四种散白蚁的分子鉴定及系统发育地位 ( 等翅目 : 鼻白蚁科 ) *

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1 Chinese Journal of Applied Entomology 207, 54(3): DOI: /j.issn 四种散白蚁的分子鉴定及系统发育地位 ( 等翅目 : 鼻白蚁科 ) * 彭一丁 ** 张成玉 肖元玺 宋安东 楚君鹏 王风芹 李 琰 苏丽娟 席玉强 *** 摘 要 目的 4 方法 4 Reticulitermes hunanensis Tsai et Peng Reticulitermes planifrons Li et Ping Reticulitermes perilucifugus Ping Reticulitermes minutus Ping et Xu 6S rdna GeneBank 4 结果 6S rdna 380 bp 720 bp AT GC 6S rdna 结论 6S rdna 6S rdna 关键词 DNA 6S Molecular identification and phylogeny of four Reticulitermes species (Isoptera: Rhinotermitidae) PENG Yi-Ding ** XIAO Yuan-Xi CHU Jun-Peng LI Yan XI Yu-Qiang 2 ZHANG Cheng-Yu SONG An-Dong WANG Feng-Qin SU Li-Juan *** (. College of Life Sciences of Henan Agricultural University, Zhengzhou , China; 2. College of Plant Protection of Henan Agricultural University, Zhengzhou , China) Abstract [Objectives] To evaluate using molecular methods to identify, and construct a phylogeny of, four Reticulitermes species, which, like other termites, have been traditionally identified on the basis of morphological features. [Methods] Mitochondrial 6S rdna and partial gene sequences of four Reticulitermes species (Reticulitermes hunanensis Tsai et Peng Reticulitermes planifrons Li et Ping Reticulitermes perilucifugus Ping and Reticulitermes minutus Ping et Xu ) were amplified, sequenced and compared. After Blast and base composition analysis, sequences were submitted to the GeneBank database and neighbor-joining trees of the phylogenetic relationships between species constructed. [Results] The four 6S rdna sequences are about 380 bp and the four sequences are about 720 bp, long. The proportion of AT is much greater than that of CG in both genes, and the genetic distances between the 6S rdna of each species are all greater than those between gene sequences. Phylogenies based on the 6S rdna and sequences were consistent. [Conclusion] That based on gene sequences more closely reflected the geographical origin of the species but 6S rdna sequences has greater nucleotide diversity than gene sequences. The gene is more applicable to research on phylogenetic differences caused by geographical location and by migration and immigration. The 6S rdna is, however, more useful than * Supported projects ** First author pengyiding68x@26.com *** Corresponding author sulijuan86@26.com Received Accepted

2 3 : 40 the gene for termite species identification. Key words mitochondrial DNA, Reticulitermes, 6S,, molecular identification, phylogeny Itakura S rdna 3% Austin 0% % Austin 2006a S Legendre 2008 Eggleton S rdna 5% Buchli 956 Reticulitermes spp. %~2% Agnarsson S rdna 6S rdna Scheffrahn 2004 Itakura NCBI GeneBank DNA 7 R. hunanensis R. planifrons R. minutus 4 6S rdna GeneBank 4

3 402 Chinese Journal of Applied Entomology 54 材料与方法. 材料 R. hunanensis R. planifrons R. minutus 3 48 N 5 24 E MEGA7 Align Dnaman8.0 MEGA7 Tajima-Nei 000 MEGA7 6S rdna Neighbor-Joining 结果与分析 200 g mol/l PBS Soil DNA Kit OMEGA American DNA.2 PCR 扩增 PCR DNA Elution Buffer μl 2 Taq Plus PCR Master Mix Tiangen China 25 μl 2 μl 6S rdna CGC CTG TTT ATC AAA AAC AT TTA CGC TGT TAT CCC TAA TCT AAT ATG GCA GAT TAG TGC GAG ACC AGT ACT TGC TTT CAG TCA TC Legendre 2008 DNA 200 ng ddh 2 O 50 μl 94 5 min s s s 50 s 72 0 min 5 μl %.3 系统发育分析 NCBI Blast GeneBank 7 6S rdna 2. PCR 结果 PCR 6S rdna bp bp NCBI GeneBank 4 6S rdna R. hunanensis KX29972 KX29977 R. planifrons KX29974 KX29978 R. minutus KT KX2998 R. perilucifugus KX29973 KX 基因碱基序列分析 GeneBank MEGA7 6S rdna 332 bp 658 bp 6S rdna A+T 6.4% G+C 38.6% A 33.3% C 20.7% G 7.9% T 28.% A+T 63.3% G+C 36.7% A 39.6% C 23.3% G 3.4% T 23.7% AT GC 6S rdna T T-G A-C

4 3 : 403 表 所选取白蚁来源及其相应基因登录号 Table Sources and GenBank accession numbers of the chosen samples Species 6S rdna Locality Reticulitermes aculabialis JX4263 JX427 Hangzhou, China Reticulitermes labralis JX4262 JX4249 Beijing, China Reticulitermes chinensis KM KM Jinmen, Hubei, China Reticulitermes speratus JX4267 JX4254 Beijing, China Reticulitermes santonensis AY80828 AY Tussac, France Reticulitermes flaviceps AY8082 AY California,USA Reticulitermes lucifugus AY80825 AY80809 Seville, Spain A-G T-C 2.3 遗传距离和差异减基数分析 6S rdna S rdna ~ ~ ~ ~ S rdna 2.4 系统发育分析 6S rdna NJ 2 R. hunanensis R. planifrons R.minutus R. hunanensis R. flavipes R. santonensis 4 R. aculabialis R. labralis R. speratus R. chinensis 2 R. planifrons R. hunanensis R. minutus 3 表 2 种散白蚁线粒体 6S rdna Tajima-Nei 遗传距离 ( 对角下 ) 和差异减基数 ( 对角上, 含缺失位点 ) Table 2 Tajima-Nei genetic distance matrix (below diagonal) and number of difference of nucleotides (above diagonal, including nucleotide deletion mutation) of Reticulitermes species mitochondrial 6S rdna No. Species *** R. hunanensis *** R. minutus *** R. planifrons *** R. labralis *** R. speratus *** R. chinensis *** R. aculabialis *** R. flavipes *** R. santonensis *** 29 R. lucifugus ***

5 404 Chinese Journal of Applied Entomology 54 表 3 种散白蚁线粒体 基因 Tajima-Nei 遗传距离 ( 对角下 ) 和差异减基数 ( 对角上, 含缺失位点 ) Table 3 Tajima-Nei genetic distance matrix (below diagonal) and number of difference of nucleotides (above diagonal, including nucleotide deletion mutation) of Reticulitermes species mitochondrial genes No. Species *** R. hunanensis *** R. minutus *** R. planifrons *** R. labralis *** R. speratus *** R. chinensis *** R. aculabialis *** R. flavipes *** R. santonensis *** 0 R. lucifugus *** Fig. 图 6S rdna 基因 Neighbor-Joining 系统发育树 Neighbor-Joining phylogenetic tree inferred from 6S rdna sequences Fig. 2 图 2 基因 Neighbor-Joining 系统发育树 Neighbor-Joining phylogenetic tree inferred from gene sequences

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