시설상추에서 문제가 되고 있는 해충에는 싸리수염진딧물, 꽃노랑총채벌레, 검은은무늬밤나방 등이 알려져 있다. 본 연구는 토양 속 꽃노랑총채벌레의 번데기를 포식하는 총채가시응애(Hypoaspis aculeifer)와 상추 잎에 서식하는 꽃노랑총채벌레의 약충과 성충을 포식하는 미끌애꽃노린재(Orius laevigatus)를 이용하여 보다 효율적으로 꽃노랑총채벌레를 방제하기 위하여 수행되었다. 꽃노랑총채벌레의 피해가 증가하는 7월~8월에 정식하는 시설상추에서 꽃노랑총채벌레의 발생초기에 총채가시응애는 1회 방사하고, 미끌애꽃노린재는 방사량, 방사횟수를 달리하여 방제효과를 조사한 결과, 7월 하순~8월 초순 중 정식하는 3작기 시설상추에서 꽃노랑총채벌레가 상추 100엽당 1~10마리 사이에 발생할 때 총채가시응애를 m2당 15.2마리(15,000마리/300평/10a) 밀도로 1회 방사한 후 예찰을 통하여 꽃노랑총채벌레가 100엽당 15~25마리 사이에 발생하면 미끌애꽃노린재를 m2당 0.8마리(750마리/300평/10a) 밀도로 15~30일 사이의 간격을 두고 2회 방사하면 90% 이상의 밀도감소 효과가 있었다. 그리고 꽃노랑총채벌레가 100엽당 25마리 이상 발생하였을 때는 총채가시응애를 ㎡당 15.2마리 밀도로 1회 방사하고 미끌애꽃노린재는 m2당 0.8마리 밀도가 되게하여 15~30일 사이의 간격을 두고 3회 방사하거나, 미끌애꽃노린재 밀도를 2배 증가시켜 m2당 1.6마리 밀도로 15~30일 사이의 간격을 두고 2회 방사하면 80% 이상의 효과를 보였다.
The toxicity of methanol extract from 28 plants toward four agricultural insect pests were examined using contact toxicity bioassay with spray methods. At a concentration of 1,000 ppm Achyranthes japonica whole body was observed strong activity against Myzus persicae. Paeonia lactiflora root and Corydalis turtschaninovii rhizome methanol extract were observed strong activity toward Tetranychus urticae. Catalpa ovata leaf and Ginkgo biloba leaf and stem methanol extracts were revealed high activity toward Nilaparvata lugens. Actium lap root methanol extract was observed moderate activity against Plutella xylostella. High or moderate insecticidal activity of plant extracts were listed in Fig.1. These plant resource, particularly plant methanol extract or insecticidal constituents, merit further as potential insect pest control agents or leads because of their great activity as a insecticide.
Insect pest monitoring program in subsistence warehouses can detect pest problems in early infestation stages. Temporal and spatial distributions of pests produced by monitoring allows facility managers and pest control operators to visualize pest problems and evaluate effectiveness of cleaning, sanitation and pest management practices. Pheromone-based insect monitoring traps were placed to study temporal and spatial distribution of Lepidopteran and Coleopteran stored product pests in two central distribution centers of United States Military Forces in the Republic of Korea, from March to December 2008. The two distribution centers receive store and issue subsistence and they are located in two different cities, Seoul and Taegu. As a result, three pests, Plodia inter punctella Hübner (Indian meal moth), Lasioderma serricorne F. (cigarette beetle), and Trogoderma variabile Ballion (warehouse beetle), were detected on the traps. P. interpunctella was most abundant, followed by L. serricorne and T. variabile. Different phenologies and spatial distribution of each species were seen from the two warehouses, that were related with internal temperature of the two warehouses and food availability. The program benefited to lead a better understandings for existence and extent of local pest problems in the two subsistence distribution centers, and also helped to find effective pest management periods and locations.
