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Plant and Soil

, Volume 444, Issue 1–2, pp 177–191 | Cite as

Proteomics combined with BSMV-VIGS methods identified some N deficiency-responsive protein species and ABA role in wheat seedling

  • Guozhang Kang
  • Yufang Wu
  • Gezi Li
  • Pengfei Wang
  • Qiaoxia Han
  • Yonghua Wang
  • Yingxin Xie
  • Wei Feng
  • Dongyun Ma
  • Chenyang Wang
  • Tiancai GuoEmail author
Regular Article
  • 111 Downloads

Abstract

Aims

Crops often encounter a soil deficiency of nitrogen (N), the most important macronutrient for plants; however, the molecular mechanism of plant responses to N deficiency remains unclear. In this study, proteome-level changes that occur in bread wheat seedlings suffering from N deficiency were investigated to identify some N deficiency-responsive protein species in bread wheat.

Methods

We utilized isobaric tagging for relative and absolute quantification (iTRAQ) to measure changes in the proteome patterns of N-deficient wheat seedlings and validated the role of abscisic acid (ABA) using the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method.

Results

A total of 1515 N deficiency–responsive protein species were successfully identified in both root and leaf tissues of wheat seedlings suffering from 8-d N deficiency. Of these, abundance of wheat zeaxanthin epoxidase (TaZEP), a key ABA synthesis-related enzyme, was significantly upregulated, and the endogenous ABA contents also markedly increased. After TaZEP gene was further silenced using BSMV-VIGS method, BSMV-VIGS-TaZEP infected wheat seedlings showed enhanced sensitivity to N deficiency, suggesting silencing of TaZEP gene decreased the tolerance to N deficiency remarkably.

Conclusion

Our results identified some N deficiency-responsive protein species and revealed the role of ABA in wheat responses to N deficiency.

Keywords

Abscisic acid BSMV-VIGS iTRAQ Nitrogen deficiency Proteome Triticum aestivum

Abbreviations

ABA

abscisic acid

BSMV-VIGS

barley stripe mosaic virus-virus induced gene-silencing

CTK

cytokinin

DW

dry weight

ETH

ethylene

FW

fresh weight

IAA

auxin

iTRAQ

isobaric tagging for relative and absolute quantification

JA

jasmonic acid

MW

molecular weights

pI

isoelectric points

qRT-PCR

quantitative real-time PCR

SA

salicylic acid

ZEP

zeaxanthin epoxidase

Notes

Acknowledgements

This study was financially supported by the Science and Technology Innovation Program for Increase in Yield and Efficiency of Food Crop (2016YFD0300105) and the National Key Technology Support Program (2015BAD26B01).

Supplementary material

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Supplementary Table S1 (DOC 33 kb)
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Supplementary Fig. S2 (DOC 409 kb)

