장내 미생물 군집은 소화 과정, 면역 시스템, 질병 발생 등 숙주의 다양한 면에 광범위한 영향을 주는 것으로 알려져 있으며, 주요 장내 미생물 종은 숙주의 생리 기능에 핵심적인 역할을 수행한다고 발표된 바 있다. 곤충의 장내 미생물 군집에 관한 연구가 최근 활발히 이루어지고 있으며, 이들 연구는 주로 장내 미생물 군집과 기생충, 병원체 간의 상호작용, 종간의 신호 전달 네트워크, 먹이의 소화 과정 등을 중심으로 이루어지고 있다. 이러한 연구들은 대부분 Illumina MiSeq을 활용하여 16S rRNA 유전자의 V1부터 V9 영역 중 선택된 특정 부분을 대상으로 짧은 서열 정보를 대상으로 진행되었다. 그러나, 최근에는 PacBio HiFi 기술이 상용화되면서 16S rRNA의 전장 분석이 가능할 수 있게 되었다. 이번 연구는 장수말벌(Vespa mandarinia)의 해부를 통해 gut과 carcass 부분을 분리한 뒤, 각 샘플을 Illumina MiSeq과 PacBio HiFi 기술을 활용하여 미생물 군집 간의 차이점을 확인하기 위하여 수행되었다.
본 연구는 국내 자생하는 진달래속(Rhododendron) 종들의 종자 휴면유형 분류 및 발아특성 구명을 목표로 하였다. 진달래속 종들의 배는 형태적휴면(MD)이 없는 완전히 발달된 직선(linear) 형태였으며, 만병초 및 꼬리진달래 종자는 탈리 시점에 이미 휴면이 없는 것으로 밝혀졌다. 반면에 털진달래 종자는 population 수준에서 부분적으로 생리적휴면(PD)을 가지고 있는 것으로 나타났다. 털진달래의 이러한 생리적휴면 (PD)은 외생 지베렐린(GA3) 1,000mg・L-1 처리를 통해 타파될 수 있었다. 그러나 4℃에서의 저온층적처리는 털진달래 종자 휴면 타파에 효과가 없는 것으로 나타났다. 종합적으로 판단 했을 때, 진달래속(Rhododendron) 종들의 적정 발아 환경조 건은 광조건・25/15℃(만병초), 암조건・20/10℃(꼬리진달래), 광 조건・25/15℃(털진달래)로 확인된다. 진달래속(Rhododendron) 에서의 종간 차이(interspecific variation)로 인해 모든 종이 종자 휴면유형 또는 발아특성에서 구별이 되었다. 본 연구는 국내 자생하는 진달래속(Rhododendron) 종들의 생리・생태 특성을 이해하는 데 이용될 수 있을 것이다.
Recycling of drained nutrient solution in hydroponic cultivation of horticultural crops is important in the conservation of the water resources, reduction of production costs and prevention of environmental contamination. Objective of this research was to obtain the fundamental data for the development of a recirculation system of hydroponic solution in semi-forcing cultivation of ‘Bonus’ tomato. To achieve the objective, tomato plants were cultivated for 110 days and the contents of inorganic elements in plant, supplied and drained nutrient solution were analyzed when crop growth were in the flowering stage of 2nd to 8th fruiting nodes. The T-N content of the plants based on above-ground tissue were 4.1% at the flowering stage of 2nd fruiting nodes (just after transplanting), and gradually get lowered to 3.9% at the flowering stage of 8th fruiting nodes. The tissue P contents were also high in very early stage of growth and development and were maintained to similar contents in the flowering stage of 3rd to 7th fruiting nodes, but were lowed in 8th node stages. The tissue Ca, Mg and Na contents in early growth stages were lower than late growth stages and the contents showed tendencies to rise as plants grew. The concentration differences of supplied nutrient solution and drained solution in NO3-N, P, K, Ca, and Mg were not significant until 5 weeks after transplanting, but the concentration of those elements in drained solution rose gradually and maintained higher than those in supplied solution. The concentrations of B, Fe, and Na in drained solution were slightly higher in the early stages of growth and development and were significantly higher in the mid to late stages of growth than those in supplied solution. The above results would be used as a fundamental data for the correction in the inorganic element concentrations of drained solution for semi-forcing hydroponic cultivation of tomato.