A genome project has been launched and aims to sequence total genome of Cotesia plutellae bracovirus. On this process, several open reading frames (ORFs) have been identified. This study was intended to clone and express protein tyrosine phosphatase genes, PTP1 and PTP6. The ORFs of these two genes consist of 900 and 891 bp, respectively. PTP1 and PTP6 are genes of a group of genes that has been implicated as important regulatory components in cell growth, differentiation and malignant transformation by certain viruses. In this work,we studied the cloning and expression patterns of these genes in Plutella xylostella, a lepidopteran host of C. plutellae. A polymerase chain reaction (PCR) produced the corresponding products of PTP1/6. These PCR products were cloned and expressed using an expression vector pBAD-TOPO, and then over-expressed using an inducer, L-arabinose. Then the purified proteins were confirmed using Western blotting (immunoblotting using V5 antibody) and the apparent molecular weights of both proteins were about 40 kDa. Expression of PTP genes were analyzed in the parasitized P. xylostella by realtime RT-PCR, which indicated late expression pattern of both PTPs during parasitization. We are pursuing to elucidate their physiological function in the parasitized host insect.
Accurate estimation of pest density is a prerequisite in achieving efficient pest management. An automatic pest detection system with image processing was installed on a robot to recognize brown marmorated stink bug (Halyomorphahalys) on leaves of paprika(Capsicumannuumvar.angulosum). The shape of pest was recognized and subsequently the robot arm was moved toward the leaves to spray pesticides. The detection system was efficient along with increasing population densities increased. The robot with image processing system was useful for estimating population densities in spatial and temporal domain efficiently.
The hemipteran whitefly Bemisia tabaci (Gennadius) is one of the most destructive pests damaging more than 600 agricultural crop species worldwide. The B and Q biotypes are most widely spread in Korea but they are not distinguishable based on morphological characters. Carboxylesterase 2 (Coe2) was determined to be 5.9 times more expressed in B biotype compared to Q biotype. Comparison of deduced amino acid sequences of Coe2 (595 a.a.) showed a total of 3.85% polymorphisms between B and Q types but no differences in major active sites. Quantitative real-time PCR revealed that both B and Q biotypes possess a single copy of coe2, suggesting that the overexpression of Coe2 in B biotype is likely due to overtranscription. To determine the putative role of Coe2 in insecticide tolerance, esterases were separated by native isoelectric focusing (IEF) and inhibited by various insecticides. The putative Coe2 band was apparently inhibited by pyrethroid and organophosphate insecticides, but not by imidacloprid. These findings suggest that overexpression of Coe2 confers chemical defense against pyrethroid and organophosphate insecticides, perhaps by sequestration.
Origanum essential oil [Origanum acutidens (Hand.-Mazz.)] showing insecticidal activity and repellency against red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae) adults was analysed by GC-MS. All constituents of the oil were identified, and the main components were carvacrol (67.2%), p-cymene (16.2%), γ-terpinene (5.5%), thymol (4.9%),and linalool(2.1%). In vapor phase fumigant assay, the origanum oil was more effective in closed condition (LD50 = 0.555 mg/cm3) than in open one (LD50 = 0.353 mg/cm3).This result suggests that the toxicity of the oil to red flour beetle is exerted largely by vapor phase. Based on 24-h LD50 values, the toxicity of caryophyllene oxide (0.00018 mg/cm3) was comparable with that of dichlorvos (0.00007mg/cm3) as a positive insecticide. In addition, thymol, camphene, α-pinene, p-cymene, and γ-terpinene gave good insecticidal activity (LD50 = 0.012 - 0.195 mg/cm3). In repellent test using 9 constituents of origanum oil, caryophyllene oxide showed 100% repellent activity at 0.0158 and 0.0032 mg/cm2. These results indicate that the origanum oil and its some components could be potential candidates as a fumigant and repellent for managing T. castaneum adults. Potential molecular target of repellents will be discussed.
