In the present study, the 17,694-bp long complete mitochondrial genome (mitogenome) of the dwarf honey bee, Apis florea (Hymenoptera: Apidae), is described with an emphasis on the noteworthy triplicated tRNAser(AGN) region and an extraordinary long A+T-rich region with repeat regions. The gene arrangement of A. florea mitogenome is identical to that of A. mellifera, but has triplicated tRNASer(AGN), each of which contains the precedent 44 bp-long and following another 64 bp-long repeats plus one complete first repeat abutting to tRNAMet. A total of 1,610-bp long two repeat regions in 1,987 bp-long A+T-rich region is composed of nearly identical 141 ~ 219-bp long five tandem repeats and 50 ~ 52-bp long 12 tandem repeats that are encompassed by three non-repeat sequences. One of the common interpretations for such repeat sequence is slipped-strand mispairing and unequal crossing-over events during DNA replication.

We previously reported Pear Pest Forecasting Management System (PPFMS) for the Improvement of pass ratio of Korean exporting pears. It is consisted of regular field forecasting by pear farmers, meteorological data obtained by automatic weather station (AWS), an internet web page (http://pearpest.jnu.ac.kr/) as information collecting and providing ground, and information providing service. Currently, we are expanding this system to the area, Cheonan and Ansung, where pear orchards are organized into exportation-specific group. Further, the information obtained from field forecasting and AWS were up-loaded to under-constructing upgraded webpage (http://www.kpear.kr), with several pest/disease-related information. We hope this pest forecasting management system increases the pass ratio of Korean exporting pears throughout establishment of farmer-oriented forecasting, inspiring farmers’ effort for the prevention and forecasting of diseases and pests occurring at pear orchards.

This study was conducted to estimate the control thresholds (CTs) at different larval densities of Oides decempunctatus Billberg (Coleoptera: Chrysomelidae) of Campbell early in the vineyard and investigated life cycle. Each stage of O. decempunctatus was sampled 18 times from May to September in 2010~2012. The seasonal occurrence of O. decempunctatus showed the highest peak in mid-late June and mid-late August. Overwintered O. decempunctatus's eggs were hatched from late May to early June. Larva period was from late May to mid July and adults appeared in mid July. The percentage of leaf damage (Y) of Campbell early inoculated by different densities of O. decempunctatus (X, no. of larvae/fruiting mother branch) for six weeks was estimated by Y= 0.498X+2.041 (R2=0.988) during vegetation period. The decreasing rate of soluble solid (Y) after grape harvest of Campbell early damaged by different densities of O. decempunctatus (X) was estimated by Y= - 0.046X+15.3 (R2=0.8543). Based of the relationships between the densities of O. decempunctatus larvae and the index of reducing soluble solid of Campbell early, the number of larvae (2nd to 3rd instar) which decreased less than 15°Bx loss of soluble solid was determined as the injury level of 7/fruiting mother branch.

본 연구는 염생식물인 퉁퉁마디를 대상으로 하여 신간척지에서 안정적인 재배법을 구명하고자 수행하였다. 먼저 퉁퉁마디의 발아특성을 알아보기 위해 염농도(NaCl 0 ~ 3% 농도) 별발아율 시험을 수행하였다. 10oC에서는 모든 염농도에서 발아속도는 느리나 15일까지 발아가 지속되어, 78 ~ 90%의 발아율을 나타내었다. 20oC이상에서는 4일 안에 최대의 발아율을 나타내었으며, NaCl 3%에서 온도가 올라갈수록 발아율이 저하되는 경향을 보였다. 퉁퉁마디의 질소시비량 별 생육특성을 알아보기 위해 온실포트실험을 실시한 결과, 퉁퉁마디 파종 후 5일 후부터 발아가 시작하였고, 퉁퉁마디의 생육은 질소시비량이 증가할수록초장 및 분지 등이 증가하는 경향을 나타내었다. 또한 염농도에 따른 생육특성을 조사하기 위해 양액 재배 조건에서 염농도에 따른 퉁퉁마디 생육을 실험하였다. 양액조건 상태에서NaCl을 처리하지 않으면 생육이 저조하여 1개월 이후에 고사하기 시작했으며, NaCl을 처리한 구에서는 NaCl 200 mM에서생육이 좋았고, NaCl 500 mM 이상의 염농도에서도 재배가 가능할 것이라 생각된다. 신간척지에서 퉁퉁마디의 어린 싹을6월에 이식하여 질소시비량이 퉁퉁마디 생육에 미치는 영향을조사하였다. 퉁퉁마디의 초장 및 분지수 등이 질소시비량이 증가할수록 유의적으로 증가하였으나, 인산을 비롯한 무기성분함량은 유의적인 차이가 없었다.