Incursions of red imported fire ant (RIFA), Solenopsis invicta into Korea have been increasing. After a first interception of a colony of S. invicta on Gamman pier, Pusan port while intensive surveillance by Animal & Plant Quarantine Agency (APQA) in September 2017, three more RIFA colonies have been found in sea port piers of Pyeongtek, Incheon and Pusan cities. The social forms and mitochondrial DNA haplotypes of the intercepted RIFA colonies were analysed by allelic discrimination assay (peptide nucleic acid probe based RT-PCR) of Gp-9 gene and mt-DNA fragment of 768 bp, which is part of the Cytochrome oxidaseⅠ gene. The colony on Gamman pier, intercepted in Sep. 2017 was previously reported as a haplotype 5 (H5) of mitochondrial DNA and a social form of polygyne. The colony on Hutchison pier of Pusan port, intercepted in June 2018 were confirmed as a H22 haplotype and a monogyne. Those different social forms show different origins of each colonies. Those on piers of Pyeongtek and Incheon ports, also found in 2018 were confirmed samely as H22 and monogyne. However, it could be putatively assumed that those two colonies were differently introduced via different container cargoes, considering those colonies were found in container yards of distantly located different sea ports. More genetic variation analyses using diverse sets of molecular markers such as microsatellites, genome-wide single nucleotide polymorphisms, etc. in nuclear gens are being proceeded for more exact introduction routes (origins).
The hypopharyngeal gland (HPG) of the honeybee worker produces royal jelly (RJ) and has a developmental cycle closely related to the division of labor.
In this study, we investigated to compare the HPG acini diameter of differently aged worker bees with high royal jelly producing colony (HRC) or less producing colony (LRC). Additionally, we also evaluated whether the fresh weight of the head is a reliable indicator of the developmental status of HPG.
The HRC showed a significantly higher RJ production about two-times as compared with those of the LRC. We measured the HG-diameters on days 1, 3, 6, 9, 12, 15. The microscopic analysis revealed that the acini size of the HRC was significantly larger than the LRC. In addition, the acini diameter of HRC was 15% longer than the LRC on the first day after emerging. It was shown that the fastest development during 3 days which is preparing for nurse the brood. The HPG acini diameters increased in both colonies in a similar fashion until day 12 and then decreased. We also compared the fresh head weight of the experimental colonies, differences were similar to the development of HPG.
Therefore, high royal jelly production may have a positive correlation between HPG acini size and the fresh head weight.
The common bed bug, Cimex lectularius, possesses a cholinesterase expressed exclusively in the salivary gland (ClSChE). In this paper, we investigated the molecular structure, tissue distribution patterns, and biochemical properties of ClSChE and showed that ClSChE exists as a soluble monomeric form or a soluble dimeric form connected by a disulfide bridge. Immunohistochemical analysis confirmed that ClSChE was expressed in the epithelial cells of both the salivary gland and the duct. In addition, the secretion of monomeric ClSChE through the proboscis during feeding was detected by western blotting using a ClSChE-specific antibody. To predict the role of ClSChE injected into the tissue of an animal host, we analyzed the extent of sequestration and hydrolysis of acetylcholine (ACh)/choline (Ch) by ClSChE by ultra-performance liquid chromatography-tandem mass spectrometry. Kinetic analysis revealed that ClSChE possesses extremely low Km (high affinity to ACh) and Vmax values. These findings suggest that ClSChE functions as a sequestering enzyme specific to ACh (not to Ch) by having a very strong affinity to ACh but an extremely long turnover time.
수경재배 시스템에 있어서 배액의 재활용은 생산비 절감 및 환경오염 방지를 위하여 중요하다. 방울토마토를 반촉성 수경재배 하면서 생육 단계별 공급액, 배액 및 잎의 무기성분 분석을 통하여 순환식 수경재배 시스템 개발을 위한 기초자료를 확보하고자 본 연구를 수행하였다. 연구목적을 달성하기 위해 주기적으로 공급액, 배액 및 식물체 잎을 채취한 후 무기물 함량을 분석하였다. 생육 초기에는 배액의 EC가 공급액과 비슷한 약 2.0dS·m-1였지만, 생육 후기로 갈수록 높아져 9화방 착과기에는 4.5dS·m-1였다. 영양생장이 왕성한 생육 초기의 pH는 6.4~6.7 범위였으나 생식생장이 강해진 생육 후기에는 5.9~6.1로 낮아지는 경향이었다. 생육 초기에는 공 급액과 배액의 NO3-N, P, K, Ca 및 Mg 농도가 비슷하였지만 생육 후기로 갈수록 공급액보다 배액의 농도가 높아지는 경향이었다. 생육 초기에 잎의 T-N 함량이 높았지만 후기로 갈수록 낮아지는 경향이었다. K와 Ca 함량은 생육 초기에는 낮았으나 후기로 갈수록 높았으며, P와 Mg 함량은 생육초기부터 후기까지 유사한 수준으로 분석되었다. 이상의 결과는 토마토 수경재배를 배액재 활용에 있어 무기원소 농도 변화를 교정하기 위한 기초 자료로 활용될 수 있을 것이다.