The diamondback moth, Plutella xylostella, is one of the most important pests of cole crops in the world and is the first insect to evolve resistance to Bt toxins in open-field populations. To search for useful molecular markers for Bt reistance monitoring, the PCR-based restriction fragment length polymorphism (RFLP) profiles of three aminopeptidase N (PxAPN1, PxAPN2 and PxAPN4) were determined for 15 representative regional field populations of P. xylostella. Most regional samples had similar RFLP patterns, whereas PxAPN1 from four regions and PxAPN4 from two regions showed different banding patterns after restriction enzyme treatment, but no differences were found in PxAPN2 among populations. The DNA sequence analysis revealed that a point mutation at the restriction site was responsible for the polymorphism of PxAPN1 but no mutations were observed in PxAPN4. Comparing amino acid sequences of PxAPNs from regional populations with reference PxAPNs (GenBank accession no. AAB70755) revealed that four regional populations possessed a point mutation in the Cry1A binding site of PxAPN1 and five regional populations possessed a deletion of eight amino acids in PxAPN4. These RFLP patterns were consistently observed in Southern regions of Korea, including Kyungsangnam-Do and Jeju-Do. The functional association of these RFLP with Bt resistance is currently under investigation
A new Bacillus thuringiensis isolate 19-22 (Bt 19-22) exhibited high anti-fungal activity against barley powdery mildew (Blumeria graminis f. sp. hordei). The cry gene content of Bt 19-22 comprised cry1Aa, cry1Ab, cry1Ac and cry1D which have high insecticidal activity against lepidopteran larvae. We tried to confer a dipteran insecticidal activity to Bt 19-22 for constructing a recombinant strain which has multiple functions, anti-fungal and dual insecticidal activity. The insecticidal cry11Aa gene of B. thuringiensis was constructed under cry1Ac promoter in an E. coli-B. thuringiensis shuttle vector (pPro11A). The plasmid, pPro11A was introduced into Bt 19-22 isolate by electroporation and four transformants which had different cry gene contents were identified by PCR with cry11Aa and cry1-type specific primers. Among them, a Bt 19-22 transformant (11A/19-22 No. 7) expressed Cry11A protein (approximately 70 kDa) successfully without change of its inherent characteristics such as Cry protein expression and antifungal activity. The insecticidal activity of 11A/19-22 No. 7 was checked against Plutella xylostella and Culex pipiens. These results suggests that the recombinant strain shows dual insecticidal activity against lepidopteran and dipteran larvae as well as antifungal activity.
Through an application of plasmid capture system (PCS) to Bacillus thuringiensis plasmid DNAs, we acquired 21 polymorphic clones of putative genomic DNA of bacteriophage. The genome size of phage 1-3 (PhBT1-3) was determined to be 46,517 base pairs (bp) with 35.43% G + C content and 83% coding region. Sixty-five putative open reading frames (ORFs) with more than 50 codons were found in the new phage genome. In accordance with this genome finding, the phage particles and its DNA were confirmed from the supernatant of B. thuringiensis 1-3. Morphological characterization and infectivity assay demonstrated that PhBT1-3 belongs to the family Siphoviridae and it showed infectivity to three B. thuringiensis type strains, galleriae, entomocidus, and morrisoni. Based on these results, we screened the existence of phages in B. thuringiensis type strains by PCR with terminase small subunit-specific primers. Ten of 67 type strains showed PCR products and the similarity of those putative amino acids was more than 70%. Furthermore, we verified the existence of various shaped phages from the supernatants of 10 B. thuringiensis type cultures. In conclusion, we characterized a putative genome of phage, PhBT1-3 from B. thuringiensis 1-3, and confirmed the distribution of phages in the group of 67 B. thuringiensis type strains.
Autographa californica nucleopolyhedrovirus (AcMNPV) has a large doublestrand DNA genome of approximately 134 kbp and comprises 156 open reading frames (ORFs). To elucidate DNA replication cascade of AcMNPV, we developed a novel baculovirus genome that can be maintained in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. This genome, named bAc-MK, contains a mini-F replicon and a kanamycin resistance marker. Using a convenient Tn7 transposon-based system, pPCS-S, which contains an ampicillin resistance gene, 56 single ORF-truncated mutants were generated by random insertion into bAc-MK genome. These single ORF-truncated mutants were independently transfected into Sf9 cells to verify viral replication. Interestingly, both lef-1 and p48 knockout mutants showed normal viral replication in infected cells, which are reported to essential for viral replication. These results suggest that these single ORF-truncated mutants are useful for elucidation of viral replication cascade.
