Study On Toxic Effect Of Petroleum Hydrocarbon Pollution To Hemicentrotus Pulcherrimus

Nowadays, in many countries, the petroleum industry is the main economic source, and the technology of exploitation transport production the technology are more and more mature. With the increasing demand of petroleum and petrole um industry development, oil spill events occurr frequently all over the world. It has a serious impact on the marine and estuarine environment. In the paper, He micentrotus pulcherrimus was selected as the test object. Molecular biology techn ology was used to find out the toxicological effects of diesel dispersed liquid on Hemicentrotus pulcherrimus. The results are as follows:(1) Sea urchin embryos was exposed in different concentrations of diesel dispersed liquid. In the low concentration, the induced effect occurred. But the higher the oil concentration was, the earlier the induced and deducted time occurred. The effects of the four enzymes on the embryos were very different, and the effects on CAT and SOD were significant than the effects on GPx and GST. In the cell growth periods that we researched, the blastocyst stage was the most sensitive.(2) In order to find out the damage on DNA of Hemicentrotus pulcherrimus tube, single cell gel electrophoresis (SCGE) was used. The results show that:in the concentrations of5,20,30and40mg-L-1, the damage on DNA wre not found, but in the concentration of50mg·L-1, the damage on DNA was significant. The longer the exposure time was, the more serious the damage was.(3) From the molecular level, SPLOX gene mutation was detected through single-strand conformation polymor-phism (SNP) technique and sequence determination. The results suggest that single base mutation has taken place in coding region of the41, with the conversion of A-G, where amino acid changed from lysine into arginine. WNT8gene mutation detection results show that single base mutation has taken place in coding region of the706, with the conversion of T-C, where amino acid changed from phenylalanine into leucine. The effects on HNF-6gene was that:single base mutation has taken place in coding region of the1119, with the conversion of G-C, where amino acid changed from serine into threonine. The globle genomic DNA was detected by high performance liquid chromatography and the longer the exposure time was, the more serious the methylation was.(4) This paper analyses the Hemicentrotus pulcherrimus’genetic diversity using the technology of RAPD. First, optimize the conditions of the test by taking some single factor designs to the4factors-Taq enzyme, primer, dNTP, DNA which will affect the result of the test to find the optimal reacting concentration. Through optimization, the writer found the optimal reacting concentration of the4factors:2U of Taq enzyme,100μmol·L-1of primer,2500μmol·L-1of dNTP, diluted80-fold of DNA. Next, the writer analyses the Hemicentrotus pulcherrimus’ genetic diversity in the sea area of Dalian in the optimal reacting conditions. Fragment PCR amplification to the DNA of45Hemicentrotus pulcherrimuses using15double primers. The maximum, minimum and average genetic distance between45individuals were0.5731,0.2569and0.3254. So, there are some diversities in the Hemicentrotus pulcherrimuses. In the concentration of30mg·L-1, the primer65, the DNA was damaged, in the concentration of40mg·L-1, the primer11, the DNA was damaged and in the concentration of50mg·L-1, the primer49and the primer67, the DNA was also damaged.