植物根际促生菌发酵液中植物激素含量的液液萃取－高效液相色谱测定法
English
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参考文献
[1] JON C S, ZOU Y L, ZHAO J H, RI H C, WANG L Y, KAW H Y, MENG L Y, SHANG H B, LI D H. Simultaneous determination of multiple phytohormones in tomato by ionic liquid-functionalized carbon fibers-based solid-phase microextraction coupled with liquid chromatography-mass spectrometry. Analytica Chimica Acta, 2020, 1137(9): 143-155.
[2] TANK J G, PANDYA R V, THAKER V S. ChemInform abstract: Phytohormones in regulation of the cell division and endoreduplication process in the plant cell cycle. ChemInform, 2014, 45(30): 12605-12613.
[3] 晁春艳, 王啸波, 李朝昉. 高效液相色谱法分离植物促生菌中的激素GA、IAA. 化学工程与装备, 2008, 34(8): 94-97. CHAO C Y, WANG X B, LI C F. Separation of GA and IAA in plant growth promoting rhizobacteria by high performance liquid chromatography. Chemical Engineering & Equipment, 2008, 34(8): 94-97.
[4] CAVITE H J M, MACTAL A G, EVANGLISTA E V, CRUZ J A. Growth and yield response of upland rice to application of plant growth promoting rhizobacteria. Journal of Plant Growth Regulation, 2020, 40(2): 1-15.
[5] 吴倩, 王璐, 吴大朋, 段春凤, 关亚风. 植物激素样品前处理方法的研究进展. 色谱, 2014, 32(4): 319-329. doi: 10.3724/SP.J.1123.2014.01039 WU Q, WANG L, WU D P, DUAN C F, GUAN Y F. Recent advances in sample preparation methods of plant hormones. Chinese Journal of Chromatography, 2014, 32(4): 319-329. doi: 10.3724/SP.J.1123.2014.01039
[6] 姚拓. 植物根际促生菌研究与应用. 北京: 中国农业出版社, 2021. YAO T. Research and Application of Plant Growth Promoting Rhizobacteria. Beijing: China Agriculture Press, 2021.
[7] 任卓然, 邵新庆, 李金升, 李慧, 何宜璇, 古维娜, 王茹颖, 杨灵婧, 刘克思. 微生物菌肥对退化高寒草甸地上生物量和土壤理化性质的影响. 草地学报, 2021, 29(10): 2265-2273. REN Z R, SHAO X Q, LI J S, LI H, HE Y X, GU W N, WANG R Y, YANG L J, LIU K S. Effects of microbial fertilizer on aboveground biomass and soil physiochemical properties of degraded alpine meadow. Acta Agrestia Sinica, 2021, 29(10): 2265-2273.
[8] BHATTACHARYYA P N, JHA D K. Plant growth-promoting rhizobacteria (PGPR): Emergence in agriculture. World Journal of Microbiology & Biotechnology, 2012, 28(4): 1327-1350.
[9] ZVI P, EDUARDO B. Hormone balance and abiotic stress tolerance in crop plants. Current Opinion in Plant Biology, 2011, 14(3): 290-295. doi: 10.1016/j.pbi.2011.02.001
[10] 肖爱华, 陈发菊, 贾忠奎, 桑子阳, 朱仲龙, 马履一. 梯度洗脱高效液相色谱法测定红花玉兰中4种植物激素. 分析试验室, 2020, 39(3): 249-254. XIAO A H, CHEN F J, JIA Z K, SANG Z Y, ZHU Z L, MA L Y. Determination of 4 plant hormones in Magnolia wufengensis by gradient elutionhigh performance liquid chromatography. Chinese Journal of Analysis Laboratory, 2020, 39(3): 249-254.
[11] LIU Y, FANG X A, CHEN G S, YE Y X, XU J Q, OUYANG G F, ZHU F. Recent development in sample preparation techniques for plant hormone analysis. Trends in Analytical Chemistry, 2019, 113(2): 224-233.