References

  1. Agrawal GK, Yamazaki M, Kobayashi M, Hirochika R, Miyao A, Hirochika H (2001) Screening of the rice viviparous mutants generated by endogenous retrotransposon Tos17 insertion. Tagging of a zeaxanthin epoxidase gene and a novel OsTATC gene. Plant Physiol 125:1248–1257PubMedPubMedCentralGoogle Scholar
  2. Borisjuk N, Kishchenko O, Eliby S, Schramm C, Anderson P, Jatayev S, Kurishbayev A, Shavrukov Y (2019) Genetic modification for wheat improvement: from transgenesis to genome editing. Biomed Res Int 2019:6216304Google Scholar
  3. Brenchley R, Spannagl M, Pfeifer M, Barker GL, D'Amore R, Allen AM et al (2012) Analysis of the bread wheat genome using whole–genome shotgun sequencing. Nature 491:705–710PubMedPubMedCentralGoogle Scholar
  4. Cai H, Lu Y, Xie W, Zhu T, Lian X (2012) Transcriptome response to nitrogen starvation in rice. J Biosci 37:731–747PubMedGoogle Scholar
  5. Cao A, Xing L, Wang X, Yang X, Wang W, Sun Y, Qian C, Ni J, Chen Y, Liu D, Wang X, Chen P (2011) Serine/threonine kinase gene Stpk-V, a key member of powdery mildew resistance gene Pm21, confers powdery mildew resistance in wheat. Proc Natl Acad Sci U S A 108:7727–7732PubMedPubMedCentralGoogle Scholar
  6. Chao WS, Doğramaci M, Horvath DP, Anderson JV, Foley ME (2016) Phytohormone balance and stress-related cellular responses are involved in the transition from bud to shoot growth in leafy spurge. BMC Plant Biol 16:47PubMedPubMedCentralGoogle Scholar
  7. Criado MV, Roberts IN, Echeverria M, Barneix AJ (2007) Plant growth regulators and induction of leaf senescence in nitrogen–deprived wheat plants. J Plant Growth Regul 26:301–307Google Scholar
  8. Cuming AC, Stevenson SR (2015) From pond slime to rain forest: the evolution of ABA signalling and the acquisition of dehydration tolerance. New Phytol 206:5–7PubMedGoogle Scholar
  9. Curci PL, Cigliano RA, Zuluaga DL, Janni M, Sanseverino W, Sonnante G (2017) Transcriptomic response of durum wheat to nitrogen starvation. Sci Rep-UK 7:1176Google Scholar
  10. Deng G, Liu L, Zhong X, Lao C, Wang H, Wang B, Zhu C, Shah F, Peng D (2014) Comparative proteome analysis of the response of ramie under N, P and K deficiency. Planta 239:1175–1186PubMedGoogle Scholar
  11. Elberse IAM, van Damme JMM, Tienderen PHV (2003) Plasticity of growth characteristics in wild barley (Hordeum spontaneum) in response to nutrient limitation. J Ecol 91:371–382Google Scholar
  12. Feldman M, Levy AA, Fahima T, Korol A (2012) Genomic asymmetry in allopolyploid plants: wheat as a model. J Exp Bot 63:5045–5059PubMedGoogle Scholar
  13. Findlay GD, MacCoss MJ, Swanson WJ (2009) Proteomic discovery of previously unannotated, rapidly evolving seminal fluid genes in Drosophila. Genome Res 19:886–896PubMedPubMedCentralGoogle Scholar
  14. Gelli M, Duo Y, Konda AR, Zhang C, Holding D, Dweikat I (2014) Identification of differentially expressed genes between sorghum genotypes with contrasting nitrogen stress tolerance by genome-wide transcriptional profiling. BMC Genomics 15:60–66Google Scholar
  15. Grobei MA, Qeli E, Brunner E, Rehrauer H, Zhang R, Roschitzki B, Basler K, Ahrens CH, Grossniklaus U (2009) Deterministic protein inference for shotgun proteomics data provides new insights into Arabidopsis pollen development and function. Genome Res 19:1786–1800PubMedPubMedCentralGoogle Scholar
  16. Grün A, Buchner P, Broadley MR, Hawkesford MJ (2018) Identification and expression profiling of Pht1 phosphate transporters in wheat in controlled environments and in the field. Plant Biol 20:374–389PubMedGoogle Scholar
  17. Guo T, Xuan H, Yang Y, Wang L, Wei L, Wang Y, Kang G (2014) Transcription analysis of genes encoding the wheat root transporter NRT1 and NRT2 families during nitrogen starvation. J Plant Growth Regul 33:837–848Google Scholar
