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Comparison of physiological and methylational changes in resynthesized Brassica napus and diploid progenitors under drought stress

  • Jinjin Jiang
  • Yi Yuan
  • Shuang Zhu
  • Tingting Fang
  • Liping Ran
  • Jian Wu
  • Youping WangEmail author
Original Article
  • 106 Downloads

Abstract

Brassica napus is a polyploid of certain research and economical value. Resynthesizing B. napus with diploid B. rapa and B. oleracea is essential for Brassica research because of the limited genetic background of B. napus. Considering that polyploids possess better agronomic traits and resistance compared with the corresponding diploids, we investigated drought tolerance after polyploidization of B. napus and revealed the epigenetic differences between polyploids and diploids. After drought stress, B. rapa and first-generation of synthesized hybrids (F1) were more wilted than B. oleracea and F2–F4 generations. However, the relative water content and water retention in F1 were better than others after drought stress. The increased number of partially opened and closed stomata in F1 was not as significant as that in F2 and F3, but stomata density in F1 was lower than F2, and the stomatal size in F1 was significantly reduced than F3. Physiological parameters varied among different generations of B. napus and diploid parents, and most of these parameters in hybrids were higher than B. rapa and lower than B. oleracea. However, the peroxidase activity in F3 and F4 was significantly higher than both parents, and the malondialdehyde content in F3 and F4 was lower than both parents, indicating that F3 and F4 might be more adaptive to oxidative stresses than other generations. DNA methylation level was decreased in F2 and F3 compared with F1, and then increased in F4. Methylation-sensitive amplified polymorphism analysis revealed that DNA methylation and demethylation broadly happened after drought stress. The methylation and demethylation level was F1 > F4 > B. oleracea > F2 > F3 > B. rapa and B. rapa > F4 > F3 > F2 > B. oleracea > F1, respectively. The epigenetic changes under drought stress might be related to the different stress tolerances during B. napus polyploidization.

Keywords

Resynthesized Brassica napus Drought stress Antioxidants DNA methylation 

Abbreviations

CMT3

Methyltransferase chromomethylase 3

MDA

Malondialdehyde

MET1

Methyltransferase 1

MS

Murashige and Skoog

MSAP

Methylation-sensitive amplified polymorphism

NBT

Nitrotetrazolium blue chloride

PBS

Phosphate-buffered saline

PEG

Polyethylene glycol

POD

Peroxidase

PVP

Polyvinyl pyrrolidone

qRT-PCR

Quantitative reverse transcriptase polymerase chain reaction

RWC

Relative water content

RWR

Relative water retention

SOD

Superoxide dismutase

TBA

Thiobarbituric acid

TCA

Trichloroacetic acid

Notes

Acknowledgements

This work was supported by the National Natural Science Foundations (31771824, 31330057, 31771825, 31401414), National Key Research and Development Program of China (2016YFD0101000, 2016YFD0102000), China Postdoctoral Science Foundation (2015T80591, 2014M561719), Jiangsu Postdoctoral Science Foundation (1401078B), and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

Supplementary material

11738_2019_2837_MOESM1_ESM.xlsx (11 kb)
Supplementary material 1 (XLSX 11 KB)

