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Transcriptional response of plasma membrane H+-ATPase genes to ammonium nutrition and its functional link to the release of biological nitrification inhibitors from sorghum roots

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Abstract

Aims

Sorghum (Sorghum bicolor) roots release biological nitrification inhibitors (BNIs) to suppress soil nitrification. Presence of NH4 + in the rhizosphere stimulates BNIs release and it is hypothesized to be functionally associated with plasma membrane (PM) H+-ATPase activity. However, whether the H+-ATPase is regulated at the transcriptional level, and if so, which isoforms of the H+-ATPases are involved in BNIs release are not known. Also, it is not clear whether the stimulation on BNIs release from roots is due to NH4 + uptake or its assimilation, which are addressed in this study.

Methods

Root exudates from intact sorghum plants were collected using aerated solutions of NH4 + or methyl-ammonium (MeA); and the BNI-activity release was determined. PM vesicles were isolated from fresh roots using a two-phase partitioning system; and the hydrolytic H+-ATPase activity was determined. All genes encoding PM H+-ATPases were searched in sorghum genome, and their expression in response to NH4 + or MeA were analyzed by quantitative RT-PCR in sorghum roots.

Results

BNIs release and PM H+-ATPase activity increased with NH4 + concentration (≤1.0 mM) in the root-exudate collection solutions, but at higher concentrations, it did not respond further or declined in case of the PM H+-ATPase activity. Twelve PM H+-ATPase genes were identified in sorghum genome; and these isoforms were designated SbA1 to SbA12. Five H+-ATPase genes were stimulated by NH4 + in the rhizosphere, and have similar expression pattern, which is consistent with the variation in H+-ATPase activity. MeA, a non-metabolizable analogue of NH4 +, had no significant effects on BNIs release, H+-ATPase activity, or expression of the H+-ATPase genes.

Conclusions

Our results suggest that the functional link between PM H+-ATPase activity and BNIs release is evident only at NH4 + levels of ≤1.0 mM in the rhizosphere. The variation in PM H+-ATPase activity by NH4 + is due to transcriptional regulation of five isoforms of the H+-ATPases. The stimulatory effect of NH4 + on BNIs release is functionally associated with NH4 + assimilation and not just with NH4 + uptake alone.

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References

  • Alsaadawi I, Al-Uqaili J, Alrubeaa A, Al-Hadithy S (1986) Allelopathic suppression of weed and nitrification by selected cultivars of Sorghum bicolor (L.) moench. J Chem Ecol 12:209–219

    Article  CAS  PubMed  Google Scholar 

  • Alvarez-Pizarro JC, Gomes-Filho E, Prisco JT, Grossi-De-Sá MF, De Oliveira-Neto OB, Da Rocha Fragoso R (2014) Plasma membrane H+-ATPase in sorghum roots as affected by potassium deficiency and nitrogen sources. Biol Plant 58:507–514

    Article  CAS  Google Scholar 

  • Arango M, Gevaudant F, Oufattole M, Boutry M (2003) The plasma membrane proton pump ATPase: the significance of gene subfamilies. Planta 216:355–365

    CAS  PubMed  Google Scholar 

  • Axelsen KB, Palmgren MG (2001) Inventory of the superfamily of P-type ion pumps in Arabidopsis. Plant Physiol 126:696–706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Axelsen K, Venema K, Jahn T, Baunsgaard L, Palmgren M (1999) Molecular dissection of the C-terminal regulatory domain of the plant plasma membrane H+-ATPase AHA2: mapping of residues that when altered give rise to an activated enzyme. Biochemistry 38:7227–7234

    Article  CAS  PubMed  Google Scholar 

  • Duby G, Boutry M (2009) The plant plasma membrane proton pump ATPase: a highly regulated P-type ATPase with multiple physiological roles. Pflugers Arch Eur J Physiol 457:645–655

    Article  CAS  Google Scholar 

  • Ermilova EV, Nikitin MM, Fernández E (2007) Chemotaxis to ammonium/methylammonium in Chlamydomonas reinhardtii: the role of transport systems for ammonium/methylammonium. Planta 226:1323–1332

