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利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响

达晓伟, 孙敏, 王馨, 庞海龙, 贾凌云, 冯汉青

达晓伟, 孙敏, 王馨, 庞海龙, 贾凌云, 冯汉青. 利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响[J]. 植物科学学报, 2022, 40(3): 390-397. DOI: 10.11913/PSJ.2095-0837.2022.30390
引用本文: 达晓伟, 孙敏, 王馨, 庞海龙, 贾凌云, 冯汉青. 利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响[J]. 植物科学学报, 2022, 40(3): 390-397. DOI: 10.11913/PSJ.2095-0837.2022.30390
Da Xiao-Wei, Sun Min, Wang Xin, Pang Hai-Long, Jia Ling-Yun, Feng Han-Qing. Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer[J]. Plant Science Journal, 2022, 40(3): 390-397. DOI: 10.11913/PSJ.2095-0837.2022.30390
Citation: Da Xiao-Wei, Sun Min, Wang Xin, Pang Hai-Long, Jia Ling-Yun, Feng Han-Qing. Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer[J]. Plant Science Journal, 2022, 40(3): 390-397. DOI: 10.11913/PSJ.2095-0837.2022.30390
达晓伟, 孙敏, 王馨, 庞海龙, 贾凌云, 冯汉青. 利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响[J]. 植物科学学报, 2022, 40(3): 390-397. CSTR: 32231.14.PSJ.2095-0837.2022.30390
引用本文: 达晓伟, 孙敏, 王馨, 庞海龙, 贾凌云, 冯汉青. 利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响[J]. 植物科学学报, 2022, 40(3): 390-397. CSTR: 32231.14.PSJ.2095-0837.2022.30390
Da Xiao-Wei, Sun Min, Wang Xin, Pang Hai-Long, Jia Ling-Yun, Feng Han-Qing. Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer[J]. Plant Science Journal, 2022, 40(3): 390-397. CSTR: 32231.14.PSJ.2095-0837.2022.30390
Citation: Da Xiao-Wei, Sun Min, Wang Xin, Pang Hai-Long, Jia Ling-Yun, Feng Han-Qing. Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer[J]. Plant Science Journal, 2022, 40(3): 390-397. CSTR: 32231.14.PSJ.2095-0837.2022.30390

利用荧光共振能量转移检测外源信号分子对拟南芥ATP水平的影响

基金项目: 

国家自然科学基金(31870246);甘肃省引导科技创新发展专项资金(2019ZX-05);西北师范大学新农村发展研究院开放基金;甘肃省重点研发计划(18YF1NA051)。

详细信息
    作者简介:

    达晓伟(1995-),女,硕士研究生,研究方向为植物生理生态(E-mail:2659708522@qq.com)。

    通讯作者:

    冯汉青,E-mail:fenghanq@nwnu.edu.cn

  • 中图分类号: Q945

Detection of the effects of exogenous signal molecules on ATP levels in Arabidopsis thaliana(L.)Heynh. using fluorescence resonance energy transfer

Funds: 

This work was supported by grants from the National Natural Science Foundation of China (31870246), Special Fund for Guiding Scientific and Technological Innovation Development of Gansu Province (2019ZX-05), Open Fund of New Rural Development Institute of Northwest Normal University, Key Research and Development Program of Gansu Province (18YF1NA051).

  • 摘要: 本研究使用ATP特异性荧光共振能量转移(Fluorescence resonance energy transfer,FRET)为基础的荧光蛋白传感器(Ateam1.03-nD/nA),分析了4种外源信号分子(细胞外ATP、Ca2+、H2O2和NO)对拟南芥(Arabidopsis thaliana(L.)Heynh.)幼苗叶绿体和细胞质中ATP水平的影响。结果显示,细胞质ATP水平整体高于叶绿体,在4种不同浓度的信号分子处理下,叶绿体Ateam1.03-nD/nA的FRET比值仅在1.2 ~ 1.8波动;细胞质Ateam1.03-nD/nA 的FRET比值仅在2.2 ~ 3.0之间波动,未产生显著变化。结果表明在以上外源信号分子的作用下,植物细胞质和叶绿体ATP均维持在较为稳定的水平。
    Abstract: We used an adenosine triphosphate (ATP) fluorescence protein-sensor (Ateam1.03-nD/nA) based on fluorescence resonance energy transfer (FRET) to analyze the effects of extracellular ATP, Ca2+, H2O2, and NO on ATP levels in the cytoplasm and chloroplasts of Arabidopsis thaliana seedlings. Results showed that ATP levels were higher in the cytoplasm than in the chloroplasts. Under treatment with different concentrations of the signal molecules, the FRET ratio of plastid-localized Ateam 1.03-nD/nA fluctuated non-significantly between 1.2 and 1.8. The FRET ratio of cytosol-localized Ateam1.03-nD/nA fluctuated non-significantly between 2.2 and 3.0. These results indicate that both cytoplasmic ATP and chloroplast ATP maintained relatively stable levels upon stimulation by signal molecules at different intensities.
  • [1]