The purpose of this study is to investigate the influence of a two-week English-language orientation program at a Korean university on first-year students’ perceptions of English. This quantitative study contributes to the local literature by documenting the changes on a preand post-course survey that measured participants’ willingness to communicate, foreign language anxiety, attitudes towards English, and perceptions of English as a global language. Key results of independent-samples t-tests indicated that student perceptions showed statistically significant changes on five of the six scales, and that the biggest influences of the program were on willingness to communicate and lessening student anxiety. The study also investigated the link between program satisfaction and experience of studying English abroad, finding that there was no relation between them. However, a one-way ANOVA and correlation analysis indicated that there was a relationship between program satisfaction and students’ self-rated proficiency, finding that the lower-proficiency students were slightly less satisfied than those rating themselves as “average” or “good” English speakers. The study concludes with recommendations for future studies and for planning English-language orientation programs in Korean universities.
In the present study, we investigated the effect of porcine follicular fluid (PFF) concentration (10% vs. 1%) and protein-free media (PFF 0%) on maturation of porcine oocytes in vitro and analysed difference in gene expression in resulting blastocysts following parthenogenetic activation. Three groups were tested; 1) 10% PFF: Tissue culture medium (TCM) 199+10% PFF; 2) 1% PFF: TCM 199+1% PFF; and 3) 0.1% PVA: TCM 199+0.1 PVA. Cumulus-oocyte-complexes were cultured in the respective media containing gonadotrophin (1 ug/ml), epidermal growth factor (10 ng/ml), cystein (0.57 mM), sodium pyruvate (0.91 mM), insulin (5 ug/ml), 9-cis retinoic acid (5 nM) for 20~22 h and then without hormonal supplements for an additional 20-22 h. Data was analyzed using statistical analysis system(SAS) program. There was no significant difference in oocyte maturation rate. However, significantly higher (p<0.05) proportions of embryos developed to the blastocyst stage when oocytes were matured in 10% PFF group (45%) than in the 1% PFF group (31.1%). The total cell numbers were not significantly different among groups (52 ± 1.3 vs. 54.6±3.1 vs. 54.4±2.5, respectively). The relative abundance (ratio to beta-actin mRNA) of gene transcripts related to apoptosis in blastocysts was measured by real- time PCR. The expression of anti-apoptotic gene (BclxL) was up-regulated and the expression of pro-apoptotic gene (Bax) was down-regulated in 10% PFF group than in the other groups. Therefore, it can be concluded that supplementation of 10% PFF during in vitro maturation improves embryo development by reduction of apoptosis. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), MKE (#10033839-2011-13), Institute for Veterinary Science, the BK21 program and TS Corporation.
Embryo transfer (ET) is the final procedure for getting pregnancy through assisted reproductive technology such as IVF (in vitro fertilization), SCNT (somatic cell nuclear transfer). In our laboratory, the porcine cloned embryos loaded in ET medium are carried for 3 hours by portable incubator because of the great distance from the laboratory to the experimental farm. Thus, before transferring into recipient, porcine cloned embryos are exposed in vitro condition for long time. Medium which is used in this process is the TALP (Tyrode’s medium supplemented with 10 mM HEPES), but it includes little nutrients for embryo. Thus, the aim of this study is to determine whether ET media containing nutrients affect the in vitro development of embryos compared to TALP. For the experiment, porcine zygote medium (PZM)-5 which has amino acids for developing embryo was chosen as ET medium containing nutrients, added 10 mM Hepes as PZM-5 does not contain buffering system. For experiment, we carried out parthenogenesis through a chemical method using Thi/DTT. Parthenogenetic embryos were cultured in PZM-5 for 2 days, and then they were randomly divided into two group; loaded in a straw with TALP or PZM-5-Hepes, respectively. They were stored in a portable incubator for 3 hours to simulate the time consumed in ET, thereafter embryos in both TALP and PZM-5-Hepes groups were respectively cultured in PZM-5 for additional 5 days. All experiments were repeated 5 times. In result, blastocyst formation rate were 22.46%±1.47 and 23.17%± 2.13, respectively and total cell number were 32.9±2.22 and 37.09±2.18, respectively. There is no significant difference between TALP and PZM-5-Hepes groups. * Further study will investigate effect of PZM-5-Hepes on in vivo development of porcine cloned embryo. This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program and TS Corporation.
