The Activity Of The Tobacco Retrotransposons Tnt1 Implanted By Low-energy Ion Beam

Ion Beam Bio-engineering technology is created by Chinese scientists first used in the biological effects of ion beam mutation breeding, cell processing and gene transfer has played an important role, which created the Ion Beam Bio-engineering precedent. After years of research, our predecessors have biological effects on ion a certain understanding of the mechanism, but its mechanism, the current study are not strong enough. Ion beam in order to promote the development of bio-engineering techniques, the mechanism of the urgent need to set out more clearly. Retrotransposons and plant genome as an important component of the composition, it is the distribution of eukaryotic cells may be the most extensive mobile genetic element. However, plant growth in most of the time of the transposon does not occur, and in silence. Reported that the occurrence of the transposition and environmental stress have a great relationship, as a result of Retrotransposons have occurred in a coercive environment characterized by transposition, we think of the time of ion implantation can lead to Retrotransposons the transposon, which caused a number of physiological characteristics of plants changes. In this paper, based on Retrotransposons of molecular markers TRAP, for the ion-beam irradiation after the injection of tobacco retrotransposons Tnt1 research. The results obtained are as follows:1. Low-energy nitrogen ions on the biological effects of test materials significantly. Different doses, the germination rate test materials show some differences. After ion implantation of tobacco seed, after a certain period of time the growth, morphological plant show relatively high, stout stem, green leaf color, leaf shapechanged. 1×10¹⁷ ions/cm2 dose, found a single one of the phenomenon of leaf curl. 1.5×10¹⁷ ions/cm2 dose, the three plants found there thick stem, leaf spacingdecreases, higher seed-setting rate.2. Tobacco RNA, protein, carbohydrates, pigments, nicotine and high polyphenol content. In the process of extracting DNA, the CTAB method were optimized. Extract to addβ-Mercaptoethanol; improve speed centrifugation; control centrifugal temperature no higher than 15℃; increase in the number of blending, so that leaves powder and extract the full mixed; increased by adding DNA solution after RNAse temperature bath time and so on. Purity of extracted DNA testing, and achieved good results.3. Preliminary results from the experiment found that after ion beam irradiation, the tobacco Tnt1 retrotransposon transposition occurred. TRAP analysis of the adoption, Tnt1 must demonstrate polymorphism. Confirmed Retrotransposons in the outside world under the influence of a coercive environment, transposition occurs in the DNA sequence changes have taken place in the location. Retrotransposons from the back of the characteristics are, by ion beam irradiation, the occurrence of transposition in tobacco plants on show in the form of some of the more obvious changes. Specific to emerging bands sequenced, the BLASTn search, no match was found with the higher rate of sequence, determine the initial sequence of unknown function.