Over-Expression of Masson Pine PmPT1 Gene in Transgenic Tobacco Confers Tolerance Enhancement to Pi Deficiency by Ameliorating P Level and the Antioxidants

Abstract

Previously, we cloned the full sequence of masson pine (Pinus massoniana) phosphate transporter gene (PmPT1) from a phosphorus (Pi) deficiency tolerant strain. To further verify whether PmPT1 presumably function in angiosperms, i.e. tobacco, as well as to generate the new germplasm with high tolerance to Pi deficiency, currently, this gene was transferred into tobacco (Nicotiana tabacum) through Agrobacterium-mediated method. PmPT1 chiefly expressed in the roots of the transgenic plants, and considerably promoted the expression of two endogenous phosphate transporter genes of tobacco (NtPT1 and NtPT2) irrespectively of Pi status. Under low Pi conditions, the total P contents of the roots and shoots increased by 33.3% and 25.5%, respectively in L7, and by 30.7% and 23.9%, respectively in L18 in comparison with those of the wild type (WT). Also, the inorganic phosphorus (Pi) content of whole plants in L7 and L18 increased by 42.9% and 42.3%, respectively compared to the WT. The dry weight, contents of chlorophyll, soluble sugar and soluble protein, as well as the activities of peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) were significantly elevated, conversely the MDA accumulation was obviously decreased in transgenic lines compared to the WT. Therefore, PmPT1, a phosphate transporter gene of Pht1 family from masson pine might function in tobacco and two overexpressed-PmPT1 transgenic lines substantially enhanced the tolerance of the transgenic tobacco to low-P stress, which was at least chiefly ascribed to the improvement of P accumulation and oxidant alleviation.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  1. Ai P, Sun S, Zhao J, Fan X, Xin W, Guo Q, Yu L, Shen Q, Wu P, Miller AJ, Xu G (2009) Two rice phosphate transporters, Ospht1;2 and Ospht1;6, have different functions and kinetic properties in uptake and translocation. Plant J 57(5):798–809. https://doi.org/10.1111/j.1365-313x.2008.03726.x

    Article  PubMed  CAS  Google Scholar 

  2. Baldwin JC, Karthikeyan AS, Cao A, Raghothama KG (2008) Biochemical and molecular analysis of the LePS2;1: a phosphate starvation induced protein phosphatase gene from tomato. Planta 228(2):273–280. https://doi.org/10.1007/s00425-008-0736-y

    Article  PubMed  CAS  Google Scholar 

  3. Bieleski RL (1973) Phosphate pools, phosphate transport, and phosphate availability. Annu Rev Plant Physiol 24(1):225–252. https://doi.org/10.1146/annurev.pp.24.060173.001301

    Article  CAS  Google Scholar 

  4. Bucher M, Rausch C, Daram P (2001) Molecular and biochemical mechanisms of phosphorus uptake into plants. J Plant Nutr Soil Sci 164(2):209–217. https://doi.org/10.1002/1522-2624(200104)164:2<209::aid-jpln209>3.0.co;2-f

    Article  CAS  Google Scholar 

  5. Cai H, Chu Q, Gu R, Yuan L, Liu J, Zhang X, Chen F, Mi G, Zhang F (2012) Identification of QTLs for plant height, ear height and grain yield in maize ( Zea mays L.) in response to nitrogen and phosphorus supply. Plant Breed 131(4):502–510. https://doi.org/10.1111/j.1439-0523.2012.01963.x

    Article  CAS  Google Scholar 

  6. Carvalhais LC, Dennis PG, Fedoseyenko D, Hajirezaei MR, Borriss R, von Wirén N (2011) Root exudation of sugars, amino acids, and organic acids by maize as affected by nitrogen, phosphorus, potassium, and iron deficiency. J Plant Nutr Soil Sci 174(1):3–11. https://doi.org/10.1002/jpln.201000085

    Article  CAS  Google Scholar 

  7. Chacónlópez A, Ibarralaclette E, Sánchezcalderón L, Gutiérrezalanis D, Herreraestrella L (2011) Global expression pattern comparison between low phosphorus insensitive 4 and WT arabidopsis reveals an important role of reactive oxygen species and jasmonic acid in the root tip response to phosphate starvation. Plant Signal Behav 6(3):382–392. https://doi.org/10.4161/psb.6.3.14160

