DOCSLIB.ORG

Discovery of Hyperactive Antifreeze Protein from Phylogenetically Distant Beetles Questions Its Evolutionary Origin

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Discovery of Hyperactive Antifreeze Protein from Phylogenetically Distant Beetles Questions Its Evolutionary Origin International Journal of Molecular Sciences Article Discovery of Hyperactive Antifreeze Protein from Phylogenetically Distant Beetles Questions Its Evolutionary Origin Tatsuya Arai 1,2,† , Akari Yamauchi 3,† , Ai Miura 1, Hidemasa Kondo 1,3 , Yoshiyuki Nishimiya 1, Yuji C. Sasaki 2,4 and Sakae Tsuda 1,3,4,* 1 Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo 062-8517, Japan; [email protected] (T.A.); [email protected] (A.M.); [email protected] (H.K.); [email protected] (Y.N.) 2 Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8561, Japan; [email protected] 3 Graduate School of Life Sciences, Hokkaido University, Sapporo 060-0810, Japan; [email protected] 4 OPERANDO Open Innovation Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8563, Japan * Correspondence: [email protected]; Tel.: +81-11-857-8912 † T.A. and A.Y. contributed equally to this work. Abstract: Beetle hyperactive antifreeze protein (AFP) has a unique ability to maintain a supercooling state of its body fluids, however, less is known about its origination. Here, we found that a popular stag beetle Dorcus hopei binodulosus (Dhb) synthesizes at least 6 isoforms of hyperactive AFP (DhbAFP). Cold-acclimated Dhb larvae tolerated −5 ◦C chilled storage for 24 h and fully recovered after warming, Citation: Arai, T.; Yamauchi, A.; suggesting that DhbAFP facilitates overwintering of this beetle. A DhbAFP isoform (~10 kDa) Miura, A.; Kondo, H.; Nishimiya, Y.; appeared to consist of 6−8 tandem repeats of a 12-residue consensus sequence (TCTxSxNCxxAx), Sasaki, Y.C.; Tsuda, S. Discovery of which exhibited 3 ◦C of high freezing point depression and the ability of binding to an entire Hyperactive Antifreeze Protein from surface of a single ice crystal. Significantly, these properties as well as DNA sequences including Phylogenetically Distant Beetles Dhb Questions Its Evolutionary the untranslated region, signal peptide region, and an AFP-encoding region of are highly Origin. Int. J. Mol. Sci. 2021, 22, 3637. similar to those identified for a known hyperactive AFP (TmAFP) from the beetle Tenebrio molitor https://doi.org/10.3390/ijms22073637 (Tm). Progenitor of Dhb and Tm was branched off approximately 300 million years ago, so no known evolution mechanism hardly explains the retainment of the DNA sequence for such a long divergence Academic Editor: Masafumi Yohda period. Existence of unrevealed gene transfer mechanism will be hypothesized between these two phylogenetically distant beetles to acquire this type of hyperactive AFP. Received: 14 March 2021 Accepted: 28 March 2021 Keywords: antifreeze protein (AFP); thermal hysteresis (TH); stag beetle; ice binding; freeze avoid- Published: 31 March 2021 ance; supercooling; tandem repeat; evolutionary origin Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- 1. Introduction iations. Stag beetles include approximately 1200 species of insects belonging to the family Lucanidae, characterized by a pair of long horns, which resemble antlers of a stag, protrud- ing from the mandibles of the male (Figure1A) [ 1,2]. Stag beetles established immense popularity in Japan [3,4] as evidenced by cartoons, trading card games, fan websites, or Copyright: © 2021 by the authors. the Japanese Samurai helmet of the 1500s, which was decorated with an ornament called Licensee MDPI, Basel, Switzerland. “Kuwagata” designed after the stag beetle (Supplementary Material, Figure S1). Among This article is an open access article them, Dorcus hopei binodulosus (Dhb) (Figure1A,B) is especially popular because of its stout distributed under the terms and large body [5]. The Dhb beetle is found in northeastern China, Korea, and Japan that have conditions of the Creative Commons Attribution (CC BY) license (https:// an icy winter season, while it uniquely possesses a long-life overwintering nature; larvae creativecommons.org/licenses/by/ live for 1–2 years until pupation in a rotten tree or underground, and the adult lives for 4.0/). 