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作物学报

作物学报 ›› 2019, Vol. 45 ›› Issue (12): 1891-1898.doi: 10.3724/SP.J.1006.2019.94016

• 耕作栽培·生理生化 • 上一篇    下一篇

单粒花生主要脂肪酸含量近红外预测模型的建立及其应用

李建国,薛晓梦,张照华,王志慧,晏立英,陈玉宁,万丽云,康彦平,淮东欣(),姜慧芳,雷永(),廖伯寿   

  1. 中国农业科学院油料作物研究所 / 农业部油料作物生物学与遗传育种重点实验室, 湖北武汉 430062
  • 收稿日期:2019-05-15 接受日期:2019-06-20 出版日期:2019-07-16 网络出版日期:2019-07-16
  • 通讯作者: 淮东欣,雷永
  • 作者简介:李建国, E-mail: 15611821765@163.com
  • 基金资助:
    本研究由玛氏-中国高油酸花生育种计划项目(MARS-China HOAP 2013-2018);国家自然科学基金项目(31671734);国家自然科学基金项目(31461143022);国家自然科学基金项目(31770250);国家自然科学基金项目(31371662);中央级科研院所基本科研业务费专项(Y2018PT52);国家现代农业产业技术体系建设专项(CARS-13)

Establishment and applicant of near-infrared reflectance spectroscopy models for predicting main fatty acid contents of single seed in peanut

Jian-Guo LI,Xiao-Meng XUE,Zhao-Hua ZHANG,Zhi-Hui WANG,Li-Ying YAN,Yu-Ning CHEN,Li-Yun WAN,Yan-Ping KANG,Dong-Xin HUAI(),Hui-Fang JIANG,Yong LEI(),Bo-Shou LIAO   

  1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences / Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, Hubei, China
  • Received:2019-05-15 Accepted:2019-06-20 Published:2019-07-16 Published online:2019-07-16
  • Contact: Dong-Xin HUAI,Yong LEI
  • Supported by:
    This study was supported by the Mars-China High Oleic Acid Peanut Breeding Project(MARS-China HOAP 2013-2018);the National Natural Science Foundation of China(31671734);the National Natural Science Foundation of China(31461143022);the National Natural Science Foundation of China(31770250);the National Natural Science Foundation of China(31371662);the Fundamental Research Funds for Central Non-profit Scientific Institution(Y2018PT52);the China Agriculture Research System(CARS-13)

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摘要：

脂肪酸组成是影响花生营养价值和货架寿命的主要因素, 高油酸花生以其营养保健价值高、化学稳定性好、耐储藏等特点, 深受广大消费者和花生加工企业的喜爱。因此, 培育高油酸品种是花生育种的重要目标, 建立快速、高效、准确检测花生中主要脂肪酸含量的无损方法是加快花生脂肪酸改良和高油酸品种选育进程的重要技术保障。本研究利用近红外光谱技术建立了可以非破坏性地快速检测单粒花生中油酸、亚油酸、棕榈酸含量的数学模型, 其中油酸模型的决定系数(R 2)为0.907, 均方差为3.463; 亚油酸模型的决定系数为0.918, 均方差为2.824; 棕榈酸模型的决定系数为0.824, 均方差为0.782。使用100粒花生验证该模型的准确性, 结果油酸、亚油酸和棕榈酸的近红外预测值与化学值的相关系数分别为0.88、0.90和0.71, 表明此模型可以准确地预测单粒花生中这3种脂肪酸的含量。本研究借助该模型建立了一种不依赖分子标记的快速、高效选育高油酸花生的方法, 并成功应用于高油酸花生育种, 选育出高油酸花生品种中花215。

关键词: 单粒花生, 脂肪酸, 近红外模型, 高油酸, 育种方法

Abstract:

