Abstract
Aims
Alfalfa is the most important forage legume but sensitive to aluminum (Al), which largely limits its growth in acid soils. To improve Al resistance in alfalfa, responses to Al toxicity were investigated for understanding of the mechanisms of Al resistance in alfalfa.
Methods
Growth performance and Al resistance in forty-two cultivars was evaluated. Organic acids synthesis and exudation as well as the key genes in response to Al were investigated.
Results
Alfalfa cultivars showed diversity in Al resistance. Compared to the sensitive cultivar ‘Magnum 801’, the Al resistant cultivar ‘WL414’ had higher relative root elongation in response to Al toxicity, with less accumulation of Al. Al activated citrate and malate exudation in alfalfa, with higher citrate exudation and concentration as well as higher levels of citrate synthase (CS) activity, MsCS, MsALMT1, and MsMATE22 transcripts in root apex in WL414 than in Magnum 801. Citrate exudation is the major mechanism in Al resistance in alfalfa.
Conclusions
Alfalfa cultivars had diversity in Al resistance. Citrate synthesis and exudation plays a key role in Al resistance in alfalfa. Higher levels of citrate concentration and exudation are associated with Al resistance in Al resistant cultivar as compared with the sensitive cultivar.
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Abbreviations
- Al:
-
Aluminum
- ALMT:
-
Al-activated malate transporter
- CS:
-
Citrate synthase
- DTNB:
-
5,5′-dithio-bis-2-nitrobenzonic acid
- HPLC:
-
High performance liquid chromatography
- ICP-OES:
-
Inductively coupled plasma optical emission spectrometer
- IAA:
-
indole-3-acetic acid
- MATE:
-
Multidrug and toxic compound extrusion
- MDH:
-
Malate dehydrogenase
- OAs:
-
Organic acids
- PEP:
-
Phosphoenolpyruvate
- PEPC:
-
Phosphoenolpyruvate carboxylase
- qPCR:
-
Quantitative real-time PCR
References
An Y, Zhou P, Xiao Q, Shi DY (2014) Effects of foliar application of organic acids on alleviation of aluminum toxicity in alfalfa. J Plant Nutr Soil Sc 177:421–430
Annicchiarico P, Nazzicari N, Li X, Wei Y, Pecetti L, Brummer EC (2015) Accuracy of genomic selection for alfalfa biomass yield in different reference populations. BMC Genomics 16:1020. https://doi.org/10.1186/s12864-015-2212-y
Barone P, Rosellini D, Lafayette P, Bouton J, Veronesi F, Parrott W (2008) Bacterial citrate synthase expression and soil aluminum tolerance in transgenic alfalfa. Plant Cell Rep 27:893–901. https://doi.org/10.1007/s00299-008-0517-x
Biazzi E, Nazzicari N, Pecetti L, Brummer EC, Palmonari A, Tava A, Annicchiarico P (2017) Genome-wide association mapping and genomic selection for alfalfa (Medicago sativa) forage quality traits. PLoS One 12:e0169234. https://doi.org/10.1371/journal.pone.0169234
Bouton JH (1996) Screening the alfalfa core collection for acid soil tolerance. Crop Sci 36:198–200
Delhaize E, Craig S, Beaton CD, Bennet R, Jagadish V, Randall P (1993) Aluminum tolerance in wheat (Triticum aestivum L.) (I. Uptake and distribution of aluminum in root apices). Plant Physiol 103:685–693. https://doi.org/10.3389/fpls.2015.00587
Delhaize E, Hebb DM, Ryan PR (2001) Expression of a Pseudomonas aeruginosa citrate synthase gene in tobacco is not associated with either enhanced citrate accumulation or efflux. Plant Physiol 125:2059–2067. https://doi.org/10.1104/pp.125.4.2059
Ding ZJ, Yan JY, Xu XY, Li GX, Zheng SJ (2013) WRKY46 functions as a transcriptional repressor of ALMT1, regulating aluminum-induced malate secretion in Arabidopsis. Plant J 76:825–835. https://doi.org/10.1111/tpj.12337
Foy CD (1988) Plant adaptation to acid, aluminum-toxic soils. Commun Soil Sci Plant Anal 19:959–987
Furukawa J, Yamaji N, Wang H, Mitani N, Murata Y, Sato K, Katsuhara M, Takeda K, Ma JF (2007) An aluminum-activated citrate transporter in barley. Plant Cell Physiol 48:1081–1091. https://doi.org/10.1093/pcp/pcm091
Gou J, Debnath S, Sun L, Flanagan A, Tang Y, Jiang Q, Wen J, Wang ZY (2018) From model to crop: functional characterization of SPL8 in M. truncatula led to genetic improvement of biomass yield and abiotic stress tolerance in alfalfa. Plant Biotechnol J 16:951–962. https://doi.org/10.1111/pbi.12841
