利用CRISPR/Cas9技术编辑BADH2-1/BADH2-2创制香米味道玉米新种质

张翔,史亚兴,卢柏山,武莹,刘亚,王元东,杨进孝,赵久然

Creation of New Maize Variety with Fragrant Rice Like Flavor by Editing BADH2-1 and BADH2-2 Using CRISPR/Cas9

ZHANG Xiang,SHI YaXing,LU BaiShan,WU Ying,LIU Ya,WANG YuanDong,YANG JinXiao,ZHAO JiuRan

电子附表1 本研究所使用引物

Table 1 Primers used in this study

引物名称 Primer name 引物序列 Primer sequence (5’-3’) 目的 Purpose Cas9-F1 TTGCAGTACGTAATGGACTACAAGGACCACGACG 构建骨架载体表达盒2 Construction of cassette2 Cas9-R1 AGTATCGGTCTCATGTCGAGTCCGATGGAATATTTCTTATCGG Cas9-F2 CTTATCGGTCTCAGACATTGGCACGAATAGCGTCG Cas9-R2 TTTCAGACTAGTCTATTTCTTCTTCTTAGCCTGTCC t35s-F TATCGTACTAGTCTGAAATCACCAGTCTCTCTCTACAA t35s-R CCACCAGGCGCGCCCCACTGGATTTTGGTTTTAGG prZmUbi-F CGTCCGCCTAGGCTGCAGTGCAGCGTGAC 构建骨架载体表达盒3 Construction of cassette3 prZmUbi-R ATTTCATGGATCCCTGCAGAAGTAACACCAAACA PMI-03-F ATTTCATGGATCCATGCAGAAGCTGATCAACA tNos-R AGCAGCCCTGCAGGGATCTAGTAACATAGATGACACCGC Target1-F TGCATCTCCGAAGAGAGCCTATTG Target1-R AAACCAATAGGCTCTCTTCGGAGA BH2-1-F GAGGAAACCTGAGCTGGCTAA ZmBADH2-1靶点突变检测 Mutation detection of ZmBADH2-1 target site BH2-1-R ATGGTGATGAGGAAAGTTATCGA BH2-2-F AGTGTGCAATTTTTTTTCCAGTTT ZmBADH2-2靶点突变检测 Mutation detection of ZmBADH2-2 target site BH2-2-R GGCCCAACAAATACGTGAAA PMI-F TTCGCCCTGACCCCCT T0代转基因阳性植株检测 T0 Transgenic plants detection PMI-R TGGCAGGATATATTGTGGTGTAA BH-2-1-F CTTGCCGATCCCTAGCGA ZmBADH2-2靶点突变检测 Mutation detection of ZmBADH2-2 target site BH-2-1-R CGAGACCTAGCGCATCAG