摘要
为了培育无腋芽或少腋芽烟草,基于CRISPR/Cas9系统,对烟草NtLS基因进行靶向基因组编辑,构建了2个特异靶向NtLS基因的CRISPR/Cas9载体cLS1和cLS2,并借助农杆菌介导的烟草转基因技术转化烤烟品种K326,获得20株抗卡那霉素的阳性烟株。10株阳性烟株经过PCR扩增、测序和比对,检测到2株发生蛋白翻译错误的T_0突变烟株。研究结果不仅为腋芽形成的分子机理研究创制了有价值的突变体,而且为培育无腋芽或少腋芽优质烤烟品系提供了遗传材料。
To breed new tobacco varieties with no or less axillary buds, based on CRISPR/Cas9 system, the DNA sequence of NtLS was chosen as the target site for designing two CRISPR/Cas9 vectors(cLS1 and cLS2).Agrobacterium tumefaciens-mediated transformation of cv. K326 was used and twenty kanamycin-resistance tobacco plants(T_0 generation) were obtained. Ten positively transformed tobacco plants were analyzed by PCR amplification, sequencing and alignment, and some mutations of NtLS protein were detected in two plants.These research results provide useful mutants to study the molecular mechanism of axillary bud formation and create genetic materials for new flue-cured tobacco varieties with no or less axillary buds.
引文
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