Honeybee (Apis mellifera) egg-yolk protein vitellogenin (Vg) plays roles in immunity, antioxidation, and life span beyond reproduction, but it also acts as an allergen Api m 12 in venom. Here we established antimicrobial and antioxidant roles of honeybee Vg in the body and venom. Using the cDNA encoding Vg identified from Asiatic honeybee (A. cerana) workers, recombinant A. cerana Vg (AcVg) protein of approximately 180 kDa was produced in baculovirus-infected insect cells. In A. cerana worker bees, AcVg was expressed in the fat body and venom gland and was present in the secreted venom. AcVg induced structural damage in microbial cell walls via binding to microbial surfaces and exhibited antimicrobial activity against bacteria and fungi. AcVg protected mammalian and insect cells against oxidative damage through direct shielding of cell membranes. Interestingly, AcVg exhibited DNA protection activity against reactive oxygen species (ROS). Furthermore, the transcript level of AcVg was upregulated in the fat body, but not in the venom gland, of worker bees with antimicrobial peptides and antioxidant enzymes in response to microbial infection and oxidative stress. Our data indicate that AcVg is involved in innate immunity upon infection and in a defense system against ROS, supporting a crucial role of honeybee Vg as an antimicrobial and antioxidant agent in the body and venom.

A recent study reported that Pleurotus ostreatus has the potential to be used as a β-glucan-based cream for supportive complementary therapy of atopic dermatitis. KH054 is a new herbal prescription consisting of P. ostreatus and Panax ginseng. The effects of atopic dermatitis-induced materials on the expression of cytokine genes in human monocytes (THP-1, EoL- 1) have been examined. Some reports demonstrated that P. ginseng augments the activity of natural killer cells, which plays an important role in innate immunity against infection and tumor development. Monocyte chemotactic protein 1 (MCP-1), interleukin (IL)-6, and IL-8 have important roles in mediating the infiltration of various cells into the skin of atopic dermatitis and psoriasis. The present study investigated whether KH054 on induced IL-6, IL-8, and MCP-1 secretion by house dust mite (Dermatophagoides pteronissinus) in THP-1 (human acute monocytic leukemia) and EoL-1(Human eosinophilic leukemia) cell. D. pteronissinus functions in the pathogenesis of allergic diseases, including atopic dermatitis and asthma. The inhibitory effect of KH054 on the induction of IL-6, IL-8, and MCP-1 secretion by D. pteronissinus extract in THP-1 and EoL-1 cells was examined. KH054 potently suppressed the elevated production of IL-6 and IL-8 induced by D. pteronissinus treatment in THP-1 and EoL-1 cells. Based on the present results, KH054 may be useful for developing functional foods to treat atopic dermatitis.

Serine protease inhibitors play a critical role in physiological processes and immune responses by regulating serine protease activities. Here we describe the molecular cloning and antimicrobial activities of a serine protease inhibitor from the mason bee, Osmia cornifrons (OcSPI). OcSPI consists of 405 amino acid residues and contains a potential reactive center loop (RCL) region in its C-terminus. Recombinant OcSPI was produced as a 64-kDa glycoprotein in baculovirus-infected insect cells and exhibited inhibitory activity against chymotrypsin. Additionally, OcSPI demonstrated inhibitory activity against microbial serine proteases, such as subtilisin A and proteinase K, but not against tissue plasminogen activator, thrombin, or plasmin. Recombinant OcSPI bound directly to Escherichia coli, Bacillus subtilis, and Beauveria bassiana and exhibited antimicrobial activity against both bacteria and fungi. Our results demonstrated the antimicrobial functions of OcSPI and suggest a role for OcSPI in the immune response of O. cornifrons.

