Effects of Co-expression of AtDXS and AtDXR on Terpene Metabolites in Tobacco (Nicotiana tabacum)
ZHANG Han1, LENG Lu1, ZU Qing-Xue2, NIE Zhong-Yang2, LIN Song2, CHENG Zhi-Jun3, LI Guo-Ming1, LU Yin-Gang1,*
1 College of Agriculture, Guizhou University, Huaxi 550025, China; 2 Kaiyang Branch of Guizhou Tobacco Company, Kaiyang 550300, China; 3 Hunan China Tobacco Industry Co., Ltd., Changsha 410014, China
Abstract 1-deoxy-d-xylulose-5-phosphate synthase (DXS) and 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) are the first and second rate limiting enzyme of the synthesis pathway of terpene precursor 1-deoxy-d-xylulose-5-phosphate (DXP), and play important regulatory roles in the secondary metabolism of tobacco (Nicotiana tabacum). In this study, based on the DXS and DXR in Arabidopsis thaliana,the CaMV 35S promoter and NOS terminator, which are the most commonly used plant gene expression, were selected to synthesize the whole gene sequences DXS-NOS and 35S-DXR,and the co-expression vector pSH737-DXS-DXR was constructed. The tobacco transgenic plants co-expressing DXS and DXR were successfully obtained through Agrobacterium tumefacien-mediated genetic transformation. GUS chemical activity assay, PCR and Southern blot showed that the fusion genes had been successfully introduced into the genome of regenerated tobacco plants and expressed. RT-PCR showed that the DXS gene expression in transgenic plants was significantly higher than that of the control (P<0.05). The results of GC-MS showed that the neophytodiene and glandular hair secretion of transgenic plants increased in varying degrees compared with the control (P<0.05). This study provides research materials for tobacco breeding by using genetic improvement technology, and provides a reference basis for further regulating the biosynthesis of tobacco terpenoids by means of metabolic engineering.
ZHANG Han,LENG Lu,ZU Qing-Xue, et al. Effects of Co-expression of AtDXS and AtDXR on Terpene Metabolites in Tobacco (Nicotiana tabacum)[J]. 农业生物技术学报, 2022, 30(8): 1480-1487.
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