Profile of fatty acid is the main factor determining the nutritional value and shelf life of peanut. High oleate peanut has been increasingly favored by customers and peanut processing enterprises, as it provides a prolonged shelf-life and a beneficial effect on human health. Therefore, creating high oleate peanut cultivar is an important objective for peanut breeding. Developing a nondestructive method which could detect the fatty acid composition in peanut rapidly, efficiently and accurately is an important technical support for accelerating the processing. In this study, models for predicting the contents of main fatty acids (oleic acid, linoleic acid and palmitic acid) in a single peanut seed were built up using near-infrared reflectance spectroscopy (NIRS). The coefficient of multiple determination (R 2) and the root mean squared error of external calibration (RMSECV) of prediction models were 0.907 and 3.463 for oleic acid, 0.918 and 2.824 for linoleic acid, 0.824 and 0.782 for palmitic acid, respectively. One hundred peanut seeds were analyzed by both NIR and gas chromatography (GC) to validate the accuracy of the prediction models. The correlation coefficients of oleic acid, linoleic acid and palmitic acid between NIR values and GC values were 0.88, 0.90, and 0.71, respectively, suggesting that these models could accurately predict the contents of the three main fatty acids in a single peanut seed. Furthermore, based on these prediction models, a breeding method of high oleate peanut without assistance of molecular marker was developed, and a high oleate peanut cultivar Zhonghua 215 was successfully bred.

Key words: single peanut seed, fatty acid, near infrared model, high oleic acid, breeding method

图1

本试验中使用的近红外检测仪Unity-SpectrastarXL"

图2

样品集的光谱分析"

表1

建立数学模型使用的单粒花生种子的油酸、亚油酸和棕榈酸含量"

棕榈酸(C16:0)含量
Palmitic acid content		 油酸(C18:1)
含量
Oleic acid
content		 亚油酸(C18:2)含量
Linoleic acid content
样本数
Sample size		 1013 1195 1202
最小值Minimum (%) 5.9 32.8 0.7
最大值Maximum (%) 16.6 80.8 43.9
均值Mean (%) 11.1 51.8 26.5

图3

本方法中油酸模型、亚油酸模型和棕榈酸模型的决定系数"

附表1

检验数学模型准确性使用的单粒花生种子的油酸、亚油酸和棕榈酸含量的NIR预测值与化学值"

