Molecular Systematics Of Some Tribes Of Pentatominae

Relationship of 18 genera of some tribes of Pentatominae was primarily studied by using 16S rRNA gene in mitochondrial DNA as molecular marks. With the PCR and DNA sequencing, 27 16S rRNA gene sequences were obtained from 22 species of 18 genera of 7 tribes. The examined species were as follows: Dolycoris baccarum (Linnaeus), Priassus spiniger Haglund, Placosternum urus St?l, Erthesina fullo (Thunberg), Graphosoma rubrolineata (Westwood), Stollia guttiger (Thunberg), Hoplistodera fergussoni Distant, Pentatoma angulata Hsiao et Cheng, Pentatoma armandi Fallou, Pentatoma distincta Hsiao et Cheng, Pentatoma mosaicus Hsiao et Cheng, Homalogonia obtuse (Walker), Dalpada nodifera Walker, Palomena viridissim (Poda), Eurydema dominulus (Scopoli), Carbula sinica Hsiao et Cheng, Carbula. obtusangula Reuter, Plautia fimbriata (Fabricius), Nezara viridula Linnaeus, Cappaea tibialis Hsiao et Cheng , Hippota dorsalis (St?l), Halyomorpha Picus (Fabricius).The genetic polymorphism in mitochondrial DNA of 27 samples of the 22 species was analyzed and 400bp was amplified with the PCR (polymerase chain reaction) and sequenced. From the consensus sequence of the two strands from each specimen, an alignment of 350 nucleotides coulde be used for comparison of the 27 specimens. The genetic polymorphism among populations and 27 haplotypes had been found by analysing genetic sequences. DNA sequences were aligned, compared and analysed by ClustalX1.8, DnaSP, Bioedit5.09 and MEGA, to calculate residue composition, polymorphic sites and genetic distance, to create Neighbor-joining (NJ) phylogenetic tree and determine phylogeny of the taxa involved in the research. The results were as follows:1. The mtDNA-16S rRNA gene sequences of Pentatominae were amplified by specific primer for the first time in China.Gene sequences of 27 pentatomine specimens from 22 species of 18 genera of 7 tribes are obtained by DNA sequencing. The gene is good for revealing the relationship of tribes or genera. 2. The spemcimens deposited in alcohol can be used to extract DNA, amplify 16S rRNA gene sequences by specific primer.3. The slight degradation of DNA specimen does not affect the results of experiment.4. In the gene , 191 nucleotide polymorphic positions(54.57%of the whole sequence) have been checked. The sequence variation was 42.75% except the sites of deletion or insertion.The average contents of A,T,C,G are 33.84ï¼…,43.24ï¼…,6.77ï¼…,16.14ï¼… respectively, and A+T 77.08ï¼… is higher than G+C 22.92ï¼… Which is consistent with other insects.5. The Bootstrap confidence values of each branch were obtained by 1000 repeats. Molecular phylogenetic tree was constructed by Neighbor-Joining (NJ), Minimum Evolution (ME) and Maximum Parsimony (MP), and show that the confidence values of NJ is the highest. 6. Molecular phylogenetic tree was constructed by using 16S rRNA gene sequences of 27 specimens. It showed that Erthesina and Eurydema are closer, and they appeared earlier at the root of the tree than the other genera. The relationship between Nezara and Palomena, Graphosoma and Dolycoris, Stollia and Carbula are closer respectively. Pentatoma and Dalpada are closer than to Homalogonia, which is different from the genera phylogeny at generic level of traditional taxonomy.