Hartel PG, Bouton JH (1989) Rhizobium meliloti inoculation of alfalfa selected for tolerance to acid, aluminum-rich soils. Plant Soil 116:283–285
Hayes JE, Ma JF (2003) Al-induced efflux of organic acid anions is poorly associated with internal organic acid metabolism in triticale roots. J Exp Bot 54:1753–1759. https://doi.org/10.1093/jxb/erg188
Hoekenga OA, Vision TJ, Shaff JE, Monforte AJ, Lee GP, Howell SH, Kochian LV (2003) Identification and characterization of aluminum tolerance loci in Arabidopsis (Landsberg erecta × Columbia) by quantitative trait locus mapping. A physiologically simple but genetically complex trait. Plant Physiol 132:936–948. https://doi.org/10.3389/fpls.2015.00587
Hufnagel B, Guimaraes CT, Craft EJ, Shaff JE, Schaffert RE, Kochian LV, Magalhaes JV (2018) Exploiting sorghum genetic diversity for enhanced aluminum tolerance: allele mining based on the AltSB locus. Sci Rep 8:10094
Johnson JF, Allan DL, Vance CP (1994) Phosphorus stress-induced proteoid roots show altered metabolism in lupinus albus. Plant Physiol 104:657–665. https://doi.org/10.1104/pp.104.2.657
Khu DM, Reyno R, Han Y, Zhao PX, Bouton JH, Brummer EC, Monteros MJ (2013) Identification of aluminum tolerance quantitative trait loci in tetraploid alfalfa. Crop Sci 53:148–163
Kochian LV, Hoekenga OA, Piñeros MA (2004) How do crop plants tolerate acid soils? Mechanisms of aluminum tolerance and phosphorous efficiency. Annu Rev Plant Biol 55:459–493. https://doi.org/10.1146/annurev.arplant.55.031903.141655
Kochian LV, Piñeros MA, Liu J, Magalhaes JV (2015) Plant adaptation to acid soils: the molecular basis for crop aluminum resistance. Annu Rev Plant Biol 66:571–598. https://doi.org/10.1146/annurev-arplant-043014-114822
Li XF, Ma JF, Matsumoto H (2000) Pattern of aluminum-induced secretion of organic acids differs between rye and wheat. Plant Physiol 123:1537–1544. https://doi.org/10.1104/pp.123.4.1537
Liao H, Wan H, Shaff J, Wang X, Yan X, Kochian LV (2006) Phosphorus and aluminum interactions in soybean in relation to aluminum tolerance. Exudation of specific organic acids from different regions of the intact root system. Plant Physiol 141:674–684. https://doi.org/10.1104/pp.105.076497
Ligaba A, Shen H, Shibata K, Yamamoto Y, Matsumoto H (2004) The role of phosphorus in aluminium-induced citrate and malate exudation from rape (Brassica napus). Physiol Plant 120:575–584. https://doi.org/10.1111/j.0031-9317.2004.0290.x
Liu MY, Lou HQ, Chen WW, Piñeros MA, Xu JM, Fan W, Kochian LV, Zheng SJ, Yang JL (2018) Two citrate transporters coordinately regulate citrate secretion from rice bean root tip under aluminum stress. Plant Cell Environ 41:809–822. https://doi.org/10.1111/pce.13150
Ma Z, Miyasaka SC (1998) Oxalate exudation by taro in response to Al. Plant Physiol 118:861–865. https://doi.org/10.1104/pp.118.3.861
Maron LG, Guimarães CT, Kirst M, Albert PS, Birchler JA, Bradbury PJ, Buckler ES, Coluccio AE, Danilova TV, Kudrna D, Magalhaes JV, Piñeros MA, Schatz MC, Wing RA, Kochian LV (2013) Aluminum tolerance in maize is associated with higher MATE1 gene copy number. Proc Natl Acad Sci U S A 110:5241–5246
Matsumoto H (2000) Cell biology of aluminum toxicity and tolerance in higher plants. Int Rev Cytol 200:1–46. https://doi.org/10.1016/s0074-7696(00)00001-2
Meyer FM, Gerwig J, Hammer E, Herzberg C, Commichau FM, Völker U, Stülke J (2011) Physical interactions between tricarboxylic acid cycle enzymes in Bacillus subtilis: evidence for a metabolon. Metab Eng 13:18–27. https://doi.org/10.1016/j.ymben.2010.10.001
Min X, Jin X, Liu W, Wei X, Zhang Z, Ndayambaza B, Wang Y (2019) Transcriptome-wide characterization and functional analysis of MATE transporters in response to aluminum toxicity in Medicago sativa L. PeerJ 7:e6302. https://doi.org/10.7717/peerj.6302
O'Rourke JA, Fu F, Bucciarelli B, Yang SS, Samac DA, Lamb JF, Monteros MJ, Graham MA, Gronwald JW, Krom N, Li J, Dai X, Zhao PX, Vance CP (2015) The Medicago sativa gene index 1.2: a web-accessible gene expression atlas for investigating expression differences between Medicago sativa subspecies. BMC Genomics 16:502. https://doi.org/10.1186/s12864-015-1718-7
