Regeneration and Agrobacterium-mediated transformation of japonica rice varieties developed for a cold region

Mingfang Feng, Jing Cang, Junhong Wang, Jian Sun, Jing Yu, Qinghua Xu, Da Zhang, Ning Yang, Qiuwei Lu and Yan Lv
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Mingfang Feng: College of Life Science and
Jing Cang: College of Life Science and
Junhong Wang: College of Life Science and
Jian Sun: College of Agriculture, Northeast Agricultural University, Harbin, P.R. China
Jing Yu: College of Life Science and
Qinghua Xu: College of Life Science and
Da Zhang: College of Life Science and
Ning Yang: College of Life Science and
Qiuwei Lu: College of Life Science and
Yan Lv: College of Life Science and

Czech Journal of Genetics and Plant Breeding, 2018, vol. 54, issue 4, 161-167

Abstract: So far, a large number of transformation systems have been established for japonica rice, but only a few have been reported for cold-region varieties. In our study, we established highly efficient tissue culture systems for two cold-region rice cultivars, Dongnong 427 and Longdao 14. Plant growth regulator (PGR) levels were optimized by an orthogonal experimental design. The culture ability, constituted by induction and differentiation rate, served as the detection index of orthogonal experiments. The optimal combinations of PGRs for callus induction and regeneration of Dongnong 427 and Longdao 14 were 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) + 2 mg/l 6-benzyladenine (BA) + 4 mg/l kinetin (KIN) + 0.2 mg/l α-naphthaleneacetic acid (NAA) and 1 mg/l 2,4-D + 4 mg/l 6-BA + 4 mg/l KIN + 0.5 mg/l NAA, respectively. Agrobacterium strain EHA 105 containing the plasmid pCAMBIA1301 was used for transformation. The frequency of transient transformation was expressed as the ratio between the number of calli showing GUS expression and the total number of calli kept for staining. The highest transformation efficiency in Dongnong 427 was obtained when calli were immersed in 0.272 OD600 (optical density determined at 600 nm) for 10 min. While it was best for Longdao 14 calli to be infected with 0.592 OD600 for 20 min. Infected calli of the two varieties were co-cultivated on two pieces of sterile filter paper moistened with 1 ml liquid co-cultivation medium for three days. The expression of the GUS gene was confirmed by PCR analysis of plants of both varieties.

Keywords: Agrobacterium; GUS expression; round-grained non-glutinous rice; tissue culture; transformation efficiency (search for similar items in EconPapers)
Date: 2018
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Persistent link: https://EconPapers.repec.org/RePEc:caa:jnlcjg:v:54:y:2018:i:4:id:86-2017-cjgpb

DOI: 10.17221/86/2017-CJGPB

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