  18. Hakeem KR, Ahmad A, Iqbal M, Gucel S, Ozturk M (2011) Nitrogen-efficient rice cultivars can reduce nitrate pollution. Environ Sci Pollut Res 18:1184–1193Google Scholar
  19. Han YL, Liao JY, Yu Y, Song HX, Rong N, Guan CY, Lepo JE, Ismail AM, Zhang ZH (2017) Exogenous abscisic acid promotes the nitrogen use efficiency of Brassica napus by increasing nitrogen remobilization in the leaves. J Plant Nutr 40:2540–2549Google Scholar
  20. He M, Zhu C, Dong K, Zhang T, Cheng Z, Li J, Yan Y (2015) Comparative proteome analysis of embryo and endosperm reveals central differential expression proteins involved in wheat seed germination. BMC Plant Biol 15:1–17Google Scholar
  21. He X, Ma H, Zhao X, Nie S, Li Y, Zhang Z et al (2016a) Comparative RNA-seq analysis reveals that regulatory network of maize root development controls the expression of genes in response to N stress. PLoS One 11:3Google Scholar
  22. He L, Zhang H, Zhang Y, Song X, Feng W, Kang G, Wang C, Guo T (2016b) Estimating canopy leaf nitrogen concentration in winter wheat based on multi-angular hyperspectral remote sensing. Eur J Agron 73:170–185Google Scholar
  23. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Calif Agric Exp Stn Circ 347:1–32Google Scholar
  24. Hu G, Jin K, Yoo MJ, Grupp K, Chen S, Wendel JF (2013) Proteomic profiling of developing cotton fibers from wild and domesticated Gossypium barbadense. New Phytol 200:570–582PubMedGoogle Scholar
  25. Hu J, Rampitsch C, Bykova NV (2015) Advances in plant proteomics toward improvement of crop productivity and stress resistance. Front Plant Sci 6:209PubMedPubMedCentralGoogle Scholar
  26. International Wheat Genome Sequencing Consortium (IWGSC) (2014) A chromosome-based draft sequence of the hexaploid bread wheat (Triticum aestivum) genome. Science 345:1251788Google Scholar
  27. International Wheat Genome Sequencing Consortium (IWGSC) (2018) Shifting the limits in wheat research and breeding using a fully annotated reference genome. Science 361:661Google Scholar
  28. Jia J, Zhao S, Kong X, Li Y, Zhao G, He W et al (2013) Aegilops tauschii draft genome sequence reveals a gene repertoire for wheat adaptation. Nature 496:91–95Google Scholar
  29. Jin X, Li W, Hu D, Shi X, Zhang X, Zhang F, Fu Z, Ding D, Liu Z, Tang J (2015) Biological responses and proteomic changes in maize seedlings under nitrogen deficiency. Plant Mol Biol Rep 33:490–504Google Scholar
  30. Kang G, Li G, Wang L, Wei L, Yang Y, Wang P, Yang Y, Wang Y, Feng W, Wang C, Guo T (2015) Hg–responsive proteins identified in wheat seedlings using iTRAQ analysis and the role of ABA in Hg stress. J Proteome Res 14:249–267PubMedGoogle Scholar
  31. Kosová K, Vítámvás P, Prášil IT, Renaut J (2011) Plant proteome changes under abiotic stress–contribution of proteomics studies to understanding plant stress response. J Proteome 74:1301–1322Google Scholar
  32. Kosová K, Vítámvás P, Urban MO, Prášil IT, Renaut J (2018) Plant abiotic stress proteomics: the major factors determining alterations in cellular proteome. Front Plant Sci 9:122Google Scholar
  33. Krapp A, Berthomé R, Orsel M, Mercey-Boutet S, Yu A, Castaings L, Elftieh S, Major H, Renou JP, Daniel-Vedele F (2011) Arabidopsis roots and shoots show distinct temporal adaptation patterns toward nitrogen starvation. Plant Physiol 157:1255–1282PubMedPubMedCentralGoogle Scholar
  34. Kuang Q, Zhang S, Wu P, Chen Y, Li M, Jiang H, Wu G (2017) Global gene expression analysis of the response of physic nut (Jatropha curcas L.) to medium- and long-term nitrogen deficiency. PLoS One 12:e0182700PubMedPubMedCentralGoogle Scholar
  35. Lee SC, Luan S (2012) ABA signal transduction at the crossroad of biotic and abiotic stress responses. Plant Cell Environ 35:53–60PubMedGoogle Scholar
  36. Li G, Peng X, Xuan H, Wei L, Yang Y, Guo T, Kang G (2013) Proteomic analysis of leaves and roots of common wheat (Triticum aestivum L.) under copper-stress conditions. J Proteome Res 12:4846–4861Google Scholar
