Function Analysis Of γ-tubulin Based On Its DsRNA Vector Construction

As the accomplishment of human genome project, more and more researches were concentrated on the area of functional genomics. One of the best methods is RNA interference. It was developed recently and would be a very potential technology on gene functional research because of its discrepancy, efficiency and universality. It can make corresponding gene inactive quickly and efficiently to cause organism behave the corresponding phenotype, thereby we can identify the function of unknown gene.γ-Tubulin is the very important component in the microtubule organizingcenter in animal cells, it exists nearly all the eukaryotic cells. As reported, it not only played very important role in the microtubule nucleation, assemble and array, but also involved in the cell spindle formation and cell cycle regulation. But its function in higher plants was still uncertain. In this paper, we got the down-regulated γ-tubulin tobacco plant of N. tabacum L. cv. Sumsun NN. by genetic engineering. The dsRNA expression vector was constructed by inserting sense and antisense partial γ-tubulin cDNA fragment and used to transfer tobacco by leaf disc method. The result showed that low expression γ-tubulin inhibited the tobacco transformants growth and development obviously. The down-regulated γ-tubulin tobacco plants grew very slowly, its plant height and leaves number were less than one half of the control plants and this phenotype was still stable after subculturing several times; The root regenerated from the low expressed γ-tubulin tobacco plants after 30 days, but it appeared brown and the root hair and transport system were undeveloped, which maybe the key factor that maybe effected on absorbing the nutrient material in the medium. Therefore, we concluded that downregulating the γ-tubulin in tobacco cells could inhibit plant growth and development obviously, but the direct mechanism of inhibiting still need to be discovered from small RNA level in the future.