새만금 간척지에서 돈분퇴비를 이용하여 수수×수단그라스 교잡종 재배 후 토양변화에 수량성을 평가하기 위한 시험연구 결과는 아래와 같았다. 시험토양은 용적밀도가 높고 공극률이 낮은 간척지토양으로돈분퇴비 시용 후 수수×수단그라스를 재배한 결과 토양경도는 1.2~3 mm까지 증가하였으며 용적밀도는 0.14~0.17Mg·m−3감소하였으나, 이는 토양수분 등 계절적인 요인에 기인한 것으로 생각된다. 토양물리성 중 토양경도와 용적밀도의 관행시비와 돈분퇴비 시용구간의 차이는 없었다. 토양pH, 토양유기물 및 유효인산은 관행시비와 돈분퇴비시용구 처리간 차이가 뚜렷하였으며, 특히 토양유기물과 유효인산은 돈분퇴비의 투입량이 증가할수록 토양 중 함유량의 증가가 뚜렷하였다. 수수×수단그라스 교잡종 건물수량은 가축분퇴비구에서는 질소의 총투입량이 증가할수록 수량이 증가하였다. 새만금 간척지내에서 돈분퇴비를 이용하여 수수×수단그라스 교잡종 재배시 돈분퇴비를 기비로 ha당 17,210 kg이상 시용하면 관행시비와 비슷한 수량이 생산되었다.

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 investigate the effects of NEAA and leptin supplemented to in vitro culture medium on the developmental competence of porcine embryos after intracytoplasmic sperm injection (ICSI), and to modify the culture condition to improve the quality and the development of ICSI-derived porcine embryos in vitro. After ICSI, the putative zygotes were then cultured in PZM-3 medium with/without NEAA or leptin. The proportion of embryos that developed to the blastocyst stage significantly increased when 1% NEAA (24.62%) was added to the medium compared with 2% NEAA and no NEAA group (17.24% and 20.24%, respectively, p<0.05). The effect of different concentration of leptin (0, 10, 100, 500 ng/ml) was evaluated on the development of porcine ICSI embryos cultured in vitro. In case of blastocyst formation, 100 ng/ml group (27.05%) showed significantly higher rate than 10, 500 ng/ml, and control group (23.45%, 17.99%, and 19.68%, respectively, p<0.05). We also evaluated the effects of different NEAA and leptin treatment time on the development of porcine embryos after ICSI. Among groups of embryos cultured in the presence of NEAA or leptin for whole 7 days (D 1-7), first 4 days (D 1-4), the subsequent 3 days (D 5-7), both NEAA (27.13%, 21.17 %, and 17.56%, respectively, p<0.05) and leptin (25.60%, 20.61%, and 16.53%, respectively, p<0.05) showed that supplementation for whole 7 days significantly increased the blastocyst formation rate compared with the other groups of D1-4 and D5-7. We further evaluated the combination effect of 1% NEAA and 100 ng/ml leptin compared with the effect of each supplementation with 1% NEAA or 100 ng/ml leptin or no supplementation on development of embryos. For blastocyst formation, combination group of NEAA and leptin (24.78%) showed significantly higher rate than other three groups (18.37%, 20.44 %, and 13.27%, respectively, p<0.05). We further evaluated the expression of proapoptosis genes such as BAX and BAK and anti-apoptosis genes, BCL-XL and BCL-2 in blastocysts cultured in the presence of 100 ng/ml leptin. RT-PCR analysis revealed that leptin supplementation significantly decreased the expression of pro-apoptosis genes as well as increased the expression of anti-apoptosis genes. These results of present study demonstrate that NEAA and leptin could improve the in vitro development of ICSI- derived porcine embryos with optimal concentration of each reagent. Furthermore, the optimal culture condition could increase the quality of ICSI-derived embryos in vitro.