    Stella N, Si LH, Ma Q, Gao Z. Optical aptasensors for adenosine triphosphate[J]. Theranostics, 2016, 6(10):1683-1702.

    [2]

    Valentina DC, Philippe F, Thomas N, Marlene E, Pao VC, et al. ATP sensing in living plant cells reveals tissue gradients and stress dynamics of energy physiology[J]. eLife, 2017, 6(6):e26770.

    [3]

    Chivasa S, Tomé DF, Hamilton JM, Slabas AR. Proteomic analysis of extracellular ATP-regulated proteins identifies ATP synthase beta-subunit as a novel plant cell death regulator[J]. Mol Cell Proteomics, 2011, 10(3).

    [4]

    Manfredi G, Yang L, Gajewski CD, Mattiazzi M. Measurements of ATP in mammalian cells[J]. Methods, 2002, 26(4):317-326.

    [5]

    Khlyntseva SV, Bazel'YR, Vishnikin AB, Andruch V. Methods for the determination of adenosine triphosphate and other adenine nucleotides[J]. J Anal Chem, 2009, 64(7):657-673.

    [6]

    Corriden R, Insel PA. Basal release of ATP:an autocrine-paracrine mechanism for cell regulation[J]. Sci Signal, 2010, 3(104):re1.

    [7]

    Conley JM, Saranya R, Valentino SA, Mathew T, Marco I. Imaging extracellular ATP with a genetically-encoded, ratiometric fluorescent sensor[J]. PLoS One, 2017, 12(11):e0187481.

    [8]

    Lmamura H, Nhat K, Togawa H, Saito K, Lino R, et al. Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators[J]. Proc Natl Acad Sci USA, 2009, 106(37):15651-15656.

    [9]

    Kotera I, Iwasaki T, Imamura H, Noji H, Nagai T. Reversible dimerization of aequorea victoria fluorescent proteins increases the dynamic range of FRET-based indicators[J]. ACS Chem Biol, 2010, 5(2):215-222.

    [10]

    Logan DC, Leaver CJ. Mitochondria-targeted GFP highlights the heterogeneity of mitochondrial shape, size and movement within living plant cells[J]. J Exp Bot, 2000(346):865-871.

    [11]

    Ando T, Imamura H, Suzuki R, Aizaki H, Suzuki T. Visua-lization and measurement of ATP levels in living cells replicating hepatitis C virus genome RNA[J]. PLoS Pathog, 2012, 8(3):e1002561.

    [12]

    Lerchundi R, Huang N, Rose CR. Quantitative imaging of changes in astrocytic and neuronal adenosine triphosphate using two different variants of ateam[J]. Front Cell Neurosci, 2020, 14(14).

    [13]

    Tang D, Wei F, Khan A, Munsif F, Zhou R, et al. Degradation of mitochondrial structure and deficiency of complexⅠ[STXFZ] were associated with the transgenic CMS of rice[J]. Biol Res, 2021, 54(1):6.

    [14]

    Luo D, Hong X, Liu Z, Guo J, Liu YG. A detrimental mitochondrial-nuclear interaction causes cytoplasmic male sterility in rice[J]. Nat Genet, 2013, 45(5):573.

    [15]

    Lang T, Deng C, Yao J, Zhang H, Deng S. A salt-signaling network involving ethylene, extracellular ATP, hydrogen peroxide, and calcium mediates K+/Na+ homeostasis in Arabidopsis[J]. Int J Mol Sci, 2020,21(22):8683.

    [16]

    Kim SY, Sivaguru M, Stacey G. Extracellular ATP in plants. visualization, localization, and analysis of physiological significance in growth and signaling[J]. Plant Physiol, 2006,142(3):984-992.