样品
Sample		 C18:1 C18:2 C16:0
NIR GC NIR GC NIR GC
1 52.9 49.9 26.0 31.9 11.1 10.4
2 80.0 74.1 1.6 9.2 6.7 6.5
3 70.3 60.0 10.1 21.6 8.5 10.5
4 73.5 69.9 8.0 14.3 7.8 7.5
5 68.5 64.6 11.4 14.3 8.5 8.1
6 67.0 49.7 13.3 29.1 8.7 8.5
7 69.2 59.9 11.5 20.8 8.5 10.3
8 64.4 39.1 14.4 35.5 9.7 13.9
9 81.2 69.7 1.3 12.3 6.1 6.4
10 83.7 69.6 -1.7 11.3 5.6 9.2
11 82.4 74.6 -0.5 9.6 5.9 7.7
12 67.8 54.9 13.5 23.5 8.7 10.6
13 54.4 49.7 24.4 29.9 11.1 9.4
14 89.5 85.4 -5.8 1.4 4.8 5.8
15 88.2 85.1 -4.9 1.2 5.1 5.1
16 57.1 44.7 22.8 32.5 10.3 15.5
17 86.2 79.8 -3.6 2.5 5.3 6.9
18 93.3 79.8 -9.2 2.4 3.9 7.0
19 55.7 38.5 23.8 37.2 10.4 15.6
20 75.0 69.6 6.4 12.8 7.1 6.1
21 91.8 79.7 -7.7 2.1 4.5 7.5
22 66.7 54.9 13.7 24.3 8.6 12.6
23 60.3 44.8 20.3 33.0 10.0 13.8
24 59.2 49.8 20.1 32.0 10.2 9.9
25 62.0 44.8 17.5 31.1 9.3 11.0
26 90.8 79.7 -6.9 2.0 5.2 6.8
27 81.1 74.8 1.1 7.0 6.3 7.6
28 68.2 59.9 12.3 21.2 9.2 8.9
29 71.8 55.0 9.6 25.8 8.2 8.8
30 77.4 69.6 4.5 11.7 7.1 6.4
31 74.7 74.2 5.9 10.6 7.7 6.8
32 85.2 84.9 -2.1 2.1 5.7 6.0
33 83.8 74.3 -1.8 8.8 6.1 5.6
34 66.1 45.0 25.3 32.3 10.5 12.9
35 71.4 59.8 9.5 20.8 8.2 7.4
36 73.3 69.8 7.6 11.9 8.1 8.0
37 61.1 54.8 18.5 26.1 10.1 8.0
38 72.2 54.9 8.5 26.1 7.7 10.6
39 90.9 79.5 -7.9 2.5 4.4 6.2
40 86.5 44.9 -4.2 34.2 5.7 11.7
41 51.3 39.5 27.2 40.6 12.2 10.6
42 48.4 38.9 29.9 41.7 12.2 10.9
43 79.9 74.4 1.7 7.6 6.6 5.9
44 54.6 45.0 23.9 36.2 11.2 10.0
45 70.3 55.0 11.8 24.0 8.2 12.7
46 64.3 54.9 16.7 25.1 9.5 11.1
47 69.3 64.9 11.4 18.4 8.6 6.5
48 74.1 65.0 6.9 14.6 7.6 7.4
49 73.0 69.6 7.9 13.6 7.5 6.6
50 58.5 49.6 20.2 31.2 10.0 9.8
51 91.2 79.7 -7.6 2.4 4.6 7.5
52 87.0 79.7 -3.6 2.3 6.0 7.8
53 72.2 64.7 8.6 16.5 7.7 9.2
54 69.4 59.9 10.7 21.4 8.3 9.7
55 49.9 45.0 28.5 36.5 11.9 10.5
56 91.8 79.9 -9.2 2.4 4.3 6.3
57 70.5 38.7 10.0 36.9 8.0 12.7
58 73.7 59.8 4.0 20.4 6.5 8.9
59 87.7 79.6 -4.8 2.3 5.4 7.1
60 67.0 60.0 13.6 22.5 8.7 10.9
61 66.1 49.8 14.7 29.7 9.2 12.8
62 82.9 85.6 -0.4 1.4 6.0 6.3
63 81.6 85.6 -0.4 1.4 6.0 6.3
64 64.4 54.8 16.1 26.8 9.5 9.2
65 78.2 74.4 3.7 10.2 7.0 8.3
66 83.6 85.3 -1.4 1.4 5.8 5.6
67 89.3 85.6 -6.4 1.1 5.0 5.3
68 59.5 44.6 19.9 33.9 10.5 11.1
69 58.9 44.9 21.1 35.4 10.0 10.3
70 45.6 39.8 32.6 40.6 13.3 10.8
71 71.6 64.9 8.6 18.2 8.3 8.4
72 72.7 64.8 7.7 16.1 7.9 9.5
73 80.2 74.2 0.8 8.8 6.4 5.9
74 84.2 85.1 -0.6 1.4 5.8 6.6
75 76.0 69.9 4.8 12.4 6.8 8.3
76 87.5 64.7 -5.0 12.5 5.1 7.9
77 88.2 85.6 -4.8 1.7 4.9 5.4
78 60.8 38.7 18.1 37.0 9.6 11.3
79 54.0 38.7 24.6 37.9 11.4 12.5
80 58.6 44.8 20.7 32.1 10.7 12.2
81 50.6 39.7 28.8 40.5 11.9 9.6
82 53.9 49.9 25.0 28.3 10.8 7.6
83 84.9 84.8 -1.8 1.6 5.4 6.5
84 56.9 50.0 23.2 30.0 10.3 9.1
85 71.1 74.2 10.2 9.4 8.2 8.0
86 75.2 69.7 6.9 14.7 7.4 7.2
87 62.6 54.8 17.5 26.4 9.5 9.2
88 68.7 64.8 11.4 17.9 8.4 7.3
89 73.7 65.0 7.5 16.7 7.8 7.6
90 72.6 59.8 9.1 20.5 7.8 11.3
91 59.7 39.8 20.5 35.4 9.8 13.6
92 59.8 49.6 19.6 28.1 10.2 11.3
93 89.8 79.9 -6.8 2.5 5.2 6.3
94 66.8 59.9 14.2 19.7 8.9 8.6
95 86.5 84.8 -4.1 1.6 5.7 6.4
96 75.0 69.8 6.1 14.5 7.4 7.4
97 67.3 54.9 13.4 24.1 9.1 12.5
98 77.4 74.1 4.6 10.4 6.8 7.0
99 69.1 50.0 12.0 29.2 8.6 11.2
100 68.4 59.9 12.4 20.3 8.8 9.3