    Article  CAS  Google Scholar 

  8. Chen A, Hu J, Sun S, Xu G (2007) Conservation and divergence of both phosphate-and mycorrhiza-regulated physiological responses and expression patterns of phosphate transporters in solanaceous species. New Phytol 173(4):817–8https://doi.org/10.1111/j.1469-8137.2006.01962.x

  9. Chiou T-J, Liu H, Harrison MJ (2001) The spatial expression patterns of a phosphate transporter (MtPT1) from Medicago truncatula indicate a role in phosphate transport at the root/soil interface. Plant J 25(3):281–293. https://doi.org/10.1046/j.1365-313x.2001.00963.x

    Article  PubMed  CAS  Google Scholar 

  10. Ding Y, Luo W, Xu G (2006) Characterisation of magnesium nutrition and interaction of magnesium and potassium in rice. Ann Appl Biol 149(2):111–123. https://doi.org/10.1111/j.1744-7348.2006.00080.x

    Article  CAS  Google Scholar 

  11. Du H, Klessig DF (1997) Identification of a soluble, high-affinity salicylic acid-binding protein in tobacco. Plant Physiol 113(113):1319–1327. https://doi.org/10.1104/pp.113.4.1319

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Gao Y, Bian L, Shi J, Xu J, Xi M, Wang G (2013) Expression of a conifer cobra-like gene clcobl1 from chinese fir (cunninghamia lanceolata) alters the leaf architecture in tobacco. Plant Physiol Biochem 70(1):483–491. https://doi.org/10.1016/j.plaphy.2013.06.013

    Article  PubMed  CAS  Google Scholar 

  13. Gaxiola RA, Edwards M, Elser JJ (2011) A transgenic approach to enhance phosphorus use efficiency in crops as part of a comprehensive strategy for sustainable agriculture. Chemosphere 84(6):840–845. https://doi.org/10.1016/j.chemosphere.2011.01.062

    Article  PubMed  CAS  Google Scholar 

  14. Guan L, Smirnova I, Irina V, Nagamoni SS, Kaback R (2006) Manipulating phospholipids for crystallization of membrane transport proteins. Proc Natl Acad Sci U S A 103(6):1723–1736. https://doi.org/10.1073/pnas.0510922103

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Guo C, Zhao X, Liu X, Zhang L, Gu J, Li X, Lu W, Xiao K (2013) Function of wheat phosphate transporter gene tapht2;1, in pi translocation and plant growth regulation under replete and limited pi supply conditions. Planta 237(4):1163–1178. https://doi.org/10.1007/s00425-012-1836-2

    Article  PubMed  CAS  Google Scholar 

  16. Hammond JP, Bennett MJ, Bowen HC, Broadley MR, Eastwood DC, May ST, Rahn C, Swarup R, Woolaway KE, White PJ (2003) Changes in gene expression in Arabidopsis shoots during phosphate starvation and the potential for developing smart plants. Plant Physiol 132(2):578–596. https://doi.org/10.1104/pp.103.020941

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Jain A, Nagarajan VK, Raghothama KG (2012) Transcriptional regulation of phosphate acquisition by higher plants. Cell Mol Life Sci 69(19):3207–3224. https://doi.org/10.1007/s00018-012-1090-6

    Article  PubMed  CAS  Google Scholar 

  18. Jia H, Ren H, Gu M, Zhao J, Sun S, Zhang X, Chen J, Wu P, Xu G (2011) The phosphate transporter gene OsPht1;8 is involved in phosphate homeostasis in Rice. Plant Physiol 156(3):1164–1175. https://doi.org/10.1104/pp.111.175240

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Kalaji HM, Jajoo A, Oukarroum A, Brestic M, Zivcak M, Samborska I, Cetner M, Łukasik I, Goltsev V, Ladle R (2016) Chlorophyll a, fluorescence as a tool to monitor physiological status of plants under abiotic stress conditions. Acta Physiol Plant 38(4):102. https://doi.org/10.1007/s11738-016-2113-y

    Article  CAS  Google Scholar 

  20. Leyla PA, Andrea DC, Nuria F, Luz MM (2018) Aluminium toxicity and phosphate deficiency activates antioxidant systems and up-regulates expression of phosphate transporters gene in ryegrass ( Lolium perenne L) plants. Plant Physiol Biochem 9(9):445–454. https://doi.org/10.1016/j.plaphy.2018.07.031

