Construction and transformation of LmPDS gene RNAi vector in Lemna minor [绿萍LmPDS基因RNAi载体的构建及遗传转化]

作者：Liu Y.; Fang Y.; Jin Y.; Du A.; Yang G.; Guo L.; Tan L.; He K.; Zhao H.^*

来源：Chinese Journal of Applied and Environmental Biology, 2020, 26(2): 280-286.

DOI：10.19675/j.cnki.1006-687x.201904004

• Genetic transformation
• Lemna minor L
• LmPDS gene
• RNA interference

The phytoene desaturase gene (PDS) encodes the key enzyme for the biosynthesis of carotenoid. Silencing or inhibition of PDS gene expression leads to the appearance of mottled, chlorosis, or albino leaves. It is of great importance to select the LmPDS gene for the construction and transformation of the LmPDS gene RNAi vector for RNA interference in Lemna minor. In this study, the 595-base pair LmPDS gene fragment was selected as the interference fragment and the cloned LmPDS interference fragment was inserted in the plant expression vector pCAMBIA2300-35S-OCST using the intermediate vector pUCCRNAi. The constructed RNAi vector was transformed into the callus of L. minor by Agrobacterium-mediated transformation. Then, using the callus resistance screen, plant regeneration, plant culture, and marker gene NPTII detection processes, 70 plants were obtained, with four being transgenic positive plants. After fluorescence quantitative polymerase chain reaction analysis, lycopene content detection, and phenotype observation, the relative expression of LmPDS in all transgenic positive plants decreased significantly compared with 28.8%-40.5% of the control plants. The lycopene content in the transgenic positive plants also decreased significantly and these plants showed an obvious albino phenotype in their leaves. In conclusion, we successfully constructed the LmPDS gene RNAi vector in L. minor and the application of RNAi can provide materials and methods for further research in gene function analysis of L. minor.

• 单位
  中国科学院大学