This study was conducted to investigate the insecticidal capacity of several recombinant baculoviruses to P. xylostella and S. exigua larvae. NeuroBactrus was constructed as follows: the cry1-5 of Bacillus thuringiensis 2385-1 was inserted into Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) genome by fusion of polyhedrin-cry1-5-polyhedrin under the control of polyhedrin gene promoter, and insect-specific neurotoxin from the scorpion Androctonus australis (AaIT) under the control of early promoter from Cotesia plutellae bracovirus was introduced by fusion of orf603 partial fragment in the opposite direction of polyhedrin gene, respectively. Other recombinant baculoviruses derived from the NeuroBactrus - NBt-DelA (deleted AaIT), NBt-Del5 (deleted cry1-5), and NBt-DelA5 (deleted AaIT and cry 1-5) - were manufactured in serial passages in vitro. The data were analyzed by SPSS. The value of LC50 was lower when P. xylostella larvae fed on cabbage coated with NeuroBactrus (4068.4) than when it fed on cabbage coated with AcMNPV (4.5x106). Survival time (ST50) of P. xylostella larvae (2.54days) was shorter when it fed on cabbage coated with NeuroBactrus than when it fed on cabbage coated with other recombinant baculoviruses (7.54days, 7.68days, and 8.26days) and AcMNPV (9.67days). S. exigua larvae presented the same results.

Strawberry, Fragaria ananassa Duchesne, is one of the important horticultural crops cultivated in greenhouses. Tetranychus urticae is one of major strawberry pests, and Neoseiulus californicus and Phytoseiulus persimilis have been used as biological control agents for control of T. urticae. The interactions between T. urticae and N. californicus and between T. urticae and P. persimilis were investigated to compare their control efficiency for T. urticae on a spatially-structured strawberry leaf disc area at different temperatures and on different treatments. The experimental arena was an array of leaf discs (3 cm diameter) placed upside down on a water-saturated cotton pad in an aluminum pan (17.4 x 21.5 cm). Twenty leaf discs (4 x 5) were placed adjacent each other for allowing dispersal of T. urticae and its predatory mites. The temperature conditions were 20, 25, and 30°C and there were six different treatments. The overall population densities of T. urticae were influenced by temperature (20, 25, and 30°C) in N. californicus treatment. In the same temperature condition, P.persimilis was more effective than N. californicus to control T. uritcae. Two predator systems were better than one predator systems to suppress the population density of T. urticae at 25°C. Some results of this study could be used to understand the spatial association of T. urticae and its predatory mites in greenhouse crops and fields.

The study was conducted to explore whether environmental differences, in this case the physical characteristics of abaxial leaf surfaces of strawberry cultivars ('Maehyang' and 'Sulhyang' cultivars), affect the functional response of adult female N. californicus preying on immature stages (egg, larva and nymph) of T. urticae. We also evaluated the functional response of N. californicus to eggs of T. urticae at different temperatures (15, 20, 25, 30 and 35℃). We conducted a logistic regression of the proportion of prey consumed as a function of initial prey density to identify functional response types, and used nonlinear squares regression and the random predator equation to estimate attack rates and handling times. The functional response of adult female N. californicus to T. urticae was not influenced by non-glandular trichomes and epicuticular waxes on the abaxial leaf but was affected by temperature. Overall, the functional response of adult female N. californicus exhibited a type 2 functional response to T.urticae. The handling time of N. californicus was highest (1.9970) against T. urticae nymphs. The attack rate did not change much at 15-30℃, but the handling time decreased linearly with increasing temperature. At 35℃, the attack rate was highest (0.1876) and the handling time was lowest (0.9296). The results of this study may be used to evaluate the potential of N. californicus to suppress T. urticae and to estimate parameters for relevant prey-predator models.

The porcine reproductive and respiratory syndrome virus (PRRSV) has six structural proteins which encoded by ORFs 2 to 7 are designated as GP2, 3, 4, 5, M and N, repectively. In this study, we determined the expression of each protein using novel transfer vector, pBmKSK4 which has the polyhedrin promoter of BmNPV and 6xHis tag. The recombinant transfer vector was co-transfected into Bm5 cells along with bBpGOZA DNA. Recombinant virus was purified by plaque assay and amplified in Bm5 cells. Expression of each protein was identified by SDS-PAGE and Western blot analysis using anti-6xHis monoclonal antibody. The expression levels of the structural proteins in Bm5 cells were stronger than the expression system using pBacPAK9 transfer vector in Sf21 cells. As expected, GP5 was expressed at low levels from its structural properties and its toxicity for cells. In addition, each recombinant protein was purified using Ni-NTA spin columns. The ability to produce each protein in the baculovirus system indicates that these could be major candidates for the development of a vaccine against PRRSV.

배추좀나방과 담배거세미나방 유충에 대한 30종 한방 식물체 메탄올 조추출몰의 살충활성 및 섭식 저해활성을 잎침지법을 이용하여 5,000ppm으로 검정하였다. 황련의 추출물은배추좀나방에 대해 살충작용을 보였다. 길경, 사삼, 세신, 오배자 및 자초의 추출물은 배추좀나방 유충에 대해, 목통 및 속새의 추출물은 담배거세미나방 유충에 대한 강한 섭식저해활성을 보였으나, 오배자와 황련의 추출물은 이들 나비목 유충 모두에 강한 섭식저해활성을 나타내었다.