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Iizumi T, Mizumoto M, Nakamura K (1998) A bioluminescence assay using Nitrosomonas europaea for rapid and sensitive detection of nitrification inhibitors. Appl Environ Microbiol 64:3656–3662

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ishiyama K, Kojima S, Takahashi H, Hayakawa T, Yamaya T (2003) Cell type distinct accumulations of mRNA and protein for NADH-dependent glutamate synthase in rice roots in response to the supply of NH4+. Plant Physiol Biochem 41:643–647

    Article  CAS  Google Scholar 

  • Kinoshita T, Shimazaki K (2002) Biochemical evidence for the requirement of 14-3-3 protein binding in activation of the guard-cell plasma membrane H+-ATPase by blue light. Plant Cell Physiol 43:1359–1365

    Article  CAS  PubMed  Google Scholar 

  • Kosola KR, Bloom AJ (1994) Methylammonium as a transport analog for ammonium in tomato (Lycopersicon esculentum L.). Plant Physiol 105:435–442

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lata JC, Durand J, Lensi R, Abbadie L (1999) Stable coexistence of contrasted nitrification statuses in a wet tropical savanna ecosystem. Funct Ecol 13:762–768

    Article  Google Scholar 

  • Lata JC, Degrange V, Raynaud X, Maron PA, Lensi R, Abbadie L (2004) Grass populations control nitrification in savanna soils. Funct Ecol 18:605–611

    Article  Google Scholar 

  • Loqué D, von Wirén N (2004) Regulatory levels for the transport of ammonium in plant roots. J Exp Bot 55:1293–1305

    Article  PubMed  Google Scholar 

  • Maudoux O, Batoko H, Oecking C, Gevaert K, Vandekerckhove J, Boutry M, Morsomme P (2000) A plant plasma membrane H+-ATPase expressed in yeast is activated by phosphorylation at its penultimate residue and binding of 14-3-3 regulatory proteins in the absence of fusicoccin. J Biol Chem 275:17762–17770

    Article  CAS  PubMed  Google Scholar 

  • Moore DRE, Waid JS (1971) The influence of washings of living roots on nitrification. Soil Biol Biochem 3:69–83

    Article  CAS  Google Scholar 

  • Ninnemann O, Jauniaux JC, Frommer WB (1994) Identification of a high affinity NH4+ transporter from plants. EMBO J 13:3464–3471

    CAS  PubMed  PubMed Central  Google Scholar 

  • Olsson A, Svennelid F, Ek B, Sommarin M, Larsson C (1998) A phosphothreonine residue at the C-terminal end of the plasma membrane H+-ATPase is protected by fusicoccin-induced 14-3-3 binding. Plant Physiol 118:551–555

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • O’Neill SD, Spanswick RM (1984) Effects of vanadate on the plasma membrane ATPase of red beet and corn. Plant Physiol 75:586–591

    Article  PubMed  PubMed Central  Google Scholar 

  • Palmgren MG (2001) Plant plasma membrane H+-ATPases: powerhouses for nutrient uptake. Annu Rev Plant Biol 52:817–845

    Article  CAS  Google Scholar 

  • Palmgren MG, Harper JF (1999) Pumping with plant P-type ATPases. J Exp Bot 50:883–893

    Article  CAS  Google Scholar 

  • Palmgren M, Sommarin M, Serrano R, Larsson C (1991) Identification of an autoinhibitory domain in the C-terminal region of the plant plasma membrane H+-ATPase. J Biol Chem 266:20470–20475

    CAS  PubMed  Google Scholar 

  • Portillo F (2000) Regulation of plasma membrane H+-ATPase in fungi and plants. Biochim Biophys Acta 1469:31–42

    Article  CAS  PubMed  Google Scholar 

  • Schubert S, Yan F (1997) Nitrate and ammonium nutrition of plants: effects on acid/base balance and adaptation of root cell plasmalemma H+-ATPase. Z Panzenphysiol Bodenkd 160:275–281