3–5 years on the ground [2,5]. Because hyperactive antifreeze protein (AFP) discovered Int. J. Mol. Sci. 2021, 22, 3637. https://doi.org/10.3390/ijms22073637 https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, x FOR PEER REVIEW 2 of 15 Int. J. Mol. Sci. 2021, 22, 3637 2 of 15 ditions of the Creative Commons At- 3–5 years on the ground [2,5]. Because hyperactive antifreeze protein (AFP) discovered in tribution (CC BY) license (http://crea- fishes,in fishes, insects, insects, and and bacteria bacteria is is known known to to faci facilitatelitate theirtheir cold-survival cold-survival [6 ],[6], we we speculated speculated tivecommons.org/licenses/by/4.0/). whetherwhether the the DhbDhb beetlebeetle produces thisthis proteinprotein in in its its body body fluids. fluids. Since Since only only four four beetles, beetles, namelynamely TenebrioTenebrio molitor molitor, Dendroides canadensiscanadensis, ,Anatolica Anatolica polita polita, and, andMicrodera Microdera punctipennis punctipennis, , areare known known to to synthesize synthesize hyperactive AFPAFP composed composed of of a consensusa consensus sequence sequence [7– [7–10]10] and and allall of of them them belong belong to to the the same superfamily Tenebrionoidea Tenebrionoidea, less, less is is known known about about the the origins origins ofof this this protein, protein, while while the the Dhb beetle belongsbelongsto to a a different different superfamily superfamilyScarabaeoidea Scarabaeoidea[2]. [2]. Therefore,Therefore, if if DhbDhb containscontains hyperactive AFP,AFP,the the discovery discovery provides provides a firsta first clue clue to to under- under- standingstanding the the origin origin of of this this protein asas wellwell asas new new resources resources (i.e., (i.e., stag stag beetles) beetles) to exploreto explore moremore new new species species of of hyperactive AFP.AFP. FigureFigure 1. A 1. stagA stag beetle beetle DorcusDorcus hopei hopei binodulosus binodulosus (Dhb(Dhb) and) and a bursting a bursting ice-crystal ice-crystal growth growth observed observed for for its its hemolymph hemolymph at at the freezingthe freezing point. point.(A) An (A adult) An adultform of form Dhb of andDhb (Band) its ( Bfinal) its instar final instar larva. larva. The adult The adult is uniquely is uniquely equipped equipped with with a pair a pairof antlers of likeantlers a stag. like (C a) A stag. photomicroscope (C) A photomicroscope image of image a single of aice single crystal ice crystalin ordinary in ordinary supercooled supercooled water water(−0.1 ° (−C),0.1 which◦C), which shows a roundedshows adisk-like rounded morphology. disk-like morphology. (D,E) The ( Ddisk,E) Theexpanded disk expanded when the when temperature the temperature was held, was implying held, implying that there that thereis no isanti- freezeno antifreeze substance. substance. (F) A photomicroscope (F) A photomicroscope image imageof a single of a single ice crystal ice crystal prepared prepared in larval in larval hemolymph, hemolymph, which which kept kept un- ° changedunchanged even evenif the if temperature the temperature was waschanged changed between between −5.0− and5.0 and 0 C 0 because◦C because of the of theice-binding ice-binding of ofhyperactive hyperactive AFPs. AFPs. The hyperactiveThe hyperactive AFP-bound AFP-bound ice crystal ice crystal is uniquely is uniquely modified modified into into a lemon-like a lemon-like