Abstract: 【Objective】Oryza officinalis Wall. has many abiotic tolerance-related genes,which are very important germplasms in rice breeding, however, it is difficult to utilize these genes in cultivated rice by interspecific hybridization due to the reproductive isolation. This study was planned to establish an efficient and quick method for transferring useful genes of O. officinalis into elite cultivated rice varieties. 【Method】Positive clones were screened by southern hybridization from the TAC library of O. officinalis with the conservative sequence of AP2/EREBP and bZIP family genes as probes. The probes were labeled with α-32P during the first screening and with Digoxigenin during the second screening. PCR was used to verify the positive clones finally, which were introduced into cultivated rice by Agrobacterium tumefaciens-mediated transformation. The transformed plants were detected by PCR amplification with the primes designed according to the conservative sequence of Hpt and SacB that located on both sides of the fragment in TAC. Southern hybridization was also used to identify the transformation plants. PEG-6000 was used to identify the drought tolerance of T2 at germination and seedling stages.【Result】A total of 1 073 clones were developed from the TAC library with AP2/EREBP and bZIP probes that were labeled with α-P32. A total of 147 clones were obtained from the 1 073 clones with Digoxigenin labeled probes, and 95 clones detected by AP2/EREBP probe and 52 clones by bZIP probe. After PCR detection, a total of 103 clones produced PCR amplified products, among them 63 clones were detected by AP2/EREBP probe and 40 clones by bZIP probe and the percentages of positive clones were 66.32% and 76.92%, respectively. Five positive clones (49R-O14, 55R-A17, 8R- A24, 22D-P2q and 52D-M16, named by the clone’s number) were successfully introduced into Nipponbare by the Agrobacterium tumefaciens-mediated transformation. The result showed that the longer inserted fragment was more difficult to be transformed into O. sativa. The results of PCR and Southern hybridization showed that foreign fragments were transferred successfully into the genome of regenerated plantlets. Further, tests of hygromycin resistance also showed the same results. The transgenic lines showed strong tolerance to drought stress, among which R12-23, R15-41, and R1-8 were more tolerance to drought stress than Nipponbare did through detection by tolerance to drought stress at budstage; M63-9 and R12-23 were better in tolerance to drought stress than Nipponbare did through detection by tolerance to drought stress at seedingstage. It was concluded that R12-23 is high tolerance to drought stress.【Conclusion】 The work demonstrated that introduction of stress-related TAC clones coupled with a transgenic validation approach is an efficient strategy to transfer agronomically important genes from O. officinalis to cultivated rice.

Key words: Oryza officinalis Wall. , TAC library , genetic transformation , drought tolerance