| There are88million hm of grasslands in Inner Mongolia where grasshoppers occur frequently and intensively. In2012, the grasshopper occurring area was457.1ha, containing217.9ha of severely-harmed grassland. The grasshoppers have seriously affected the quality of ecological environment on the grasslands. As a biological indicator of ecological environment, the grasshopper has a high research value in terms of biodiversity. The main results are as following:1. Four levels of five factors (Taq DNA polymerase, DNA, dNTPs concentration, primers concentration and Mg2+concentration) in SSR-PCR system were selected by L16(45) orthogonal design. A suitable SSR-PCR system (20μL) was established, including1.5U/20uL Taq DNA polymerase,1.50mmol/L Mg2+,0.2mmol/L dNTPs,0.2μmol/L primers and180ng/20μL DNA.2. The grasshopper DNA was extracted by using kit. After optimizing PCR system, six microsatellite DNA markers were selected to study the genetic diversity of11grasshopper species in Inner Mongolia.The number of alleles for11grasshopper species were from19to37. The effective number of alleles was between15.8134and27.4494. Shannon’s information index was from5.2021to9.875. The polymorphic information content was between0.9396and0.94519. The site MwGTD9was the lowest and the site Ata68was the highest. All PICs were>0.5, which indicated that they were highly polymorphic locis. Genetic distances of11populations were from0.2220to0.7657. The genetic distance between Bryodemella holdereri holdereri and Bryodemella tuberculatum dilutum was the smallest (0.2220). The genetic distance between Calliptamus abbreviatus and Dasyhippus barbipes was the largest (0.7657).11grasshopper species were cluster analyzed based on UPGMA method and clustered into three groups. Bryodemella tuberculatum dilutum and Bryodemella holdereri holdereri, and Bryodema luctuosum luctuosum and Angaracris rhodopa were clustered into the first group.The second group consisted of Pararcyptera microptera meridionalis and Dayhops barbipes as well as Myrmeleotettix palpalis and Oedaleus asiaticus. The third group included Angaracris barabensis, Calliptamus barbarus and Calliptamus abbreviatus.3. We also analyzed the mitochondrial16S rDNA sequences of11grasshopper species in Inner Mongolia. forming a phylogenetic tree with11species. The genetic relationships of the grasshoppers were analyzed on the molecular level.156polymorphic loci were detected in505bp base, accounting for30.9%of the total number of bases, and including50single variation polymorphic sites and106parsimony informative sites in156polymorphic sites. Coefficient of genetic differentiation among different species was from0.0000to0.9927. The highest value of FST was0.9927among B. holdereri holdereri and M. palpalis.28haplo-types were identified in the54sequences, including2shared haplotypes and26exclusive haplotypes. UPGMA method was used to construct a molecular phylogenetic tree. It could be divided into five major branches. The first branch included two groups. The first one included A. rhodopa, B. holdereri holdereri and A. barabensis. The other one consisted of B. luctuosum and B. tuberculatum dilutum. The second branch included C. abbreviatus and C. barbarus. The third branch made up of D. barbipes and M. palpalis. The fourth and fifth branch were P. microptera meridionalis and O. asiaticus, respectively.In summary, mitochondrial16S rDNA sequences reflect the phylogenetic relationships of grasshoppers whereas SSR is not suitable for grasshopper kinship. |
