| Cyt b gene and COII gene in mitochondrial DNA of partial species belonging to Adelphocoris Reuter were analyzed by means of specific primer PCR and sequencing. Eight 433bp sequences of Cyt b gene of 6 species (Ad. suturalis (Jakovlev),Ad. melanocephalus Reuter,Ad. luridus Reuter,Ad. rufescens Hsiao,Ad. fasciaticollis Reuter,Ad. lineolatus (Goeze)) and two 381bp sequences of COII of 2 speceis (Ad. lineolatus (Goeze), Ad. ponghvariensis Josifov ) were obtained.The following results were showed: 1. Both pure ethanol fresh specimens and dried ones are used in extracting DNA , but not any PCR products have been produced successfully from the latter. Those good PCR products are purified and sequenced. 2. Eight 433bp sequences of Cyt b gene and two 381bp sequences of COII gene of Adelphocoris species were obtained and used in analysis. The methods and results of gene sequences analysis reflected directly the variation of the genetic materials and its essences of the taxa involved in the research. They gave the most objective basis for classification and phylogeny. So they can be used to identify species, analyze on intraspecific populations and determine phylogenetic relationship between the taxa. 3. The purposes of this research are phylogeny of interspecific. The obtained sequences data are compared, aligned and analysed by DNA sequence analysis software package DnaStar,DnaSP and Mega,to calculated residue composition,polymorphic sites and genetic distance,to consider the types and rates of residus substitution and to create NJ, ME, UPGMA phylogenetic trees as well as determine phylogeny of the classification taxa involved in the research. 4. In the composition of 432bp sequences of Cyt b gene of Adelphocoris bugs, A+T average high about 69.8%,whereas C+G relatively low about 30.2%. A+T of the three sites average respectively 60.3%,67.4%,81.7%,with the 3rd site averaging the highest. Majority of polymorphic site are located on the 3rd site of triplet code,average 66.79% of total polymorphic sites, and then on the 1st site, average 24.64%. The 2nd site is the most conservative one, only average 9.57%. Location of the informative site is also under the same rule. Informative sites on the 3rd, 1st and 2nd site of the triplet code average respectively 62.13%, 22.28% and 15.59%.Majority of the variant residues are produced by transformation and all kinds of resides substitution are existing. In the residue composition of 381bp sequences of COII gene of Ad. lineolatus and Ad. ponghvariensis , A+T average high about 68.45%, whereas C+G relatively low about 31.55%. A+T of the 1st site average 77.7 % , the highest,2nd and 3rd average 64.6%, 63.1%, a little different. Majority of polymorphic sites and informative sites of are on the 3rd site of triplet code, average 50%, whereas the 2nd is the most conservative. The ammonia acid variation is 0.048 between this two species. 5. NJ, ME and UPGMA trees base on the Cyt b 432bp sequences are the same expect confidence of the branches on the molecular phylogentic tree. The phylogenetic relationship determined based on the molecular phylogenetic tree of the involved species is showed as:((((Ad.rufescens,Ad. luridus)Ad. melanocephalus)Ad. suturalis)(Ad. fasciaticollis,Ad lineolatus))。The phylogenetic relationship of three different population as:((Ad.lineolatus3 (Ad.lineolatus1,Ad. lineolatus2). 6. Based on calculation, comparation and analysis on residues composition and rate of variation of the Cyt b 432bp sequences, no variation is found between Ad. rufescens and Ad. luridus. It should be further studied on their classification positions. Based on the analysis of the COII gene sequences, different between Ad. ponghvariensis and Ad. lineolatus are showed, the rate of variation is 0.048. Although there are many middle types between the two, they should be further determined for being independent species . |
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