Osmia cornifrons is a cavity-nesting solitary species used as an apple pollinator in Korea. To elucidate the developmental characteristics of O. cornifrons, we investigated its development from the egg to adulthood, including a dormant prepupal phase and mating through indoor rearing (25 °C, 65% R.H.). The egg durations of the female and male bees were 3.6 ± 0.8 days and 3.1 ± 1.3 days, respectively. During larval development, the head widths of the 1st to 5th instars ranged from 0.7 ± 0.1 mm to 1.3 ± 0.1 mm. The peak of the growth in head width was the 2nd instar. The larval lengths ranged from 3.7 ± 0.6 mm to 13.6 ± 1.3 mm. The peak of growth was the 4th instar. The larval weights ranged from 4.5 ± 1.2 mg to 78.3 ± 16.1 mg. The peak of growth was the 3rd instars. The total larval durations of from the 1st to 5th instars for the females and males were 14.0 ± 6.0 days and 13.2 ± 5.8 days, respectively. The spinning durations of the females and males were 2.2 ± 0.7 days and 2.3 ± 0.8 days, the prepupation durations were 55.5 ± 5.9 days and 55.8 ± 2.9 days, and the pupation durations were 26.4 ± 2.1 days and 25.3 ± 2.3 days, respectively. The average longevity of the female adults and male adults was 21.8 ± 8.7 days and 24.4 ± 12.4 days, respectively. The total duration of from the egg to an adult bee of the O. cornifrons females and males was 123.5 days and 124.1 days, respectively. Mating consisted of the three following phases: the precopulatory (courtship and attempting copulation), copulation and postcopulatory phases. The mating times of the precopulatory, copulation and postcopulatory phases were 159.6 ± 288.9, 8.4 ± 7.1, 12.9 ± 4.5, and 198.8 ± 69.8 seconds.

The honeybee inhibitor cysteine knot (ICK) peptide acts as an antifungal peptide and insecticidal venom toxin. However, the ICK peptide from bumblebees has not been characterized. Here, we report the molecular cloning and antifungal activity of a bumblebee (Bombus ignitus) ICK peptide (BiICK). We identified a BiICK that contains an ICK fold. The BiICK was expressed in the epidermis, fat body, and venom gland of B. ignitus worker bees. A 6.7-kDa recombinant BiICK peptide was expressed in baculovirus-infected insect cells. Recombinant BiICK peptides directly bound to Beauveria bassiana, Ascosphaera apis, and Fusarium graminearum, but they did not bind to Escherichia coli, Paenibacillus larvae, or Bacillus thuringiensis. Consistent with this finding, BiICK exhibited antifungal activity against fungi. These results demonstrate that BiICK acts as an antifungal peptide.

The Osmia cornifrons plays an important role in pollinating fruit trees, such as apple trees. To better understand diapause and oviposition in O. cornifrons, we investigated the correlation between the ovarian development and secretion level of OcVg protein in hemolymph. During ovarian development in wintering the number of oocytes progressively increased in comparison with the length of the ovaries and the oocytes. After emergence, the oocyte and ovary sizes developed until 6 days after emergence and declined after 6 days, but the number of oocytes decreased gradually. The secretion level of OcVg protein in hemolymph revealed that during wintering, the secretion level increased from 1 month to 2 months and then stagnated after 2 months. After diapause, the secretion level increased gradually until day 6 of the newly emerged adult from cocoon stage, and thereafter gradually declined, remaining detectable until day 30 of the adult stage. The correction analysis between ovarian development and OcVg secretion level in hemolymph found that in wintering, the number of oocytes was positively correlated to OcVg secretion level. After diapause, the ovary and first oocyte lengths and the number of oocytes showed significant changes in OcVg secretion level and positively correlated with OcVg secretion level, respectively. These results suggest that there is a significant interaction between ovarian development and the secretion level of OcVg protein and the pattern of ovarian development and secretion of OcVg protein are stage-specific in the O. cornifrons female.