[12] 张占畅, 孙志红, 郑小能, 刘怀锋. 高效液相色谱法测定库尔勒香梨幼果中的4种内源激素. 新疆农业科学, 2017, 54(5): 886-892. doi: 10.6048/j.issn.1001-4330.2017.05.012 ZHANG Z C, SUN Z H, ZHENG X N, LIU H F. Determination of four endogenous hormones in the young fruit of korla feagrant pear by high performance liquid chromatography (HPLC). Xinjiang Agricultural Sciences, 2017, 54(5): 886-892. doi: 10.6048/j.issn.1001-4330.2017.05.012
[13] WANG L, XU J Y, WEI J, CHEN Z, XU Z C. Chloride salinity in a chloride-sensitive plant: Focusing on photosynthesis, hormone synthesis and transduction in tobacco. Plant Physiology and Biochemistry, 2020, 153(5): 119-130.
[14] SUNAYNA C, PAL K I, JAI M. Development and validation of a novel, rapid gradient HPLC method for simultaneous estimation of bioactive marker compounds in a mixture of convolvulus pluricaulis, withania somnifera and bacopa monnieri extracts. Journal of Chromatographic Science, 2020, 57(10): 920-930.
[15] 綦艳, 李锦清, 陈满英, 伍尚森, 唐楚林, 刘德群. 高效液相色谱法同时测定生活用纸及纸制品中19种有机抗菌剂. 分析测试学报, 2021, 40(7): 1018-1024. doi: 10.3969/j.issn.1004-4957.2021.07.006 QI Y, LI J Q, CHEN M Y, WU S S, TANG C L, LIU D Q. Simultaneous determination of 19 organic antibacterial agents in tissue paper and paper products by high performance liquid chromatography. Journal of Instrumental Analysis, 2021, 40(7): 1018-1024. doi: 10.3969/j.issn.1004-4957.2021.07.006
[16] LAO F, GIUSTI M M. Quantification of purple corn (Zea mays L.) anthocyanins using spectrophotometric and HPLC approaches: Method comparison and correlation. Food Analytical Methods, 2016, 9(5): 1367-1380. doi: 10.1007/s12161-015-0318-0
[17] 刘婷, 姚拓, 陈建纲, 刘欢. 固相萃取-高效液相色谱法测定植物根际促生菌发酵产物中3种植物激素的含量. 分析科学学报, 2017, 33(2): 201-206. LIU T, YAO T, CHEN J G, LIU H. Determination of plant hormones in bacterial fermentation products of plant growth promoting rhizobacteria by solid phase extraction-high perdormance liquid chromatography. Journal of Analytical Science, 2017, 33(2): 201-206.
[18] OMER Z S, OMBOLINI R, BROBERG A, GERHARDSON B. Indole-3-acetic acid production by pink-pigmented facultative methylotrophic bacteria. Plant Growth Regulation, 2004, 43(1): 93-96. doi: 10.1023/B:GROW.0000038360.09079.ad
[19] NAUTIYAL C S. An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiology Letters, 1999, 170(1): 265-270. doi: 10.1111/j.1574-6968.1999.tb13383.x
[20] 王璐. 基于基质固相分散萃取－高效液相色谱－串联质谱的植物激素分析. 合肥: 中国科学技术大学博士学位论文, 2014. WANG L. Determination of plant hormones by matrix solid－phase dispersion－high performance liquid chromatography-tandem mass spectrometry. PhD Thesis. Hefei: University of Science and Technology of China, 2014.
[21] ABBAS M B, HASHEM HANAA M A, ALPHONSE M M, TAREK M S. Synchronized determination of sacubitril and valsartan with some co-administered drugs in human plasma via UPLC-MS/MS method using solid phase extraction. Biomedical Chromatography, 2021, 35(11): 5203-5214.
