苹果愈伤组织基因编辑技术的建立  

Establishment of Apple Callus Gene Editing Technology

作　　者：董超华[1,2,3] 于家惠 刘瑛双 王南 曲衍杰 孙欣 柏素花 张玉刚[2] DONG Chaohua;YU Jiahui;LIU Yingshuang;WANG Nan;QU Yanjie;SUN Xin;BAI Suhua;ZHANG Yugang(College of Life Sciences,Qingdao Agricultural University,Qingdao 266109,China;College of Horticulture,Qingdao Agricultural University/Qingdao Key Laboratory of Genetic Improvement and Breeding of Horticultural Plants;University Key Laboratory of Plant Biotechnology of Shandong Province,Qingdao 266109,China)

机构地区：[1]青岛农业大学生命科学学院,山东青岛266109 [2]青岛农业大学园艺学院/青岛市园艺植物遗传改良与育种重点实验室 [3]山东省高校植物生物技术重点实验室,山东青岛266109

出　　处：《青岛农业大学学报（自然科学版）》2022年第3期157-162,170,共7页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家现代苹果产业技术体系(CARS-27);国家重点研发计划(2021YFE0104200);青岛农业大学研究生创新项目(QNYCX21087;QNYCX20073)。

摘　　要：基因编辑技术能够打破传统的基因突变方式,通过碱基的缺失、插入和替换等对目标基因进行定点改造,提高基因突变的效率和准确性。基因编辑技术在拟南芥、水稻和番茄等植物中应用广泛,如调控植物株型、提高抗性,但在苹果中应用还不成熟,需要进一步完善。以苹果‘Orin’愈伤组织为试验材料,基于成簇规律间隔短回文重复序列(clustered regulatory interspaced short palindromic repeat,CRISPR)基因编辑技术在愈伤组织中突变MdVPE1基因。通过测序及本生烟瞬时转化进行靶序列验证,结果表明MdVPE1基因已编辑成功。通过对野生型及MdVPE1基因编辑前后的愈伤组织接种轮纹病病原菌,发现MdVPE1基因编辑后能够提高苹果愈伤组织的抗病性。CRISPR/Cas9基因编辑技术在苹果愈伤组织中的成功应用,为在苹果中利用CRISPR/Cas9基因编辑技术研究基因功能奠定了重要基础。Gene editing technology can break the traditional way of gene mutation by modifying the target gene through base deletion,insertion and replacement to improve the efficiency and accuracy of gene mutation.Gene editing technology is widely used in plants such as Arabidopsis,rice and tomato for regulating plant architecture and increasing resistance,but its application in apple is immature and needs to be further improved.Using callus as,the MdVPE1 gene was mutated in the test material of apple‘Orin’callus based on the clustered regulatory interspaced short palindromic repeat(CRISPR)gene editing technology.The target sequence was verified by DNA sequencing and transient transformation.The results showed that the MdVPE1 gene was edited successfully.By inoculating the callus of wild type and callus of before and after MdVPE1 gene editing with rot pathogen,it was found that MdVPE1 gene editing can improve the disease resistance of apple callus.The successful application of CRISPR/cas9 gene editing technology in apple callus has laid an important foundation for the study of apple gene function.

关 键 词：基因编辑 苹果 CRISPR/Cas9 MdVPE1 

分 类 号：S661.1[农业科学—果树学]