大豆脂肪氧化酶缺失种质生化鉴定方法比较及其营养品质分析  

Comparison of Biochemical Identification Methods and Nutritional Quality Analysis for Lipoxygenase Mutants in Soybean

作　　者：马彩优 张晟瑞 李斌[2] 李静[2] 杜维俊[1] 孙君明[2] MA Cai-you;ZHANG Sheng-rui;LI Bin;LI Jing;DU Wei-jun;SUN Jun-ming(College of Agriculture,Shanxi Agricultural University,Jinzhong 030801,China;Institute of Crop Sciences,Chinese Academy of Agricultural Sciences/National Engineering Research Center of Crop Molecular Breeding/MARA Beijing Key Laboratory of Soybean Biology,Beijing 100081,China)

机构地区：[1]山西农业大学农学院,山西晋中030801 [2]中国农业科学院作物科学研究所/作物分子育种国家工程研究中心/农业农村部北京大豆生物学重点实验室,北京100081

出　　处：《大豆科学》2022年第2期150-158,共9页Soybean Science

基　　金：中国农业科学院农业科技创新工程(2060302-2);山西农业大学省部共建有机旱作农业国家重点实验室自主研发项目(202105D121008-3-8);山西农业大学育种工程项目(YZGC096)。

摘　　要：大豆脂肪氧化酶(LOX)是一种重要的豆腥味因子,为了筛选适宜的LOX鉴定方法和优异的LOX缺失育种亲本,本研究以大豆LOX缺失的近等基因系(Century和Suzuyutaka)和6个低豆腥味品种为材料,采用比色法、变性聚丙烯酰胺凝胶电泳(SDS-PAGE)和等点聚焦聚丙烯凝胶电泳(IEF-PAGE)技术鉴定LOX缺失情况,比较3种鉴定方法的优缺点;同时,采用近红外光谱(NIRS)、超高液相色谱(UPLC)、气相色谱(GC)和高效液相色谱(HPLC)等方法分析LOX缺失种质的主要营养品质组分(蛋白质、油分、可溶性糖、脂肪酸、异黄酮、维生素E和类胡萝卜素)含量差异。结果表明:比色法简便快捷,试剂需要现配现用;SDS-PAGE方法简单且成本低,但分辨率较差;而IEF-PAGE方法检测准确性较高,但成本也较高。品质分析发现,Century近等基因系的不同营养品质组分差异显著;而Suzuyutaka近等基因系的蛋白质、油分和类胡萝卜素组分差异显著。综上,针对大量种质材料筛选可先采用比色法进行快速初筛,然后结合IEF-PAGE法进行精准鉴定,既可加速低豆腥味大豆育种进程,同时可降低检测成本。筛选出具有不同营养品质组分的LOX缺失的优异种质,可为低豆腥味大豆品种选育提供可靠的育种和亲本材料。Soybean lipoxygenase(LOX)is the important off-flavor factor in soybean.In order to screen suitable LOX identification methods and excellent parents of missing sports species,we selected two near-isogenic line populations(NIL,Century and Suzuyutaka)with LOX mutants and six cultivars with low beany flavor as materials,and determined the LOX mutants using colorimetry,sodiumdodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)and isopoint focusing polyacrylamide gel electrophoresis(IEF-PAGE)biochemical technique,and compared their advantages and disadvantages among three methods in this study.Moreover,we also analyzed the main nutritional quality components,including protein,oil,soluble sugar,fatty acid,isoflavone,vitamin E and carotenoids in the LOX mutants using NIRS,UPLC,GC and HPLC methods.The results showed that the colorimetric method was a simple and fast technique,however,its reagents need to be prepared and used in time,the SDS-PAGE method was a simple and low-cost technique with poor resolution,and the IEF-PAGE had the highest detection accuracy with high cost.The quality analysis showed that the main nutritional quality components were significantly different in the NILs of Century,while the protein,oil and carotenoid contents were significantly different in the NILs of Suzuyutaka.It can conclude that in a large-scale of breeding lines,we can use the colorimetric method to rapidly screen LOX mutants firstly,and then employed the IEF-PAGE to validate the results,which can not only speed up the breeding progress of soybean cultivars with low beany flavor,but also reduce the detection cost.Otherwise,the cultivars with various elite quality traits selected can provide the key parents for low off-flavor soybean breeding in the future.

关 键 词：大豆脂肪氧化酶 比色法 SDS-PAGE IEF-PAGE 营养品质 低豆腥味 

分 类 号：S565.1[农业科学—作物学]