    Article  CAS  Google Scholar 

  21. Liu LZ, Gong ZQ, Zhang YL, Li PJ (2011) Growth, cadmium accumulation and physiology of Marigold (Tagetes erecta L.) as affected by arbuscular mycorrhizal Fungi. Pedosphere 29(3):319–327. https://doi.org/10.1016/s1002-0160(11)60132-x

    Article  CAS  Google Scholar 

  22. Loth-Pereda V, Orsini E, Courty PE, Lota F, Kohler A, Diss L, Blaudez D, Chalot M, Nehls U, Bucher M, Martin F (2011) Structure and expression profile of the phosphate Pht1 transporter gene family in mycorrhizal Populus trichocarpa. Plant Physiol 156(4):2141–2154. https://doi.org/10.1104/pp.111.180646

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Misson J, Thibaud MC, Bechtold N, Raghothama K, Nussaume L (2004) Transcriptional regulation and functional properties of Arabidopsis Pht1;4, a high affinity transporter contributing greatly to phosphate uptake in phosphate deprived plants. Plant Mol Biol 55(5):727–741. https://doi.org/10.1007/s11103-004-1965-5

    Article  PubMed  CAS  Google Scholar 

  24. Misson J, Raghothama KG, Jain A, Jouhet J, Block MA, Bligny R, Ortet P, Creff A, Somerville S, Rolland N, Doumas P, Nacry P, Herrerra-Estrella L, Nussaume L, Thibaud MC (2005) A genome-wide transcriptional analysis using Arabidopsis thaliana Affymetrix gene chips determined plant responses to phosphate deprivation. Proc Natl Acad Sci U S A 102(102):11934–11939. https://doi.org/10.1073/pnas.0505266102

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Mittova V, Guy M, Tal M, Volokita M (2002) Response of the cultivated tomato and its wild salt-tolerant relative lycopersicon pennellii to salt-dependent oxidative stress: increased activities of antioxidant enzymes in root plastids. Free Radic Res 36(2):195–202. https://doi.org/10.1080/10715760290006402

    Article  PubMed  CAS  Google Scholar 

  26. Morcuende R, Bari R, Gibon Y, Zheng W, Pant BD, Bläsing O, Usadel B, Czechowski T, Udvardi MK, Stitt M, Scheible WR (2007) Genome-wide reprogramming of metabolism and regulatory networks of Arabidopsis in response to phosphorus. Plant Cell Environ 30(1):85–112. https://doi.org/10.1111/j.1365-3040.2006.01608.x

    Article  PubMed  CAS  Google Scholar 

  27. Mouradov A, Hamdorf B, Teasdale RD, Kim JT, Winter KU, Theissen G (1999) A def/glo-like mads-box gene from a gymnosperm: Pinus radiata contains an ortholog of angiosperm b class floral homeotic genes. Dev Genesis 25(3):245–252. https://doi.org/10.1002/(SICI)1520-6408(1999)25:3<245::AID-DVG7>3.0.CO;2-N

    Article  CAS  Google Scholar 

  28. Mudge SR, Rae AL, Diatloff E, Smith FW (2002) Expression analysis suggests novel roles for members of the Pht1 family of phosphate transporters in Arabidopsis. Plant J 31(3):341–353. https://doi.org/10.1046/j.1365-313X.2002.01356.x

    Article  PubMed  CAS  Google Scholar 

  29. Nagy R, Karandashov V, Chague V, Kalinkevich K, Tamasloukht M, Xu G, Jakobsen I, Levy AA, Amrhein N, Bucher M (2005) The characterization of novel mycorrhiza-specific phosphate transporters from Lycopersicon esculentum and Solanum tuberosum uncovers functional redundancy in symbiotic phosphate transport in solanaceous species. Plant J 42(2):236–250. https://doi.org/10.1111/j.1365-313X.2005.02364.x

    Article  PubMed  CAS  Google Scholar 

  30. Ni Z, Kim ED, Ha M, Lackey E, Liu J, Zhang Y, Sun Q, Chen ZJ (2009) Altered circadian rhythms regulate growth vigor in hybrids and allopolyploids. Nature 457(7227):327–331. https://doi.org/10.1038/nature07523