shape. shape. (G (G) )Bursting Bursting ice ice growth occurredoccurred from from a tip of the lemon crystal when it further lowered the temperature to freezing point (−5.1 °C). (H) The bursting ice growth a tip of the lemon crystal when it further lowered the temperature to freezing point (−5.1 ◦C). (H) The bursting ice growth continues to show a vein-like pattern, which is also a known phenomenon for hyperactive AFPs. continues to show a vein-like pattern, which is also a known phenomenon for hyperactive AFPs. AqueousAqueous solutions solutions thatthat remain remain in in liquid liquid phase phase by loweringby lowering the temperature the temperature below below the themelting melting point point are are called called supercooled supercooled solutions, solutions, in which in which numerous numerous “embryo” “embryo” ice crystals ice crys- talsare are present present dispersively dispersively [11 [11,12].,12]. When When the the temperature temperature is further is further lowered lowered and and thermal thermal movementmovement of of embryo embryo ice ice crystals decreases,decreases, they they aggregate aggregate rapidly rapidly to to form form a large a large cluster cluster toto nucleate nucleate numerous numerous ice ice particles, therebythereby changingchanging the the water water molecules molecules into into a multi-a multi- crystallinecrystalline state, state, which which is the general iceice crystalcrystal structure. structure. Note Note that that a substancea substance other other than than waterwater molecules molecules sometimes sometimes catalyzed thethe aggregationaggregation process process [11 [11].]. The The overwintering, overwintering, cold-adaptedcold-adapted insects insects have long beenbeen knownknown for for their their ability ability to to stabilize stabilize such such a supercooleda supercooled statestate of of their their body body fluids fluids at at subzero subzero temperatures,temperatures,
Load more

Recommended publications
  • Distribution and Functional Characterisation of Antifreeze Proteins in Polar Diatoms
    Living inside Sea Ice - Distribution and Functional Characterisation of Antifreeze Proteins in Polar Diatoms Dissertation zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften - Dr. rer. Nat. - am Fachbereich 2 (Biologie/Chemie) der Universit¨atBremen vorgelegt von Christiane Uhlig Bremen Oktober, 2011 1. Pr¨ufer: Prof. Kai Bischof 2. Pr¨ufer: Prof. Ulrich Bathmann F¨urmeine Mutter Monika Was Menschen in die Polargebiete trieb, war die Macht des Unbekannten ¨uber den men- schlichen Geist. Sie treibt uns zu den verborgenen Kr¨aftenund Geheimnissen der Natur, hinab in die unermesslich kleine mikroskopische Welt und desgleichen hinaus in die uner- forschten Weiten des Universums. Sie l¨asstuns keine Ruhe, bis wir den Planeten, auf dem wir leben, von der tiefsten Tiefen des Ozeans bis zu den h¨ochsten Schichten der Atmosph¨arekennen. Fridtjof Nansen Danksagung Ich danke Prof. Ulrich Bathmann und Prof. Kai Bischof f¨urdie Begutachtung dieser Arbeit. Prof. Rudolf Amann danke ich, dass er sich kurzfristig bereit erkl¨arthat die Aufgabe des 3. Pr¨uferzu ¨ubernehmen. Bei Prof. Kai Bischof und Andreas Krell bedanke ich mich f¨urdie Hilfe zur Einwerbung des Stipendiums, welches diese Arbeit ¨uberhaupt erst erm¨oglicht hat. Der Studienstiftung des deutschen Volkes danke ich f¨ur die finanzielle Unterst¨utzung. Ich bedanke mich ganz besonders bei der Meereis-Gruppe, die mehr ist als nur eine Ar- beitsgruppe. Besonders danke ich Gerhard Dieckmann, dass ich in Deiner Arbeitsgruppe meine Arbeit durchf¨uhrendurfte und f¨ur Deine Unterst¨utzungin allen m¨oglichen organ- isatorischen und pers¨onlichen Dingen. Klaus Valentin danke ich f¨urdie Unterst¨utzung, kreativen Titelvorschl¨ageund den Einsatz daf¨ur,dass ich noch eine Weile am AWI bleiben kann.