[22] 林舒婷, 丁青青, 张文敏, 张兰, 卢巧梅. 固相萃取吸附材料在植物激素样品前处理中的应用新进展. 色谱, 2021, 39(12): 1281-1290. doi: 10.3724/SP.J.1123.2021.03045 LIN S T, DING Q Q, ZHANG W M, ZHANG L, LU Q M. Novel adsorption material for solid phase extraction in sample pretreatment of plant hormones. Chinese Journal of Chromatography, 2021, 39(12): 1281-1290. doi: 10.3724/SP.J.1123.2021.03045
[23] 蔡保东. 植物激素分析方法研究. 武汉: 武汉大学博士学位论文, 2016. CAI B D. Analytical methods for determination of phytohormones in plant. PhD Thesis. Wuhan: Wuhan University, 2016.
[24] 卢玉秋, 宋阿琳, 唐治玉, 李艳玲, 董炜灵, 王恩召, 唐治喜, 范分良. UPLC-MS/MS同时测定土壤中19种植物激素方法的建立和验证. 植物营养与肥料学报, 2019, 25(6): 953-962. doi: 10.11674/zwyf.18129 LU Y Q, SONG A L, TANG Z Y, LI Y L, DONG W L, WANG E Z, TANG Z X, FAN F L. Establishment and validation of UPLC-MS/MS method for simultaneous determination of 19 phytohormones in soil. Journal of Plant Nutrition and Fertilizer, 2019, 25(6): 953-962. doi: 10.11674/zwyf.18129
[25] 朱梅青, 崔蓉. 高效液相色谱法测定小鼠肝脏中的两种苯并三唑类紫外线吸收剂. 分析科学学报, 2021, 37(1): 93-97. doi: 10.2116/analsci.20SAR06 ZHU M Q, CUI R. Determination of two benzotriazole ultraviolet absorbents in mouse liver by highperformance liquid chromatography. Journal of Analytical Science, 2021, 37(1): 93-97. doi: 10.2116/analsci.20SAR06
[26] 纪晓娜, 吴文博, 任志敏, 陈国军, 李延君. 高效液相色谱法检测蔬菜中甲萘威残留. 分析科学学报, 2022, 38(1): 122-124. doi: 10.13526/j.issn.1006-6144.2022.01.022 JI X N, WU W B, REN Z M, CHEN G J, LI Y J. Detection of crbarayl rsidue in vegetables by high performance liquid cromatohgraphy. Journal of Analytical Science, 2022, 38(1): 122-124. doi: 10.13526/j.issn.1006-6144.2022.01.022
[27] WEILER E W. Radioimmunoassays for trans-zeatin and related cytokinins. Planta, 1980, 149(2): 155-162. doi: 10.1007/BF00380877
[28] 文静, 孔维军, 罗红梅, 王建, 杨美华. 植物内源激素检测方法新进展. 中南药学, 2014, 12(1): 47-52. doi: 10.7539/j.issn.1672-2981.2014.01.011 WEN J, KONG W J, LUO H M, WANG J, YANG M H. Progress in detection of plant endogenous hormones. Central South Pharmacy, 2014, 12(1): 47-52. doi: 10.7539/j.issn.1672-2981.2014.01.011
[29] 张丹, 孙萍, 陈思瑾, 幸华, 芮仕立, 牛早霞, 栗孟飞. 液相色谱-串联质谱联用法同时测定6种植物激素. 甘肃农业大学学报, 2020, 55(4): 98-103. ZHANG D, SUN P, CHEN S J, XING H, RUI S L, NIU Z X, LI M F. Simultaneous determination of six plant hormones by liquid cromatohgraphy-tandem mass spectrometry. Journal of Gansu Agricultural University, 2020, 55(4): 98-103.
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图 1 混合激素标准溶液的色谱图
t-Z：玉米素；GA3：赤霉素；IAA：吲哚乙酸；下图同。
Figure 1. Chromatogram of the standard mixture of hormones
t-Z: trans-Zeatin; GA3: gibberellin A3; IAA: indole-3-acetic acid; this is applicable for the following figures as well.
