[en] Cereals high in amylose content (AC) and resistant starch (RS) offer potential health benefits. Previous studies using chemical mutagenesis or RNA interference have demonstrated that starch branching enzyme (SBE) plays a major role in determining the fine structure and physical properties of starch. However, it remains a challenge to control starch branching in commercial lines. Here, we use CRISPR/Cas9 technology to generate targeted mutagenesis in SBEI and SBEIIb in rice. The frequencies of obtained homozygous or bi-allelic mutant lines with indels in SBEI and SBEIIb in T0 generation were from 26.7 to 40%. Mutations in the homozygous T0 lines stably transmitted to the T1 generation and those in the bi-allelic lines segregated in a Mendelian fashion. Transgene-free plants carrying only the frame-shifted mutagenesis were recovered in T1 generation following segregation. Whereas no obvious differences were observed between the sbeI mutants and wild type, sbeII mutants showed higher proportion of long chains presented in debranched amylopectin, significantly increased AC and RS content to as higher as 25.0% and 9.8%, respectively, and thus altered fine structure and nutritional properties of starch. Taken together, our results demonstrated for the first time the feasibility to create high-amylose rice through CRISPR/Cas9-mediated editing of SBEIIb.
Francis, Frédéric ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Gestion durable des bio-agresseurs
Jiao, Guiai
Liu, Zupei
Zhang, Xin
Li, Jingying
Guo, Xiuping
Du, Wenming
Du, Jinlu
Zhao, Yunde
Language :
English
Title :
Generation of High-Amylose Rice through CRISPR/Cas9-Mediated Targeted Mutagenesis of Starch Branching Enzymes
Publication date :
March 2017
Journal title :
Frontiers in Plant Science
eISSN :
1664-462X
Publisher :
Frontiers Research Foundation, Lausanne, Switzerland
Volume :
8
Pages :
298
Peer reviewed :
Peer Reviewed verified by ORBi
Funding text :
This work is partly funded by the Ministry of Science and Technology of China (grant no. 2016YFD0100500) and the Ministry of Agriculture of China (grant no. 2016ZX08010-003). YS is supported by a GSCAAS-ULg Joint Ph.D. Program.
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