连作烟草根系分泌物鉴定及潜在化感物质的筛选研究

doi:10.13304/j.nykjdb.2023.0034

摘要/Abstract

摘要：

为探索长期连作状态下烟草根系分泌物内含有的化感物质组成成分，设置石英砂栽培和连作土壤栽培2种烟草培养处理，对2种烟草不同生长时期的根系分泌物进行提取，利用超高效液相色谱串联质谱（ultra-high performance liquid chromatography tandem mass spectrometry， UHPLC-MS/MS）对提取的根系分泌物进行分析，通过数据库比对、差异分析和时间聚类分析等方法筛选连作烟草根系分泌物中潜在的化感物质。结果表明，共筛选出11类潜在化感物质，分别为生物碱、萜类、胺类、有机酸、苯酸及其衍生物、脂肪酸、酚类、氨基酸和多肽、香豆素、肉桂酸及其衍生物和其他。生物碱、萜类、胺类和有机酸4类化合物相对含量占所有化合物的73.3%。L-烟碱、熊果酸、β-氨基丙腈、5-羟基吲哚乙酸、早熟素Ⅱ、单乙基己基邻苯二甲酸、己内酰胺、姜酚、L-乳酸、邻苯二甲酸二乙酯和2，3-二氢-3，5-二羟基-2-氧代-3-吲哚乙酸甲酯是连作烟草根系分泌物中相对含量较高的潜在化感物质。研究结果为烟草根系化感物质的化感作用研究及缓解烟草连作障碍方法研究提供理论基础。

关键词: 烟草, 根系分泌物, 化感物质, 化感作用, 连作障碍, UHPLC-MS/MS

Abstract:

To explore the components of allelopathic substances contained in tobacco root exudates under long-term continuous cropping， quartz sand cultivation and continuous soil cultivation 2 tobacco culture experiments were set up， root exudates of non-continuous cropping and continuous cropping tobacco at different growth stages were extracted， and the extracted root exudates were identified and analyzed by ultra-high performance liquid chromatography tandem mass spectrometry（UHPLC-MS/MS）， then potential allelopathic substances in the root exudates of continuous cropping tobacco were screened by database comparison， difference analysis and time cluster analysis. The results showed that a total of 11 potential allelopathic substances were screened， namely alkaloids， terpenes， amines， organic acids， benzic acids and derivatives， fatty acids， phenols， amino acids and peptides， coumarins， cinnamic acid and derivatives and others. The relative content of alkaloids， terpenes， amines and organic acids accounted for 73.3% of all compounds. L-nicotine， ursolic acid， β?-aminopropionitrile， 5-hydroxyindoleacetic acid， precocene Ⅱ， monoethylhexylphthalic acid， caprolactam， gingerol， L-lactic acid， diethyl phthalate， and methyl 2，3-dihydro-3，5-dihydroxy-2-oxo-3-indoleacetate were potential allelopathic substances with relatively high levels in continuous tobacco root exudates. This study provided a theoretical basis for the study of allelopathic substances in tobacco roots and the research on methods to alleviate tobacco continuous cropping disorders.

Key words: tobacco, root exudates, allelopathic substances, allelopathy, continuous cropping disorder, UHPLC-MS/MS

中图分类号:

S181

周喜新, 袁世林, 杨柳, 夏滔, 张毅, 范伟. 连作烟草根系分泌物鉴定及潜在化感物质的筛选研究[J]. 中国农业科技导报, 2024, 26(7): 136-146.

Xixin ZHOU, Shilin YUAN, Liu YANG, Tao XIA, Yi ZHANG, Wei FAN. Identification of Continuous Cropping Tobacco Root Exudates and Screening of Potential Allelopathic Substances[J]. Journal of Agricultural Science and Technology, 2024, 26(7): 136-146.