Baculovirus chitinase gene (ChiA) is a late gene and is essential for liquefying host insect at the late stage of infection for its hydrolyzing chitin function. In previous report, baculovirus ChiA can offer many interseting new opportunities for pest control. Recently, a putative chitinase gene (ChiA) was identified in the Spodopter litura nucleopolyhedorvirus (SlMNPV-K1) genome. The open reading frame (ORF) contains 1,692 nucelotides (nt) and encodes a protein of 563 amino acids (aa) with a predicted molecular weight of 62.62 kDa. To conform the insecticidal activity of ChiA from SlMNPV-K1, we constructed a baculovirus transfer vector, pBac-SlChiA, and this transfer vector was co-transfected with the bApGOZA DNA into sf9 cell to generate corresponding recombinant viru which designed Ap-SlChiA. Western blot analysis indicate that SlMNPV-K1 ChiA was successfully expressed. We found the chitinase activity of recombinant virus was enhanced 53% than wide type AcMNPV by chitinase assay, and the recombinant virus showed higher evidently insecticidal activity against 3rd instar larvae of Spodotera exigua than wide type AcMNPV (4.5 time). These results suggested that the chitinase gene from SlMNPV-K1 could be successfully applied to improve pathogenicity of bauclovirus
Bacillus thuringiensis 1-3 (Bt 1-3) which was isolated from a Korean soil sample showed high insecticidal activity against Aedes aegypti as well as Plutella xylostella. The isolate was determined to belong to ssp. aizawai (H7) type by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins with a molecular weight of 130 kDa. PCR analysis with cry gene specific primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2A gene, differing from spp. aizawai (reference strain) which contains cry1Aa, cry1Ab, cry1C and cry1D. We modified the plasmid capture system (PCS) to clone plasmid from Bt 1-3 through in vitro transposition. Fifty-three clones were acquired and their sizes were approximately 10 kb. Based on the sequence analysis, they were classified according to similarities with four known Bt plasmids, pGI3, pBMB175, pGI1 and pGI2, respectively. One of pGI3-like clones, named as pBt1-3, was fully sequenced and its 20 putative open reading frames (ORFs), Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified. The structure of pBt1-3 showed high similarity with pGI3 which is one of rolling-circle replication (RCR) group VI family.
Entomopathogenic fungi are widely available as biological control agents for controlling insect pests in agriculture and forestry. The fungal culture broth contains various pathogenesis-related components such as blastospores, mycelium and insecticidal enzymes such as chitinase, Pr1- and Pr2-proteases, which have been reported to play an important role in penetrating insect cuticles. In this study, we tried to evaluate the utility of culture broth from Beauveria bassiana SFB-205 to control lepidopteran pests. High level of insecticidal activity correspond to over 90% of mortality were observed when the culture broth of B. bassiana SFB-205 was inoculated to the Spodoptera litura larvae together with the B. thuringiensis K1. The freeze-dried culture broth showed synergistic effects in insecticidal activity against larvae of S. exigua and S. litura when treated with corresponding baculoviruses, SeNPV and SlNPV. Active ingredient of the B. bassiana SFB-205 culture broth was identified to chitinase, which have truncated form by insertional mutation compared to previously reported chitinases.
Electroantennogram techniques (EAGs) were employed to record olfactory responses in the antennae of the adult female stable fly, Stomoxys calcitrans (Diptera: Muscidae), to Zanthoxylum piperitum pericarp steam distillate (ZP-SD), Z. armatum seed oil (ZA-SO) and their 29 volatile constituents alone as well as binary mixture of the ZP-SD and ZA-SO with 1-octen-3-ol. The electrophysiological responses of the test materials were compared with those of DEET and 1-octen-3-ol. At concentration of 10-1 (v/v) in mineral oil, ZP-SD, ZA-SO, and all volatiles elicited EAG responses in the fly antennae except for DEET, which is extremely low volatile. ZP-SD, ZA-SO, and some of the test volatiles elicited EAG responses rather lower than 1-octen-3-ol, which showed vapor phase repellency and toxicity to the fly in our previous behavioral research. This suggests that the stable fly possesses olfactory receptor neurons responding to given repellent compounds per se. In binary mixture with 1-octen-3-ol at concentration of 10-1, ZP-SD and ZA-SO elicited 16±0.55 and 18±0.63 mV while ZP-SD, ZA-SO, and 1-octen-3-ol alone elicited responses of 14±0.45, 15.8±0.37 and 16.2±0.58 mV, respectively. Among the volatile compounds, terpinen- 4-ol, β-myrcene, α-phellanderene, citronellal, and limonene oxide elicited 80 – 96% relative EAG amplitude compared to 1-octen-3-ol as a reference (100%). Based on structure-activity relationships, constituents having aldehyde moiety elicited bigger EAG responses than those of alcohol or ether one. The perception in the fly antenna to plant volatile compounds exhibited complicated patterns of olfactory responses. Current and future directions of this study on sensory processing mechanisms underlying repellent behaviors will be discussed.
