Upland Rice Transformation Of The Soybean Ferritin Gene

Through gene engineering approach, ferritin gene has been transformed into upland rice, which not only help to breed transgenic rice varieties with ferritin, but also may help to solve the problem of dietary iron deficiency.The ferritin gene was transferred into upland rice by Microprojectile Bombardment and by Agrobacterium-mediated transformation. In both of two transformations, the calli induced from mature and immature embryos was viewed as experimental material. The integration and expression of extraneous gene in rice genome were studied, and at the same time, after the factors influencing transformation had been discussed carefully, a highly efficient transformation system was established.After screening for basta resistance, pre-regeneration and regeneration, 86 transgenic plants with ferritin gene had been obtained, which included 76 transgenic plants of 297 upland rice and 10 transgenic plants of 277 upland rice. 70 positive plants and 8 positive plants were ascertain by PCR analysis, respectively. The results of PCR and Southern blotting analysis demonstrated that ferritin gene had integrated into upland rice genome successfully. It conferred endosperm-specific expression of the ferritin gene by reverse transcription PCR. Iron levels hi mature seeds of transgenic plants varied from 19 to 62.8 ug/gDW. Given that the iron contents in seeds of nontransformants is 17.2 ug/gDW, we estimated a 2.5 times increase in iron content of seeds from the hightest-expressing transgenic upland rice lines.