Population Genetic Diversity Of Pardosa Pseudoannulata Based On ISSR And SRAP Analysis

This paper investigated the genetic diversity of the wolf spider (Pardosa pseudoannulata Bo|¨Senberget et strand 1906) by using the ISSR and SRAP reaction systems and analyzed the effect which ecological factor carried on genetic diversity.The wolf spider was collected from five geographical regions in Hunan,Jiangxi,Hubei and Yunnan province of China.(1) The establishment and optimization of ISSR and SRAP.systems for population genetics of wolf spiderThe ISSR and SRAP system were applied in Arachnology in this paper,to study genetic diversity of the wolf spider in different geographical regions.The results indicated that ISSR and SRAP are sensitive and reliable molecular marker technique for studying genetic diversity of the wolf spider.The eight primers pairs would generate accurate bands whose polymorphism is high.(2) The wolf spider sample which was preserved by alcohol for a long time is not a suitable material for DNA extractionIn the experiment,the DNA of the wolf spider which was preserved by alcohol for a long time was extracted in order to get more material for PCR amplification.The results showed that the alcohol of the wolf spider sample must be replaced to get the DNA,and the DNA bands were smaller and illdefined.So the sample is not a suitable material for DNA extraction because the genome DNA had been degraded.(3) The natural population of the wolf spider has abundant genetic diversity and there were difference in the genetic diversity of the population from different geographical regionsBy the optimal reaction conditions,ISSR and SRAP system got bands of DNA.It showed abundant genetic diversity that the 63.5%-74% of total fragments were polymorphic fragment.The size of fragments ranged from 300bp to 500bp.The similar result that the genetic diversity of the wolf spider in Leifeng town is highest and the Fugong is lowest came from ISSR and SRAP system.According to the parameters such as Shannon's index and Nei's gene diversity index,the wolf spider had a high genetic diversity either in species or population level and obvious genetic differentiation.From the population variation source,24.62-27.99%of total genetic variation came from intra-population and 75.38-72.01%came from inter-population.(4) The ecological factor affected genetic diversity of the wolf spider in different environmentThe matrix showed that in SRAP system,the genetic distance is farthest between Fugong population and Jishou populantion and nearest between Wuhan population and Changsha population.In ISSR system, the genetic distance is farthest between Fugong and Wuhan population and nearest between Wuhan population and Changsha population.So, different habitat made the wolf spider have genetic differentiation.The correlation analysis of ecological factors and population genetic diversity showed that there was obviously negative correlation between the genetic diversity and the altitude,but there was positive correlation between the genetic diversity and the accumulated temperature.It means that the polymorphic loci,Shannon's diversity index and Nei's gene diversity index of population decreased with the hoist of altitude,but increased with the augment of accumulated temperature.Because of the rich rainfall in the south and south-west of China,the correlation between genetic diversity and rainfall was not significant.The cluster analysis indicated that there was distinct genetic differentiation between Fugong population in high altitude regions and other population.It showed that high altitude may be the obstacle of spider distributing and may lead to the generation of new specie.(5) The Variation Source of the wolf spider population by ISSR system is similar with which of SRAP system,but the result of ISSR is higher than SRAPThe result of ISSR is higher than SRAP such as the polymorphism bands,percentage of polymorphism bands,effective allele,polymorphic loci,Shannon's diversity index and Nei's gene diversity index.The ISSR and SRAP system were both suitable for the analysis of genetic diversity of the wolf spider and other spiders.