Bumblebee venom serine protease inhibitors have been shown to inhibit plasmin activity. In this study, a bumblebee (Bombus ignitus) venom serine protease inhibitor (BiVSPI) that acts as an antimicrobial factor was identified. BiVSPI is a 55-amino acid mature peptide with ten conserved cysteine residues and a P1 methionine residue. BiVSPI was expressed in the venom gland and was present as an 8-kDa peptide in venom. Recombinant BiVSPI expressed in baculovirusinfected insect cells exhibited inhibitory activity against chymotrypsin, but not trypsin. BiVSPI also exhibited inhibitory activity against microbial serine proteases, such as subtilisin A (Ki 6.57 nM) and proteinase K (Ki 7.11 nM), indicating that BiVSPI acts as a microbial serine protease inhibitor. In addition, BiVSPI was also shown to bind directly to Bacillus subtilis, B. thuringiensis, and Beauveria bassiana, but not to Escherichia coli. Consistent with these results, BiVSPI exhibited antimicrobial activity against Gram-positive bacteria and fungi. These findings provide novel evidence for the antimicrobial function of this bumblebee venom serine protease inhibitor.

1. 야콘의 덩이뿌리는 ‘땅속의 과일’ 또는 ‘올리고당의 왕’으로 불리우며 기능성 성분으로 프락토올리고당을 함유하고있다.2. 야콘의 프락토올리고당은 난소화성의 당질로서 주요 성분으로 케스토즈(kestose), 니스토즈(nystose) 등이 있다.3. 야콘의 프락토올리고당은 프리바이오틱스(prebiotics)로서장내 유산균인 비피더스 균 증식, 미네랄 흡수 촉진, 정장 기능, 지질대사 개선, 항비만 효과, 심혈관계 질환 예방 및 중성지방 합성 감소 역할을 한다.4. 야콘의 식품학적 기능성 및 우수성을 다른 뿌리작물과 비교하고자 프락토올리고당 함량을 조사한 결과, 야콘 8.1%, 마늘 3.3%, 우엉 2.0% 순으로서, 야콘이 가장 우수하였다.5. 또한 저장기간 변화에 따른 프락토올리고당 함량 변화 양상을 조사한 결과 저장기간이 길어질수록 그 함량이 감소하였으며, 저장 1개월부터 그 함량이 8.1%에서 4.8%로 59% 이상급격히 감소하였다. 따라서 저장조건 개선연구가 필요할 것으로 사료된다. 6. 이러한 결과를 바탕으로 야콘의 기능성 식품소재로의 다양한 연구가 가능할 것이다.

땅속의 과일로 알려진 야콘은 라틴 아메리카 안데스 지역이 원산인 국화과 식물로 고구마처럼 생겼으며, 생으로 먹으면 아삭아삭한 배 맛을 느낄 수 있다. 다년생 초본으로 분류되지만 초겨울에 동상해가 자주 발생하는 곳에서는 식물체가 고사하기 쉬우므로 일년생으로 재배하기도 한다. 최근 세계 야콘 생산동향을 살펴 본 결과를 요약하면 다음과 같다. 1. 안데스 지역에서 천년 이상 재배되어 온 야콘의 최근 기능성 성분이 밝혀지면서 전세계적으로 생산 수요가 급증하였다. 2. 야콘은 원산지인 안데스를 벗어나 최근 30년간 전세계적으로 확대되어 일본, 미국, 체코, 독일, 뉴질랜드, 및 우리나라에서 적응성 및 생산성이 검토되었다. 3. 야콘의 생산성은 ha 당 체코 24톤, 에콰도르 41톤, 페루 27 ~ 52톤, 브라질 32 ~ 100톤, 뉴질랜드 40톤, 일본 49톤, 대한민국 19~55톤으로 국가별로 많은 차이를 보인다. 4. 국내에서는 야콘의 기능성 성분이 알려지면서 최근 10년간 야콘의 재배면적과 재배농가 수는 1999년 8 ha, 10여 농가에서 2009년에는 166 ha, 447농가로 각각 20배, 45배 증가하였다. 5. 우리나라에서 친환경 재배 가능한 작물인 야콘의 수요가 점진적으로 증가할 것으로 판단된다.