Chicken Insulin-like Growth Factor-1 (cIGF-1), one of the most important hormone for regulating physiological function includes body growth, muscle volume, bone density, chicken cell development and metabolism. In order to find in vitro Knokdown expression of cIGF-1, this study introduced tetracycline inducible RNA interference expression system (TetRNAi system). Tet system can inductively control high expression of extrinsic genes and expression of intrinsic genes. So it has advantages such as minimized physiological side-effects any cell and low cytotoxicity. RNAi system is proving to be a powerful experimental tool for inhibition of gene expression and post-transcriptional mechanism of gene silencing. RNAi is mediated by small interfering RNA (siRNA) consisting of 19- to 23- nucleotide double-stranded RNA duplexes that promote specific endonucleolytic cleavage of mRNA targets through an RNA-induced silencing. Then, this study RNAi-based gene knockdown can be achieved by retroviral-based expression systems. Stable integration of our inducible siRNA vector allowed the production of siRNA on doxycycline induction, followed by specific down regulation of chicken IGF-1 gene. Analyses of Real-time PCR to determine expression of the cIGF-1 gene showed successful from chicken embronic fibroblast (CEF) cells with the reduced rate of an approximately 92%. Our results demonstrate the successful regulation of cIGF-1 knockdown expression in CEF cells and support the application of an tetracycline inducible RNAi expression system in transgenic Mini chicken production. This research was supported by Bio-industry Technology Development Program, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio- technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being.

벼 담수표면산파재배시 주요 품종들의 적응성을 평가하기위하여 1996~1998년에 국립식량과학원 벼맥류부 벼 시험포장에서 공시품종들의 중요한 형질인 입모율, 도복 및 수량성 등을 검토한 결과를 요약하면 다음과 같다. 1. 입모율이 80% 이상인 품종은 동안벼, 호안벼, 동진벼,80% 이하인 품종은 화영벼, 팔공벼, 대산벼 등 이었다. 2. 도복이 발생하지 않았거나 경미한 품종은 대산벼, 동안벼,금남벼, 안산벼, 주안벼, 안중벼, 화남벼, 금남벼, 오봉벼, 도복발생이 심했던 품종은 금오벼, 화영벼, 동진벼, 낙동벼 이었다. 3. 당 수수와 수당립수는 조생종이 중생종과 중만생종에 비하여 현저히 적었고, 등숙비율은 중생종과 중만생종간에는 차이가 없었으나 조생종은 다소 낮았다. 4. 쌀 수량은 중만생종과 중생종간에는 별 차이가 없었으나,조생종은 4품종 모두 중생종과 중만생종 품종보다 0.79~0.85MT/ha정도 낮았다. 5. 따라서 벼 담수표면산파재배시 주요 품종들의 입모, 도복 및 수량성 등을 평가해 본 결과 적응성이 양호한 품종은 동안 벼, 대산벼, 호안벼 이었다.

새만금 간척지에서 동계 사료작물의 재배 가능성을 검토하기 위하여 2009년 가을부터 2011년 봄까지 염분농도가 다양한 토양조건에서 청보리, 호밀, 밀, 트리티케일, 귀리 등을 공시하여 생육특성,생초 및 건물수량 등을 검토한 결과를 요약하면 다음과 같다. 1. 출현율과 입모율은 토양에 염분이 거의 없는 시험구에서 84%와 79%인 반면, 토양 염농도 0.2% 시험구에서는 73%와 64%, 토양 염농도 0.3% 시험구에서는 52%와 40%, 토양 염농도 0.4% 시험구에서는 37%와 22%이었다. 2. 사료작물별 출현율과 입모율은 호밀, 밀, 트리티케일 및 청보리는 비슷하였으며, 귀리는 상대적으로 낮았다. 3. 초장은 토양 염농도가 높을수록 현저히 단축되었으며, 토양 염농도 0.3% 이상 시험구에서는 단축정도가 매우 컸다. 4. 생초 및 건물수량은 토양에 염농도가 거의 없는 시험구 대비 토양 염농도 0.2% 시험구에서 66~77% 수준이었으나, 토양염농도 0.3% 이상 시험구에서는 30~61% 수준이었다. 5. 사료작물별 생초수량은 호밀 >밀 >청보리 >트리티케일 >귀리 순으로 많았으며, 건물수량은 밀> 호밀=청보리> 트리티케일> 귀리 순으로 많았다. 6. 따라서 새만금 간척지에서 동계 사료작물의 재배가 가능한 토양 염농도는 공시작물의 생육특성, 생초 및 건물수량 등을 종합해 볼 때 0.2% 이하이고, 염류토양에 대한 적응력이 높은사료작물은 호밀, 밀, 청보리, 트리티케일 이었다.

