中国农业科学 ›› 2021, Vol. 54 ›› Issue (2): 239-247.doi: 10.3864/j.issn.0578-1752.2021.02.001

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦籽粒黄色素含量检测方法的改良与应用

翟胜男(),刘爱峰,李法计,刘成,郭军,韩冉,訾妍,汪晓璐,吕莹莹,刘建军()   

  1. 山东省农业科学院作物研究所,济南 250100
  • 收稿日期:2020-04-27 接受日期:2020-07-06 出版日期:2021-01-16 发布日期:2021-02-03
  • 通讯作者: 刘建军
  • 作者简介:翟胜男,E-mail: zsn19870322@163.com
  • 基金资助:
    国家自然科学基金(31701420);山东省农业良种工程(2019LGC001);山东省农业科学院农业科技创新工程(CXGC2018E01);山东省重大科技创新工程项目(2018YFJH0602)

Improvement and Application of the Method for Determining Yellow Pigment Content in Wheat Grain

ZHAI ShengNan(),LIU AiFeng,LI FaJi,LIU Cheng,GUO Jun,HAN Ran,ZI Yan,WANG XiaoLu,LÜ YingYing,LIU JianJun()   

  1. Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100
  • Received:2020-04-27 Accepted:2020-07-06 Online:2021-01-16 Published:2021-02-03
  • Contact: JianJun LIU

摘要:

【目的】小麦籽粒黄色素含量是影响面制品颜色和营养品质的重要因素,为快速、准确、简便地检测小麦籽粒黄色素含量,对传统小麦籽粒黄色素测定方法AACC 14-50进行改良,为面制品颜色性状和营养品质遗传改良提供技术支撑。【方法】将酶标仪应用于小麦籽粒黄色素含量测定,对传统AACC 14-50法进行改良,分析改良方法的准确性和稳定性,探讨不同提取时间和保存时间对面粉黄色素含量的影响,利用新改良方法测定283份国内外小麦品种籽粒黄色素含量,进一步验证改良方法的有效性,发掘面制品颜色性状和营养品质优异的种质资源。【结果】改良方法与传统AACC 14-50法所测黄色素含量呈极显著正相关,相关系数达0.994(P<0.001),方差分析显示,2种方法所测黄色素含量在品种间差异均极显著(P<0.01)。应用改良方法对144份小麦品种籽粒黄色素含量进行重复测定,3次重复两两间的相关系数分别为0.997、0.998和0.998。藁优5218、冀师02-2和郑麦366面粉黄色素含量在提取时间为30 min时达到最大值,分别为0.93、1.24和1.53 μg·g-1,且提取时间为30—120 min时,黄色素含量保持在稳定水平(0.94—0.97、1.30—1.33和1.59—1.62 μg·g-1)。在室温保存0—20 d的藁优5218、冀师02-2和郑麦366的面粉,其黄色素含量没有显著变化(0.93—0.97、1.29—1.33和1.60—1.64 μg·g-1;P>0.05)。283份国内外小麦品种籽粒黄色素含量变异范围广,不同环境间相关系数为0.73—0.98。其中,黄淮麦区小麦品种黄色素含量平均值为1.15 μg·g-1,变异范围为0.51—2.42 μg·g-1;北部冬麦区平均值为1.57 μg·g-1,变异范围为0.90—2.52 μg·g-1;长江中下游麦区平均值为1.07 μg·g-1,变异范围为0.56—2.54 μg·g-1;国外品种平均值为1.61 μg·g-1,变异范围为0.94—2.48 μg·g-1。筛选到24份黄色素含量较低和26份黄色素含量较高的品种,可作为亲本材料用于面制品颜色性状和营养品质遗传改良。与传统方法相比,改良方法显著降低了工作量,缩短检测时间,效率提高12—15倍,且样本、试剂用量仅为原来的1/16,大大节约了成本。【结论】建立了一个准确稳定、经济高效、操作简便的小麦籽粒黄色素含量检测方法,可代替传统AACC 14-50法用于大规模样本黄色素含量测定。

关键词: 小麦, 面制品颜色, 营养品质, 酶标仪, 遗传改良

Abstract:

【Objective】The yellow pigment content (YPC) in wheat grain is an important factor affecting the color and nutritional quality of flour end-use products. To quickly, accurately and simply detect YPC in wheat grain, the traditional method for YPC determination, AACC 14-50, was improved, providing a technical support for genetic improvement of the color and nutritional quality of flour end-use products.【Method】In this study, improvements have been made to the AACC 14-50 method using a Multiskan Spectrum microplate reader. The accuracy and stability of the modified method were analyzed, and the influence of extraction time and preservation time on flour YPC was dissected. The YPC of 283 domestic and foreign wheat varieties were determined by the improved method to further verify its effectiveness and explore excellent resources for genetic improvement of the color and nutritional quality of flour end-use products. 【Result】 The results showed that YPC determined by the modified method was significantly and positively correlated with those by the traditional AACC 14-50 method, with a correlation coefficient of 0.994 (P<0.001), and analysis of variance showed that YPC measured by the two methods was significant among varieties. The YPC of 144 wheat varieties were measured repeatedly using the modified method, and the correlation coefficients of YPC among three replicates were 0.997, 0.998 and 0.998, respectively. The flour YPC of Gaoyou 5218, Jishi 02-2 and Zhengmai 366 reached the maximum value (0.93, 1.24 and 1.53 μg·g-1) at the extraction time of 30 min, and remained stable during 30-120 min (0.94-0.97 μg·g-1, 1.30-1.33 μg·g-1 and 1.59-1.62 μg·g-1). No significant changes were observed in the flour YPC of Gaoyou 5218, Jishi 02-2 and Zhengmai 366 stored at room temperature for 0-20 days (0.93-0.97 μg·g-1, 1.29-1.33 μg·g-1 and 1.60-1.64 μg·g-1; P>0.05). A wide range of YPC variation was identified in 283 domestic and foreign wheat varieties, with correlation coefficients ranging from 0.73 to 0.98 among different environments. Briefly, the mean YPC of varieties in the Huang-Huai wheat region was 1.15 μg·g-1, ranging between 0.51-2.42 μg·g-1. The average value of YPC in varieties from the Northern winter wheat region was 1.57 μg·g-1, ranging from 0.90 to 2.52 μg·g-1. The average value of YPC in the Middle-Lower reaches of the Yangtze River wheat region was 1.07 μg·g-1, and the variation ranged from 0.56 to 2.54 μg·g-1. The average YPC of foreign varieties was 1.61 μg·g-1, ranging from 0.94 to 2.48 μg·g-1. Twenty-four varieties with lower YPC and 26 with higher YPC were identified, which can be used as parents in wheat breeding programs to improve the color and nutritional quality of flour end-use products. Compared with the traditional AACC 14-50 method, the modified one reduces the workload, decreases the detection time, and increases the detection efficiency by about 12-15 times, and the consumption of samples and reagents is only 1/16 of those in the traditional method, which greatly reduces the cost. 【Conclusion】 The modified method for determining YPC in wheat grain established in this study is accurate, reliable, economical, efficient and simple, which can replace the traditional AACC 14-50 method for a large-scale determination of YPC.

Key words: Triticum aestivum, color of flour end-used products, nutritional quality, multiskan spectrum microplate reader, genetic improvement