천적의 성공적인 정착과 산란력은 천적의 능력을 판단하는 중요한 기준이 다. 점박이응애를 방제하기 위해 사막이리응애를 사용함에 있어 천적과 해충사이의 적절한 밀도관리를 통한 해충 발생억제기작이 장기간 유지되는 것이 이상적이라 할 수 있다. 본 연구에서는 실험실에서 강낭콩과 장미, 딸기를 기주로 하여 IOBC quality control guideline에 따라 산란력 및 생존율을 조사하여 점박이응애의 생물적 방제인자로서 사막이리응애의 적용 가능성을 확인하고자 하였다. 온도 20, 25, 30℃, 상대습도 70%RH, 광조건 16L:8D에서 각 작물별 산란력을 조사한 결과, 사막이리응애는 20℃에서 산란수는 적고 생존율은 높았으며, 30℃에서 산란수는 증가고 생존율이 급속히 감소하여 산란력과 생존율 간에 반비례의 관계를 보였다. 작물별로는 온도에 관계없이 장미에서 산란수 및 생존율이 높았으며, 강낭콩과 딸기에서의 산란수 및 생존율은 비슷하였다. 따라서 연중 20~28℃의 온도 조건에서 재배되는 장미에 사막이리응애를 사용했을 때 좋은 방제효과를 얻을 수 있을 것으로 기대되며, 딸기의 경우에도 주간 17~20℃, 야간 10℃ 내외로 서늘하여 사막이리응애의 방사밀도를 조절하면 높은 방제 효과를 기대할 수 있을 것으로 생각된다.
최근 DDA, FTA 협상 등으로 인해 국가간 경제교류와 통합 등이 활발하게 진행되고 있으며 국제적으로 통용될 수 있는 우수농산물관리제도(Good Agricultural Practices, GAP)등의 농산물의 식품안전성 문제가 대두되고, 농산물을 안전하게 생산하는 것은 건강 유지뿐만 아니라 환경에 미치는 위해요소를 제거하여 생태계 보존 및 청결한 과원 관리가 절실하게 요구되고 있다. 오미자는 주로 해발 300m이상 되는 고랭지 지역에서 재배되고 있는 덩굴성 식물로 수명은 8-12년 정도 되는 우리나라 약용작물 중의 하나다. 특히 최근에 오미자의 맛과 효능이 소비자로부터 인정을 받으면서 재배면적이 전북 동부산악권 (무주, 진안, 장수, 남원, 순창)을 중심으로 날로 증가하고 있는 실정이다. 그러나 오미자 재배면적이 집단화 규모화 되면서 병해충의 발생양상도 다르게 나타나- 오미자에 대한 주요 병해충을 파악하여 방제의 기초자료 제공 및 병해충의 위험도를 평가하고자 이에 대한 실태를 조사 한 결과 오미자에 주로 발생하는 해충은 오미자 식나무깍지벌레가 피해가 많았으나 오미자 식재년수가 늘어나고 노후화 과원이 증가하면서 포도유리나방 및 신종 해충으로서 검은점애바구미 (Dendrobaris maculata(Roelofs, 1879))가 발생하였으며, 그밖에도 거위벌레, 노린재류, 자나방, 주머니나방, 열매를 가해하는 깍지벌레가 발생함을 알 수 있었다. 또한 해충이 아니었던 검거세미나방도 돌발해충으로 발생되어 많은 잎을 가해하기도 했다. 따라서 오미자 신종해충인 검은점애바구미의 사진자료와 피해증상을 보고하고, 오미자 줄기를 가해하는 주요 해충에 대한 정보를 공유하며, 병해충별 정확한 생리생태와 방제방법, 친환경 방제자재 선발 등을 연구함으로서 농가의 안정생산에 기여하고자 한다.
