인 메틸바이올로젠(methyl viologen, MV) 100 μM과 200 μM 처리에서 교잡후대들은 그들의 교배 모본 SOD2 유전자나 NDPK2 유전자가 단독으로 도 입된 형질전환 계통이나 비형질전환체 보다 메틸바이 올로젠에 의한 피해를 적게 받았다. 이는 SOD2 유전 자나 NDPK2 유전자가 단독으로 도입된 형질전환 계 통간 교잡에 의해 획득된 후대들이 그들의 교배모본 (SOD2 유전자나 NDPK2 유전자가 단독으로 도입된 형질전환 계통)이나 비형질전환체 보다 산화적 스트레 스에 대한 저항성이 증진되었음을 증명해 준다고 할 수 있다. 이들 교잡후대들은 초장 등 11종류의 양적형질의 특성이 비형질전환체에 비해 약간 길거나 짧긴 하였지 만 비형질전환체와 거의 유사하였으며, 꽃 색갈이나 모양 또한 그들의 교배모본 (SOD2 유전자나 NDPK2 유전 자가 단독으로 도입된 형질전환 계통)이나 비형질전환 체와 차이가 없었다.
Propylea japonica is small ladybug(approximately 4-5mm) and is met with everywhere in Korea. When there was no aphid, the survival rate of P. japonica was 0% among another instar bugs and 26.7~32.8% among same instar bugs after 48 hours. When there were sufficient aphids, the cannibalism of P. japonica was low, so the survival rate of P. japonica was 78.6~95.8% among another instar bugs and 80.6~100% among same instar bugs after 48 hours. In the comparison of the number of eggs at different oviposition site, P. japonica preferred the corrugated cardboard. In experiment using T-tube, the rate of decoy was 81% at cotton aphid compared with pepper leaf and 62% at cotton aphid compared with artificial diet. In the artificial diet using shrimp and chicken liver, the developmental period from egg to adult was 17days and the survival rate from egg to adult was 66.7% but the vitality was not good. These data suggest that P. japonica can be used effective natural enemy for control of aphids and is needed to research about artificial diet, mass rearing system and control effect in field.
Background : Capsaicin is an active component of chili peppers, which are plants belonging to the genus Capsicum. Research reported capsaicin has antioxidant and anti-inflamatory activity and osteoclast lineages are very susceptible to oxidative stress, as osteoclasts are produced by increased-generation of intracellular ROS and osteoclasts are activated by ROS. Therefore, our study was evaluated the influence of intracellular oxidative stress such as increased ROS level on RANKL-mediated osteoclast differenciation. Methods and Results : Capsaicin showed a good free radical scavenging activity at in-vitro antioxidant activity. The inhibitory effect of osteoclast differentiation on capsaicin was confirmed. Osteoclast differentiation from murine macrophage RAW 264.7 cells was induced by RANKL. The effect of capsaicin on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation was demonstrated using a tartrate-resistant acid phosphatase (TRAP) assay and TRAP staining. Capsaicin showed an inhibitory effect on TRAP activity. The TRAP staining showed that the number of TRAP positive osteoclasts was reduced in capsaicin-treated cells. Conclusion : Capsaicin revealed an inhibitory effect in osteoclast differentiation induced by RANKL. These results suggest that capsaicin may have a beneficial effect for the prevention or treatment of osteoclast caused bone diseases such as osteoporosis.
Background : Plants of Taraxacum spp. are widely used in medicine, but some of them have some problems related to propagation, such as strong dormancy and inactive germination. This study investigated the effects of temperature, gibberellic acid (GA3), and potassium nitrate (KNO3) on seed germination in Taraxacum ohwianum. Methods and Results : The seeds (NIBRGR0000135524) were exalbuminous, and their length and width were 4.25 ± 0.118 mm and 0.89 ± 0.062 mm, respectively. Among various temperatures (15, 20, 25, and 30℃), the optimum temperature for germination was found to be 20℃ (31.3%). High temperature (30℃) induced off-type in seedlings (thickened and crumpled cotyledons, and restricted root system). GA3 treatments increased germination percentage and speed, but germination percentage was higher with KNO3 treatment. Under KNO3 treatments (50 to 200 mg·L-1), germination percentage exceeded 80% after 12 days, with 50 mg·L-1 KNO3 being notably effective (91.2%, after 15 days). Conclusion : Seeds of T. ohwianum showed germination ability at low temperature. The best method for germination was pre-soaking in 50 mg·L-1 KNO3 (4℃, dark, 24 h) and incubating at 20℃ for 15 days.
