국화에서 줄기괴저바이러스와 토마토반점위조바이러스를 매개하는 것으로 알려진 꽃노랑총채벌레 약충과 성충이 잎, 꽃, 과실의 흡즙을 통해 확산시킨다. 최근 무분별한 농약 사용으로 약제저항성 계통 출현이 지속적으로 보고되고 있다. 총채벌레류는 식물바이러스 매개하는 시간이 짧아 상대적으로 효과가 낮은 약제로는 총채벌레 방제효과를 기대하기가 어렵다. 시판되는 국화 총채벌레류 등록 살충제 10종을 꽃노랑총채벌레 발육단계별(약충, 번데기, 성충)로 살포한 결과, 유효성분 피롤계 클로르페나피르, 카바메이트계 벤퓨라카브와 스피노신계 스피네토람이 함유된 약제가 우수한 살충력을 보였다. 총채벌레류 화학적 방제를 위해서는 위 성분이 함유된 살충제를 교호로 사용하는 것이 효과적일 것으로 보여진다.

This study was carried out to confirm the predatory and developmental features of N. stenoferus. We determine the host range of N. stenoferus. As a result, it was confirmed that Aphis gssypii, Myzus persicae, Planococcus citri, and Frankliniella occidentalis. N. stenoferus is thought to be able to feed on other micro pests. The test for a developmental period of N. stenoferus at 25℃ showed that the egg period was about 10 days. The nymphal period was about 18 days. Each nymphal period from 1st instar to 3rd instar nymphs was about 3 days. And the nymphal period of 4th and 5th were about 3.5 and 6 days, respectively. The female adult laid eggs in stem tissue or on leaves, and sometimes on the soaked cotton for water supply.

국화를 가해하는 총채벌레류로 꽃노랑총채벌레, 대만총채벌레, 미나리총채벌레, 파총채벌레, 오이총채벌레, 고사 리그물총채벌레(Hercinothrips femoralis)가 보고되었다. 특히 꽃노랑총채벌레는 토마토반점위조바이러스(TSWV)와 국화줄기괴저바이러스(CSNV)를 매개, 확산시켜 국화 품질저하, 생산성 감소의 주요 원인 중 하나이다. 한편 고사리그 물총채벌레는 국화과, 장미과, 백합과, 사초과, 앵초과, 협죽도과, 후추과, 두릅나무과, 쐐기풀과, 천남성과, 꿀풀과, 선인장과 등 여러 식물을 가해하는 하는 것으로 국외에서는 이미 보고되었으나 국내에서는 아직 연구되고 있지 않다. 2018년 전북 완주군 이서면 국립원예특작과학원의 국화 시험온실(무농약)에서 꽃노랑총채벌레와 고사리그물총 채벌레가 발견되었다. 해충이 없는 국화에 꽃노랑총채벌레와 고사리그물총채벌레를 각각 정착시켜 피해 증상을 비교한 결과, 2종의 총채벌레가 가해한 국화 잎에서 각각 다른 피해 양상이 나타났다.

뽕나무깍지벌레는 복숭아, 매실, 뽕나무 등 31종의 기주식물을 가해하는 기주범위가 광범위한 난방제 해충으로 알려져 있다. 2017년도에 뽕나무깍지벌레는 5월 상순부터 부화를 시작하여 암컷성충 1마리가 75.5개(47~159개)의 알을 낳으며 모든 알이 부화하는데 약 19일이 소요되었고, 부화율은 약 87%였다. 반면에 2018년도에는 4월 하순부터 부화를 시작하여 암컷성충 1마리가 49.4개(12~71개)의 알을 낳으며 모든 알이 부화하는데 13일이 소요되었으며, 부화율은 약 72%를 보였다. 부화한 약충들은 이동 후 5월 중순부터 고착약충으로 되고, 6월 중순부터 2세대 성충이 활동하기 시작하였다. 부화약충기에 약제를 살포하여 살충효과를 조사한 결과, 살충률이 100%인 반면에, 고착약충기 에는 살충율은 2.7%로 살충효과가 낮은 것을 확인하였다.