    [Show full text]
  • Lessons from Genome Skimming of Arthropod-Preserving Ethanol Benjamin Linard, P
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Archive Ouverte en Sciences de l'Information et de la Communication Lessons from genome skimming of arthropod-preserving ethanol Benjamin Linard, P. Arribas, C. Andújar, A. Crampton-Platt, A. P. Vogler To cite this version: Benjamin Linard, P. Arribas, C. Andújar, A. Crampton-Platt, A. P. Vogler. Lessons from genome skimming of arthropod-preserving ethanol. Molecular Ecology Resources, Wiley/Blackwell, 2016, 16 (6), pp.1365-1377. 10.1111/1755-0998.12539. hal-01636888 HAL Id: hal-01636888 https://hal.archives-ouvertes.fr/hal-01636888 Submitted on 17 Jan 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. 1 Lessons from genome skimming of arthropod-preserving 2 ethanol 3 Linard B.*1,4, Arribas P.*1,2,5, Andújar C.1,2, Crampton-Platt A.1,3, Vogler A.P. 1,2 4 5 1 Department of Life Sciences, Natural History Museum, Cromwell Road, London SW7 6 5BD, UK, 7 2 Department of Life Sciences, Imperial College London, Silwood Park Campus, Ascot 8 SL5 7PY, UK, 9 3 Department
    [Show full text]
  • Effects of Antifreeze Protein III on Sperm Cryopreservation of Pacific Abalone, Haliotis Discus Hannai
    International Journal of Molecular Sciences Article Effects of Antifreeze Protein III on Sperm Cryopreservation of Pacific Abalone, Haliotis discus hannai Shaharior Hossen 1 , Md. Rajib Sharker 1,2, Yusin Cho 1, Zahid Parvez Sukhan 1 and Kang Hee Kho 1,* 1 Department of Fisheries Science, College of Fisheries and Ocean Sciences, Chonnam National University, 50 Daehak-ro, Yeosu 59626, Jeonnam, Korea; [email protected] (S.H.); [email protected] (M.R.S.); [email protected] (Y.C.); [email protected] (Z.P.S.) 2 Department of Fisheries Biology and Genetics, Faculty of Fisheries, Patuakhali Science and Technology University, Patuakhali 8602, Bangladesh * Correspondence: [email protected]; Tel.: +82-616-597-168; Fax: +82-616-597-169 Abstract: Pacific abalone (Haliotis discus hannai) is a highly commercial seafood in Southeast Asia. The aim of the present study was to improve the sperm cryopreservation technique for this valuable species using an antifreeze protein III (AFPIII). Post-thaw sperm quality parameters including motility, acrosome integrity (AI), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), DNA integrity, fertility, hatchability, and mRNA abundance level of heat shock protein 90 (HSP90) were determined to ensure improvement of the cryopreservation technique. Post-thaw motility of sperm cryopreserved with AFPIII at 10 µg/mL combined with 8% dimethyl sulfoxide (DMSO) (61.3 ± 2.7%), 8% ethylene glycol (EG) (54.3 ± 3.3%), 6% propylene glycol (PG) (36.6 ± 2.6%), or 2% glycerol (GLY) (51.7 ± 3.0%) was significantly improved than that of sperm cryopreserved without AFPIII. Post-thaw motility of sperm cryopreserved with 2% MeOH and 1 µg/mL of AFPIII was also improved than that of sperm cryopreserved without AFPIII.
    [Show full text]
  • Iiillllllllli'lllll | US005627051A United States Patent [191 [11] Patent Number: 5,627,051 Duman [45] Date of Patent: May 6, 1997
    llilll|||l|l|||||ll||||||i|lllllllllllllllllllllllllllll.nIiillllllllli'lllll | US005627051A United States Patent [191 [11] Patent Number: 5,627,051 Duman [45] Date of Patent: May 6, 1997 [54] NUCLEIC ACID SEQUENCES ENCODING LG. Duman et al.. ‘Thermal hysteresis proteins”. Advances DENDROIDES ANTIFREEZE PROTEINS in Low Temperature Biology, vol. 2. pp. 131-182, (1993). [75] Inventor: John G. Duman. South Bend. Ind. D. Tursman et al., “Cryoprotective eifects of Thermal Hys teresis Protein on Survivorship of Frozen Gut Cells from the [73] Assignee: University of Notre Dame du Lac, Freeze Tolerant Centipede Lithobius for?catus”, Journal of Notre Dame, Ind. Experimental Zoology. vol. 272, pp. 249-257 (1995). D.W. Wu et al., “Activation of antifreeze proteins from [21] Appl. No.: 485,359 larvae of the beetle Dendroides canadensis”, Comp. Physiol. [22] Filed: Jun. 7, 1995 B, vol. 161, pp. 279-283 (1991). [51] Int. Cl.6 ........................... .. C12P 21/02; C07H 21/04 D.W. Wu et al., “Puri?cation and characterization of anti [52] US. Cl. .. 435/69.1; 536/235; 536/2431 freeze proteins from larvae of the beetle Dendroides [58] Field of Search .............................. .. 435/3201, 69.1; canadensis”, Comp. Physiol. B, vol. 161, pp. 271-278 536/235. 24.31 (1991). [56] References Cited Primary Examiner—Dian C. Jacobson PUBLICATIONS Assistant Examiner-Kawai Lau J.G. Duman et al., “Hemolymph proteins involved in insect Attorney, Agent, or Firm—Barnes & Thomburg subzero tolerance: Ice nucleators and antifreeze proteins”, In Insects at Low Temperatures, Chapman and Hall (N .Y.), pp. [57] ABSTRACT 94-127 (1991). The present invention is directed to nucleic acid sequences D.W.
