低磷胁迫下黄瓜木质部与韧皮部汁液的代谢物变化

doi:10.3864/j.issn.0578-1752.2022.08.012

摘要/Abstract

摘要：

【背景】木质部汁液负责为地上部分运输水和营养物质,韧皮部汁液则主要负责光合产物的运输,在植株受到非生物胁迫时,它们会发生不同的变化。【目的】研究低磷胁迫下黄瓜木质部与韧皮部汁液内各种代谢物含量变化,探究黄瓜在低磷胁迫下的代谢变化规律,为黄瓜磷匮乏状态诊断提供新思路与依据。【方法】采集正常磷水平（1.25 mmol∙L^-1）与低磷胁迫（0.3125 mmol∙L^-1）下黄瓜植株的木质部与韧皮部汁液,经三甲基硅烷化衍生化处理后用气相色谱质谱联用技术（GC-MS）进行检测和定性定量分析。【结果】低磷胁迫下木质部汁液内糖类没有发生显著变化,多数氨基酸的含量与正常情况相比明显上升,有机酸中的苹果酸呈显著下降趋势,软脂酸、油酸等则显著高于正常水平;低磷胁迫下的韧皮部汁液中,糖类与氨基酸多数低于正常水平,草酸、琥珀酸等有机酸的含量也明显降低。此外,为筛选黄瓜木质部与韧皮部汁液内潜在的生物标志物,使用OPLS-DA与独立样本T检测方法对GC-MS试验数据进行进一步分析,结果表明,在黄瓜木质部汁液检出8种低磷胁迫下的潜在生物标志物（VIP>1,P<0.05）,分别为尸胺、苹果酸、四氨基丁酸、软脂酸、肌醇、纤维二糖、N, N-二甲基十二酰胺与单硬脂酸甘油酯;在黄瓜韧皮部汁液中检出7种低磷胁迫下的潜在生物标志物（VIP>1,P<0.05）,分别为丝氨酸、亚油酸、10, 13-环氧-11-甲基十八碳-10, 12-二烯酸、脱氢枞酸、N, N-二甲基十二酰胺、2, 2'-亚甲基双（6-叔丁基-4-甲基苯酚）、L-2-哌啶酸。【结论】低磷胁迫下,黄瓜的木质部与韧皮部汁液内的糖类、氨基酸和有机酸均发生了变化,但是变化趋势并不相同。分别在木质部和韧皮部汁液中筛选出8种和7种生物标志物,可为黄瓜磷匮乏状态诊断提供新思路与依据。生物标志物中,苹果酸、四氨基丁酸、软脂酸、丝氨酸与亚油酸的检测手段已经较为成熟,以其为检测对象的黄瓜磷匮乏状态诊断方法具有较好的实用性。

关键词: 黄瓜, 木质部, 韧皮部, 磷胁迫, 代谢组学, 生物标志物

Abstract:

【Background】 Xylem sap is responsible for the transport of water and nutrients for the aerial part, while phloem sap is mainly responsible for the transport of photosynthetic products. When plants are subjected to abiotic stress, the sap of xylem and phloem undergoes different changes. 【Objective】 The aim of this study was to investigate the content changes of various metabolites in cucumber xylem and phloem sap under low phosphorus stress, and to explore the metabolic changes of cucumber under low phosphorus stress, so as to provide new ideas and basis for the diagnosis of cucumber phosphorus deficiency. 【Method】 The xylem and phloem sap were collected from cucumber plants under normal phosphorus level (1.25 mmol∙L^-1) and low phosphorus stress (0.3125 mmol∙L^-1). These sap samples were analyzed qualitatively and quantitatively by GC-MS after trimethylsilyl derivatization. 【Result】In xylem sap under low phosphorus stress, there was no significant change in carbohydrates, and the content of most amino acids increased significantly. Among organic acids, the malic acid showed a significant downward trend, while others, such as palmitic acid and oleic acid, were significantly higher than the normal levels. In phloem sap under low phosphorus stress, most carbohydrates and amino acids were lower than the normal level, and the contents of organic acids, such as oxalic acid and succinic acid, were also significantly reduced. Further, in order to screen potential biomarkers in cucumber xylem and phloem sap, the detection data were further analyzed by OPLS-DA and Student's t test. Eight potential biomarkers (VIP>1, P<0.05) under low phosphorus stress were detected in cucumber xylem sap, which were cadaverine, malic acid, 4-aminobutyric acid, palmitic acid, myo-inositol, cellobiose acid, N, N-dimethyldodecamide, and glycerol monostearate. Seven potential biomarkers (VIP>1, P<0.05) under low phosphorus stress were detected in cucumber phloem sap, which were serine, linoleic acid, 9-(3-methyl-5-pentylfuran-2-yl) nonanoic acid, dehydroabietic acid, N, N-dimethyldodecamide, 2, 2'-methylenebis (6-tert-butyl-4-methylphenol), and L-2-piperic acid. 【Conclusion】 Under low phosphorus stress, the carbohydrates, amino acids and organic acids in the xylem and phloem sap of cucumber changed, but the change trends were not the same. Eight and seven biomarkers were screened from xylem and phloem sap, which could provide new ideas and basis for the diagnosis of cucumber phosphorus deficiency. Among the biomarkers, the detection methods of malic acid, 4-aminobutyric acid, palmitic acid, serine and linoleic acid have been relatively mature, and the cucumber phosphorus deficiency diagnosis method based on them was more practical.

Key words: cucumber, xylem, phloem, phosphorus stress, metabolomics, biomarkers

李青林,张文涛,徐慧,孙京京. 低磷胁迫下黄瓜木质部与韧皮部汁液的代谢物变化[J]. 中国农业科学, 2022, 55(8): 1617-1629.

LI QingLin,ZHANG WenTao,XU Hui,SUN JingJing. Metabolites Changes of Cucumber Xylem and Phloem Sap Under Low Phosphorus Stress[J]. Scientia Agricultura Sinica, 2022, 55(8): 1617-1629.

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磷水平 Phosphorus level	株高增长量 Height growth (cm)		茎粗增长量 Diameter growth (mm)
磷水平 Phosphorus level	第一周 Week 1	第二周 Week 2	第一周Week 1	第二周 Week 2
100%	28.07±2.56a	31.96±3.50a	0.77±0.09a	0.90±0.07a
25%	25.47±2.02a	22.31±3.87b	0.64±0.19a	0.71±0.12b

序号 Sequence	组别名 Group alias	位置 Position	营养液中磷离子浓度 Phosphorus ion concentration in nutrient solution (mmol∙L^-1)
1	P25X	木质部汁液 Xylem sap	0.3125
2	P25P	韧皮部汁液 Phloem sap	0.3125
3	P100X	木质部汁液 Xylem sap	1.25
4	P100P	韧皮部汁液 Phloem sap	1.25