Genetic Diversity And Molecular Phylogeny Of Main Groups Of Wolf Spiders (Lycosidae) In China

Based on the molecular markers, including random amplified polymorphic DNA (RAPD) and analysis of DNA sequences, the genetic diversity and molecular phylogeny of the main groups of wolf spiders from China were studied by using the principle and methods of group genetics and analysis of the DNA sequences. The results were summarized as follows:1. The method of applicability of RAPD in study of genetic diversity of spiders was discussed. The appendix of spiders can be employed to extract their genomic DNA; The production and purity of genomic DNA of spiders, on the basis of the method, could completely satisfy RAPD research. The conditions of polymerase chain reaction (PCR) were optimized for the highly clear and repeatable RAPD bands in study of genetic diversity of spiders. The RAPD bands of raised without food for above a month Trochosa wuchangensis' cephaiothorax compared with its own appendix from random primers S92 amplified polymorphic DNA-PCR(P>0.05). The results showed:The appendix and cephaiothorax of spiders can be, as the stuff of extracted genomic DNA, both used to study their genetic diversity.But applicability of genomic DNA of appendix as RAPD-PCR models in study of genetic diversity of spiders is better than that of their own cephaiothorax, because of no interference from foreign DNA.2. The intraspecific variants of wolf spiders Trochosa wuchangensis were studied by structural features of genitalia combined with RAPD molecular mark technique. The results show that the intraspecific variants exist in the natural population of Trochosa wuchangensis. There are several kinds ofintraspecific variants in Trochosa wuchangensis, even in the same geographic population. The intraspecific variants do not totally result from the different geological zone. Three types that were found in the epigynum of Trochosa wuchangensis population by structural features of genitalia are nearly the same as the types detected by RAPD technique. According to the structural features of genitalia , 3 different types of the epigynum were found as follows: ( i )The two hoods pear-formed, the lengththe width ;The inner shorter than the galea of spermatheca . (iii) The two hoods like gall bladder, the lengthHhe width;the galea slightness clearly. Type ii , according to analysis on variance from the genetic similarity index both typeii and type iii of the individual differences, is unsiglificant buttype i is siglificant (P<0.05). 253 bp is the specific size of the DNA band of male spiders Trochosa wuchangensis in the RAPD patterns by the primer S92. The different growth periods of wolf spiders Trochosa wuchangensis show the intraspecific variants. The DNA bands of the different growth periods of Trochosa wuchangensis are different from the RAPD patterns by the primer S82 and S266. The statistic data showed that the types of RAPD patterns of the adults are stable (less variant), but the types of RAPD patterns of the young spiders are unstable (more variant).3. Molecular response mechanism of predominance populations of the wolf spiders under long-period pesticide force were studied based on the analysed mitochondrial 12S rRNA gene sequence data. Comparing the third domain mitochondrial 12S rRNA gene sequences, the results show that: i .The thirddomain mitochondrial 12S rRNA gene sequences of wolf siders Pirata subpiraticus were changed obviously under long-period pesticide force, For example, 315 bp of the base pair composition of 12S rRNA gene failed into 300 bp, transisions/transversions was 17/5, the precent of nucleotide variance (P-distance = sequence difference/sequence size) was 0. 095, homology was only 85.8%. ii.The third domain mitochondrial 12S rRNA gene sequences of the wolf siders Pardosa pseudoannulata were also changed under long-period pesticide force, For example, 301 bp of 12S rRNA gene dropped into 299 bp, transisions/transversions was 11/10,the precent of nucleotide varia...