    [17]

    Lu Y, Li N, Sun J, Hou P, Jing X, et al. Exogenous hydrogen peroxide, nitric oxide and calcium mediate root ion fluxes in two non-secretor mangrove species subjected to NaCl stress[J]. Tree Physiol, 2013,33(1):81-95.

    [18]

    Wei Z, Miao Q, Sun D, Yang G, Wu C, et al. The mitochondrial phosphate transporters modulate plant responses to salt stress via affecting ATP and gibberellin metabolism in Arabidopsis thaliana[J]. PLoS One, 2012, 7(8):e43530.

    [19]

    Noriyuki H, Vadim PK, Hiromi I, Hiroyuki N, Takeharu N. Changes in cytosolic ATP levels and intracellular morpho-logy during bacteria-induced hypersensitive cell death as revealed by real-time fluorescence microscopy imaging[J]. Plant Cell Physiol, 2012,53(10):1768.

    [20]

    Elsa M, Sun J, Wang L, Bhat MG, Mohammad-Sidik AB, et al. DORN1/P2K1 and purino-calcium signalling in plants; making waves with extracellular ATP[J]. Ann Bot, 2020,124(7):1227-1242.

    [21]

    Voon CP, Guan X, Sun Y, Sahu A, Lim BL. ATP compartmentation in plastids and cytosol of Arabidopsis thaliana revealed by fluorescent protein sensing[J]. Proc Natl Acad Sci USA, 2018, 115(45):E10778-E10787.

    [22]

    Tripathi D, Tanaka K. A crosstalk between extracellular ATP and jasmonate signaling pathways for plant defense[J]. Plant Signal Behav, 2018,13(5):511-523.

    [23]

    Choi J, Tanaka K, Cao Y, Yue Q, Jing Q, et al. Identification of a plant receptor for extracellular ATP[J]. Science, 2014, 343(6168):290-294.

    [24]

    Riveras E, Alvarez JM, Vidal EA, Oses C, Vega A, Gutiérrez RA. The calcium ion is a second messenger in the nitrate signaling pathway of Arabidopsis[J]. Plant Physiol, 2015, 169(2):1397-404.

    [25]

    Huang F, Luo J, Ning T, Cao W, Jin X, et al. Cytosolic and nucleosolic calcium signaling in response to osmotic and salt stresses are independent of each other in roots of Arabidopsis seedlings[J]. Front Plant, 2017, 8(8):1648.

    [26]

    Suzuki N. Alleviation by calcium of cadmium-induced root growth inhibition in Arabidopsis seedlings[J]. Plant Biotechnol, 2005, 22(1):19-25.

    [27]

    Clark G, Roux S. Role of Ca2+in mediating plant responses to extracellular ATP and ADP[J]. Int J Mol Sci, 2018, 19(11):3590.

    [28]

    Hieno A, Naznin HA, Inaba-Hasegawa K, Yokogawa T, Hayami N, et al. Transcriptome analysis and identification of a transcriptional regulatory network in the response to H2O2[J]. Plant Physiol, 2019,180(3):1629-1646.

    [29]

    Ara N, Nakkanong K, Yang JH, Hu ZY, Zhang MF. Dissecting the heat stress-induced alterations in the leaf ultrastructure and some antioxidant network components in interspecific (Cucurbita maxima×Cucurbita moschata) inbred line of squash'Maxchata'as to its parents posses-sing variable heat toleranc[J]. Plant Growth Regul, 2015,76(3):289-301.

    [30]

    Jagadis GK, Aprajita K, Igor FS, Fernie AR, Igamberdiev AU. Interaction of nitric oxide with the components of the plant mitochondrial electron transport chain[J]. J Exp Bot, 2018,69(14):14.

    [31]

    Tiwari BS, Belenghi B, Levine A. Oxidative stress increased respiration and generation of reactive oxygen species, resulting in ATP depletion, opening of mitochondrial permeability transition, and programmed cell death[J]. Plant Physiol. 2002,128(4):1271-1281.

    [32]

    Fuente IMDL, Cortés JM, Valero E, Desroches M, Rodrigues S, et al. On the dynamics of the adenylate energy system:homeorhesis vs homeostasis[J]. PLoS One, 2014, 9(10):e108676.

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出版历程
  • 收稿日期:  2021-10-11
  • 修回日期:  2021-12-17
  • 网络出版日期:  2022-10-31
  • 发布日期:  2022-06-27

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