http://www.paper.edu.cn

Molecular Phylogenetics and Evolution 34 (2005) 448–451

Short communication The Bayesian phylogenetic analysis of the 18S rRNA sequences from the main lineages of Trichophora (Insecta: : )

Qiang Xie, Wenjun Bu¤, Leyi Zheng

Insect Molecular Systematic Lab, Institute of Entomology, College of Life Science, Nankai University, No. 94 Weijin Road, 300071 Tianjin, People’s Republic of China

Received 22 December 2003; revised 19 October 2004

Communicated by Rob Desalle

Pentatomomorpha is the sub-largest one in the seven gave the topology (( + ) infraorders of Heteroptera (Insecta) and one of the + (Idiostoloidea + )), with the position of most phylogenetically distal group, along with Cimico- inside the branches of Lygaeoidea. morpha, among them (Schuh, 1979; Wheeler et al., Currently, there is no molecular detail about the phy- 1993). It is constituted by Trichophora along with Ara- logeny of Trichophora. Our goal in this work is to use doidea. Up to now, the phylogenetic hypothesis the sequences of 18S rDNA to study the evolutionary ( + ( + the remainder of relationships among the main lineages of Eutrichophora Trichophora)) of the infraorder has been supported by and to determine the position of Piesmatidae in Tricho- Leston (1958) and Henry (1997) and is accepted by phora. In this report, we have analyzed 26 representative most researchers. Among those superfamilies, only Ara- species, which belong to 13 diVerent families of most doidea and Pentatomoidea are consistently recognized major Trichophora groups (Table 1). Among the as monophyletic groups (Henry, 1997). And, the phylo- sequences we used, eight of them (AY252224, genetic relationship of the superfamilies within the AY252229, AY252262, AY252322, AY252323, remaining Trichophora, for which we raised the name AY252410, AY252422, X89495) were attained from Eutrichophora here, has been debated and is still uncer- GenBank directly. tain. About a decade ago, most researchers agreed on Samples were stored in 70% ethanol immediately the 5-superfamily scheme of Eutrichophora (Henry and after the collection. DNA was extracted from single indi- Froeschner, 1988; Slater, 1982) which included vidual of each species. The PCR primers were designed Coreoidea, Idiostoloidea, Lygaeoidea, Piesmatoidea, in reference to Campbell et al. (1994), Campbell et al. and Pyrrhocoroidea. Nowadays, the general view is a 3- (1995) and Sorensen et al. (1995), with slight modiWca- superfamily one with Idiostolidae and Piesmatidae tion. CLUSTAL X was used to obtain multiple align- belonging to Lygaeoidea (Schuh and Slater, 1995) or a ments. The secondary structure of rRNA was 4-superfamily one with only Piesmatidae belonging to determined by the software RNAstructure 4.0 (Mathews Lygaeoidea (Henry, 1997). Leston (1958) raised the et al., 2004) and was employed to correct the positional hypothesis of ((Coreoidea +Pyrrhocoroidea) + homology. The 18S rRNA secondary structure model of (Piesmatoidea + Lygaeoidea)). As Schuh (1986) noted, ’s provided by Ouvrard et al. (2000) was also Piesmatidae is a group of . Henry (1997) used to conWrm some details. Modeltest 3.06 (Posada and Crandall, 1998), MrBayes 3.0 b4 (Huelsenbeck and * Corresponding author. Fax: +86 22 23508800. Ronquist, 2001) and PAUP 4.0 b10 (SwoVord, 1998) E-mail address: [email protected] (W. Bu). were employed to deduce evolutionary relationships.

1055-7903/$ - see front matter  2004 Elsevier Inc. All rights reserved. doi:10.1016/j.ympev.2004.10.015

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Q. Xie et al. / Molecular Phylogenetics and Evolution 34 (2005) 448–451 449

Table 1 Sequences of representative species being used Species a,b GenBank Accession No. Lengthc signatus L: AY665544, AY665533 1518 Phaenacantha bicolor L: AY665542, AY665532 1519 Macropes exilis L: : Blissinae AY665538, AY665529 1520 Cymus tumescens L: Lygaeidae: Cyminae: Cymini AY665534, AY665526 1516 Ninomimus Xavipes L: Lygaeidae: Cyminae: Ninini AY665539, AY264352 1522 Geocoris pallidipennis L: Lygaeidae: Geocorinae AY665536, AY665527 1512 Henestaris oschanini L: Lygaeidae: Henestarinae AY324853 1517 Kleidocerys sp. L: Lygaeidae: Ischnorhynchinae AY252410 1517 Emphanisis fuscus L: Lygaeidae: Lygaeinae AY665535, AY264349 1520 Oxycarenus brunneus L: Lygaeidae: Oxycareninae AY665540, AY665530 1512 Pachygrontha antennata L: Lygaeidae: Pachygronthinae AY324852 1520 Gyndes pallicornis L: Lygaeidae: : Myodochini AY665537, AY665528 1520 Plinthisus sp. L: Lygaeidae: Rhyparochrominae: Plinthisini AY252422 1513 Panaorus albomaculatus L: Lygaeidae: Rhyparochrominae: Rhyparochromini AY665541, AY665531 1517 Udeocoris nigroaeneus L: Lygaeidae: Rhyparochrominae: Udeocorini AY252262 1518 maculata L: Piesmatidae AY665543, AY264356 1514 Riptortus linearis C: AY665547, AY264357 1514 grossipes C: AY665546, AY264355 1516 latus C: AY665545, AY264348 1513 Physopelta cincticollis Py: AY665548, AY264354 1518 Melamphaus rubrocinctus Py: AY665549, AY264351 1508 sp. Pe: AY252322 1514 sp. Pe: Canopidae AY252229 1513 Allocoris sp. Pe: AY252323 1512 Megymenum sp. Pe: AY252224 1512 Rhaphigaster nebulosa Pe: X89495 1514 a Refers to Schuh and Slater (1995). b L, Lygaeoidea; C, Coreoidea; Py, Pyrrhocoroidea; and Pe, Pentatomoidea. c Total length used.

