基于代谢组学分析‘秋香’核桃贮藏期间脂肪酸的变化

doi:10.16420/j.issn.0513-353x.2020-0731

摘要/Abstract

摘要：

以‘秋香’核桃为试材,设置常温贮藏1年（CS）、4 ℃低温贮藏1年（LS）和未贮藏（WS）3个处理,利用超高效液相色谱/质谱（Ultra high performance liquid chromatography/mass spectrometry,LC/MS）定性定量分析核仁脂肪酸。结果表明:常温贮藏与未贮藏比较组样品共筛选到差异代谢物969个,而低温贮藏与未贮藏比较组共筛选到975个;通过变量权重值筛选（VIP > 1）,与未贮藏相比,常温贮藏核桃差异代谢物有35个,而低温贮藏的有31个,其中硬脂酸含量均降低,亚油酸和α-亚麻酸在常温贮藏核桃中明显降低,但油酸含量却稍有增加,在低温贮藏中油酸、亚油酸和α-亚麻酸无明显变化差异,此外,在常温贮藏核桃中还检测到泛醌类代谢物下调,而在低温贮藏中角质类物质增加。KEGG通路分析发现差异代谢物主要参与不饱和脂肪酸的合成及α-亚麻酸和亚油酸的代谢,还有部分参与角质和泛醌类物质的合成。代谢物之间相关性分析表明,不饱和脂肪酸与泛醌类物质存在正相关关系,硬脂酸与角质存在负相关关系。综上,常温贮藏核桃泛醌类物质降低,抗氧化能力下降,α-亚麻酸和亚油酸氧化减少,而低温贮藏可抑制α-亚麻酸和亚油酸氧化减少,并提高角质含量适应低温环境。

关键词: 核桃, 代谢组学, 贮藏温度, 脂肪酸, 不饱和脂肪酸

Abstract:

The‘Qiuxiang’walnuts were set up three treatments:climate storage for one year（CS）,4 ℃ low-temperature storage for one year（LS）and without storage（WS）. Then the fatty acids of walnut kernels were analyzed qualitatively and quantitatively using LC/MS. The results show that a total of 969 differential metabolites were obtained in the CS_WS comparison group,and a total of 975 differential metabolites were obtained in the LS_WS comparison group. Through further screening according to variable weight value（VIP > 1）,the CS_WS and LS_WS have 35 and 31 different metabolites,respectively. Comparing with WS,the stearidonic acid is reduced in CS and LS,linoleic acid and α-linolenic acid are significantly reduced in CS,the oleic acid is slightly increased in CS,however,in LS,oleic acid,linoleic acid and α-linolenic acid have no significant difference. In addition,ubiquinone is down-regulated in CS,while keratinous substances are increased in LS. The analysis of KEGG pathway shows that differential metabolites are mainly involved in the synthesis of unsaturated fatty acids and the metabolism of α-linolenic acid and linoleic acid. In addition,some differential metabolites are involved in the synthesis of keratin and ubiquinone. The correlation analysis among differential metabolites shows that there is a positive correlation between unsaturated fatty acids and ubiquinone,and a negative correlation between stearidonic acid and keratin. In summary,the ubiquinone is decreased and the antioxidant capacity is reduced of walnut in natural storage,thereby leading to reduced α-linolenic acid and linoleic acid,but the walnut in cold storage at 4 ℃ can inhibit the reduction of α-linolenic acid and linoleic acid,in addition,it also increase the keratin content to adapt to low-temperature environment.

Key words: walnut, metabolomics, storage temperature, fatty acid, unsaturated fatty acid

中图分类号:

S664.1

许海峰, 陈新, 张士刚, 相昆, 张美勇, 徐颖, 王贵芳. 基于代谢组学分析‘秋香’核桃贮藏期间脂肪酸的变化[J]. 园艺学报, 2021, 48(11): 2161-2170.

XU Haifeng, CHEN Xin, ZHANG Shigang, XIANG Kun, ZHANG Meiyong, XU Ying, WANG Guifang. Change of Fatty Acid During Storage Period of‘Qiuxiang’Walnut Based on Metabolomics Analysis[J]. Acta Horticulturae Sinica, 2021, 48(11): 2161-2170.

图/表 7

图1 核桃常温贮藏与未贮藏相比和低温贮藏与未贮藏相比差异代谢物火山图。 FC:差异倍数;P -value:差异显著性。

Fig. 1 The Volcano plot of differential metabolites between CS_WS and LS_WS FC:Fold change;P-value:significance of difference.

