CRISPR/Cas9介导的番茄SlMAPK6突变对植株形态的影响

CRISPR/Cas9-Mediated Mutagenesis of SlMAPK6 in Tomato Affect Plant Morphology

促分裂原活化蛋白激酶(MAPK)级联信号参与调控植物的多种逆境、生长发育以及花青苷的合成通路。为探究SlMAPK6在番茄中的功能,本研究利用CRISPR/Cas9技术设计3个靶标位点定点编辑SlMAPK6,并获得突变体CRISPR-3和CRISPR-7。PCR和测序结果发现SlMAPK6敲除载体构建成功。通过对T1植株进行卡那霉素基因筛选、靶位点扩增测序及行脱靶效应分析,结果发现矮番茄植株成功突变且未脱靶,与野生型对照表型相比,所有突变植株表现出根系更发达、侧枝增多的表型,表明SlMAPK6在番茄植株形态建成的过程中发挥了重要作用,为进一步探究SlMAPK6在番茄生长发育中的作用奠定了基础。

MAPK cascade signals are involved in the regulation of a variety of stresses,growth and development,anthocyanin synthesis pathways in plants.In order to know the function of SlMAPK6 in tomato,three target sites were designed for fixed-point editing SlMAPK6 by using CRISPR/Cas9,and mutant CRISPR-3 and CRISPR-7 were obtained The results of PCR and sequencing showed that the SlMAPK6 knockout vector was successfully constructed.Through kanamycin gene screening,target site amplification,sequencing and off-target effect analysis of T1 plants,the dwarf tomato plants were mutated successfully and did not miss the target.Through phenotypic observation and measurement,compared with the wild type control,all the mutants showed more flourished root system and lateral branches.The results showed that SlMAPK6 played an important role in the process of tomato plant morphogenesis,which laid a foundation for further exploring the role of SlMAPK6 in tomato growth and development.