基于RNA干扰初步探究光受体对三孢布拉霉β-胡萝卜素合成的影响

o analyze the effect of interfering with the expression of btwc-1 (1a, 1b, 1c) and btwc-2 (2a, 2b, 2c, 2d) on β-carotene synthesis in Blakeslea trispora. The RNA interference vectors of btwc-1 and btwc-2 genes were constructed, and the efficiency of shRNA interference sequences on target gene expression was measured by real-time fluorescence quantitative PCR. The β-carotene content and the expression of β-carotene structural genes carB and carRA were measured in RNA-interfered strains and control strains at different times of continuous darkness and blue light pulses for 30 min. The expression of carB and carRA genes was determined. The results showed that btwc-1 and btwc-2 were transcriptionally activated under light-induced conditions; the maximum efficiency of interfering with the expression of btwc-1a, btwc-1b, btwc-1c , btwc-2a, btwc-2b, btwc-2c and btwc-2d was 72%, 67%, 63%, 66%, 51%, 86% and 61%; The accumulation of β-carotene and the expression of carB and carRA in the interference strains of btwc-1a, btwc-1c, btwc-1a, btwc-2a, and btwc-2c at different time points of the optical pulse mode were basically lower than those in the control group. Therefore, the synthesis of β-carotene in Blakeslea trispora is affected by the expression of btwc-1a, btwc-1c, btwc-1a, btwc-2a, and btwc-2c, and interference with the above gene expression can reduce the transcription level of β-carotene structural genes and inhibit the synthesis of β-carotene.