    Article  CAS  Google Scholar 

  • Shen H, He LF, Sasaki T, Yamamoto Y, Zheng SJ, Ligaba A, Yan XL, Ahn SJ, Yamaguchi M, Sasakawa H (2005) Citrate secretion coupled with the modulation of soybean root tip under aluminum stress. Up-regulation of transcription, translation, and threonine-oriented phosphorylation of plasma membrane H+-ATPase. Plant Physiol 138:287–296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Subbarao GV, Ishikawa T, Ito O, Nakahara K, Wang HY, Berry WL (2006) A bioluminescence assay to detect nitrification inhibitors released from plant roots: a case study with Brachiaria humidicola. Plant Soil 288:101–112

    Article  CAS  Google Scholar 

  • Subbarao GV, Rondon M, Ito O, Ishikawa T, Rao IM, Nakahara K, Lascano C, Berry WL (2007a) Biological nitrification inhibition (BNI)—is it a widespread phenomenon? Plant Soil 294:5–18

    Article  CAS  Google Scholar 

  • Subbarao GV, Wang HY, Ito O, Nakahara K, Berry WL (2007b) NH4 + triggers the synthesis and release of biological nitrification inhibition compounds in Brachiaria humidicola roots. Plant Soil 290:245–257

    Article  CAS  Google Scholar 

  • Subbarao GV, Nakahara K, Hurtado MP, Ono H, Moreta DE, Salcedo AF, Yoshihashi AT, Ishikawa T, Ishitani M, Ohnishi-Kameyama M, Yoshida M, Rondon M, Rao IM, Lascano CE, Berry WL, Ito O (2009) Evidence for biological nitrification inhibition in Brachiaria pastures. Proc Natl Acad Sci U S A 106:17302–17307

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Subbarao GV, Sahrawat K, Nakahara K, Ishikawa T, Kishii M, Rao I, Hash C, George T, Srinivasa Rao P, Nardi P (2012a) Biological nitrification inhibition—a novel strategy to regulate nitrification in agricultural systems. Adv Agron 114:249

    Article  CAS  Google Scholar 

  • Subbarao GV, Sahrawat K, Nakahara K, Rao I, Ishitani M, Hash C, Kishii M, Bonnett D, Berry W, Lata J (2012b) A paradigm shift towards low-nitrifying production systems: the role of biological nitrification inhibition (BNI). Ann Bot 112:297–316

    Article  PubMed  PubMed Central  Google Scholar 

  • Subbarao GV, Nakahara K, Ishikawa T, Ono H, Yoshida M, Yoshihashi T, Zhu Y, Zakir H, Deshpande S, Hash C (2013) Biological nitrification inhibition (BNI) activity in sorghum and its characterization. Plant Soil 366:243–259

    Article  CAS  Google Scholar 

  • Subbarao GV, Yoshihashi T, Worthington M, Nakahara K, Ando Y, Sahrawat KL, Rao IM, Lata J, Kishii M, Braun H (2015) Suppression of soil nitrification by plants. Plant Sci 233:155–164

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  • Tesfamariam T, Yoshinaga H, Deshpande SP, Srinivasa Rao P, Sahrawat KL, Ando Y, Nakahara K, Hash CT, Subbarao GV (2014) Biological nitrification inhibition in sorghum: the role of sorgoleone production. Plant Soil 379:325–335

    Article  CAS  Google Scholar 

  • Ullrich WR, Larsson M, Larsson CM, Lesch S, Novacky A (1984) Ammonium uptake in Lemna gibba G 1, related membrane potential changes, and inhibition of anion uptake. Physiol Plant 61:369–376

    Article  CAS  Google Scholar 

  • Van Beusichem ML, Kirkby EA, Baas R (1988) Influence of nitrate and ammonium nutrition on the uptake, assimilation, and distribution of nutrients in Ricinus communis. Plant Physiol 86:914–921

    Article  PubMed  PubMed Central  Google Scholar 

  • Walker N, Beilby M, Smith F (1979) Amine uniport at the plasmalemma of charophyte cells: I. Current–voltage curves, saturation kinetics, and effects of unstirred layers. J Membr Biol 49:21–55

    Article  CAS  Google Scholar 

  • Xu GH, Fan XR, Miller AJ (2012) Plant nitrogen assimilation and use efficiency. Annu Rev Plant Biol 63:153–182

    Article  CAS  PubMed  Google Scholar 

  • Yamashita K, Kasai M, Ezaki B, Shibasaka M, Yamamoto Y, Matsumoto H, Sasakawa H (1995) Stimulation of H+ extrusion and plasma membrane H+-ATPase activity of barley roots by ammonium-treatment. Soil Sci Plant Nutr 41:133–140

