• Title/Summary/Keyword: Vector insects

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Transmission of Paulownia witches'-broom virus by tobacco leaf bug, Cyrtopeltis tenuis REUTER (담배장님노린재 (Cyrtopeltis tenuis REUTER)에 의한 오동나무천구 소병(도깨비집병) 바이러스의 매개전염)

  • La Y. J.;Pyun B. H.;Shim K. J.
    • Korean journal of applied entomology
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    • v.5_6
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    • pp.1-7
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    • 1968
  • The present investigation was initiated to find out the possible insect vector of the paulownia witches'-broom virus disease. 1. Among a number of piercing-sucking mouth type insects that were feeding on the witches'-broom infected Paulownia leaves, the tobacco leaf bug, Cyrtopeltis tenuis REUTER, was most abundant. Eggs, nymphs and adults of Cyrtopeltis tenuis were observed from the mid-August through the end of October. 2. It was found that Paulownia witches'-broom virus is transmitted by Cyrotopeltis tenuis. 3. On young Pnulownia plants, first symptom appeared twenty days after exposure to virus-bearing colonies of Cyrtopeltis tenuis. Forty days after exposure to these viruliferous insects, the characteristic symptom of Paulownia witches'-broom was apparent. 4. Paulownia witches' -broom virus was also transmitted by Cyrtopeltis tenuis to calendula and morning glory. No app"rent symptom was observed from tomato, carrot. aster, zinnia, cosmos, soybean and plantago at least until sixty days after exposure to the viruliferous colonies of Cyrtopeltis tenuis. 5. Calendula and morning glory appear to be highly suitable test plants for Paulownia witches'-broom vims because of their relatively short incubation period and distinct symptom appearance.

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Baculovirus Expression and Biochemical Characterization of the Bombyx mori Protein Disulfide Isomerase (bPDI)

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Park, Kwang-Ho;Hwang, Jae-Sam;Kwon, O-Yu;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.2
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    • pp.127-131
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    • 2003
  • Protein disulfide isomerase (PDI) found in the endoplasmic reticulum (ER) catalyzes disulfide bond exchange and assists in protein folding of newly synthesized proteins. PDI also functions as a molecular chaperone and has been found to be associated with proteins in the ER. In addition, PDI functions as a subunit of two more complex enzyme systems: the prolyl-4-hydroxylase and the triacylglycerol transfer proteins. A cDNA that encodes protein disulfide isomerase was previously isolated from Bombyx mori (bPDI), in which open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and an ER retention signal of the KDEL motif at its C-terminal, and we report its functional characterization here. This putative bPDI cDNA is expressed in insect Sf9 cells as a recombinant proteins using baculovirus expression vector system. The bPDI recombinant proteins are successfully recognized by antirat PDI antibody, and shown to be biologically active in vitro by mediating the oxidative refolding of reduced and scrambled RNase. This suggests that bPDI may play an important role in protein folding mechanism of insects.

Construction of shuttle vectors expressing the cry11Aa gene and their mosquitocidal activity (모기살충성 cry11Aa 유전자를 발현하는 벡터의 구축과 모기살충효과)

  • Lee, Dae-Weon;Kim, Ho-San;Je, Yeon-Ho;Kim, Joo-Up;Yu, Hyo-Sok;Kang, Seok-Kwon
    • The Korean Journal of Pesticide Science
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    • v.2 no.1
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    • pp.91-96
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    • 1998
  • For the expression of the cry11Aa gene highly toxic to dipteran insects, we constructed two cyanobacteria-Escherichia coli(E. coli) shuttle vectors, pCYASK5-l and pCYASK5-2. These vectors were transformed into E. coli and selected with kanamycin. The expression of the cry11Aa gene in E. coli was characterized by SDS-polyacrylamide gel electrophoresis and Western blot analysis. Two E. coli transformants harboring pCYASK5-1 and pCYASK5-2 expressed the cry11Aa gene in size of 72 kDa and 64 kDa, respectively and showed over 89% mortality against Culex pipiens larvae.