References

  1. Adams KL, Cronn R, Percifield R, Wendel JF (2003) Genes duplicated by polyploidy show unequal contributions to the transcriptome and organ-specific reciprocal silencing. Proc Natl Acad Sci 100:4649–4654CrossRefGoogle Scholar
  2. Ahmad J, Bashir H, Bagheri R, Baig A, Al-Huqail A, Ibrahim MM, Qureshi MI (2017) Drought and salinity induced changes in ecophysiology and proteomic profile of Parthenium hysterophorus. PLoS One 12(9):e0185118CrossRefGoogle Scholar
  3. Akram NA, Iqbal M, Muhammad A, Ashraf M, Al-Qurainy F, Shafiq S (2017) Aminolevulinic acid and nitric oxide regulate oxidative defence and secondary metabolisms in canola (Brassica napus L.) under drought stress. Protoplasma.  https://doi.org/10.1007/s00709-017-1140-x CrossRefPubMedGoogle Scholar
  4. Albertin W, Alix K, Balliau T, Brabant P, Davanture M, Malosse C, Valot B, Thiellement H (2007) Differential regulation of gene products in newly synthesized Brassica napus allotetraploids is not related to protein function nor subcellular localization. BMC Genom 8:56–70CrossRefGoogle Scholar
  5. Anssour S, Krugel T, Sharbel TF, Saluz HP, Bonaventure G, Baldwin IT (2009) Phenotypic, genetic and genomic consequences of natural and synthetic polyploidization of Nicotiana attenuate and Nicotianaobtusifolia. Ann Bot 103(8):1207–1217CrossRefGoogle Scholar
  6. Ascencio-Ibáñez JT, Sozzani R, Lee TJ, Chu TM, Wolfinger RD, Cella R, Hanley-Bowdoin L (2008) Globle analysis of Arabidopsis gene expression uncovers a complex array of changes impacting pathogene response and cell cycle during geminivirus infection. Plant Physiol 148:436–454CrossRefGoogle Scholar
  7. Barrs H, Weatherley P (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15:413–428CrossRefGoogle Scholar
  8. Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 103(4):551–560CrossRefGoogle Scholar
  9. Chen ZJ (2007) Genetic and epigenetic mechanisms for gene expression and phenotypic variation in plant polyploids. Annu Rev Plant Biol 58:377–406CrossRefGoogle Scholar
  10. Choudhury FK, Rivero RM, Blumwald E, Mittler R (2017) Reactive oxygen species, abiotic stress and stress combination. Plant J 90:856–867CrossRefGoogle Scholar
  11. Del Pozo JC, Ramirez-Parra E (2014) Deciphering the molecular bases for drought tolerance in Arabidopsis autotetraploids. Plant Cell Environ 37:2722–2737CrossRefGoogle Scholar
  12. Enjalbert JN, Zheng S, Johnson JJ, Mullen JL, Byrne PF, McKay JK (2013) Brassicaceae germplasm diversity for agronomic and seed quality traits under drought stress. Ind Crop Prod 47:176–185CrossRefGoogle Scholar
  13. Foster J, Luo B, Nakata PA (2016) An oxalyl-CoA dependent pathway of oxalate catabolism plays a role in regulating calcium oxalate crystal accumulation and defending against oxalate-secreting phytopathogens in Medicago truncatula. PLoS One 11(2):e0149850CrossRefGoogle Scholar
  14. Gaeta RT, Pires JC, Iniquez-Luy F, Leon E, Osborn TC (2007) Genomic changes in resynthesized Brassica napus and their effect on gene expression and phenotype. Plant Cell 19(11):3403–3417CrossRefGoogle Scholar
  15. Giannopolitis CN, Ries SK (1977) Superoxide dismutase I. Occurrence in higher plants. Plant Physiol 59:309–314CrossRefGoogle Scholar
  16. Gleason C, Huang S, Thatcher LF, Foley RC, Anderson CR, Carroll AJ, Millar AH, Singh KB (2011) Mitochondrial complex II has a key role in mitochondrial-derived reactive oxygen species influence on plant stress gene regulation and defense. Proc Natl Acad Sci US 108(26):10768–10773CrossRefGoogle Scholar
  17. Guo YM, Samans B, Chen S, Kibret KB, Hatzig S, Turner NC, Nelson MN, Cowling WA, Snowdon RJ (2017) Drought-tolerant Brassica rapa shows rapid expression of gene networks for general stress responses and programmed cell death under simulated drought stress. Plant Mol Biol Report 35(4):416–430CrossRefGoogle Scholar
  18. Hennig A, Kleinschmit JR, Schoneberg S, Loffler S, Jansen A, Polle A (2015) Water consumption and biomass production of protoplast fusion lines of poplar hybrids under drought stress. Front Plant Sci 6:330–343CrossRefGoogle Scholar
  19. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611CrossRefGoogle Scholar
  20. Jian H, Wang J, Wang T, Wei L, Liu L (2016) Identification of rapeseed microRNAs involved in early stage seed germination under salt and drought stresses. Front Plant Sci 7:658–672PubMedPubMedCentralGoogle Scholar
  21. Jiang J, Shao Y, Du K, Ran L, Fang X, Wang Y (2013) Use of digital gene expression to discriminate gene expression differences in early generations of resynthesized Brassica napus and its diploid progenitors. BMC Genom 14:72–82CrossRefGoogle Scholar
  22. Kong F, Mao S, Jiang J, Wang J, Fang X, Wang Y (2011) Proteomic changes in newly synthesized Brassica napus allotetraploids and their early generations. Plant Mol Biol Rep 29:927–935CrossRefGoogle Scholar
  23. Kumar D, Hassan MA, Naranjo MA, Agrawal V, Boscaiu M, Vicente O (2017) Effects of salinity and drought on growth, ionic relations, compatible solutes and activation of antioxidant systems in oleander (Nerium oleander L.). PLoS One 12(9):e0185017CrossRefGoogle Scholar
  24. Meng H, jiang S, Hua S, Lin X, Li Y, Guo W, Jiang L (2011) Comparison between a tetraploid turnip and its diploid progenitor (Brassica rapa L.): the adaptation to salinity stress. Agr Sci China 10:363–375CrossRefGoogle Scholar
  25. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498CrossRefGoogle Scholar
  26. Muscolo A, Sidari M, Anastasi U, Santonoceto C, Maggio A (2014) Effect of PEG-induced drought stress on seed germination of four lentil genotypes. J Plant Interact 9:354–363CrossRefGoogle Scholar
  27. Nir I, Moshelion M, Weiss D (2014) The Arabidopsis METHYL TRANSFERASE 1 suppresses gibberellin activity, reduces whole-plant transpiration and promotes drought tolerance in transgenic tomato. Plant Cell Environ 37:113–123CrossRefGoogle Scholar
  28. Niu S, Wang Y, Zhao Z, Deng M, Cao L, Yang L, Fan G (2016) Transcriptome and degradome of microRNAs and their targets in response to drought stress in the plants of a diploid and its autotetraploid Paulownia australis. PLoS One 11(7):e0158750CrossRefGoogle Scholar
  29. Osakabe Y, Osakabe K, Shinozaki K, Tran LSP (2014) Response of plants to water stress. Front Plant Sci 5:86–93CrossRefGoogle Scholar
  30. Qaderi MM, Kurepin LV, