应用CRISPR/Cas9基因编辑技术获得高直链淀粉水稻种质

CRISPR/Cas9 Technology-generated High-amylose Rice Varieties

【目的】通过CRISPR/Cas9基因编辑技术遗传改良水稻种质,创制高直链淀粉新材料。【方法】以水稻品种中花11为试验材料,利用CRISPR/Cas9编辑系统对水稻淀粉分支酶(Starch branching enzymes,SBE)基因OsSBE3进行靶向编辑,利用PCR技术鉴定无标记纯合突变体,并测定其淀粉含量。【结果】T_(0)代获得10株突变体株系,T1代获得5个无标记纯合突变株系,其中4个株系(sbe3-22-6、sbe3-25-3、sbe3-25-4、sbe3-25-6)的直链淀粉含量和淀粉直支比显著高于野生型。【结论】本研究创制了高直链淀粉含量的水稻新种质,为水稻品质改良提供了参考。

【Objective】A series of new rice germplasms with high content of amylose derived from Zhonghua 11 using the CRISPR/Cas9 technology was generated.【Method】The gene of starch branching enzyme in rice,OsSBE3,was targeted for the genetic editing by CRISPR/Cas9.Homozygous T-DNA-free mutants were verified by PCR with starch content measured.【Result】Ten mutant lines were obtained from the T_(0) generation.From the T1 generation,5 homozygous T-DNA-free lines were obtained that included 4 lines,i.e.,sbe3-22-6,sbe3-25-3,Sbe3-25-4,and sbe3-25-6,showing significantly increased amylose content and amylose/amylopectin ratio over the wild type.【Conclusion】A series of new rice germplasms with high amylose content was created.