2009년 5월에 전라북도농업기술원 수경재배 비닐하우스에 가지를 정식하고 황색 등 8종의 색상지에 점착액을 도포하여 담배가루이 유인효과를 살펴보았으며, 트랩 점착액에 포도당, 과당, 올리고당, 물엿, 설탕 등을 첨가하여 당류 첨가에 따른 유인효과를 살펴보았다. 또한, 점착트랩을 지제부 30cm 위, 중간부, 정단부, 정단 30cm 위에 설치하고 담배가루이 유인 효과를 조사한 결과 색상지에 따른 담배가루이 유인효과는 황색이 100cm2 당 87.9마리로 가장 많았고, 연두>주황> 녹색>검정>적색>청색>백색순이었다. 점착액 당류 첨가효과는 포도당과 과당이 100cm2 당 각각 159.3, 159.0마리로 유인량이 가장 많았고, 올리고당>물엿>설탕 순이었으며, 설치위치에 따라서는 정단부가 100㎠ 당 103.3마리로 가장 많고, 지제부 30cm 위 지점이 85.0마리였으며, 정단부 30cm 위와 중간부는 각각 51.9마리, 55.4마리로 가장 적었다. 따라서 담배가루이 유인을 위한 점착트랩은 포도당 또는 과당을 첨가한 점착액을 황색지에 도포하고 정단부와 지제부 30cm 위 지점에 설치하는 것이 좋을 것으로 판단된다.
To control Thrips tabaci in Korean leek and green onion which grow at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control. In general treatment group, 120 mesh gauze was installed on the side window after planting(May, 2nd) and, in spring, Orius strigicollis (1.0/m2) was grazed 3 times every 7 days from mid of May which is early development stage and In the mid of June, which is the peak stage, plant extracts were sprayed one time. In autumn, O. strigicollis was grazed 3 times every 7 days from mid of September which is early development stage, and in the mid of October, which is the peak stage, plant extracts were sprayed one time. The result shows that the leaf damage ratio was decreased by 22% in Korean leek and by 27% in green onion compared to the untreatment. And the control value of Thrips tabaci shows 78.7% in korean leak and 90.6% in green onion. The density control effect of Thrips tabaci was significantly controlled under max 6.2/plant in general treatment group compared max 25/plant in the untreatment and this result was similar in green onion. The yields by general treatment was increased by 85% in Korean leek and 56% in green onion, compared with non-treatment yields, which was 900kg/10a in Korean leek and 1,287kg/10a in green onion.
To control pest of Brassicaceae leafy vegetable(leaf broccoli, Red Mustard Leaf, Tatsoi) which grows at vinyl house in IKSAN, Jeollabuk-do, the non-treatment was set as negative control, and treatment was divided into Parasitic natural treatment group and general treatment group. And incidence density of pest was surveyed and control effect was analyzed. As a result, in case of spring plants, the leaf damage ratio was decreased by 31% in Leaf broccoli, 30% in Red mustard leaf and 27% in Tatsoi compared to untreatment. In case of autumn plants, it was decreased by 32%, 41% and 17% respectively. The key pests were Plutella xylostella, Pieris rapae, Myus persicae Sulzer, Thrips palmi and Striped cabbage flea-beetle. Compared with the untreatment, the incidence density of each was significantly controlled. Other pests include Spodoptera exigua, Macdunnoughia purissima, Macdunnoughia purissima which showed high incidental density sometimes. In case of spring plants, the number by treatment was increased by 117% in Leaf broccoli, 85% in Red mustard leaf and 1,000% in Tatsoi. In autumn plants, it was increased by 132%, 257% and 1,077% respectively. The used Parasitic natural and eco-friendly materials were Cotesia glomerata, lacewing, ladyburg, Orius laevigatus, Encarsia formosa, Entomopathogenic nematode, Chungjigi and Togkaki. During early development of each pest, the Parasitic naturals were grazed 2~3 times at the interval of 7~10days. During the peak time, eco-friendly materials were sprayed 1~2 times. Based on this, comprehensive management model was drafted by period for each Brassicaceae vegetables pest.