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

摘要:
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OsSBE 3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 T₀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。

Abstract:
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 T₀ 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.