    [Show full text]
  • A New Genus of Fire-Colored Beetles (Coleoptera: Pyrochroidae: Pyrochroinae) from the Sunda Shelf, with a Key to the Three Species Daniel K
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Center for Systematic Entomology, Gainesville, Insecta Mundi Florida 2014 Sundapyrochroa: A new genus of Fire-Colored Beetles (Coleoptera: Pyrochroidae: Pyrochroinae) from the Sunda Shelf, with a key to the three species Daniel K. Young University of Wisconsin - Madison Follow this and additional works at: http://digitalcommons.unl.edu/insectamundi Young, Daniel K., "Sundapyrochroa: A new genus of Fire-Colored Beetles (Coleoptera: Pyrochroidae: Pyrochroinae) from the Sunda Shelf, with a key to the three species" (2014). Insecta Mundi. 846. http://digitalcommons.unl.edu/insectamundi/846 This Article is brought to you for free and open access by the Center for Systematic Entomology, Gainesville, Florida at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Insecta Mundi by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. INSECTA MUNDI A Journal of World Insect Systematics 0341 Sundapyrochroa: A new genus of Fire-Colored Beetles (Coleoptera: Pyrochroidae: Pyrochroinae) from the Sunda Shelf, with a key to the three species Daniel K. Young Department of Entomology 445 Russell Laboratories University of Wisconsin Madison, Wisconsin 53706-1598 USA Date of Issue: January 31, 2014 CENTER FOR SYSTEMATIC ENTOMOLOGY, INC., Gainesville, FL Daniel K. Young Sundapyrochroa: A new genus of Fire-Colored Beetles (Coleoptera: Pyrochroidae: Pyrochroinae) from the Sunda Shelf, with a key to the three species Insecta Mundi 0341: 1-18 ZooBank Registered: urn:lsid:zoobank.org:pub:994D7E0D-D7DF-49A3-9A25-25EE0B3D7F22 Published in 2014 by Center for Systematic Entomology, Inc. P. O. Box 141874 Gainesville, FL 32614-1874 USA http://centerforsystematicentomology.org/ Insecta Mundi is a journal primarily devoted to insect systematics, but articles can be published on any non-marine arthropod.