图/表 9

图1 质控样本UHPLC-MS/MS检测总离子流A：负离子流；B：正离子流

Fig. 1 Total ion flow of quality control samples by UHPLC-MS/MS assayA： Negative ion flow； B： Positive ion flow

图2 根系分泌物在Super Class层级不同分类中所占比重A：正离子模式；B：负离子模式

Fig. 2 Share of root secretions in the different classifications of the Super Class hierarchyA： Positive ion mode； B： Begative ion mode

图3 不同生长时期2种培养方式烟草根系分泌物的正交偏最小二乘判别分析

Fig. 3 OPLS-DA of tobacco root secretions from two cultures at different growth periods

图4 不同生长时期2种培养方式烟草根系分泌物的OPLS-DA模型交叉验证结果

Fig. 4 Cross-validation results of the OPLS-DA model for tobacco root secretions from two cultures at different growth periods

图5 不同生长时期两种培养方式烟草根系分泌物的差异表达火山图注：红色节点代表Q<0.05且log2FC>2的上调代谢物；蓝色节点代表Q<0.05且log2FC<-2的下调代谢物。

Fig. 5 Differential expression of tobacco root exudates between two culture methods at different growth stagesNote： Red node represents the upregulated metabolite of Q<0.05 and log2FC>2； blue node represents the downregulated metabolite with Q<0.05 and log2FC<-2.

表1 不同时期土培烟草根系分泌物上调的化合物

Table 1 Compounds upregulated by root exudates of soil tobacco at different times

培养时间 Cultivation time/d	代谢物名称 Metabolite name	相对含量 Relative content
30	利克香豆素 Liqcoumarin	0.018 1
	β-水芹烯 β-phellandrene	0.174 6
	（+）-α-蒈烯（+）-α-carene	0.025 5
	γ-萜品烯 γ-terpinene	0.020 4
	L-谷氨酸 L-glutamic acid	0.004 7
	N-γ-谷氨酰-S-丙基半胱氨酸 N-γ-glutamyl-S-propylcysteine	0.014 0

图6 烟草根系分泌物时间序列聚类分析

Fig. 6 Time series clustering analysis of tobacco root exudates in continuous soil cultivation

图7 烟草根系分泌物潜在化感物质相对含量

Fig. 7 Relative levels of potential allelopathic substances in tobacco root exudates

表2 轮作烟草根系分泌物潜在化感物质

Table 2 Potential allelopathic substances in tobacco root exudates in crop rotation

化合物类别 Compound class	代谢物名称 Metabolite name	分子式 Molecular formula	相对含量 Relative content
香豆素 Coumains	利克香豆 Liqcoumarin	C₁₂H₁₀O₄	0.002 1
	2，3-环氧黄曲霉毒素B1 2，3-epoxyaflatoxin B1	C₁₇H₁₂O₇	0.010 6
	东莨菪内酯 Scopoletin	C₁₀H₈O₄	0.007 7
脂肪酸 Fatty acid	1，11-十一烷二羧酸 1，11-undecanedicarboxylic acid	C₁₃H₂₄O₄	0.012 9
	15-脱氧-D-12，14-PGJ2 15-deoxy-d-12，14-PGJ2	C₂₀H₂₈O₃	0.010 5
	20-羟基-PGF2a 20-hydroxy-PGF2a	C₂₀H₃₄O₆	0.026 9
	5，6-二氢乙二醇 5，6-DHET	C₂₀H₃₄O₄	0.012 1
	16-羟基十六烷酸 16-hydroxy hexadecanoic acid	C₁₆H₃₂O₃	0.019 4
	二十二碳五烯酸 Docosapentaenoic acid （22n-3）	C₂₂H₃₄O₂	0.005 9
	前列腺素I2 Prostaglandin I2	C₂₀H₃₂O₅	0.007 9
	3-羟基癸酸 3-hydroxydecanoic acid	C₁₀H₂₀O₃	0.008 7
	13-L-氢过氧亚油酸 13-L-hydroperoxylinoleic acid	C₁₈H₃₂O₄	0.007 4

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