烟草单倍体基因编辑系统的建立

Establishment of gene editing system for haploid in tobacco

【背景和目的】单倍体材料作为转化受体具有不受同源染色体联会配对影响、转化效率较高、外源基因在后代基因组中稳定性较好、加倍后即可获得纯合转化植株等优点,结合基因编辑CRISPR/Cas9系统,可大大加快烟草品种选育进程。【方法】通过花粉离体培育获得单倍体,利用CRISPR/Cas9系统编辑八氢番茄红素脱氢酶基因位点,产生白化单倍体植株。【结果】(1)经过4℃处理4~6天的实验组,花药愈伤组织形成率最高可达11%;(2)获得转化后单倍体植株120株,其中白化86株(白化率71.67%),红花大金元51株,其中白化24株(白化率47.06%)。【结论】(1)适当低温处理可提高花药培育单倍体效率;(2)以单倍体为受体的基因编辑效率有所提高。

[Background]As transformation receptors,haploid has the advantages of not being affected by the pairing of homologous chromosomes,high transformation efficiency,good stability of exogenous genes in the offspring genome,and homozygous transformed plants that can be obtained after doubling.Combined with gene editing CRISPR/Cas9 system,the breeding process of tobacco varieties can be greatly accelerated.[Methods]Haploid was obtained by pollen culture in vitro,and the gene locus of hydrolycopene dehydrogenase was edited by CRISPR/Cas9 system to produce albino haploid plants.[Results]Test data show:1)The callus formation of anther rate was up to 11%after 4℃treatment for 4~6 days;2)120 transformed haploid plants were obtained,of which 86 were albino(albino rate 71.67%),51 were transformed honghuadajinyuan,and 24 were albino(album rate 47.06%).[Conclusion]1)A proper low temperature treatment can improve the efficiency of anther culture for haploid;2)The efficiency of gene editing with haploid receptor was improved.