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图 2 测定实际样品促生菌MBM5和PAM12的色谱图
Figure 2. Chromatograms of the MBM5 strain and PAM12 strain
表 1 植物激素的线性方程、线性范围、相关系数及检出限和定量限
Table 1 Linear equations, linear ranges, correlation coefficients, limits of detection (LOD), and limits of quantitative (LOQ) of plant hormones
植物激素
Plant
hormones相关系数
Correlation
coefficients (R2)线性方程
Linear
equations线性范围
Linear
ranges/(μg·mL−1)检出限
LOD/
(μg·mL−1)定量限
LOQ/
(μg·mL−1)赤霉素 Gibberellin A3 0.997 9 y = 0.908 2x − 3.538 3 26.68～111.11 0.75 2.50 吲哚乙酸 Indole-3-acetic acid 0.997 8 y = 11.957 0x − 19.494 0 10.67～44.44 0.01 0.03 玉米素 Trans-Zeatin 0.997 5 y = 34.182 6x − 47.536 7 10.67～44.44 0.03 0.09 
下载: 导出CSV
表 2 植物激素的添加回收率和相对标准偏差(n = 3)
Table 2 Recovery and relative standard deviation of 3 plant hormones (n = 3)
菌株编号
Strains
code植物激素
Plant
hormones添加量
Added/
(μg·mL−1)平均回收率
Average
recovery/%相对标准偏差
Relative standard
deviation/%MBM5 赤霉素
Gibberellin A310.00 95.01 5.71 10.00 10.00 吲哚乙酸
Indole-3-acetic
acid4.00 93.04 6.51 4.00 4.00 玉米素
Trans-Zeatin4.00 95.66 0.02 4.00 4.00 PAM12 赤霉素
Gibberellin A310.00 108.40 2.29 10.00 10.00 吲哚乙酸
Indole-3-acetic
acid4.00 88.02 1.39 4.00 4.00 玉米素
Trans-Zeatin4.00 105.96 4.85 4.00 4.00
下载: 导出CSV
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[1] JON C S, ZOU Y L, ZHAO J H, RI H C, WANG L Y, KAW H Y, MENG L Y, SHANG H B, LI D H. Simultaneous determination of multiple phytohormones in tomato by ionic liquid-functionalized carbon fibers-based solid-phase microextraction coupled with liquid chromatography-mass spectrometry. Analytica Chimica Acta, 2020, 1137(9): 143-155.
[2] TANK J G, PANDYA R V, THAKER V S. ChemInform abstract: Phytohormones in regulation of the cell division and endoreduplication process in the plant cell cycle. ChemInform, 2014, 45(30): 12605-12613.
[3] 晁春艳, 王啸波, 李朝昉. 高效液相色谱法分离植物促生菌中的激素GA、IAA. 化学工程与装备, 2008, 34(8): 94-97. CHAO C Y, WANG X B, LI C F. Separation of GA and IAA in plant growth promoting rhizobacteria by high performance liquid chromatography. Chemical Engineering & Equipment, 2008, 34(8): 94-97.
[4] CAVITE H J M, MACTAL A G, EVANGLISTA E V, CRUZ J A. Growth and yield response of upland rice to application of plant growth promoting rhizobacteria. Journal of Plant Growth Regulation, 2020, 40(2): 1-15.
[5] 吴倩, 王璐, 吴大朋, 段春凤, 关亚风. 植物激素样品前处理方法的研究进展. 色谱, 2014, 32(4): 319-329. doi: 10.3724/SP.J.1123.2014.01039 WU Q, WANG L, WU D P, DUAN C F, GUAN Y F. Recent advances in sample preparation methods of plant hormones. Chinese Journal of Chromatography, 2014, 32(4): 319-329. doi: 10.3724/SP.J.1123.2014.01039
[6] 姚拓. 植物根际促生菌研究与应用. 北京: 中国农业出版社, 2021. YAO T. Research and Application of Plant Growth Promoting Rhizobacteria. Beijing: China Agriculture Press, 2021.
