Reaserch On Using NiR Gene To Increase The Regeneration Capacity Of Rice And Bamboo

The study of rice showed that the NiR activity of callus has a positive correlation with regeneration capacity. And then, NiR gene from variety with high regeneration ability was transeformed into rice with low regeneration ability can improve its regeneration ability. In this paper, we confirmed the relationship between NiR activity and bamboo regenerative capacity by measuring NiR activity of callus from different bamboo with diverse regeneration ability; Then we cloned NiR gene and analysed the fuction of D. hamiltonii which has high regeneration ability, it improve the understanding the mechanism of DhNiR how to affect of plant regeneration ability; Last not least, we transeformed DhNiR into rice and bamboo to verify its impact on the regenerative capacity. This paper laid the theoretical and practical foundation for the establishment of bamboo regeneration systems, the main findings are as follows:(1) We selected six kinds clumping bamboo of good regeneration capacity, B. polymorpha, D. hamiltonii, G. brevisvagina, T. siamensis, G. tekserah, G. felix and a kind scattered bamboo of poor regeneration capacity, P. edulis to measure the NiR activity of callus from different bamboo with diverse regeneration ability. Studies have shown that there is a positive correlation tendency between NiR activity of bamboo callus and its regeneration ability.(2) We got the NiR ORF sequence of D.ham which length was 1779 bp and coding 591 amino acids. The protein encoded by DhNiR is hydrophilic. There is a nitrite and sulfite reductase iron-sulfur/heme binding site on DhNiR amino acid. By using the in-fusion technology to construct a binary expression vector of DhNiR.(3) We transform the binary expression vector of DhNiR and empty vector respectively into rice(Zhonghua 11 and Nipponbare) and bamboo(G. tekserah and G. felix). After callus transferred to differentiation medium 30 days, the green callus frequency of Nipponbare transferred DhNiR and empty vector is 27% and 17%, respectively. On the other hand, the green callus frequency of Zhonghua 11 transferred DhNiR and empty vector is 37% and 30%, respectively. Finally, we got plants transferred DhNiR gene Zhonghua 11 and Nipponbare 12, 18 and obtained plants transferred empty vector gene Zhonghua 11 and Nipponbare 5, 2. And that on the rice differentiation of transferred DhNiR was one week ealier than that transferred empty vector, it supported the view that DhNiR play an active role on improving the regeneration of rice.