알락수염노린재(Dolycoris baccarum L.)는 구북구(Palearctic region)에 널리 분포하는 다식성 곤충으로서 국내에서는 콩, 단감, 유자 등 다양한 작물의 해충이다. 이 종은 다른 노린재과(Pentatomidae) 곤충과 마찬가지로 수컷이 집합페로몬 을 방출하는 것으로 알려져 있다. 우리는 수컷의 집합페로몬 성분을 동정하기 위하여, 고상미량추출(SPME) 방법을 이용하여 수컷과 암컷 성충에서 방출되는 휘발성 물질을 포집하여 화학구조를 분석하였다. 그 결과, 수컷 특이적인 휘발물질로서 α-bisabolol, α-bergamotene, β-bisabolene을 동정하였고, 3가지 성분이 100:15:5 비율로 존재함을 밝혔다. 야외에서 알락수염노린재 수컷과 암컷 성충 모두가 주성분인 α-bisabolol을 처리한 트랩에 유인되었다. 나머지 2가지 성분(α-bergamotene과 β-bisabolene)의 경우, 그 자체로는 유인 효과가 없었으나 주성분인 α-bisabolol에 추가되면 보강효과가 발휘되었다. 곤충에서 α-bisabolol이 동정된 것은 이것이 처음이며, 이번에 밝혀진 알락수염노린재 집합페 로몬은 이 해충의 발생예찰과 친환경 방제에 유용할 것이다.

This study was carried out to confirm the parasitic and developmental features of A. japonica and D. suzkii was used as a parasitic natural enemy. A. japonica attacked the D. suzukii larvae and the emergence of adults were observed from D. suzukii pupae. Black spots were observed in parasitized D. suzukii larvae. Mortality of parasitized larvae, rate of parasitic and developmental feature were investigated according to developmental stages of host, D. suzukii. Mortality and rate of parasitic of D. suzukii larvae were the highest when second instar larvae were attacked. Developmental period of parasitized D. suzukii larvae showed differences to developmental stages, but there was no significant difference in developmental stage of pupal period.

Glutathione S-transferase (GST) is a key gene involved in multiple stress tolerance in all living organisms, though it is still to be disclosed the gene function in teff grass [Eragrostis tef (Zucc.)Trotter].The objectives of this study were to clone and molecular characterization of GST gene in teff grass. We characterized GST1 from teff grass (EtGST1), it composed of a 645-bp open reading frame (ORF) that encoded 195 amino acid residue. Further, we transformed EtGST1 in E.coli BL21 (DE3) cells. This recombinant EtGST1 in E.coli BL21(DE3) induced at 37°C temperature. In addition, Growth of cells overexpressing EtGST1 rapidly increased in the presence of polyethylene glycol (5%), heat (46°C), NaCl (0.6%), and arsenic (1 mM) than that of cells harboring an empty vector. These results suggest that EtGST1 would be suitable candidate for improving tolerance in forages and/or grasses species against multiple abiotic stresses.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

The specific genetic modification in porcine somatic cells by gene targeting has been very difficult because of low efficiency of homologous recombination. To improve gene targeting, we designed three kinds of knock-out vectors with α1,3-galactosyltransferase gene (α1,3-GT gene), DT-A/pGT5’/neo/pGT3’, DT-A/NLS/pGT5’/neo/pGT3’ and pGT5’/neo/ pGT3’/NLS. The knock-out vectors consisted of a 4.8-kb fragment as the 5’ recombination arm (pGT5’) and a 1.9-kb fragment as the 3’ recombination arm (pGT3’). We used the neomycin resistance gene (neo) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. These vectors have a neo gene insertion in exon 9 for inactivation of α1,3-GT locus. DT-A/pGT5’/neo/pGT3’ vector contain only positive-nega-tive selection marker with conventional targeting vector. DT-A/NLS/pGT5’/neo/pGT3’ vector contain positive-negative selection marker and NLS sequences in upstream of 5’ recombination arm which enhances nuclear transport of foreign DNA into bovine somatic cells. pGT5’/neo/pGT3’/NLS vector contain only positive selection marker and NLS sequence in downstream of 3’ recombination arm, not contain negative selectable marker. For transfection, linearzed vectors were introduced into porcine ear fibroblasts by electroporation. After 48 hours, the transfected cells were selected with 300 μg/ml G418 during 12 day. The G418-resistant colonies were picked, of which 5 colonies were positive for α1,3-GT gene disruption in 3´ PCR and southern blot screening. Three knock-out somatic cells were obtained from DT-A/NLS/ pGT5’/neo/pGT3’ knock-out vector. Thus, these data indicate that gene targeting vector using nuclear localization signal and negative selection marker improve targeting efficiency in porcine somatic cells.