图2 基于变量权重值（VIP）进行核桃常温贮藏与未贮藏（A）、低温贮藏与未贮藏（B）间差异代谢物热图分析

Fig. 2 Thermal map analysis of CS_WS（A）and LS_WS（B）based on variable important in projection（VIP）

表1 核桃常温贮藏（CS）与未贮藏（WS）脂肪酸氧化酸败相关的差异代谢物相对含量

Table 1 Relative content of differential metabolites related to fatty acid oxidative rancidity in CS and WS

序号 No.	保留时间/min Retention time	物质 Metabolites	相对含量Relative content		差异倍数值 FC	类型 Type
序号 No.	保留时间/min Retention time	物质 Metabolites	CS	WS	差异倍数值 FC	类型 Type
1	13.90	油酸 Oleic acid	2 863.41 ± 404.45 a	1 356.97 ± 169.74 b	2.11	上调Up
2	6.64	异茉莉酸 Isojasmonic acid	348.91 ± 50.87 a	83.04 ± 3.76 b	4.20	上调Up
3	7.27	创伤酸 Traumatic acid	14.89 ± 3.50 a	5.79 ± 0.52 b	2.57	上调Up
4	10.89	硬脂酸 Stearidonic acid	291.35 ± 21.75 b	2 386.86 ± 142.79 a	0.12	下调Down
5	12.59	α-亚麻酸 alpha-Linolenic acid	77.98 ± 9.12 b	376.83 ± 26.26 a	0.21	下调Down
6	14.17	亚油酸 Linoleic acid	1 247.45 ± 36.64 b	7 803.43 ± 301.45 a	0.16	下调Down
7	9.36	泛醌 Ubiquinone	4.65 ± 0.56 b	16.48 ± 1.63 a	0.28	下调Down

表2 核桃低温贮藏（LS）与未贮藏（WS）脂肪酸氧化酸败相关的差异代谢物相对含量

Table 2 Relative content of differential metabolites related to fatty acid oxidative rancidity in LS and WS

序号 No.	保留时间/min Retention time	物质 Metabolites	相对含量Relative content		差异倍数值 FC	类型 Type
序号 No.	保留时间/min Retention time	物质 Metabolites	LS	WS	差异倍数值 FC	类型 Type
1	13.90	油酸 Oleic acid	1 283.41 ± 404.45 a	1 356.97 ± 169.74 a	0.95	无差异 Not-significant
2	6.64	异茉莉酸 Isojasmonic acid	89.43 ± 5.88 a	83.04 ± 3.76 a	1.08	无差异 Not-significant
4	10.89	硬脂酸 Stearidonic acid	1 467.52 ± 38.48 b	2 386.86 ± 142.79 a	0.61	下调 Down-regulated
5	12.59	α-亚麻酸 alpha-Linolenic acid	358.76 ± 30.14 a	376.83 ± 26.26 a	0.95	无差异 Not-significant
6	14.17	亚油酸 Linoleic acid	7 659.17 ± 415.63 a	7 803.43 ± 301.45 a	0.98	无差异 Not-significant
7	9.36	泛醌 Ubiquinone	15.79 ± 1.46 a	16.48 ± 1.63 a	0.96	无差异 Not-significant
8	10.45	十六烷酸 Hexadecanoic acid	229.82 ± 13.30 a	91.41 ± 6.86 b	2.51	上调 Up-regulated
9	11.86	10,16-二羟基十六烷酸 10,16-Dihydroxyhexadecanoic acid	382.03 ± 35.59 a	151.87 ± 4.65 b	2.52	上调 Up-regulated

图3 核桃常温贮藏与未贮藏（A）和低温贮藏与未贮藏（B）差异代谢物相关性分析红色表示正相关,蓝色表示负相关。

Fig. 3 Correlation analysis of differential metabolites in CS and WS（A）,LS and WS（B） Red indicates positive correlation,blue indicates negative correlation.

图4 核桃常温贮藏与未贮藏（A）和低温贮藏与未贮藏（B）差异代谢物KEGG分析图中富集因子越大,说明富集程度越大;圆点越大,说明富集到该通路上的代谢物数越多。

Fig. 4 KEGG analysis of differential metabolites in CS and WS（A）,LS and WS（B） The greater the enrichment factor,the greater the enrichment degree;the larger the round dot,the more metabolites enriched to the pathway.

图7 不同贮藏温度下对核桃脂肪酸氧化代谢通路示意图

Fig. 7 Effect of different temperature on oxidative metabolic pathway of fatty acid in walnut

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