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9:00-9:30-10:00 10:00-11:00 Lee Bok Luel Host different responses toward pathogenic and gut symbiotic bacteria in insects 11:00-11:30 11:30-12:00 12:00-13:30 13:30-14:30 14:30-15:00 PGRP 15:00-15:30 15:30-15:50 15:50-16:20 3
16:20-16:50 Wolbachia 16:50-17:30 17:30-20 http://staff.aist.go.jp/t-fukatsu/brainhome.html http://staff.aist.go.jp/t-fukatsu/brainopensymp.html http://www.naro.affrc.go.jp/brain/index.html http://www.a.u-tokyo.ac.jp/yayoi/index.html 4
20 (1) (2) EST (3) (4) (5) http://staff.aist.go.jp/t-fukatsu/brainhome.html 2012 4 24 http://www.aist.go.jp/aist_j/press_release/pr2012/pr20120424/pr20120424.html Kikuchi Y., Hayatsu M., Hosokawa T., Nagayama A., Tago K., Fukatsu T. (2012) Symbiont-mediated insecticide resistance. Proc Natl Acad Sci USA 109: 8618-8622. 2009 12 22 http://www.aist.go.jp/aist_j/press_release/pr2009/pr20091222/pr20091222.html Hosokawa T., Koga R., Kikuchi Y., Meng X.-Y., Fukatsu T. (2010) Wolbachia as a bacteriocyte-associated nutritional mutualist. Proc Natl Acad Sci USA 107: 769-774. 5
基 調 講 演 1 Host different responses toward pathogenic and gut symbiotic bacteria in insects Lee Bok Luel Global Research Laboratory of Insect Symbiosis, College of Pharmacy Pusan National University, Jangjeon Dong, Kumjeong Ku, Busan 609-735, Korea E-mail address: brlee@pusan.ac.kr Due to the absence of acquired immunity in invertebrate, insects are good model systems to study the molecular mechanisms of innate immunity. During last 15 years, we have performed biochemical studies to elucidate how holometabolous insects recognize invading pathogenic bacteria and what kinds of molecules are involved in the recognition and activation of insect s innate immune responses. Using coleopteran insect, Tenebrio molitor, we determined the detailed molecular mechanisms of recognition and activation of Toll signaling cascade, which is a typical innate immune response for the elimination of pathogenic bacteria from host. Recently, our laboratory has been interested in symbiosis where symbiotic bacteria escape from host innate immunity, and especially in the gut symbiotic bacteria harboring in hemimetabolous insect, the bean bug Riptortus pedestris. This insect acquires a specific bacterium, Burkholderia, as a gut symbiont from the environment every generation. Because the biology of Burkholderia-bean bug system was well studied by Drs. Fukatsu and Kikuchi group of AIST, we adapted this Burkholderia-bean bug system to study the biochemical mechanism of how gut symbiont regulates host innate immunity and development. When we characterized the immune phenotypes of symbiotic bean bugs with gut Burkholderia symbiont or apo-symbiont bean bug, apo-symbiont insects are more susceptible against pathogenic bacteria infection compared to symbiotic insects, suggesting that gut symbiont enhances the basal level of host innate immunity. Based on these observations, three antimicrobial peptides are purified to homogeneity from E. coli-injected symbiotic bean bugs and then the biological roles of these three peptides are examined. Next, we tried to understand the biochemical mechanisms of how gut symbiont can regulate and affect host development. We identified a novel biomarker protein in the insect blood, which is specifically expressed in the presence of gut Burkholderia symbiotic bacteria. The biological function and significance of this bio-marker protein are examined in vitro and in vivo. Taken together, I want to present host different responses toward pathogenic and symbiotic bacteria using two insect system. 6
500 Kikuchi, Y., M. Hayatsu, T. Hosokawa, A. Nagayama, K. Tago, T. Fukatsu 2012. Symbiont-mediated insecticide resistance. Proc. Natl. Acad. Sci. U. S. A. 109:8618 8622. 7
Burkholderia Burkholderia Burkholderia Burkholderia EST library defensin c-type lysozyme cathepsin L cysteine rich protein RNAseq RNAseq EST Peptidoglycan recognition protein serine proteinase inhibitor Burkholderia 8
Plasmodium berghei Anopheles stephensi Serratia marcescens HB3 HB18 HB3 HB18 HB3 HB18 9
bacteriome flavobacterium -proteobacteium Sulcia Nasuia Rickettsia Bacteriome Nc_Sulcia, Nasuia Sulcia 192 kb Sulcia 244 277 kb Sulcia Nc_Sulcia DNA C Nasuia 114 kb UGA 1.56 Mb EST PGRP Bacteriome EST peptidoglycan recognition protein PGRP bacteriome bacteriome EST 160 PGRP RNA-Seq PGRP 300 PGRP 12 PGRP bacteriome PGRP PGRP bacteriome #35gene RNAi PGRP bacteriome PGRP PGRP PGRP bacteriome 10
[1-3] 1) 2) [4] 2 [5] 138 35 indel 2 indel 1. Fukatsu and Hosokawa (2002) Appl Environ Microbiol 68, 389 2. Hosokawa et al. (2005) FEMS Microbiol Ecol 54, 471 3. Hosokawa et al. (2006) PLoS Biol 4, e337 4. Hosokawa et al. (2007) Proc R Soc B 274, 1979 5. Nikoh et al. (2011) Genome Biol Evol 3, 702 11
EST 6 RACE 829 PMDP PMDP 6 PMDP 4 PMDP PMDP RNAi RNAi 3 PMDP 1/100 2 RNAi PMDP PMDP 12
16S rrna Wolbachia Wolbachia B Wolbachia B Wolbachia Wolbachia Wolbachia B Wolbachia Hosokawa T., Koga R., Kikuchi Y., Meng X.-Y., Fukatsu T. (2010) Wolbachia as a bacteriocyte-associated nutritional mutualist. Proc. Natl. Acad. Sci. USA 107: 769-774. 13
` Acyrthosiphon pisum Rickettsiella PCR Rickettsiella Rickettsiella Rickettsiella Rickettsiella 3 Rickettsiella RNA-seq 54 Rickettsiella 14