Breeding of Marker-free Fragrant Rice Via Transgenic Approach

doi:10.7668/hbnxb.2015.05.015

Abstract

Abstract: In order to generate marker-free fragrant transgenic rice, O_SBADH2-RNAi and hygromycin phosphotransferase gene(HPT)were transferred to calli derived from a japonica rice cultivar,Wuyujing 3,mediated with a Agrobacterium strain EHA105 harboring a single twin T-DNA binary vector.After selected and regenerated on the media with hygromycin,54 independent transgenic lines were generated.According to PCR results,we got 36 co-transformation lines harboring both HPT gene and O_SBADH2 gene RNAi structure.The frequency of co-transformation was 66.7%.After two generation selections,we got 25 HPT gene free T₂ lines harboring O_SBADH2 gene RNAi structures.The frequency was 69.4%.15 fragrance lines were screened from these transgenic lines using four kind methods detecting fragrance.Real-time quantitative PCR results showed that O_SBADH2 gene expression level was reduced significantly in these transgenic fragrance lines.We found that there was no obvious difference between the transgenic lines and the wild type,in the main agronomy characteristics,including plant height,1000-grain weight,grain number of per spike,and seed setting rate.

Key words: Rice, Fragrant, O_SBADH2, RNAi, Marker-free

CLC Number:

Q78
S511.03

FENG Meng-shi, YE Jun, HOU Li-li, LI Ming, TANG Ding, ZHANG Jian-xiang, WANG Wei-yun, XU Zhi-hao, LI Ya-fei, GU Ming-hong, CHENG Zhu-kuan, YU Heng-xiu. Breeding of Marker-free Fragrant Rice Via Transgenic Approach[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(5): 92-96. doi: 10.7668/hbnxb.2015.05.015.

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