[7] 任卓然, 邵新庆, 李金升, 李慧, 何宜璇, 古维娜, 王茹颖, 杨灵婧, 刘克思. 微生物菌肥对退化高寒草甸地上生物量和土壤理化性质的影响. 草地学报, 2021, 29(10): 2265-2273. REN Z R, SHAO X Q, LI J S, LI H, HE Y X, GU W N, WANG R Y, YANG L J, LIU K S. Effects of microbial fertilizer on aboveground biomass and soil physiochemical properties of degraded alpine meadow. Acta Agrestia Sinica, 2021, 29(10): 2265-2273.
[8] BHATTACHARYYA P N, JHA D K. Plant growth-promoting rhizobacteria (PGPR): Emergence in agriculture. World Journal of Microbiology & Biotechnology, 2012, 28(4): 1327-1350.
[9] ZVI P, EDUARDO B. Hormone balance and abiotic stress tolerance in crop plants. Current Opinion in Plant Biology, 2011, 14(3): 290-295. doi: 10.1016/j.pbi.2011.02.001
[10] 肖爱华, 陈发菊, 贾忠奎, 桑子阳, 朱仲龙, 马履一. 梯度洗脱高效液相色谱法测定红花玉兰中4种植物激素. 分析试验室, 2020, 39(3): 249-254. XIAO A H, CHEN F J, JIA Z K, SANG Z Y, ZHU Z L, MA L Y. Determination of 4 plant hormones in Magnolia wufengensis by gradient elutionhigh performance liquid chromatography. Chinese Journal of Analysis Laboratory, 2020, 39(3): 249-254.
[11] LIU Y, FANG X A, CHEN G S, YE Y X, XU J Q, OUYANG G F, ZHU F. Recent development in sample preparation techniques for plant hormone analysis. Trends in Analytical Chemistry, 2019, 113(2): 224-233.
[12] 张占畅, 孙志红, 郑小能, 刘怀锋. 高效液相色谱法测定库尔勒香梨幼果中的4种内源激素. 新疆农业科学, 2017, 54(5): 886-892. doi: 10.6048/j.issn.1001-4330.2017.05.012 ZHANG Z C, SUN Z H, ZHENG X N, LIU H F. Determination of four endogenous hormones in the young fruit of korla feagrant pear by high performance liquid chromatography (HPLC). Xinjiang Agricultural Sciences, 2017, 54(5): 886-892. doi: 10.6048/j.issn.1001-4330.2017.05.012
[13] WANG L, XU J Y, WEI J, CHEN Z, XU Z C. Chloride salinity in a chloride-sensitive plant: Focusing on photosynthesis, hormone synthesis and transduction in tobacco. Plant Physiology and Biochemistry, 2020, 153(5): 119-130.
[14] SUNAYNA C, PAL K I, JAI M. Development and validation of a novel, rapid gradient HPLC method for simultaneous estimation of bioactive marker compounds in a mixture of convolvulus pluricaulis, withania somnifera and bacopa monnieri extracts. Journal of Chromatographic Science, 2020, 57(10): 920-930.
[15] 綦艳, 李锦清, 陈满英, 伍尚森, 唐楚林, 刘德群. 高效液相色谱法同时测定生活用纸及纸制品中19种有机抗菌剂. 分析测试学报, 2021, 40(7): 1018-1024. doi: 10.3969/j.issn.1004-4957.2021.07.006 QI Y, LI J Q, CHEN M Y, WU S S, TANG C L, LIU D Q. Simultaneous determination of 19 organic antibacterial agents in tissue paper and paper products by high performance liquid chromatography. Journal of Instrumental Analysis, 2021, 40(7): 1018-1024. doi: 10.3969/j.issn.1004-4957.2021.07.006
[16] LAO F, GIUSTI M M. Quantification of purple corn (Zea mays L.) anthocyanins using spectrophotometric and HPLC approaches: Method comparison and correlation. Food Analytical Methods, 2016, 9(5): 1367-1380. doi: 10.1007/s12161-015-0318-0
[17] 刘婷, 姚拓, 陈建纲, 刘欢. 固相萃取-高效液相色谱法测定植物根际促生菌发酵产物中3种植物激素的含量. 分析科学学报, 2017, 33(2): 201-206. LIU T, YAO T, CHEN J G, LIU H. Determination of plant hormones in bacterial fermentation products of plant growth promoting rhizobacteria by solid phase extraction-high perdormance liquid chromatography. Journal of Analytical Science, 2017, 33(2): 201-206.
