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Journal of Integrative Agriculture  2021, Vol. 20 Issue (5): 1193-1203    DOI: 10.1016/S2095-3119(20)63328-7
Special Issue: 油料作物合辑Oil Crops
Crop Science Advanced Online Publication | Current Issue | Archive | Adv Search |
Establishment and application of an accurate identification method for fragrant soybeans
ZHANG Yong-fang1, 2, 3, ZHANG Chun-yan3, ZHANG Bo4, YIN Man3, HONG Hui-long3, YU Li-li3, GAO Hua-wei3, GU Yong-zhe3, LIU Zhang-xiong3, LI Fu-heng1, QIU Li-juan3 
1 College of Life Sciences, Northeast Agricultural University, Harbin 150030, P.R.China
2 College of Life Sciences, Shanxi Datong University, Datong 037009, P.R.China
3 National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI)/Key Laboratory of Germplamsm Utilization, Ministry of Agriculture and Rural Affairs/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
4 School of Plant and Environmental Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24060, USA
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为筛选大豆香味种质,建立大豆叶片中香味特征化合物2-乙酰基-1-吡咯啉(2-acetyl-l-Pyrroline,2AP)的鉴定方法。本研究通过单因素及三因素四水平(L9 (34)的正交试验,以峰形、总峰面积及检测样品时间为考察指标,建立了利用气质联用仪(GC-MS)快速检测香味的方法,明确了仪器运行最佳参数包括:柱温70℃,进样口温度180℃,以及样品最优萃取时间条件(酒精含量1ml、NaCl含量0.1g,超声时间10min,萃取时间为1h)。该检测方法重复性好、简单快速、样本试剂消耗少,可精准快速测定2AP含量。利用该方法对不同地理来源的101个大豆基因型进行了分析筛选。结果表明, 2-AP平均含量为0.29ppm,变幅为0.094ppm到1.816ppm,遗传多样性指数为0.54。可被划分为3个等级,其中,1级香型大豆有7份,包括中龙608、黑农88、哈13-2958、红面豆、黑农82、黄毛豆、吉育21。本研究建立的方法及筛选的优异种质为大豆香味育种和基因发掘提供了技术和材料支撑。

In order to screen the aroma characteristics of soybean, a new method was established which can quickly quantify the content of 2-acetyl-1pyrroline (2-AP), an important compound related to soybean aroma, using gas chromatography-mass spectrometry (GC-MS).  Based on peak profile, total peak area and retention time as test indexes, an accurate identification method for fragrant soybeans was established.  The optimum parameters of the protocol consisted of column temperature 70°C, sample injector temperature 180°C, optimum extraction alcohol content 1 mL, NaCl content 0.1 g, ultrasonication time 10 min, and extraction time 1 h, which were established by using the orthogonal test of single factors and three factors with four levels (L9(3)4).  2-AP content of leaves had significant correlations with seeds, which were easier to measure.  The protocol was simple and easy to carry out, consumed only small amounts of reagents, and provided accurate and reliable results with good reproducibility.  A total of 101 soybean genotypes from different geographical sources were analyzed using this protocol.  The results showed that the average content of 2-AP was 0.29 mg L–1, ranging from 0.094 to 1.816 mg L–1, and the genetic diversity index was 0.54.  Among all genotypes-tested, they were classified into three grades, including seven elite genotypes identified as “grade one fragrant soybeans”, which were Zhonglong 608, Heinong 88, Ha13-2958, Hongmiandou, Heinong 82, Huangmaodou, and Jiyu 21.  These results provide both an identification technique and several elite aroma genotypes for gene discovery and good quality breeding in soybean.
Keywords:  soybean        2-acetyl-1pyrroline        GC-MS        quantification method        germplasm  
Received: 02 March 2020   Accepted:
Fund: This work was supported by the National Key R&D Program for Crop Breeding (2016YFD0100201) and Scientific Innovation Program of of the Chinese Academy of Agricultural Sciences (CAAS).
Corresponding Authors:  Correspondence QIU Li-juan, Tel: +86-10-82105843, Fax: +86-10-82105840, E-mail:; LI Fu-heng, Tel/Fax: +86-451-55190002, E-mail: lifuheng1963@126. com    
About author:  ZHANG Yong-fang , E-mail:;

Cite this article: 

ZHANG Yong-fang, ZHANG Chun-yan, ZHANG Bo, YIN Man, HONG Hui-long, YU Li-li, GAO Hua-wei, GU Yong-zhe, LIU Zhang-xiong, LI Fu-heng, QIU Li-juan. 2021. Establishment and application of an accurate identification method for fragrant soybeans. Journal of Integrative Agriculture, 20(5): 1193-1203.

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