NtCPS2基因对顺-冷杉醇与赤霉素合成代谢途径的影响

Effects of NtCPS2 gene on the anabolic pathways of cis-abienol and gibberellin

为探讨顺-冷杉醇合成基因NtCPS2对萜类代谢途径的影响,利用基因编辑技术CRISPR/Cas 9编辑烟草顺-冷杉醇合成基因NtCPS2,并筛选出纯合突变植株,命名为‘8306’编辑系;比较‘8306’编辑系(T_(2)-26、T_(2)-45、T_(2)-48)与野生型(WT)在顺-冷杉醇、赤霉素含量及合成关键基因的生物信息学方面的差异。结果表明:‘8306’编辑系(T_(2)-26、T_(2)-45、T_(2)-48)顺-冷杉醇含量平均减少86.20%,赤霉素含量平均增加27.05%;顺-冷杉醇合成途径中的关键基因表达量显著减小,赤霉素合成途径中正调控关键基因GA20ox表达量显著增加,负调控关键基因GA2ox表达量显著减小,并且GGPP(牻牛儿基牻牛儿基焦磷酸)合成的关键基因DXR表达量增加。NtCPS2基因不仅影响赖百当类二萜代谢途径,有效降低顺-冷杉醇的含量,并且影响赤霉素萜类代谢,增加赤霉素的含量。

To investigate the effect of cis-abienol synthesis gene NtCPS2 on terpenoid metabolic pathways,CRISPR/Cas 9 gene editing technology was used to edit the tobacco cis-abienol synthesis gene NtCPS2.The homozygous mutant plants were screened,and named as’8306’gene-editing series.The content and the key synthesis genes of cis-abienol and gibberellin were compared between the’8306’gene-editing series(T_(2)-26,T_(2)-45,T_(2)-48)and the wild type(WT).Compared with WT,the cis-abienol content in the’8306’gene-editing series was decreased by 86.20%,the gibberellin content was increased by 27.05%on average.The key gene′s expression in the synthesis pathway of cis-abienol was significantly reduced.In the gibberellin synthesis pathway,the expression level of GA20 ox,the key gene of positive regulation,was significantly increased.At the same time,the expression level of GA2 ox,the key gene of negative regulation,was significantly decreased.Moreover,the expression level of DXR,the key gene of GGPP synthesis,was increased.In conclusion,the NtCPS2 gene not only affected the labdane diterpenes metabolic pathway and effectively reduced cis-abienol content,but also affected the gibberellin metabolism and increased gibberellin content.