[18] OMER Z S, OMBOLINI R, BROBERG A, GERHARDSON B. Indole-3-acetic acid production by pink-pigmented facultative methylotrophic bacteria. Plant Growth Regulation, 2004, 43(1): 93-96. doi: 10.1023/B:GROW.0000038360.09079.ad
[19] NAUTIYAL C S. An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiology Letters, 1999, 170(1): 265-270. doi: 10.1111/j.1574-6968.1999.tb13383.x
[20] 王璐. 基于基质固相分散萃取－高效液相色谱－串联质谱的植物激素分析. 合肥: 中国科学技术大学博士学位论文, 2014. WANG L. Determination of plant hormones by matrix solid－phase dispersion－high performance liquid chromatography-tandem mass spectrometry. PhD Thesis. Hefei: University of Science and Technology of China, 2014.
[21] ABBAS M B, HASHEM HANAA M A, ALPHONSE M M, TAREK M S. Synchronized determination of sacubitril and valsartan with some co-administered drugs in human plasma via UPLC-MS/MS method using solid phase extraction. Biomedical Chromatography, 2021, 35(11): 5203-5214.
[22] 林舒婷, 丁青青, 张文敏, 张兰, 卢巧梅. 固相萃取吸附材料在植物激素样品前处理中的应用新进展. 色谱, 2021, 39(12): 1281-1290. doi: 10.3724/SP.J.1123.2021.03045 LIN S T, DING Q Q, ZHANG W M, ZHANG L, LU Q M. Novel adsorption material for solid phase extraction in sample pretreatment of plant hormones. Chinese Journal of Chromatography, 2021, 39(12): 1281-1290. doi: 10.3724/SP.J.1123.2021.03045
[23] 蔡保东. 植物激素分析方法研究. 武汉: 武汉大学博士学位论文, 2016. CAI B D. Analytical methods for determination of phytohormones in plant. PhD Thesis. Wuhan: Wuhan University, 2016.
[24] 卢玉秋, 宋阿琳, 唐治玉, 李艳玲, 董炜灵, 王恩召, 唐治喜, 范分良. UPLC-MS/MS同时测定土壤中19种植物激素方法的建立和验证. 植物营养与肥料学报, 2019, 25(6): 953-962. doi: 10.11674/zwyf.18129 LU Y Q, SONG A L, TANG Z Y, LI Y L, DONG W L, WANG E Z, TANG Z X, FAN F L. Establishment and validation of UPLC-MS/MS method for simultaneous determination of 19 phytohormones in soil. Journal of Plant Nutrition and Fertilizer, 2019, 25(6): 953-962. doi: 10.11674/zwyf.18129
[25] 朱梅青, 崔蓉. 高效液相色谱法测定小鼠肝脏中的两种苯并三唑类紫外线吸收剂. 分析科学学报, 2021, 37(1): 93-97. doi: 10.2116/analsci.20SAR06 ZHU M Q, CUI R. Determination of two benzotriazole ultraviolet absorbents in mouse liver by highperformance liquid chromatography. Journal of Analytical Science, 2021, 37(1): 93-97. doi: 10.2116/analsci.20SAR06
[26] 纪晓娜, 吴文博, 任志敏, 陈国军, 李延君. 高效液相色谱法检测蔬菜中甲萘威残留. 分析科学学报, 2022, 38(1): 122-124. doi: 10.13526/j.issn.1006-6144.2022.01.022 JI X N, WU W B, REN Z M, CHEN G J, LI Y J. Detection of crbarayl rsidue in vegetables by high performance liquid cromatohgraphy. Journal of Analytical Science, 2022, 38(1): 122-124. doi: 10.13526/j.issn.1006-6144.2022.01.022
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