Background : This study was conducted to determine the optimal conditions of growth medium, temperature, and light quality for efficient propagation of Pyrrosia petiolosa spores. Methods and Results : P. petiolosa spores were collected from Andong-si area in February 2015, and the spores were subjected to a careful selection process. The selected spores were stored at the Korea National Wild Plant Seed Bank (4℃, RH 40%) and used as our experimental materials (GR0000182302). The spore germination rate and prothalium development were high in Knop media (24.5%), which had low mineral content. The germination rate increased with the increase in temperature and reached its maximum, 24.5%, at 25°C. The germination rate was also measured under different light conditions, and the highest rate of 28.7% was observed under LED blue light compared to fluorescent lights (24.5%). The germinated spores developed into heart-shaped prothallia under LED blue light; however, 3 weeks after sowing, prothallium development decreased compared to fluorescent lights. Conclusion : In conclusion, the optimal P. petiolosa In vitro propagation method include spore germination on Knop medium at 25°C under LED blue light, and the change in light source to fluorescent light 3 weeks after sowing for prothallium formation and proliferation.
Genetically modified (GM) papaya (Carica papaya L.) line 55-1 (55-1), which is resistant to papaya ringspot virus infection, has been marketed internationally. Many countries such as the European Union, Japan, and Korea have a mandatory safety assessment, approval and labeling regulations for GM foods. Thus, there is a need for specific methods for detecting 55-1. In this study, we established a real-time PCR detection method applicable to 55-1 for a variety of papaya products. The limit of detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% (weight per weight [w/w]) for homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The 55-1 event-specific detection method observed parallelism (r2>0.99) between the concentration of line 55-1 cultivars and Ct values obtained in amplification plots at concentrations of 0.005-10% for SunUp DNA and 0.01-10% for Rainbow DNA. The method was applicable to the qualitative detection in various types of processed products (cocktail fruit, dried fruit, juice, etc.) containing papaya as a main ingredient. Monitoring papaya products for the presence of GM papaya were demonstrated using a P35S and T-nos real-time PCR detection method but no amplification signals were detected.
The effects of ovulation induction in ussurian bullhead, Leiocassis ussuriensis, were investigated by treating ussurian bullhead with hCG, LHRHa, GnRHa, ovaprim, and pimozide. hCG was injected to ussurian bullhead at 0.75% NaCl, 5,000, 10,000, 20,000, and 30,000 IU, respectively. The ovulation inducement rates were 100% in 20,000 and 30,000 IU. Fertilization rates were 82.7% and 79.8%. Hatching rates were 59.4% and 57.2%. Ovulation time was between 16-19 hr The concentrations of LHRHa injected were 0.75 NaCl, 50, 100, 200, 300, and . The ovulation inducement rates were 100% in 300 and . Fertilization and hatching rates were 84.9% and 68.4% at . The times to ovulation were between 23 hr and 34 hr. Ovaprim of 0.75% NaCl, 1.0, 1.5, 2.0, 2.5 and 3.0 ml/kg were injected to the abdominal cavity. The ovulation inducement rate was highest at 2.0 and 3.0 ml/kg to 92% and ovulation time was between 27-38 hr. LHRHa concentrations of 0.75% NaCl, 50, 100, 200, 300 and were injected with pimozide (). Ovulation inducement rate was 100% from 200 to 400 IU with pimozide. Ovulation time was 22-36 h. Fertilization and hatching rates were 88.9% and 70.4% in with pimozide.