    [Show full text]
  • Structure and Application of Antifreeze Proteins from Antarctic Bacteria Patricio A
    Muñoz et al. Microb Cell Fact (2017) 16:138 DOI 10.1186/s12934-017-0737-2 Microbial Cell Factories RESEARCH Open Access Structure and application of antifreeze proteins from Antarctic bacteria Patricio A. Muñoz1*, Sebastián L. Márquez1,2, Fernando D. González‑Nilo3, Valeria Márquez‑Miranda3 and Jenny M. Blamey1,2* Abstract Background: Antifreeze proteins (AFPs) production is a survival strategy of psychrophiles in ice. These proteins have potential in frozen food industry avoiding the damage in the structure of animal or vegetal foods. Moreover, there is not much information regarding the interaction of Antarctic bacterial AFPs with ice, and new determinations are needed to understand the behaviour of these proteins at the water/ice interface. Results: Diferent Antarctic places were screened for antifreeze activity and microorganisms were selected for the presence of thermal hysteresis in their crude extracts. Isolates GU1.7.1, GU3.1.1, and AFP5.1 showed higher thermal hysteresis and were characterized using a polyphasic approach. Studies using cucumber and zucchini samples showed cellular protection when samples were treated with partially purifed AFPs or a commercial AFP as was determined using toluidine blue O and neutral red staining. Additionally, genome analysis of these isolates revealed the presence of genes that encode for putative AFPs. Deduced amino acids sequences from GU3.1.1 (gu3A and gu3B) and AFP5.1 (afp5A) showed high similarity to reported AFPs which crystal structures are solved, allowing then generating homology models. Modelled proteins showed a triangular prism form similar to β-helix AFPs with a linear distribution of threonine residues at one side of the prism that could correspond to the putative ice binding side.
    [Show full text]
  • Replacement of the Antifreeze-Like Domain of Human N-Acetylneuraminic Acid Phosphate Synthase with the Mouse Antifreeze-Like
    Replacement of the antifreeze-like domain of human N-acetylneuraminic acid phosphate synthase with the mouse antifreeze-like domain impacts both N-acetylneuraminic acid 9-phosphate synthase and 2-keto-3-deoxy-D-glycero-D- galacto-nonulosonic acid 9-phosphate synthase activities Marshall Louis Reaves1,2, Linda Carolyn Lopez1 & Sasha Milcheva Daskalova1,* 1The Biodesign Institute, Arizona State University, Tempe Arizona, 85287, 2Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA Human NeuNAc-9-P synthase is a two-domain protein with In prokaryotes, the final stage of formation of N-ace- ability to synthesize both NeuNAc-9-P and KDN-9-P. Its tyl-D-neuraminic acid (NeuNAc) the most common sialic mouse counterpart differs by only 20 out of 359 amino acids acid in nature involves condensation of N-Acetyl-D-mannos- but does not produce KDN-9-P. By replacing the AFL domain amine (ManNAc) with phosphoenol pyruvate (PEP). The proc- of the human NeuNAc-9-P synthase which accommodates 12 ess, when irreversible, is catalyzed by the enzyme NeuNAc of these differences, with the mouse AFL domain we examined synthase (E.C. 4.1.3.19). The first bacterial gene, neuB gene, its importance for the secondary KDN-9-P synthetic activity. coding for the E. coli enzyme was identified in 1995 (2) and The chimeric protein retained almost half of the ability of the this later allowed overexpression and purification of sufficient human enzyme for KDN-9-P synthesis while the NeuNAc-9-P amount of protein for detailed characterization (3). Using the production was reduced to less than 10%.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2014/0274885 A1 Cong Et Al
    US 20140274885A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0274885 A1 Cong et al. (43) Pub. Date: Sep. 18, 2014 (54) PHI-4 POLYPEPTIDES AND METHODS FOR (52) U.S. Cl. THEIR USE CPC .............. A0IN 43/40 (2013.01); C07K 14/195 (71) Applicant: PIONEER HI-BRED (2013.01) INTERNATIONAL, INC., Johnston, IA USPC ............................ 514/4.5; 530/350; 536/23.7 (US) (57) ABSTRACT (72) Inventors: Ruth Cong, Palo Alto, CA (US); Jingtong Hou, San Pablo, CA (US); Compositions and methods for controlling pests are provided. Zhenglin Hou, Ankeny, IA (US); Phillip The methods involve transforming organisms with a nucleic Patten, Menlo Park, CA (US): Takashi acid sequence encoding an insecticidal protein. In particular, Yamamoto, Dublin, CA (US) the nucleic acid sequences are useful for preparing plants and (73) Assignee: PIONEER HI-BRED microorganisms that possess insecticidal activity. Thus, INTERNATIONAL, INC, Johnston, IA transformed bacteria, plants, plant cells, plant tissues and (US) seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use (21) Appl. No.: 13/839,702 in the construction of expression vectors for Subsequent trans (22) Filed: Mar 15, 2013 formation into organisms of interest, as probes for the isola tion of other homologous (or partially homologous) genes. Publication Classification The insecticidal proteins find use in controlling, inhibiting (51) Int. Cl. growth or killing lepidopteran, coleopteran, dipteran, fungal, AOIN 43/40 (2006.01) hemipteran, and nematode pest populations and for produc C07K I4/95 (2006.01) ing compositions with insecticidal activity. Patent Application Publication Sep.