Adipose tissue-derived mesenchymal stem cells (ASCs) are very interesting in several laboratory animals and humans because they are easy to harvest and expand to generate millions of cells from a small quantity of fat. ASCs are known as useful materials for clinical applications in human cell therapy and as a donor cell in somatic cell nuclear transfer (SCNT). Here, we investigated if 1) minipig ASCs can be isolated, self-renewed and differentiated into multiple tissue lineages, 2) ASCs can be a suitable donor cell type for generation of cloned pig. In order to isolate ASC, adipose tissues were collected from inguinal region of a 6-year-old female minipig. The ASCs were attached to the culture dish with a fibroblast-like morphology. They expressed cell-surface marker characteristics of stem cell, underwent osteogenic, adipogenic, myogenic, neurogenic and chondrogenic differentiation when exposed to specific differentiation-inducing conditions. To investigate its potential as donor cell for cloning, we respectively carried out SCNT using ASC, adult skin fibroblast (ASF) and fetal fibroblast (FF) derived from same minipig. The ratio of blastocysts to 2-cell embryos and total cell number of blastocysts were monitored as experimental parameters. In results, cleavage and developmental competence to blastocysts rate showed no significant difference among the three groups. On the other hand, total cell numbers of blastocysts derived from ASC and FF were significantly higher than in ASF (89±7.9 and 105±5.5 vs. 57.5±5.2, respectively). Our results demonstrated that ASC have potential compared to ASF and FF in terms of the in vitro development and blastocyst formation ability. In further study, we will investigate the in vivo developmental ability of ASC as donor cell for pig cloning. * This study was supported by IPET (#311011-05-1-SB010), RNL Bio (#550-20120006), Institute for Veterinary Science, the BK21 program, TS Corporation and Optifarm Solution.
We analyzed molecular and enzymatic properties of three cholinesterases (ChEs; ClAChE1, ClAChE2 and ClSChE) from Cimex lectularius. The ClAChE1 and ClAChE2 were generally present as a membrane-anchored dimeric insoluble form in the brain and ganglia. In the case of ClSChE, monomeric and dimeric soluble forms were observed. To investigate enzymatic properties, three ChEs were functionally expressed using baculovirus expression system. ClAChE1 revealed a significantly higher activity than ClAChE2 to acetylthiocholine iodide (ATChI) substrate. Kinetic analysis using two choline substrates (ATChI and butyrylthiocholine iodide) demonstrated that ClAChE2 had higher catalytic efficiency but lower substrate specificity than ClAChE1. Inhibition assay was conducted by using three inhibitors (BW284C51, eserine, Iso-OMPA) and two insecticides (chlorpyrifos-methyl and carbaryl). Two ClAChEs revealed high sensitivities to BW284C51, eserine, chlorpyrifos-methyl and carbaryl, but were not sensitive to Iso-OMPA. This inhibition profile confirmed that both ClAChEs are categorized as ChEs. Interestingly, the salivary specific cholinesterase did not show any measurable activities to choline substrates, confirming its non-synaptic function in C. lectularius
Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.
국화 6 품종을 대상으로 감마선(35 Gy) 조사에 의해 유기 된 화색변이체를 선발하고, 식물체 재생을 위한 배지내 식물생 장조절제 적정농도를 구명하여 신품종 육성 가능성을 검토하고 자 수행하였다. 고유품종과 다른 꽃잎의 화색변이율은 ‘SP-410’ 이 15.7%로 가장 높았으며, ‘예스스타’가 10.6%, ‘무지개’가 10.3%순 이었다. ‘무지개’는 이중색(황색/적색, Y7D/R40A) 에서 적색(R42B)으로, ‘예스라인’은 자주색(RP74C)에서 진자 주색(RP63B)으로, ‘예스스타’는 황색/적색(Y4A/RP59A) 이 중색에서 적자주색(RP64A)으로, ‘SP-410’은 백색(W155C)에서 황색(Y2C)으로, ‘예스투게더’의 화심색은 백색(W155C)에서 연보라색(P76B)로, ‘예스우리’의 화심색은 연녹색(YG149A)에 서 짙은 녹색(GY1C)으로 화색이 변한 변이체를 선발하였다. 화색변이체 꽃잎의 신초형성을 위한 배지내 식물생장조정제 의 최적조합은 BA 1.0 mg/L와 NAA 0.5 mg/L, BA 2.0 mg/L 와 NAA 0.5 mg/L의 2조합이였으며, 신초형성율은 품종간에 차이가 있어 ‘예스라인’이 91.6%로 가장 높았다. 감마선 조 사에 의한 화색 변이체의 꽃잎배양 유래 재생식물체는 영양 번식으로 유지되었으며, 3년동안 재배한 결과 화색과 꽃잎의 형태적 특성 차이가 없었다.