생체고분자물질은 수자원환경에서 점토, 미생물, 바이오매스 등 부유입자들을 응집시키고, 침전, 퇴적시키는 역할을 한다. 본 연구는 다양한 수질화학 조건이 생체고분자물질에 의한 부유입자 응집에 미치는 영향을 파악하고자, 수질화학 조건을 제어하여 응집실험을 수행하였다. 각 응집실험은 이온강도, 2가 양이온 농도, 휴믹물질 분율이 제어된 실험조건에서 Kaolinite 현탄액에 생체고분자물질인 Xanthan Gum을 주입하여 수행하였다. 수체가 가지는 응집능은 응집체 크기 및 잔류 고형물 농도를 측정을 통하여 평가하였다. 본 연구에서, 이온강도 증가는 점토입자 및 생체고분자물질 간 정전기적 반발력을 감소시키고 생체고분자물질이 점토 입자 간 가교를 형성하여 응집을 증대시킨 것으로 파악되었다. 이온강도가 0.001에서 0.1 M NaCl로 증대될 경우, 응집을 증진시켜 응집체 크기는 약 3배 이상 증 대되고 부유고형물농도는 약 2.5배 이상 저감되었다. 또한, 2가 양이온이 수체에 존재하는 경우, 점토입자-생체고분자물질 혹은 생체고분자물질 상호 간 가교를 형성하여, 즉 점토-Ca2+-고분자 또는 고분자-Ca2+-고분자 가교를 형성하여, 생체고분자물질에 의한 부유입자 응집을 증대시켰다. 수체에 Ca2+가 낮은 농도라도 존재 할 경우, 응집을 크게 증진시켜 부유고형물농도가 원주입농도에 비하여 20배 이상 저감되는 것으로 나타났다. 하지만, 휴믹물질이 존재하는 경우, 점토입자 표면에 흡착되어 점토입자의 정전기적 반발력을 증대시켜 생체고분자물질의 흡착을 방해하고 응집을 감소시켰다. 수체에 휴믹물질이 존재할 경우, 응집을 저감시켜 부유고형물농도는 저감되지 않고 원 주입농도와 유사하게 나타났다. 본 연구의 결과는 수자원환경에서 부유입자 및 퇴적물 거동을 이해하고 수질 및 퇴적물에 대한 최적 관리 방안을 도출하기 위한 기초 자료로 활용될 수 있으리라 기대된다.

Citrus canker caused by Xanthomonas citri is a notorious disease affecting a decrease in fruit productivity and quality. Citrus export to USA is also prohibited by the disease. Therefore, development of citrus canker resistant variety is essential and exploitation of markers for molecular breeding is urgent. To develop DNA molecular markers, we performed whole genome resequencing for 8 varieties: 4 citrus canker resistant varieties including C. hybrid ‘Kioymi’ and 4 citrus canker susceptible varieties including C. iyo ‘Miyauchiiyokan’. In total, 642 polymorphic SNPs were detected between resistant and susceptible varieties. Of the 642 SNPs, 50 SNPs were preferably selected based on integrative genomics viewer. To apply the markers in a broad range of citrus variety, we performed genotyping with 6 other varieties very well known as citrus canker resistant and susceptible varieties in addition to previous mentioned 8 varieties. Three of the 50 SNPs were identified as a marker to distinguish citrus canker resistant varieties from susceptible varieties. Secondly, we developed molecular markers to apply for F1 lines crossed by ‘Kiyomi’ and ‘Miyauchiiyokan’. Of the 50 SNPs, we identified 2 SNP markers to distinguish between F1 resistant and susceptible lines. One of them is a resistance gene that plays a role in plant defense mechanism. In this study, we developed 5 molecular marker candidates possible to apply for molecular breeding to develop citrus canker resistant variety. We are working on development of candidate markers related to citrus canker.

Citrus canker caused by Xanthomonas citri pv. citri is one of economically important diseases in the citrus industry. The devastating bacterial disease results in unattractive quality and a significant reduction in fruit production. Citrus growers and industry in Korea has been struggling with the serious disease causing the prohibition of export market. Korea also became the top import market for oranges. The development of markers linked to citrus canker resistance is strongly needed. In this study, we investigated molecular markers between ‘Kiyomi’ (Citrus unshiu ｘ C. sinensis), a resistant cultivar, and Natsudaidai (C. natsudaidai), a susceptible cultivar. To develop markers, we focused on structural variation (copy number variation, CNV, and presence/absence variation, PAV). It has been well documented that CNV and PAV of defense-related genes are associated with resistant cultivars. Using a read depth approach following next-generation sequencing, we performed genome-wide analysis of CNV and PAV in two varieties. As a result, 633 genes showing at least two times difference between the mapping reads from two varieties and 61 genes showing presence of the mapping reads in only either one of them were screened. Visual inspection using the Integrative Genomics Viewer (IGV) was performed and experimental validation is being investigated. Interestingly, one of PAV candidates showed polymorphism in ‘Kiyomi’ and ‘Natsudaidai’ as well as other resistant and susceptible cultivars. Our results suggest a necessity for the detection of structural variation and indicate that the candidates may be useful for molecular breeding for citrus canker resistance and understanding disease resistance mechanism.