图4

检验使用的单粒花生种子的油酸(A)、亚油酸(B)、棕榈酸(C)近红外预测值与化学值相关性 C18:1: 油酸; C18:2: 亚油酸; C16:0-棕榈酸。"

附表2

单粒近红外光谱检测油酸含量在70%以上的F2花生种子"

样品
Sample		 油酸
C18:1 (%)		 样品
Sample		 油酸
C18:1 (%)
6-1 82.9 4-13 73.8
2-11 82.2 9-10 73.4
10-6 81.2 5-12 73.3
8-12 80.0 3-16 73.2
3-3 79.7 9-20 73.2
11-31 79.2 5-1 73.2
6-2 78.8 4-12 72.8
7-18 78.3 3-16 72.6
8-8 77.7 10-15 72.5
11-42 76.8 9-11 72.2
4-11 76.2 11-8 72.1
4-14 75.5 4-7 72.1
4-2 75.5 8-19 71.8
5-7 75.1 9-21 71.5
8-4 74.8 4-15 71.3
1-11 74.7 1-16 70.7
7-3 74.4 6-29 70.5
3-11 74.2 2-17 70.3
12-21 74.0 10-7 70.1

附表3

单粒近红外光谱检测油酸含量在75%以上的F3花生种子"

样品
Sample		 油酸
C18:1 (%)		 样品
Sample		 油酸
C18:1 (%)		 样品
Sample		 油酸
C18:1 (%)		 样品
Sample		 油酸
C18:1 (%)
2-11-2 86.1 8-8-4 83.3 7-3-30 81.6 7-3-4 79.7
9-10-14 85.6 8-8-12 83.2 7-18-22 81.5 8-12-31 79.6
8-12-4 85.6 7-18-2 83.2 7-18-1 81.5 6-2-17 79.5
8-8-14 85.6 3-16-5 83.1 8-8-5 81.5 4-13-2 79.5
2-11-11 85.4 6-1-3 83.1 4-11-1 81.4 3-11-13 79.5
8-12-3 85.3 4-14-23 83.1 11-42-59 81.3 5-12-10 79.4
3-11-4 85.1 8-4-4 83.0 12-21-14 81.3 4-2-8 79.3
9-10-4 85.1 5-12-32 83.0 5-12-40 81.3 4-11-12 79.3
5-12-3 84.9 6-1-22 82.9 3-16-16 81.3 11-31-27 79.3
11-42-2 84.8 9-10-1 82.9 8-4-10 81.3 7-18-32 79.3
10-6-4 84.8 3-3-3 82.9 9-10-20 81.2 4-11-25 79.3
1-11-5 84.7 6-1-23 82.8 11-42-25 81.1 4-13-4 79.1
3-3-2 84.7 4-11-4 82.7 3-16-22 81.1 4-13-7 79.0
5-12-1 84.6 6-2-13 82.7 4-13-14 81.1 8-12-9 78.9
12-21-15 84.6 9-10-5 82.7 4-13-3 81.0 4-13-33 78.9
2-11-5 84.6 11-42-12 82.7 9-10-25 81.0 3-16-45 78.8
10-6-3 84.5 5-12-2 82.6 7-3-1 81.0 3-16-30 78.7
6-1-1 84.4 11-31-22 82.5 4-13-30 81.0 4-13-1 78.7
1-11-4 84.4 4-11-7 82.5 8-12-14 80.9 5-7-21 78.6
4-11-10 84.3 8-12-23 82.4 4-2-1 80.9 7-3-9 78.5
4-2-5 84.2 9-10-33 82.4 10-6-10 80.9 7-3-27 78.5
9-20-4 84.2 7-18-12 82.3 11-31-2 80.8 3-11-2 78.0
1-11-3 84.2 9-21-1 82.3 9-10-27 80.8 11-31-4 78.0
8-8-1 84.2 4-14-5 82.3 9-10-15 80.7 3-11-1 78.0
2-11-3 84.1 11-31-7 82.2 4-2-9 80.7 9-10-26 77.9
2-11-1 84.1 7-18-10 82.2 8-12-7 80.7 3-16-35 77.6
14-14-2 84.1 4-13-24 82.2 11-31-26 80.7 4-14-4 77.6
8-8-17 84.1 5-12-8 82.2 11-31-14 80.7 3-16-24 77.4
4-14-15 84.1 6-2-2 82.2 11-42-19 80.5 11-31-30 77.3
11-31-15 84.1 6-2-9 82.1 3-3-8 80.5 5-12-27 76.5
1-11-1 84.1 9-10-10 82.1 11-31-8 80.5 4-2-42 76.4
2-11-4 84.0 9-10-6 82.1 10-6-18 80.5 4-14-31 76.4
8-8-2 84.0 4-11-5 82.1 2-11-28 80.5 5-7-19 76.3
3-3-5 83.9 6-2-6 82.0 4-11-41 80.3 11-31-28 76.1
9-20-2 83.9 6-1-10 82.0 6-1-7 80.3 8-8-18 75.9
5-7-2 83.9 4-11-3 81.9 4-2-2 80.3 4-13-17 75.8
6-1-4 83.9 6-1-27 81.9 4-13-10 80.3 12-21-10 75.8
10-6-2 83.8 8-4-8 81.9 4-13-34 80.1 6-2-32 75.6
1-11-2 83.7 4-14-14 81.8 9-10-9 80.1 3-3-31 75.6
11-31-3 83.7 8-8-3 81.8 11-42-16 80.1 9-10-16 75.6
9-10-12 83.7 7-3-2 81.7 11-42-8 80.1 3-11-24 75.6
9-10-3 83.6 11-42-1 81.7 12-21-29 80.0 11-31-12 75.5
9-20-3 83.5 7-3-10 81.7 7-18-5 79.9 3-3-19 75.5
12-21-2 83.4 4-14-3 81.7 11-31-17 79.9 5-12-24 75.4
8-12-5 83.4 12-21-1 81.7 4-14-16 79.8 11-42-54 75.2
3-3-1 83.4 11-42-7 81.7 4-14-10 79.8 9-10-37 75.2
6-1-2 83.3 12-21-18 81.6 4-2-35 79.7 9-10-38 75.1
12-21-4 83.3 8-12-34 81.6 4-13-18 79.7 9-10-22 75.0
4-14-1 83.3 6-2-5 81.6 7-3-20 79.7 1-11-9 75.0

