: 16S rdna. ,, stylidae, Plataspidae, Cydnidae, Acant hosomatidae, Pentatomidae, Scutelleridae, Dinido ridae, Tessarato midae, U ro2

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1 Journal of Huazhong Agricultural University Vol. 25 No. 5 Oct. 2006, Ο2421 (2006) 05Ο0507Ο05 16S rdna ( : ) 3 1) 2) 1) ( 1), ; 2), ) 16S rdna, 8 23, : 16S rdna, ;, ; MP ML, NJ 2 ;, ; 16S rdna ; ; ; ; Q A Pentatomoidea Hemiptera Heteroptera Pentatomomorp ha,,, stylidae, Plataspidae, 4 100, L ygaeidae Coreoidae, 360,, ,,, [1 ] 1. 1, [ 2, 3 ], GenBank ( 1), b Henry (1997) [ 9 ] [4 ] ; L ygaeidae S pilostet hus hos pes Coreoi2 16S rdna [ 5 ] ; dae Ri ptort us pedest ris [ 10 ] CO II 1. 2 DNA [ 6 ] ; 18S rdna CO X1 Acid Isolation Kit ( Omega Bio2tek) DNA [7 ], 18S rdna [8 ],, : mg, ; ML1 Buffer, 16S rdna :2006Ο03Ο29 ; :2006Ο09Ο08 3 ( ) Cydnidae, Acant hosomatidae, Pentatomidae, Scutelleridae, Dinido ridae, Tessarato midae, U ro , ; 16S rdna DNA EaZy Nucleic Omega K, 60 ; (24 1),,,1971,,. :, ; EΟmail com

2 DNA Table 1 The sources of samples and DNA data of Pentatomoidea species 1) Current family classification Taxa GenBank 2) Accession No. ( GenBank) Code Pentatomoidea Scutelleridae Tessaratomidae Plataspidae Pentatomidae Acant hosomatidae Cydnidae Dinidoridae Urostylidae Coreoidea Coreidae Lygaeoidea Lygaeidae Coleotichus costatus Poecilocoris latus (Dallas) Choerocoris variegates Tessaratoma papillosa (Drury) Peltocopta crassivent ris Megacopta cribraria ( Fabricius) Plataspis coccinelloi des Erthesina f ullo ( Thunberg) Tholosanus sp. WCW22003 H y pogom phus sp. WCW22003 B rochy mena sp. WCW22003 Ocirrhoe sp. WCW22003 Elasmostet hus sp. WCW22003 Elasmostethus scotti Renter Stauralia com puncta S tauralia chloracant ha A nischys sp. WCW22003 A nischys luteovarius A m phaces sp. WCW22003 A llocoris sp. WCW22003 Megy menum sp. WCW22003 Urost y lis sp. Urostylis westwoodi Ri ptortus pedest ris ( Fabricius) S pilostet hus hos pes ( Fabricius) A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y A Y ScuCole ScuPo1 ScuCho TesTe2 TesPel PlaMe3 PlaPla PenEr5 Pen Tho Pen Hyp PenBro PenOci Aca Elas Aca Elas33 AcaStau AcaStauCh AcaAni AcaAniL AcaAmp CydAll DinMegy UroUroD UroUros CoAlyR12 LygS10 1) Henry (1997) Classification follows Henry (1997) ; 2) 3 : Wheeler &. Schuh (2004) GenBank Direct submission to NCBI GenBank database by Wheeler and Schub r/ min ; DNA, r/ min, ; 1. 3 DNA DNA, RNase A (20 mg/ ml) ; ML2 Buffer, tems), GenBank ; HiBind DNA, r/ min ; DNA Wash Buffer, r/ min ;, 16S rrna 3 [13 ] DNA,8 000 r/ min, DNA, 16S rdna 16S rdna,, Simon [11 ] L R2J (16Sbr) : 5 2CCG GT T T GA ACT CA G A TC A T G23,L R2 N (16Sar) : 5 2CGC CT G T T T AAC AAA AAC A T G23 25 L : 2 6 L DNA, 0. 5 U T aq DNA, 1. 5 L 10 mol/ L 1, 2. 0 L 2. 5 mmol/ L dn TPs, 2. 5 L 10 buffer ( neighbor2joining, NJ ), 95 5 min, 35 : s, s, 72 (maximum likelihood,ml) 1. 5 min, 1,72 8 min PCR, DNA MrModeltest, (Q IA GEN ) PE 377 SEQ ED (applied biosys2 Clustal2X [12 ] 16S rdna, ht2 tp :/ / life. zsu. edu. cn/ insect _ 16S rdna/ M EGA 2. 0 / ; p ( ) Kimura ( 2 P + ) PAU P b (maximum parsimony,mp) [ 14 ] PAU P b

3 5 : 16S rdna ( : ) 509 MrModeltest, f reqa = , freqc = , f req G = , freq T = ; I = ; = PAU P b ML 4 : 16S rdna (A Y A Y986801,A Y A Y986806), L yg2 aeidae S pilostet hus hos pes Coreoidae Ri ptort us pedest ris 16S rdna ( A Y A Y986811) ; 17 16S rdna 1 NJ ( 12C) ML GenBank,, ( ) ( ), 16S rdna 3, 1 ( ) L R2J ( 16Sbr) L R2N ( 16Sar), 16S rrna 75 (helix75),, [15 ], 16S rdna,, [13 ], 477 bp, 252 (, %), 184 (, %) ( 2 P + ) : T %,, C %, A %, G %, [3 ], Gap ud [ 2 ], A, T (A + T = %), DNA, p ( ) Kimura, Gap ud [2 ] ( 2 P + ), [3 ] MP ML, NJ,2 ( ) S rdna (MP), [ 16 ] (ML) (NJ ) [3 ] ( 1), ( ), 6 (Lengt h = 871,CI = ,RI = ), 12A 3,,,,,,, 1,,,, 1 (ML) ( 12B) 6, MP 3,, MP ( ), ( ), A llocoris sp. WCW ( ), Tessaratom a p a pillosa 1 ( ),, Peltocopt a crassi vent ris 3 16S rdna [1, 7, 8 ] MP ML NJ, Kumar [1 ],,, 16S rdna ; 3 1, ;,