    Article  PubMed  CAS  Google Scholar 

  31. Nussaume L, Kanno S, Javot H, Marin E, Pochon N, Ayadi A, Nakanishi TM, Thibaud MC (2011) Phosphate import in plants: focus on the PHT1 transporters. Front Plant Sci 2(83):83. https://doi.org/10.3389/fpls.2011.00083

    Article  PubMed  PubMed Central  Google Scholar 

  32. O'Rourke JA, Yang SS, Miller SS, Bucciarelli B, Liu J, Rydeen A, Bozsoki Z, Uhde-Stone C, Tu ZJ, Allan D, Gronwald JW, Vance CP (2013) An RNA-Seq transcriptome analysis of orthophosphate-deficient white lupin reveals novel insights into phosphorus acclimation in plants. Plant Physiol 161(2):705–724. https://doi.org/10.1104/pp.112.209254

    Article  PubMed  CAS  Google Scholar 

  33. Paszkowski U, Kroken S, Roux C, Briggs SP (2002) Rice phosphate transporters include an evolutionarily divergent gene specifically activated in arbuscular mycorrhizal symbiosis. Proc Natl Acad Sci 99(20):13324–13329. https://doi.org/10.1073/pnas.202474599

    Article  PubMed  CAS  Google Scholar 

  34. Peñaloza E, Santiago M, Cabrera S, MuñozL G, Corcuera J, Silva H (2016) Characterization of the high-affinity phosphate transporter PHT1;4, gene promoter of Arabidopsis thaliana, in transgenic wheat. Biol Plant 61(3):1–10. https://doi.org/10.1007/s10535-016-0672-9

    Article  CAS  Google Scholar 

  35. Remy E, Cabrito TR, Batista RA, Teixeira MC, Sá-Correia I, Duque P (2012) The Pht1; 9 and Pht1; 8 transporters mediate inorganic phosphate acquisition by the Arabidopsis thaliana root during phosphorus starvation. New Phytol 195(2):356–371. https://doi.org/10.1111/j.1469-8137.2012.04167.x

    Article  PubMed  CAS  Google Scholar 

  36. Ren F, Zhao CZ, Liu CS, Huang KL, Guo QQ, Chang LL, Xiong H, Li XB (2014) A Brassica napus PHT1 phosphate transporter, BnPht1;4, promotes phosphate uptake and affects roots architecture of transgenic Arabidopsis. Plant Mol Biol 86(6):595–607. https://doi.org/10.1007/s11103-014-0249-y

    Article  PubMed  CAS  Google Scholar 

  37. Rosolem CA, Tavares CA (2006) Phosphorus deficiency symptoms in soybean. Rev Bras Cienc Solo 30(2):385–389. https://doi.org/10.1590/S0100-06832006000200018

    Article  CAS  Google Scholar 

  38. Schünmann PH, Richardson AE, Smith FW, Delhaize E (2004) Characterization of promoter expression patterns derived from the Pht1 phosphate transporter genes of barley (Hordeum vulgare L.). J Exp Bot 55(398):855–865. https://doi.org/10.1093/jxb/erh103

    Article  PubMed  CAS  Google Scholar 

  39. Shin H, Shin HS, Dewbre GR, Harrison MJ (2004) Phosphate transport in Arabidopsis: Pht1;1 and Pht1;4 play a major role in phosphate acquisition from both low- and high-phosphate environments. Plant J 39(4):629–642. https://doi.org/10.1111/j.1365-313X.2004.02161.x

    Article  PubMed  CAS  Google Scholar 

  40. Smith FW, Rae AL, Hawkesford MJ (2000) Molecular mechanisms of phosphate and sulphate transport in plants. BBA-Biomembranes 1465(1):236–245. https://doi.org/10.1016/s0005-2736(00)00141-3

    Article  PubMed  CAS  Google Scholar 

  41. Smith SE, Smith FA, Jakobsen I (2003) Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses. Plant Physiol 133(1):16–20. https://doi.org/10.1104/pp.103.024380

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Thibaud MC, Arrighi JF, Bayle V, Chiarenza S, Creff A, Bustos R, Paz-Ares J, Poirier Y, Nussaume L (2010) Dissection of local and systemic transcriptional responses to phosphate starvation in Arabidopsis. Plant J 64(5):775–789. https://doi.org/10.1111/j.1365-313X.2010.04375.x