Sasaki T, Yamamoto Y, Ezaki B, Katsuhara M, Ahn SJ, Ryan PR, Delhaize E, Matsumoto H (2004) A wheat gene encoding an aluminum-activated malate transporter. Plant J 37:645–653. https://doi.org/10.1111/j.1365-313x.2003.01991.x
Silva IR, Smyth TJ, Raper CD, Carter TE, Rufty TW (2001) Differential aluminum tolerance in soybean: An evaluation of the role of organic acids. Physiol Plant 112:200–210. https://doi.org/10.1034/j.1399-3054.2001.1120208.x
Sun LL, Liang CY, Chen ZJ, Liu PD, Tian J, Liu GD, Liao H (2014) Superior aluminium (Al) tolerance of Stylosanthes is achieved mainly by malate synthesis through an Al-enhanced malic enzyme, SgME1. New Phytol 202:209–219. https://doi.org/10.1111/nph.12629
Sun X, Chen J, Liu L, Rosanoff A, Xiong X, Zhang Y, Pei T (2018) Effects of magnesium fertilizer on the forage crude protein content depend upon available soil nitrogen. J Agric Food Chem 66:1743–1750. https://doi.org/10.1021/acs.jafc.7b04028
Tesfaye M, Temple SJ, Allan DL, Vance CP, Samac DA (2001) Overexpression of malate dehydrogenase in transgenic alfalfa enhances organic acid synthesis and confers tolerance to aluminum. Plant Physiol 127:1836–1844
Vance CP (1997) The molecular biology of N metabolism. In: DT Dennis, DH Turpin, DD Lefebrre, DB Layzell, eds, Plant Metabolism, Ed 2 Longman Scientific, London, pp 449-477
Voigt PW, Morris DR, Godwin HW (1997) A soil-on-agar method to evaluate acid-soil resistance in white clover. Crop Sci 37:1493–1496
Wang SY, Ren XY, Huang BR, Wang G, Zhou P, An Y (2016) Aluminium-induced reduction of plant growth in alfalfa (Medicago sativa) is mediated by interrupting auxin transport and accumulation in roots. Sci Rep 6:30079. https://doi.org/10.1038/srep30079
Wang J, Hou Q, Li P, Yang L, Sun X, Benedito VA (2017a) Diverse functions of multidrug and toxin extrusion (MATE) transporters in citric acid efflux and metal homeostasis in Medicago truncatula. Plant J 90:1–17. https://doi.org/10.1111/tpj.13471
Wang SY, Yuan SL, Su LT, Lv A, Zhou P, An Y (2017b) Aluminum toxicity in alfalfa (Medicago sativa) is alleviated by exogenous foliar IAA inducing reduction of Al accumulation in cell wall. Environ Exp Bot 139:1–13
Wu XL, Shi HF, Guo ZF (2018) Overexpression of a NF-YC gene results in enhanced drought and salt tolerance in transgenic seashore paspalum. Front Plant Sci 9:1355. https://doi.org/10.3389/fpls.2018.01355
Xu HW, Ji XM, He ZH, Shi WP, Zhu GH, Niu JK, Li BS, Peng XX (2006) Oxalate accumulation and regulation is independent of glycolate oxidase in rice leaves. J Exp Bot 57:1899–1908. https://doi.org/10.1093/jxb/erj131
Yang ZM, Sivaguru M, Horst WJ, Matsumoto H (2010) Aluminium tolerance is achieved by exudation of citric acid from roots of soybean (Glycine max). Physiol Plant 110:72–77
Zhang P, Zhong K, Zhong Z, Tong H (2019) Mining candidate gene for rice aluminum tolerance through genome wide association study and transcriptomic analysis. BMC Plant Biol 19:490. https://doi.org/10.1186/s12870-019-2036-z
Zheng SJ, Ma JF, Matsumoto H (1998) High aluminum resistance in buckwheat. I Al-induced specific secretion of oxalic acid from root tips. Plant Physiol 117:745–751. https://doi.org/10.1104/pp.117.3.745
Zhou P, Yang F, Ren XY, Huang BR, An Y (2014) Phytotoxicity of aluminum on root growth and indole-3-acetic acid accumulation and transport in alfalfa roots. Environ Exp Bot 104:1–8
Zhou P, Su L, Lv A, Wang S, Huang B, An Y (2016, 2016) Gene expression analysis of alfalfa seedlings response to acid-aluminum. Int J Genomics. https://doi.org/10.1155/2016/2095195
Zhu HF, Wang H, Zhu YF, Zou JW, Zhao FJ, Huang CF (2015) Genome-wide transcriptomic and phylogenetic analyses reveal distinct aluminum-tolerance mechanisms in the aluminum-accumulating species buckwheat (Fagopyrum tataricum). BMC Plant Biol 15:16. https://doi.org/10.1186/s12870-014-0395-z
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Grant numbers 31672481) and the Chinese Agriculture Research System- Green Manure (CARS-22-G-04).
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Sun, G., Zhu, H., Wen, S. et al. Citrate synthesis and exudation confer Al resistance in alfalfa (Medicago sativa L.). Plant Soil 449, 319–329 (2020). https://doi.org/10.1007/s11104-020-04490-8
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DOI: https://doi.org/10.1007/s11104-020-04490-8