表2

单粒近红外光谱检测花生F4种子中的油酸含量(%)及其混样气相色谱检测结果"

2-11-3a 2-11-28 4-11-7 4-14-1 6-2-6 8-8-3 9-10-9 10-6-4 11-31-15 12-21-18
1b 87.5 86.7 77.7 81.2 78.8 78.7 76.5 76.9 75.8 77.0
2 80.3 78.2 76.5 76.7 81.2 76.7 78.7 78.0 79.2 76.6
3 84.2 76.2 76.8 76.4 75.5 76.1 75.4 81.5 79.7 82.9
4 80.6 83.4 80.0 79.0 78.3 80.0 85.8 75.4 76.0 75.8
5 78.1 76.2 77.6 80.8 75.1 78.2 83.7 77.5 86.1 77.3
6 76.3 78.0 79.0 75.9 82.5 76.6 77.8 75.9 75.4 76.5
7 82.0 83.9 77.9 76.7 78.0 81.5 77.5 76.4 76.0 79.7
8 76.7 76.2 77.3 77.5 77.8 77.7 75.2 77.6 81.2 76.6
9 76.1 82.5 82.6 84.1 75.0 79.3 77.5 84.7 83.6 82.9
10 85.1 78.9 80.1 78.3 77.8 76.2 77.5 75.0 79.5 82.2
GCc 82.0 80.1 80.4 81.7 81.0 81.3 79.7 79.8 80.5 81.2

表3

各F4株系AhFAD2的基因型"

F4株系 F4 lines 基因型Genotype
2-11-3 aabb
2002/11/28 aabb
2004/11/7 aabb
4-14-1 aabb
2006/2/6 aabb
2008/8/3 aabb
2009/10/9 aabb
2010/6/4 aabb
11-31-15 aabb
12-21-18 aabb
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