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Studies on the Dwarf Disease of the Mulberry Tree(Transmission by Grafting and Insects) (뽕나무 위축병에 관한 연구(접목전염 및 곤충전염))

  • 장병호;김종진
    • Journal of Sericultural and Entomological Science
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    • v.13 no.1
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    • pp.17-22
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    • 1971
  • A series of experiments were conducted from 1968 to 1970 to find the transmission effects of the dwarf disease of mulberry tree in connection with inlay grafting and insect vector with the following conclusions: 1. The inlay-grafting of sound scions on diseased roots showed a 30 per cent transmission rate among the successful grafts. The inlay-grafting of diseased scions on sound roots showed a 14 per cent transmission rate among the successful grafts ir the case of scions cut and stored before wintering but no transmission effect in the case of scions prepared after wintering, There were some stocks, among the unsuccesful grasfts of diseased scions upon sound roots, that sprouted out new buds, no transmission effects were found in them, 2. Leafhoppers (Hishimonus disciguttus Walker) collected from areas with no dwarf disease infection were grouped into three plots and let to feed on mulberry tree severely infected by dwarf disease for 7 days, 14 days and 21 days respectively. After these preliminary treatment, the leafhoppers were recollected to be fed on the sound seedling divided into three plots for 10 days, 15 days and 20 days respectively to find the transmission effects. The results are that the 7 day plot showed no transmission effect whereas the 14 day plot showed a 22 per cent disease rate and the 21 day plot 61 per cent rate indicating that the longer the feeding period is the higher the transmission rate.

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Experimental colonization of Culicoides arakawae in laboratory (Culicoides arakawae의 실험실내 colonization)

  • Choi, sang-ho;Joo, Hoo-don;Wee, Sung-hwan;Kim, Ki-seok;Park, Keun-sik
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.471-478
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    • 1993
  • Culicoides arakawae is a kind of the main blood sucking insects of domestic fowls and serves as a vector of Leukocytozoom caulleryi, the causative protozoon of chicken leukocytozoonosis. In this study, the complete life history of C arakawae was cycled by laboratory colonization. Adult midges were collected from various poultry farm by light trap. The laboratory colonization was performed under the conditions of constant temperature of $25{\pm}1^{\circ}C$ and relative humidity of 80% or above. The hatched larvae were cultured in larval medium consisted of rice field mud and activated charcoal powder. The surface of medium was continuously flowed with biologically conditioned water. The fine powder meal composed of pellet feed for mice and equal mount of yeast was supplied for feeding larvae at every 72 hours. The life cycle completed at $25^{\circ}C$ in 35~35 days ; the period of preoviposition, egg. larval and pupal stage was 2~3, 3~4, 28~30 and 3 days, respectively. The measurements of the eggs, the lst instar larvae, the 4th instar larvae and pupae was $36.28{\mu}m{\pm}1.95$, $13.58{\mu}m{\pm}0.72$, $4000{\mu}m{\pm}1.47$ and $219.95{\mu}m{\pm}6.25$ in $mean{\pm}S.D.$, respectively. In order to confirm experimental colonization of C arakawae in laboratory, the colonized adult midges were allowed to suck blood from chicken infected with L caulleryi. The oocysts and sporozoites could be identified in midguts and salivary grands of engorged midges at 72 hours after blood sucking.

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Pollination Mechanism of Bupleurum latissimum (Apiaceae) (멸종위기종인 섬시호(Bupleurum latissimum, 산형과)의 수분기작)

  • So, Soonku;Han, Kyeongsuk;Kim, Muyeol;Park, Hyerim;Seo, Eunkyoung;Kim, Yang-Pyo;Kim, Tae-Heung
    • Korean Journal of Plant Taxonomy
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    • v.38 no.1
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    • pp.43-50
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    • 2008
  • The pollination system of Bupleurum latissimum Nakai (Apiaceae) was investigated in the natural population of Korea. The various insects of 19 species, 11 families, 5 orders visited the flowers of B. latissimum. Episyrphus balteatus and Lasioglossum occidens were considered as the most effective pollen vector which have associated specially with B. latissimum. The visitation frequency peaked at 10 AM - 13 PM and no visitor was recognized during night time. The flowers of B. latissimum last during only three days and they are protandry. It is also confirmed that the flower of B. latissimum is self-compatible and cross-pollination by vectors is critical for successful seed setting.