4 S rdna MP (A) ML (B) NJ (C) Fig. 1 The phylogenetic trees of Pentatomomorpha according to 16S rdna sequence s(a) using Maxium Parsimony (MP),using Maximum Likelihood (B) and using neighbor2joining (NJ ) (C) Boot strap, Henry (1997) Boot strap values are shown above nodes,current superfamily and family taxa following Henry (1997) are indicated on t he right,,, DNA! [ 1 ] SCHU H R T,SLA TER J A. True bugs of t he world ( Hemip2 tera : Heteroptera) Classification and natural history[ M ]. N Y: Cornell University Press,1995. [ 2 ] GAPUD V P. A generic revision of t he subfamily Asopinae,

5 5 : 16S rdna ( : ) 511 with consideration of its phylogenetic position in the family tionships among higher groups of Heteroptera : congruence Pentatomidae and superfamily Pentatomoidea ( Hemiptera2 between morphological and molecular data sets [ J ]. Entomo2 Heteroptera) [J ]. Philipp Entomol,1991,8 (3) : logica Scandinavica,1993,24 : [3 ]. [J ]. [ 11 ] SIMON C, FRA TI F, BECKENBACH A, et al. Evolution,,1993,19 (2) : [ 4 ],. b [J ].,2004,25 (5) : weighting, and phylogenetic utility of mitochondrial gene se2 quence and a compilation of conserved polymerase chain reac2 tion primers[j ]. Ann Entomol Soc Am, 1994, 87 : [ 5 ],. mtdna216s rdna [ 12 ] T HOMPSON J D, GIBSON T J, PL EWNIA K F, et al. The [J ].,2004,26 (2) : [6 ],,. CO ClustalX windows interface : flexible strategies for multiple sequence alignment aided by quality analysis tools[j ]. Nucleic ( : ) [J ].,2006,31 (2) : Acids Research,1997,25 : [ 13 ] BUCKL EY T R, SIMON C, FLOO K P K, et al. Secondary [ 7 ] L I H M,DEN G R Q,WAN GJ W,et al. A preliminary phylog2 structure and conserved motifs of the trequently sequenced do2 eny of the Pentatomomorpha ( Hemiptera : Heteroptera) based on nuclear 18S rdna and mitochondrial DNA sequences [J ]. Mol Phylogenet Evol,2005,37 : mains IV and V of t he insect mitochondrial large subunit rrna gene[j ]. Insect Molecular Biology,2000, 9 : [ 14 ] FELSENSTEIN J. Confidence limits on phylogenies : an ap2 [ 8 ],,. 18S rdna ( : ) [J ]., 2006,27 (3) : proach using the bootstrap [J ]. Evolution,1985,39 : [ 15 ] OR TI G, Petry P, Porto J IR, et al. Patterns of nucleotide [ 9 ] HENR Y T J. Phylogenetic analysis of family groups wit hin t he infraorder Pentatomomorpha ( Hemiptera : Heteroptera), with emphasis on t he Lygaeoidea [J ]. Ann Entomol Soc Am,1997, 90 : change in mitochondrial ribosomal RNA genes and the phylog2 eny of piranhas[j ]. J Mol Evol,1996,42 : [ 16 ] KUMAR R. Morpho2taxonomical studies on the genitalis and salivery glands of some Pentatomoidea [J ]. Entomol Ts Arg, [ 10 ] WHEEL ER W C, SCHU R T, BAN GR R. Cladistic rela2 1962,83 : Phylogenetic Relationships of the Pentatomoidea Based on the Mitochondrial 16S rd NA Sequences( Heteroptera :Pentatomomorpha) L I Hong2mei 1) WAN G Xun2zhang 2) L IN Jin2tian 1) ( 1) De p artment of Pl ant Protection, Zhongk ai U ni versit y of A g ricult ure and Technolog y, Guangz hou , Chi na; 2) College of L i f e S ciences, S un Yat2sen Uni versit y, Guangz hou , Chi na) Abstract Sequences of mitochondrial 16S rdna gene were used to generate a molecular p hylogeny for t he Pentato moidea based o n 23 species rep resenting 8 p utative families. Phylogenetic analyses using M P ( maximum parsimo ny), ML ( maximum likelihood) and NJ ( neighbor2joining) showed t hat 16S rdna was a proper marker to reconstruct t he p hylogeny of Pentatomoidea. The result s strongly suppor2 ted that t he Pentatomoidea lineage was monop hyly. The position of Cydnidae was not certain, which would locate at the middle in t he evolution process of the Pentatomoidea families. In MP tree and ML tree,cydnidae and Pentatomidae were sister group formed a close relationship in NJ tree. Urostylidae appeared to fo rm an early clade and be basal to all sampled Pentato moidea. The st udy supported t he view t hat Scutelleridae, Dinidoridae and Tessarato midae sho uld be separated f ro m family Pentato midae and be three valid families. Key words Pentatomoidea ; 16S rdna ; molecular p hylogeny ; Pentatomidae ; Cydnidae ; Urostyli2 dae ( : )

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