| CYP81A6 the cytochrome P450 which was coded by the rice gene Bel has resistance to herbicide bentazon, the further research to this P450 gene can settle the foundation of using it to enhancing the economic crop resistant to herbicide. Further more, knock out the gene Bel using the gene targeting in rice can obtain the bentazon susceptive rice. This paper trans the gene Bel to tobacco and tomato, from the phenotype and RNA level to analyze the expression of different fastness transgene plant. Otherwise, construct the vector pINA134-Phpt can be used in gene Bel targeting. The mainly result as follows:1. Trans the expression vector pKYL71-Bel has been constructed in our lab to tobacco and tomato genomic. Among 70 putative Bel-transformants of tobacco 54 were PCR-positive, trangene efficient up to 77%. Among 50 putative Bel-transformants of tomato 37 were PCR-positive. Trangene efficient up to 74%.2. With the bentazon leaf testing and individual testing, serial number 1 and 13 transegene tobacco has resistance to bentazon concentration of 0.25% , serial number 3,23 and 29 transegene tomato has resistance to bentazon concentration of 0.05%.3. With RT-PCR amplification to study the expression of gene Bel in transgene plant withβ-actin as standard, the result shows that transformants with different resistant to bentazon correspond to the expression in mRNA.4. Optimizing the culture medium of rice C418 inducement of embryo callus and explant differentiation, when 2,4-D 3 mg/L, yeast powder 6 g/L, Casein hydrolysate 0.6 g/L, the C418 integrate cultivate number is the highest, 2,4-D has the sinificant effect to rice callus(P<0.05), Casein hydrolysate and yeast powder have no notable effect to rice callus.5. Constructed the rice gene Bel targeting vector pINA134-Phpt with the left Bel homology length of 891bp and right Bel homology length of 695bp, it also contains the hpt and DT-A as positive and negative selection marker.
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