基于CRISPR/Cas9技术的大白菜内源硫化氢生成酶LCD基因突变体构建  

Construction of Hydrogen Sulfide Producing Enzyme LCD Mutants in Chinese Cabbage Based on CRISPR/Cas9 Technology

作　　者：马晓丽[1,2] 景巧丽[1] 裴雁曦[1] Ma Xiaoli;Jing Qiaoli;Pei Yanxi(shool of Life Science,Shanxi University,Taiyuan 030006,China;College of Biological Science and Technology,Jinzhong University,Jinzhong 030600,China)

机构地区：[1]山西大学生命科学学院,太原030006 [2]晋中学院生物科学与技术学院,晋中030600

出　　处：《中国细胞生物学学报》2018年第11期1799-1805,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31372085)资助的课题~~

摘　　要：硫化氢(hydrogen sulfide, H2S)是植物细胞内源信号分子,对植物的生长、发育和抗逆性具有重要的调节作用。L-半胱氨酸脱巯基酶(L-cysteine desulfydrase, LCD)是合成内源H2S的关键酶。大白菜(Brassica campestris)是我国北方主要的蔬菜。为研究H2S信号在大白菜中的生理作用,并为育种工作提供新材料,该研究采用CRISPR/Cas9基因编辑技术对白菜LCD基因进行了敲除。利用农杆菌(LBA4404)介导的遗传转化法将基因编辑载体pYAO-LCD成功转化大白菜"中白60",获得基因编辑的遗传转化植株4株。DNA测序表明,基因编辑后植株在靶标位点的碱基发生了相应的基因编辑。基因编辑植株中内源H2S含量有不同程度的降低。Hydrogen sulfide(H2S) is an important gasotransmitter in plant cells and plays an important role in regulating plant growth, development and stress resistance. L-cysteine desulfydrase(LCD) is the key enzyme to produce endogenous H2S. Chinese cabbage(Brassica campestris) is one of the main vegetable crops in northern China. In order to study the physiological function of H2 S signal in Chinese cabbage and provide new materials for breeding, the CRISPR/Cas9 gene editing technique was used to knock out the LCD gene. Gene editing vector pYAO-LCD was transformed into ‘Zhongbai 60’ by genetic transformation mediated by Agrobacterium tumefaciens(LBA4404), and 4 plants of genetic transformation were obtained by gene editing. DNA sequencing showed that the target site was correspondingly edited. The endogenous H2S content in different editors decreased to varying degrees.

关 键 词：大白菜 硫化氢信号 L-半胱氨酸脱巯基酶 CRISPR/Cas9技术 遗传转化 

分 类 号：Q943.2[生物学—植物学]