Molecular Cloning and Characterization of Attacin from the Swallowtail Butterfly, Papilio xuthus

  • Kim, Seong-Ryul;Hwang, Jae-Sam;Park, Seung-Won;Goo, Tae-Won;Kim, Ik-Soo;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.2
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    • pp.231-238
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    • 2011
  • Attacin is an insect antibacterial protein that plays an important role in immune response to injury and infection. In this report, we have isolated and characterized of cDNA encoding for the attacin from the immunized larvae of swallowtail butterfly, $Papilio$ $xuthus$. A full length cDNA of $P.$ $xuthus$ attacin was obtained by employing annealing control primer (ACP)-based differential display PCR and 5' RACE. The complete $P.$ $xuthus$ attacin cDNA was comprised of 949 bp encoding a 250 amino acid precursor. It contains a putative 18 amino acid signal peptide sequence, a 42 amino acid propeptide sequence, and a 190 amino acid mature protein with a theoretical molecular mass of 19904.01 and a pI of 9.13. The putative mature protein of $P.$ $xuthus$ attacin showed 48-52% and 24-30% identity in amino acid sequences with that of lepidopteran and dipteran insects, respectively. Semiquantitive RT-PCR results revealed that the transcript of $P.$ $xuthus$ attacin gene was up-regulated at significant levels after injection with bacterial lipopolysaccharide (LPS). We sub-cloned cDNA fragment encoding mature $P.$ $xuthus$ attacin into the expression vector, highly expressed in $E.$ $coli$ BL21 cells, and its antibacterial activity was analyzed. Recombinant $P.$ $xuthus$ attacin evidenced considerably antibacterial activity against Gram-negative bacteria, $E.$ $coli$ ML 35 and $Klebsiella$ $pneumonia$.

Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

RNA Interference of Chitinase Gene in Spodoptera litura (담배거세미나방(Spodoptera litura) Chitinase gene의 RNA interference)

  • Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.202-209
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    • 2014
  • RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

Dual infections of Tomato mosaic virus (ToMV) and Tomato yellow leaf curl virus (TYLCV), or Tomato mosaic virus (ToMV) and Tomato chlorosis virus (ToCV), detected in tomato fields located in Chungcheongnam-do in 2017

  • Choi, Go-Woon;Kim, Boram;Ju, Hyekyoung;Cho, Sangwon;Seo, Eunyoung;Kim, Jungkyu;Park, Jongseok;Hammond, John;Lim, Hyoun-Sub
    • Korean Journal of Agricultural Science
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    • v.45 no.1
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    • pp.38-42
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    • 2018
  • Demand for tomatoes has been increasing every year as people desire more healthy food. In Korea, tomatoes are mainly grown in the Chungnam, Chunnam and Kyungnam provinces. Recently, reports of whitefly-transmitted viral diseases have increased due to newly emerging whitefly pressures caused by climate change in Korea. Specifically, in 2017, the main tomato growing areas, Buyeo and Nonsan in Chungnam, showed damage typical of viral infection; therefore, we investigated viral diseases in these areas. We collected samples with virus-like symptoms and found that not only whitefly transmitted Tomato yellow leaf curl virus (TYLCV) and Tomato chlorosis virus (ToCV) were detected but also Tomato mosaic virus (ToMV, for which no specific vector is known) and Tomato spotted wilt virus (TSWV, transmitted by thrips). The ToMV-infected samples were mostly co-infected with either TYLCV or ToCV. Mixed infections of different combinations of TYLCV, ToCV and ToMV were detected with the mixed infection of two whitefly-transmitted viruses (TYLCV and ToCV) causing the most severe symptoms. According to the CP sequence of each virus, the 100% identities were shown to be Mexico/ABG73017.1 (TYLCV), Greece/CDG34553.1 (ToCV), China/AKN79752 (TSWV), and Australia/NP078449.1 (ToMV). Based on the sequence data, we presumed that these tomato infecting viruses were transmitted through insects and seeds introduced from neighboring countries.