  37. Li A, Liu D, Wu J, Zhao X, Hao M, Geng S, Yan J, Jiang X, Zhang L, Wu J, Yin L, Zhang R, Wu L, Zheng Y, Mao L (2014) mRNA and small RNA transcriptomes reveal insights into dynamic homoeolog regulation of allopolyploid heterosis in nascent hexaploid wheat. Plant Cell 26:1878–1900PubMedPubMedCentralGoogle Scholar
  38. Li G, Wu Y, Liu G, Xiao X, Wang P, Gao T, Xu M, Han Q, Wang Y, Guo T, Kang G (2017) Large-scale proteomics combined with transgenic experiments demonstrates an important role of jasmonic acid in potassium deficiency response in wheat and rice. Mol Cell Proteomics 16:1889–1905PubMedCentralGoogle Scholar
  39. Liang C, Tian J, Liao H (2013) Proteomics dissection of plant responses to mineral nutrient deficiency. Proteomics 13:624–636PubMedGoogle Scholar
  40. Ling HQ, Zhao S, Liu D, Wan J, Sun H, Zhang C et al (2013) Draft genome of the wheat A-genome progenitor Triticum urartu. Nature 496:87–90PubMedGoogle Scholar
  41. Liu G, Wu Y, Xu M, Gao T, Wang P, Wang L, Guo T, Kang G (2016) Virus-induced gene silencing identifies an important role of the TaRSR1 transcription factor in starch synthesis in bread wheat. Int J Mol Sci 17:1557PubMedCentralGoogle Scholar
  42. Ma TL, Wu WH, Wang Y (2012) Transcriptome analysis of rice root responses to potassium deficiency. BMC Plant Biol 12:161PubMedPubMedCentralGoogle Scholar
  43. Ma C, Zhou J, Chen G, Bian Y, Lv D, Li X, Wang Z, Yan Y (2014) iTRAQ-based quantitative proteome and phosphoprotein characterization reveals the central metabolism changes involved in wheat grain development. BMC Genomics 15:1–20Google Scholar
  44. Marioni JC, Mason CE, Mane SM, Stephens M, Gilad Y (2008) RNA-Seq: An assessment of technical reproducibility and comparison with gene expression arrays. Genome Res 18:1509–1517PubMedCentralGoogle Scholar
  45. Meise P, Jozefowicz AM, Uptmoor R, Mock HP, Ordon F, Schum A (2017) Comparative shoot proteome analysis of two potato (Solanum tuberosum L.) genotypes contrasting in nitrogen deficiency responses in vitro. J Proteome 166:68–82Google Scholar
  46. Møller ALB, Pedas P, Andersen B, Svensson B, Schjoerring JK, Finnie C (2011) Responses of barley root and shoot proteomes to long-term nitrogen deficiency, short-term nitrogen starvation and ammonium. Plant Cell Environ 34:2024–2037PubMedGoogle Scholar
  47. Nawaz MA, Chen C, Shireen F, Zheng Z, Sohail H, Afzal M, Ali MA, Bie Z, Huang Y (2018) Genome-wide expression profiling of leaves and roots of watermelon in response to low nitrogen. BMC Genomics 19:456PubMedPubMedCentralGoogle Scholar
  48. Nazir M, Pandey R, Siddiqi TO, Ibrahim MM, Qureshi MI, Abraham G, Vengavasi K, Ahamad A (2015) Nitrogen-deficiency stress induces protein expression differentially in low-N tolerant and low-N sensitive maize genotypes. Front Plant Sci 7:298Google Scholar
  49. Oka M, Shimoda Y, Sato N, Inoue J, Yamazaki T, Shimomura N, Fujiyama H (2012) Abscisic acid substantially inhibits senescence of cucumber plants (Cucumis sativus) grown under low nitrogen conditions. J Plant Physiol 169:789–796PubMedGoogle Scholar
  50. Qin L, Walk TC, Han P, Chen L, Zhang S, Li Y, Hu X, Xie L, Yang Y, Liu J, Lu X, Yu C, Tian J, Shaff JE, Kochian LV, Liao X, Liao H (2019) Adaption of roots to nitrogen deficiency revealed by 3D quantification and proteomic analysis. Plant Physiol 179:329–347PubMedGoogle Scholar
  51. Quan X, Zeng J, Ye L, Chen G, Han Z, Munawar J, Zhang G (2016) Transcriptome profiling analysis for two Tibetan wild barley genotypes in responses to low nitrogen. BMC Plant Biol 16:30PubMedPubMedCentralGoogle Scholar
  52. Ristova D, Carré C, Pervent M, Medici A, Kim GJ, Scalia D et al (2016) Combinatorial interaction network of transcriptomic and phenotypic responses to nitrogen and hormones in the Arabidopsis thaliana root. Sci Signal 9:451Google Scholar