    [Show full text]
  • Development and Characterization of Eleven Microsatellite Markers for A
    Int. J. Indust. Entomol. 35(2) 97-99 (2017) IJIE ISSN 1598-3579, http://dx.doi.org/10.7852/ijie.2017.35.2.97 Development and characterization of eleven microsatellite markers for a popular pet stag beetle, Dorcus hopei (Coleoptera, Lucanidae) using paired-end Illumina shotgun sequencing Taeman Han, Seung-Hyun Kim, In Gyun Park, and Haechul Park* Applied Entomology Division, Department of Agricultural Biology, National Institute of Agricultural Science, Nongsaengmyeong-ro 166, Iseo-myeon, Wanju-gun, Jeollabuk-do, 55365, Republic of KOREA Abstract Eleven polymorphic microsatellite loci were developed and characterized for Dorcus hopei Received : 12 Sep 2017 in this study. The number of alleles varied from 2 to 21. The observed heterozygosity Revised : 29 Sep 2017 and expected heterozygosity ranged from 0.1058 to 0.9744 and 0.0997 to 0.8941, Accepted : 17 Oct 2017 respectively. Two loci showed low polymorphism, while the rest were highly polymorphic. Keywords: Six loci deviated from Hardy-Weinberg Equilibrium. The set of markers will provide Dorcus hopei, effective tools for examining the population genetic structures and be helpful for managing Lucanidae, wild population in D. hopei. microsatellite, © 2017 The Korean Society of Sericultural Sciences genetic diversity, Int. J. Indust. Entomol. 35(2), 97-99 (2017) Korea Introduction Materials and Methods A stag beetle, Dorcus hopei belonging to the family Genomic DNA (gDNA) was extracted from thorax muscle Lucanidae is a famous insect pet and widely reared by many tissue of a wild male (voucher no. 8421, collecting date: 9th April insect fanciers between Korea and Japan. In Japan, to increase 2013) using a DNeasy Blood and Tissue kit (QIAGEN).
    [Show full text]
  • Romanian Species of Lucanids (Coleoptera: Scarabaeoidea: Lucanidae) in the Collections of “Grigore Antipa” National Museum of Natural History MELANIA STAN
    Travaux du Muséum National d’Histoire Naturelle © 30 décembre «Grigore Antipa» Vol. LVI (2) pp. 173–184 2013 DOI: 10.2478/travmu-2013-0013 ROMANIAN SPECIES OF LUCANIDS (COLEOPTERA: SCARABAEOIDEA: LUCANIDAE) IN THE COLLECTIONS OF “GRIGORE ANTIPA” NATIONAL MUSEUM OF NATURAL HISTORY MELANIA STAN Abstract. The seven species of stag beetles of the Romanian fauna are present in the coleopteran collection of the Museum: Aesalus scarabaeoides scarabaeoides (Panzer), Ceruchus chrysomelinus (Hochenwarth), Sinodendron cylindricum (Linnaeus), Lucanus cervus cervus (Linnaeus), Platycerus caraboides caraboides (Linnaeus), Platycerus caprea (De Geer) and Dorcus parallelipipedus (Linnaeus). Information on the collecting data and distribution maps are given for each species. We present the male and female habitus for the two species of Platycerus. Résumé. Les sept espèces de lucanes de la faune de Roumanie sont présentes dans la collection des coléoptères du muséum: Aesalus scarabaeoides scarabaeoides (Panzer), Ceruchus chrysomelinus (Hochenwarth), Sinodendron cylindricum (Linnaeus), Lucanus cervus cervus (Linnaeus), Platycerus caraboides caraboides (Linnaeus), Platycerus caprea (De Geer) et Dorcus parallelipipedus (Linnaeus). On donne des informations sur les données de la capture et les cartes de distribution pour chaque espèce. Nous présentons les photos de l’habitus mâle et femelle pour les deux espèces de Platycerus. Key words: Coleoptera, Lucanidae, Romania, collections, “Grigore Antipa” National Museum of Natural History. INTRODUCTION From the 17 stag beetle species and subspecies of Europe, in the Romanian fauna there are only seven species: Aesalus scarabaeoides scarabaeoides (Panzer), Ceruchus chrysomelinus (Hochenwarth), Sinodendron cylindricum (Linnaeus), Lucanus cervus cervus (Linnaeus), Platycerus caraboides caraboides (Linnaeus), Platycerus caprea (De Geer) and Dorcus parallelipipedus (Linnaeus), included in four subfamilies, according to the Catalogue of the Palaearctic Coleoptera (Bartolozzi & Sprecher-Uebersax, 2006).