    Article  PubMed  CAS  Google Scholar 

  43. Tian L, Zhou X, Ma L, Xu S, Nasir F, Tian C (2017) Root-associated bacterial diversities of oryza rufipogon and oryza sativa and their influencing environmental factors. Arch Microbiol 199(4):563–571. https://doi.org/10.1007/s00203-016-1325-2

    Article  PubMed  CAS  Google Scholar 

  44. Ticconi CA, Abel S (2004) Short on phosphate: plant surveillance and countermeasures. Trends Plant Sci 9(11):548–555. https://doi.org/10.1016/j.tplants.2004.09.003

    Article  PubMed  CAS  Google Scholar 

  45. Wang G, Gao Y, Wang J, Yang L, Song R, Li X, Shi J (2011) Overexpression of two cambium-abundant chinese fir (cunninghamia lanceolata) α-expansin genes Clexpa1 and Clexpa2 affect growth and development in transgenic tobacco and increase the amount of cellulose in stem cell walls. Plant Biotechnol J 9(4):486–502. https://doi.org/10.1111/j.1467-7652.2010.00569.x

    Article  PubMed  CAS  Google Scholar 

  46. Wasaki J, Yamamura T, Shinano T, Osaki M (2003) Secreted acid phosphatase is expressed in cluster roots of lupin in response to phosphorus deficiency. Plant Soil 248(1–2):129–136. https://doi.org/10.1023/a:1022332320384

    Article  CAS  Google Scholar 

  47. Wu H, Echt CS, Popp MP, Davis JM (1997) Molecular cloning, structure and expression of an elicitor-inducible chitinase gene from pine trees. Plant Mol Biol 33(6):979–987. https://doi.org/10.1371/journal.pone.0126186

    Article  PubMed  CAS  Google Scholar 

  48. Ye Y, Yuan J, Chang X, Yang M, Zhang L, Lu K, Lian X (2015) The phosphate transporter gene ospht1;4 is involved in phosphate homeostasis in rice. PLoS One 10(5):e0126186. https://doi.org/10.1371/journal.pone.0126186

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Yuan H, Liu D (2008) Signaling components involved in plant responses to phosphate starvation. J Integr Plant Biol 50(7):849–859. https://doi.org/10.1111/j.1744-7909.2008.00709.x

    Article  PubMed  CAS  Google Scholar 

  50. Zhang F, Wu XN, Zhou HM, Wang DF, Jiang TT, Sun YF, Cao Y, Pei WX (2014) Overexpression of rice phosphate transporter gene Ospt6 enhances phosphate uptake and accumulation in transgenic rice plants. Plant Soil 384(1–2):259–270. https://doi.org/10.1007/s11104-014-2168-8

    Article  CAS  Google Scholar 

  51. Zhang T, Wen XP, Ding GJ (2017) Ectomycorrhizal symbiosis enhances tolerance to low phosphorous through expression of phosphate transporter genes in masson pine (pinus massoniana). Acta Physiol Plant 39(4):101. https://doi.org/10.1007/s11738-017-2392-y

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The project was supported by grants from National Key R & D Plan, P. R. China (2017YFD060030304), the Provincial Fundation of Guizhou Province, P. R. China (2019-1014), as well as the Opening Foundation of Key Laboratory of Educational Ministry (2018-474).

Author information

Affiliations

Authors

Contributions

TZ, YH, XPW designed the experiments. TZ, YH performed the experiments. TZ, YH, XPW computed, analyzed data. TZ, YH, XPW wrote the paper. All authors read and approved the manuscript.

Corresponding author

Correspondence to XiaoPeng Wen.

Ethics declarations

Conflict of Interest

The authors declared that no conflicts of interest to this work. We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Key message

A masson pine phosphate transporter gene (PmPT1) was further transferred into tobacco using the Agrobacterium-mediated method. The transgenic plantlets were able to exhibit confers tolerance enhancement to Pi deficiency, which was at least chiefly ascribed to the improvement of expression of phosphate transporter genes, P accumulation and oxidant alleviation.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, T., Hong, Y. & Wen, X. Over-Expression of Masson Pine PmPT1 Gene in Transgenic Tobacco Confers Tolerance Enhancement to Pi Deficiency by Ameliorating P Level and the Antioxidants. Plant Mol Biol Rep 38, 238–249 (2020). https://doi.org/10.1007/s11105-019-01170-5

Download citation

Keywords

  • PmPT1
  • Genetic transformation
  • Tobacco
  • Low-phosphorus stress
  • Antioxidant