  53. Schlüter H, Apweiler R, Holzhütter HG, Jungblut PR (2009) Finding one's way in proteomics: a protein species nomenclature. Chem Cent J 3:11PubMedPubMedCentralGoogle Scholar
  54. Scofield SR, Huang L, Brandt AS, Gill BS (2005) Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway. Plant Physiol 138:2165–2173PubMedPubMedCentralGoogle Scholar
  55. Shi W, Xu W, Li S, Zhao X, Dong G (2010) Responses of two rice cultivars differing in seedling–stage nitrogen use efficiency to growth under low–nitrogen conditions. Plant Soil 326:291–302Google Scholar
  56. Song J, Jiang L, Jameso PE (2012) Co-ordinate regulation of cytokinin gene family members during flag leaf and reproductive development in wheat. BMC Plant Biol 12:78PubMedPubMedCentralGoogle Scholar
  57. Torabi S, Wissuwa M, Heidari M, Naghavi MR, Gilany K, Hajirezaei MR, Omidi M, Yazdi-Samadi B, Ismail AM, Salekdeh GH (2009) A comparative proteome approach to decipher the mechanism of rice adaptation to phosphorus deficiency. Proteomics 9:159–170PubMedGoogle Scholar
  58. Tufan HA, Stefanato FL, McGrann GRD, MacCormack R, Boyd LA (2011) The barley stripe mosaic virus system used for virus-induced gene silencing in cereals differentially affects susceptibility to fungal pathogens in wheat. J Plant Physiol 168:990–994PubMedGoogle Scholar
  59. Vogel C, Marcotte EM (2012) Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet 13:227–232PubMedPubMedCentralGoogle Scholar
  60. Wang X, Bian Y, Cheng K, Zou H, Sun S, He J (2012) A comprehensive differential proteomic study of nitrate deprivation in Arabidopsis reveals complex regulatory networks of plant nitrogen responses. J Proteome Res 11:2301–2315PubMedGoogle Scholar
  61. Wei Z, Zeng X, Qin C, Wang Y, Bai L, Xu Q et al (2016) Comparative transcriptome analysis revealed genes commonly responsive to varied nitrate stress in leaves of tibetan hulless barley. Front Plant Sci 7:298Google Scholar
  62. Wisniewski JR, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Methods 6:359–362PubMedGoogle Scholar
  63. Wu H, Sparks C, Amoah B, Jones HD (2003) Factors influencing successful Agrobacterium-mediated genetic transformation of wheat. Plant Cell Rep 21:659–668PubMedGoogle Scholar
  64. Yang W, Yoon J, Choi H, Fan Y, Chen R, An G (2015) Transcriptome analysis of nitrogen-starvation responsive genes in rice. BMC Plant Biol 15:31PubMedPubMedCentralGoogle Scholar
  65. Zhang N, Huo W, Zhang L, Chen F, Cui D (2016) Identification of winter-responsive proteins in bread wheat using proteomics analysis and virus-induced gene silencing. Mol Cell Proteomics 15:2954–2969PubMedPubMedCentralGoogle Scholar
  66. Zhang Y, Zhou Y, Chen S, Liu J, Fan K, Li Z, Liu Z, Lin W (2019) Gibberellins play dual roles in response to phosphate starvation of tomato seedlings, negatively in shoots but positively in roots. J Plant Physiol 234–245:145–153Google Scholar
  67. Zhao T, Zhao S, Chen H, Zhao Q, Hu Z, Hou B, Xia G (2006) Transgenic wheat frogeny resistant to powdery mildew generated by Agrobacterium inoculumto the basal portion of wheat seedling. Plant Cell Rep 25:1199–1204PubMedGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Guozhang Kang
    • 1
  • Yufang Wu
    • 1
  • Gezi Li
    • 2
  • Pengfei Wang
    • 1
    • 2
    • 3
  • Qiaoxia Han
    • 2
  • Yonghua Wang
    • 1
    • 2
  • Yingxin Xie
    • 2
  • Wei Feng
    • 3
  • Dongyun Ma
    • 3
  • Chenyang Wang
    • 3
  • Tiancai Guo
    • 1
    Email author
  1. 1.Collaborative Innovation Center of Henan Food CropsHenan Agricultural UniversityZhengzhouPeople’s Republic of China
  2. 2.National Engineering Research Centre for WheatHenan Agricultural UniversityZhengzhouPeople’s Republic of China
  3. 3.The National Key Laboratory of Wheat and Maize Crop ScienceHenan Agricultural UniversityZhengzhouPeople’s Republic of China

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