基于转录组测序的油菜花粉BpFLS基因克隆与分析

Cloning and analysis of BpFLS gene from rape pollen based ontranscriptome sequencing

【目的】青海门源油菜耐寒冷、抗性强,其花粉积累了丰富的类黄酮物质,该研究可为提高黄酮醇含量的基因工程育种提供参考依据。【方法】研究采用转录组测序技术从青海门源油菜花粉和河南郑州油菜花粉筛选差异表达的黄酮醇合成酶基因,并对BpFLS基因进行生物信息学分析和RACE-PCR克隆。【结果】(1)筛选出6个差异表达的油菜花粉黄酮醇合成酶基因,BpFLS1-1基因在青海门源油菜花粉中差异最大且表达上调。(2)BpFLS1-1基因编码的氨基酸序列具有黄酮醇合成酶活性和黄烷酮-3-羟化酶活性。(3)BpFLS1-1基因可能含有新的蛋白质编码区序列,其长度为1 170 bp。【结论】BpFLS1-1基因在门源油菜花粉积累丰富的类黄酮物质及油菜抗逆性方面发挥重要功能。

[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 1170 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.