甘蓝型油菜不同花色转录组分析

Transcriptome Analysis of Brassica napus with Different Flower Colors

为了找出甘蓝型油菜不同花色差异基因,利用4种不同花色(白色,土黄色,黄色以及金黄色)甘蓝型油菜为试材,采用紫光-可见光分光光度计测出不同花色甘蓝型油菜花色色素成分的种类和吸光度值差异,最后利用高通量测序技术,对盛开期的油菜花瓣进行转录组测序,然后再根据4种不同花色的甘蓝型油菜基因表达谱的差异,筛选出与甘蓝型油菜花色变异相关基因。结果表明,扫描得出的RGB值发现主要由于B值决定花瓣颜色,其中白花的B值最大(>200),金黄色花的B值最小(<5)。用色素提取液无水乙醇:丙酮:水(4.5∶4.5∶1)提取花瓣的色素,测量200～600 nm波长吸光度值,发现在400～500 nm间变化有规律,可以得出是类胡萝卜素含量差异造成。对4个花色材料进行转录组分析,进一步说明造成其差异的基因位于类胡萝卜素合成通路上。本研究对后续优质油菜的选育提供理论指导。

In order to find out the differential gene in Brassica napus with different colors, four Brassica napus varieties with different flower colors （white, light yellow, yellow, and golden yellow） were selected as test materials. Purple-visible spectrophotometer was used to determine the species and absorbance value differences of the pigment species in Brassica napus with different colors. Finally, the high-throughput sequencing technology was used to sequence the transcriptomes of petals of flowering rapeseed, and then according to the differences of four Brassica napus varieties in gene expression profiles, the genes related to flower color variation in Brassica napus were screened out. The results showed that the B value of the RGB values obtained from the scan mainly determined the color of the petal, in which the B value of the white flower was the largest （〉200） and of the golden flower was the smallest （〈5）. The pigment of the flower petals was extracted by pigment extraction solution with the ratio of absolute ethanol： acetone： water of 4.5：4.5：1, and the absorbance at 200～600 nm wavelength was measured. It was found that there was a regular change between 400 nm and 500 nm, which was caused by the difference in carotenoid content. A transeriptome analysis was performed on the four flower color materials to further demonstrate that the genes responsible for the differences were located on the carotenoid synthesis pathway. The study could provide theoretical guidance for the subsequent breeding of high quality rapeseed.