The objective of this study was to evaluate in vitro production of bovine embryos in Hanwoo. Oocytes were collected by ovum pick up (OPU) from ovaries of genetically high-value Hanwoo or by needle puncture from ovaries of slaughtered cattle. OPU was done every 3 4 days duing experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, We compared the in vitro maturation rate in two groups (Experiment 1). 545 oocytes were recoverd from 4 females by 32 trials of OPU and then 433 oocytes were shown MⅡ stage after in vitro maturation (79.4%). In case of needle puncture group, 1905 oocytes were collected and then 1420 oocytes were matured to MⅡ stage during in vitro culture(74.5%). Second, we compared the developmental rate to blastocyst in two groups (Experiment 2). 1420 oocyte by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24 48 h after in vitro fertilization (IVF) was 88.6% and blastocyst development rate was 20.5% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (84.8%), blastocyst development rate was higher compared with needle puncture group (26.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio-technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being. This research was suppoted by Imsil-gun agricultural technology service center.

The black-veined white, Aporia crataegi (Lepidoptera: Papilionoidea), is nearly extinct in South Korea, although substantial numbers of dried specimens are available. One of the common practices for such species is to launch re-introduction program after proper amount of genetic information are analyzed from donor and donee populations. In this study, we sequenced complete mitochondrial genome (mitogenome) of A. crataegi to design species-specific primers for subsequent population works and to further understand the mitogenome evolution in lepodiopteran Papilionoidea. The 15,140-bp long A. crataegi mitogenome that has typical sets of 37 genes is smallest among true butterfly species with overall slightly smaller size in genes and regions throughout the genome. Arrangement of the genome is identical to those of other lepidopteran mitogenomes, in which tRNA cluster located between the A+T-rich region and ND2 gene is translocated into tRNAMet, tRNAIle, and tRNAGln from ancestral arrangement, tRNAIle, and tRNAGln, tRNAMet. The A/T content of the genome at 81.3% is the highest in Pieridae, but lower than that of lycaenid species (81.7% ~ 82.7%) The high A/T content in the genome is also reflected in codon usage, accounting for 41.69% of A/T-composed codons (TTA, ATT, TTT, and ATA). Unlikely the diversified or modified usage of anticodon for tRNASer(AGN) the species of Pieridae including A. crataegi all unanimously have GCT that has been hypothesized as ancestral for Lepidoptera. A total of 111 bp of non-coding sequences are dispersed in 13 regions, ranging in size from 1–49 bp. Among them relatively longer ones (≥ 16 bp) all have relatively higher sequence identity to other regions of the genome, suggesting partial duplication of the sequences during A. crataegi evolution. As has been reported in some species of Lepidoptera, the A. crataegi A+T-region also has typically found conserved sequences (e.g., poly-T stretch, ATAGA motif, ATTTA element, microsatellite-like A/T sequence, and poly-A stretch) and one tRNA-like sequence, and this feature was commonly found in true butterfly species.

The phylogenetic relationships among the Nymphalidae (Lepidoptera: Papilionoidea) have been controversial in several perspective. The present study sequenced a total of ~ 3,500 bp from cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-1 alpha (EF-1α) in 80 nymphalid species belonging to seven subfamilies (Linmenitidinae, Heliconiinae, Nymphalinae, Apaturinae, Libytheinae, Satyrinae, and Danainae), along with those of six lycaenid species as outgroups. Phylogenetic analyses via Bayesian Inference (BI) and Maximum Likelihood (ML) algorithms concordantly supported the subfamilial relationships of (((((Linmenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Libytheinae) + Satyrinae) + Danainae), with high nodal support for monophyletic subfamilies and tribes. This result is largely consistent with a previous study performed with a substantially large sequence information and morphological characters, except for the position of Libytheinae that has previously been placed as the sister to all reminder of Nymphalidae.