Cloning and analysis of BpFLS gene from rape pollen based on transcriptome sequencing

Abstract

Abstract [Objective] Menyuan rapeseed in Qinghai Province is cold resistant and has strong resistance. Its pollen accumulates rich flavonoids. This study aims to provide reference for genetic engineering breeding to improve flavonol content. [Methods] This study used transcriptome sequencing technology to screen differentially expressed flavonol synthase genes from rapeseed pollen in Menyuan of Qinghai Province and Zhengzhou of Henan Province, and performed bioinformatics analysis and RACE-PCR cloning of the Bp- FLS gene. [Results] (1) Six differentially expressed flavonol synthase genes in rape pollen were identified, and BpFLS1-1 was the most differentially expressed and up-regulated in the pollen of Menyuan rapeseed. (2) The amino acid sequence encoded by BpFLS1-1 had both flavonol synthase activity and flavanone-3- hydroxylase activity. (3) BpFLS1-1 gene might contain a new coding region sequence with a length of 1 170 bp. [Conclusion] Results suggest that the BpFLS1-1 gene plays an important role in the accumulation of flavonoids in Menyuan rapeseed pollen and the stress resistance of rapeseed.

Keywords

• menyuan rapeseed
• pollen
• transcriptome
• bpfls gene
• bioinformatics analysis
• cloning