Study of mutation in tomato genomic DNA using low energy ion implantation or soybean DNA by RAPD

Abstract

To analyze mutation in tomato genomic DNA by RAPD technology, certain factors influencing RAPD amplification, such as, DNA template, Taq DNA polymerase, Mg²⁺, dNTPs and primer, were studied to optimize RAPD amplification reaction system. RAPD amplification of tomato genomic DNA after low energy ion implantation along with soybean DNA was successfully performed. Compared to the control, genomic DNA of tomato implanted with N⁺ or Ar⁺ ion beam displayed mutation. Further, total number of amplification band, number of differential band and rate of differential band increased when implantation dose of ion beam increased. The rate of differential band caused by Ar⁺ ion beam was higher compared to that by N⁺ ion beam under the same implantation dose. In addition, soybean DNA also had effect on tomato genomic DNA. In comparison to whole genomic DNA of soybean, <span style="mso-fareast-font-family:SimHei" lang="EN-GB">the number and rate of differential band increased for random fractured genomic DNA of soybean, indicating that it might be easier for DNA fragment to enter into cell and integrate into genomic DNA of tomato as compared to the whole genomic DNA. Collectively, the rate of differential band was positively correlated with the implantation dose of ion beam, and the effect of soybean DNA fragment on tomato genomic DNA was higher compared to whole genomic DNA of soybean, which would provide basis for application of ion beam technology in tomato breeding.

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