The alignment result suggested there exist three E23_3 and 49 are showed in Fig. 1. After revising the length variable regions in the sequences. According to positional homology in the paired regions, the modiWed the 18S rRNA secondary structure model of Hemiptera residue ranges in the bulges between the paired regions (Ouvrard et al., 2000), they locate on the helix E23_3, 43, are realigned to eliminate the possible artiWcial changes and 49, respectively. The secondary structure of helix 43 in the process of emendation. in the existed model is applicable and need no further The evolutionary model for the sequences was calcu- calculation. The detailed models according to helices lated with Modeltest and it suggested the proper one is E23_3 and 49 are determined from the consensus results GTR +I + . The Bayesian analysis, approximately cal- deduced by RNAstructure. The stable paired regions, culated with MCMC algorithm, in MrBayes and the which are the foundation for alignment correction, in algorithms MP and ML in PAUP* 4.0 were applied in

Fig. 1. The local 18S rRNA secondary structures of the Wrst (A) and the third (B) length variable regions in the sequences. These two segments are the according parts of Rhopalus latus 18S rDNA sequences. The thick lines beside the structures show the stable helices regions. 中国科技论文在线 http://www.paper.edu.cn

450 Q. Xie et al. / Molecular Phylogenetics and Evolution 34 (2005) 448–451

the phylogenetic inference. The parameters used in (Henry, 1997; Shaefer, 1993), may be a paraphyletic MCMC and ML came from the result of Modeltest. The group and the super- taxon scheme of Eutricho- search strategy used in MP and ML was heuristic. phora may be revised further due to the changing views According to the original alignment, the three algo- on Coreoidea phylogeny, not only those on Lygaeoidea. rithms came to similar results and congruously sug- Herein, we raised the name Neotrichophora for gested that the relationships among the lineages of (Coreoidea + Lygaeoidea). And, methodologically, the Eutrichophora are (Pyrrhocoroidea + (Coreoidea + results suggested that, when the secondary structure Lygaeoidea)), with the relationship among the lineages information is used to correct the alignment, more than of Coreoidea and Lygaeoidea unresolved. The clado- one algorithm should be employed in the process of phy- gram obtained with Bayesian method is showed in Fig. 2. logeny inference to display the possible diVerence. Although taxonomists have reached no agreement on There are some common results in all of the six analy- the relationships between Pyrrhocoroidea, Coreoidea, sis mentioned above and they may be the clues to resolve and Lygaeoidea until now, nearly all of the existed phy- some phylogenetic problems of Lygaeoidea in dispute. logenetic hypothesis suggested it be ((Pyrrhocoroidea + All of the six cladograms suggested that, Lygaeidae is a Coreoidea) + Lygaeoidea) (Henry, 1997; Leston, 1958). paraphyletic group, the classiWcation of its higher groups So it is an interesting result that the node support for needs adjustment; the position of Piesmatidae is located (Coreoidea + Lygaeoidea) is very high. inside the lineages of Lygaeoidea, so its taxonomic status As to the corrected alignment, MCMC and ML came should be a family, not a superfamily; Rhyparochromi- to the similar results as above. However, MP led to 28 nae is a paraphyletic group, the tribe Plinthisini should cladograms with tree length 959 and the major-rule con- be taken out to make Rhyparochrominae monophyletic; sensus tree of them is an interesting result. It suggested monophyletic “malcid line” (Henry, 1997; Ktys, 1967) that the phylogenetic hypothesis of Eutrichophora is may not exist, especially that it should not include Colo- (Pyrrhocoroidea + (Rhopalidae + ((Coreidae + Alydidae) bathristidae in Schuh, 1986, the close relatives of Colo- + Lygaeoidea))). Alternatively, Coreoidea, which was bathristidae are Geocorinae, Henestarinae, and traditionally considered as a monophyletic group Rhyparochrominae. Among these results, the Wrst and

Fig. 2. The phylogenetic relationships imposed by the Bayesian analysis of 1.5 kb long 18S rRNA sequences. The parameters used are generations D 1000,000, nst D 6, rates D invgamma, nchains D 4. The Wrst 25,000 generations (burnin) were discarded. 中国科技论文在线 http://www.paper.edu.cn

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