    Article  CAS  Google Scholar 

  • Yan F, Feuerle R, Schaffer S, Fortmeier H, Schubert S (1998) Adaptation of active proton pumping and plasmalemma H+-ATPase activity of corn roots to low root medium pH. Plant Physiol 117:311–319

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yan F, Zhu Y, Muller C, Zorb C, Schubert S (2002) Adaptation of H+-pumping and plasma membrane H+ ATPase activity in proteoid roots of white lupin under phosphate deficiency. Plant Physiol 129:50–63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yuan L, Loque D, Kojima S, Rauch S, Ishiyama K, Inoue E, Takahashi H, von Wiren N (2007) The organization of high-affinity ammonium uptake in Arabidopsis roots depends on the spatial arrangement and biochemical properties of AMT1-type transporters. Plant Cell 19:2636–2652

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zakir HAKM, Subbarao GV, Pearse SJ, Gopalakrishnan S, Ito O, Ishikawa T, Kawano N, Nakahara K, Yoshihashi T, Ono H, Yoshida M (2008) Detection, isolation and characterization of a root-exuded compound, methyl 3-(4-hydroxyphenyl) propionate, responsible for biological nitrification inhibition by sorghum (Sorghum bicolor). New Phytol 180:442–451

    Article  CAS  PubMed  Google Scholar 

  • Zeng HQ, Liu G, Kinoshita T, Zhang RP, Zhu YY, Shen QR, Xu GH (2012) Stimulation of phosphorus uptake by ammonium nutrition involves plasma membrane H+ ATPase in rice roots. Plant Soil 357:205–214

    Article  CAS  Google Scholar 

  • Zeng H, Feng X, Wang B, Zhu Y, Shen Q, Xu G (2013) Citrate exudation induced by aluminum is independent of plasma membrane H+-ATPase activity and coupled with potassium efflux from cluster roots of phosphorus-deficient white lupin. Plant Soil 366:389–400

    Article  CAS  Google Scholar 

  • Zhu YY, Di TJ, Xu GH, Chen X, Zeng HQ, Yan F, Shen QR (2009) Adaptation of plasma membrane H+-ATPase of rice roots to low pH as related to ammonium nutrition. Plant Cell Environ 32:1428–1440

    Article  CAS  PubMed  Google Scholar 

  • Zhu YY, Zeng HQ, Shen QR, Ishikawa T, Subbarao GV (2012) Interplay among NH4 + uptake, rhizosphere pH and plasma membrane H+-ATPase determine the release of BNIs in sorghum roots—possible mechanisms and underlying hypothesis. Plant Soil 358:131–141

    Article  CAS  Google Scholar 

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Acknowledgments

The research presented here is supported through JIRCAS invitation fellowship program to co-authors (Drs. Houqing Zeng, Tingjun Di and Prof. Yiyong Zhu), and is funded by grant-in-Aid for scientific research from Ministry of Agriculture, Forestry and Fisheries of Japan (MAFF) to JIRCAS under BNI project. Funding support also came from Natural Science Foundation of China (NSFC 31172035) and Program of New Century Excellent Talent in Universities (NCET-11-0672).

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Authors and Affiliations

  1. Crop, Livestock & Environment Division, JIRCAS, 1-1 Ohwashi, Tsukuba, Ibaraki, 305-8686, Japan

    Houqing Zeng, Tingjun Di & Guntur Venkata Subbarao

  2. College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, 310036, People’s Republic of China

    Houqing Zeng

  3. College of Resource and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Yiyong Zhu

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  1. Houqing Zeng
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  2. Tingjun Di
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  4. Guntur Venkata Subbarao
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Correspondence to Guntur Venkata Subbarao.

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Responsible Editor: Ad C. Borstlap.

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Zeng, H., Di, T., Zhu, Y. et al. Transcriptional response of plasma membrane H+-ATPase genes to ammonium nutrition and its functional link to the release of biological nitrification inhibitors from sorghum roots. Plant Soil 398, 301–312 (2016). https://doi.org/10.1007/s11104-015-2675-2

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