The large bumblebee, Bombus terrestris, which is indigenous to Europe and is used extensively for high-value crop pollination, has been artificially introduced throughout the world. Here we show the interspecific cross-mating between bumblebee species, B. terrestris and B. ignitus under laboratory conditions. The mating and oviposition rates of interspecific cross-mating of B. terrestris queen with B. ignitus male are higher than those of intraspecific matings of B. ignitus. Furthermore, the competitive copulation experiment indicates that cross-mating of a B. ignitus male with B. terrestris queen is 1.8-fold more frequent than with a B. ignitus queen. The cross-mated B. ignitus queen with B. terrestris male produced B. ignitus workers and B. ignitus male offspring, and the cross-mated B. terrestris queen with B. ignitus male produced both B. terrestris males and B. ignitus male offspring. The genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid offspring indicate that mitochondrial DNA was maternally inherited. Our results indicate that interspecific cross-mating occurred between B. ignitus and B. terrestris, which suggests that the cross-mating is not an exclusive force against the negative impact of competition and genetic contamination on native bumblebees.

Background : Recently, with the growth of traditional medicine industry, management of the quality of herbal medications as a raw material for functional food and medicine becomes more important. One of the most difficult problems of quality control of medicinal plants is management in storage insects. The problem of storage insects is a matter that must be resolved in order to safely obtain raw materials in herbal sector. But the basic prevention measures as well as grasp of the exact situation in the field are not yet. In this study, we investigated to see the problems and solutions through a review of storage pest management system of medicinal plants and the relevant provisions in herbal sector. Methods and Results : For this study, we examined the Status of domestic distribution, storage pest control techniques, news, domestic and import regulations. Storage pests that occur in herbal medications have dropped commercial value and it leads to consumer mistrust. A report said that consumers who had found at least once a pest and microbial contamination when they buy a herbal medications is more than 50% (2004, Ministry of Food and Drug Safety). In the past, those who want to solve these problems had tried through the sulfur fumigation. However, due to concerns about the risk to humans nowadays it was banned in many countries. According to the latest editions of Korean Pharmacopoeia (KP), it is allowed to use a suitable fumigant. but the domestic and imported medicinal plants during distribution can not be used it. Because currently registered fumigant (aluminium phosphide) is allowed only imported medicinal plants in the quarantine process. Chemical fumigation is inappropriate for use in foods and pharmaceuticals so the introduction of eco-friendly control methods is urgent. Eco-friendly methods of pest control that is currently being developed include a cold storage, packaging, high temperature and frozen method etc. Conclusion : These methods should be considered potential cost and Feasibility and it should meet the condition that stated “"Do not change the therapeutic effects of the herbal medications.” in the KP. Also supporting policy for expanding low temperature storage facilities in distribution area will raise possibility of success.

Background : Angelica gigas N., one of the herbs has been used most frequently with Glycyrrhiza uralensis. It is usually cultivated in semi-alpine of more than 400m in North Central province of South Korea. However its production is becoming more unstable due to the climate change especially abnormally high temperature. In particular, the environmental change may cause not only a change in production but also a change in active ingredient of Angelica gigas N. This study was performed to investigate these problems. Methods and Results : This study was carried out in temperature gradient tunnel(TGT) in Pyeongchang, Gangwon province. Temperature conditions were maintained the ambient+0~2℃ (T1), ambient+2~4℃(T2), ambient+4~5℃(T3) throughout the year. We planted and grown one year old seedlings of Angelica gigas N. from mid-April to early November. As a result, there was no difference in early growth of Angelica gigas N. among the temperature range but there was a difference between each treatment region from the survey in June. Average length of soot in T1, T2, T3 was the largest(36cm), medium(27cm), smallest(19cm) respectively. This trend was similar to a survey in September and November. Each of death rate was 0%(T1), 19%(T2), 71%(T3). There was no bolting according to the temperature. The yield of a plant of Angelica gigas N. was 476g(T1), 76g(T2), 26g(T3), respectively. Production of Angelica gigas N. is foreseen 84.7% and 94.5% reduction if the temperature rises to 3±1℃ and 4±1℃ more than it is at present based on temperature in Pyeongchang 2015. Total amount of Decursin was reduced at higher temperatures. Conclusion : The results of this study show that quality and production of Angelica gigas N. is expected to decline significantly if the temperature rise than the current. Therefore development of various techniques is required to response to climate change.