    [Show full text]
  • Integrating Cultural Tactics Into the Management of Bark Beetle and Reforestation Pests1
    DA United States US Department of Proceedings --z:;;-;;; Agriculture Forest Service Integrating Cultural Tactics into Northeastern Forest Experiment Station the Management of Bark Beetle General Technical Report NE-236 and Reforestation Pests Edited by: Forest Health Technology Enterprise Team J.C. Gregoire A.M. Liebhold F.M. Stephen K.R. Day S.M.Salom Vallombrosa, Italy September 1-3, 1996 Most of the papers in this publication were submitted electronically and were edited to achieve a uniform format and type face. Each contributor is responsible for the accuracy and content of his or her own paper. Statements of the contributors from outside the U.S. Department of Agriculture may not necessarily reflect the policy of the Department. Some participants did not submit papers so they have not been included. The use of trade, firm, or corporation names in this publication is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the U.S. Department of Agriculture or the Forest Service of any product or service to the exclusion of others that may be suitable. Remarks about pesticides appear in some technical papers contained in these proceedings. Publication of these statements does not constitute endorsement or recommendation of them by the conference sponsors, nor does it imply that uses discussed have been registered. Use of most pesticides is regulated by State and Federal Law. Applicable regulations must be obtained from the appropriate regulatory agencies. CAUTION: Pesticides can be injurious to humans, domestic animals, desirable plants, and fish and other wildlife - if they are not handled and applied properly.
    [Show full text]
  • Enhancement of Insect Antifreeze Protein Activity by Solutes of Low Molecular Mass
    The Journal of Experimental Biology 201, 2243–2251 (1998) 2243 Printed in Great Britain © The Company of Biologists Limited 1998 JEB1557 ENHANCEMENT OF INSECT ANTIFREEZE PROTEIN ACTIVITY BY SOLUTES OF LOW MOLECULAR MASS NING LI, CATHY A. ANDORFER AND JOHN G. DUMAN* Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA *Author for correspondence (e-mail: [email protected]) Accepted 20 May; published on WWW 14 July 1998 Summary Antifreeze proteins (AFPs) lower the non-equilibrium Glycerol is the only one of these enhancing solutes that is freezing point of water (in the presence of ice) below the known to be present at these concentrations in melting point, thereby producing a difference between the overwintering D. canadensis, and therefore the freezing and melting points that has been termed thermal physiological significance of most of these enhancers is hysteresis. In general, the magnitude of the thermal unknown. The mechanism(s) of this enhancement is also hysteresis depends upon the specific activity and unknown. concentration of the AFP. This study describes several low- The AFP used in this study (DAFP-4) is nearly identical molecular-mass solutes that enhance the thermal hysteresis to previously described D. canadensis AFPs. The mature activity of an AFP from overwintering larvae of the beetle protein consists of 71 amino acid residues arranged in six 12- Dendroides canadensis. The most active of these is citrate, or 13-mer repeats with a consensus sequence consisting of which increases the thermal hysteresis nearly sixfold from Cys-Thr-X3-Ser-X5-X6-Cys-X8-X9-Ala-X11-Thr-X13, where 1.2 °C in its absence to 6.8 °C.
    [Show full text]