不同颜色大豆种皮成分及抗氧化性分析

Analysis of Seed Coat Composition and Antioxidant Activity of Soybean with Different Colors

为分析不同种皮颜色大豆种皮花色苷、总酚、总黄酮含量及抗氧化性差异,探讨类黄酮相关基因表达量与种皮颜色的关系,促进高抗氧化性的大豆种质的培育和筛选鉴定,以重组自交系(RIL)群体(♀中豆41×♂柘城小红豆)中4种纯合种皮颜色大豆及其亲本为试验材料,利用高效液相色谱法测定花色苷种类,利用分光光度计测定总酚和总黄酮含量,测定铁离子还原能力(FRAP)、DPPH·清除能力和ABTS^(+)·清除能力,评价体外抗氧化能力差异,对比大豆种子着色前后转录组,分析类黄酮途径相关基因的表达量变化,并探讨不同颜色种皮黄酮类物质含量与抗氧化能力及基因表达的关系。结果表明:黑色大豆种皮中花色苷含量最多且种类最丰富,种皮颜色主要与天竺葵素-3-葡萄糖苷含量有关,矢车菊-3-葡萄糖苷和矢车菊-3-半乳糖苷含量与黑色和紫红色素沉积有关。黑色大豆的总酚、总黄酮、总花青素、铁离子还原能力(FRAP)、DPPH·清除能力和ABTS^(+)·清除能力均最高,且均表现为黑色>紫红色>褐色>黄色,仅DPPH·清除能力在其他种皮颜色中差异不显著,说明黑色大豆种质在花色苷和抗氧化性方面更为优良,具有保健产品开发价值。总酚、总黄酮、总花青素含量与FRAP、DPPH·和ABTS^(+)·清除能力呈极显著正相关。CHS基因的表达与酚类物质含量和抗氧化能力呈极显著正相关。黑色种皮着色前后主要CHS基因表达显著升高,褐色种皮着色前后MYB转录因子表达显著升高,F3′5′H基因和F3′H基因的表达显著下降。不同种皮颜色着色沉积时类黄酮途径CHS基因的表达可以用来衡量大豆种皮的抗氧化能力。

In order to analyze the differences in anthocyanins,total phenols,total flavonoids and antioxidant activity of soybean seed coat colors with different seed coat colors,explore the relationship between as the expression of flavonoid-related genes and seed coat color,and promote the cultivation,screening and identification of soybean germplasm with high antioxidant resistance to reorganize.With four kinds of homozygous seed coat color soybean and parents of the recombinant inbred lines(RIL)population(♀Zhongdou 41×♂Zhechengxiaohongdou)as test materials,and the species of anthocyanins were determined by high performance liquid chromatography,and the total phenol and total flavonoid content was determined by spectrophotometer.Iron reduction capacity(FRAP),DPPH·clearing ability and ABTS^(+)·clearing ability were determined to evaluate the difference of antioxidant capacity in vitro.Compared the transcriptome before and after soybean seed coloration,analyzed the expression changes of flavonoid pathway-related genes,and to explore the relationship between flavonoid content and antioxidant capacity and gene expression in different color coats.The results showed that the content of anthocyanins in the seed coat of black soybean was the largest and the most abundant,and the color of the seed coat was mainly related to the content of geranium-3-glucoside and cornflower-3-glucoside and cornflower-3-galactoside content are associated with black and purplish pigment deposition.The contents of total phenols,total flavonoids,total anthocyanins,ferritin reducing ability(FRAP),DPPH·clearing ability and ABTS^(+)·clearing ability of black soybeans were the highest,and all manifested as black>purplish red>brown>yellow.DPPH·clearing ability was not significant in other coat colors.These results indicated that black soybean germplasm had better anthocyanins and antioxidant properties,and was valuable for developing health products.The contents of total phenols,total flavonoids and total anthocyanins were positively correlated with scavenging ability of FRAP,DPPH·and clearing ability,and the expression of CHS gene was positively correlated with phenolic content and antioxidant ability.The expression of main CHS gene was significantly increased before and after staining of black coat,while the expression of MYB transcription factor was significantly increased before and after staining of brown coat,and the expression of F3′5′H and F3′H genes were significantly decreased.The expression of CHS gene of flavonoid pathway could be used to measure the antioxidant capacity of soybean seed coat.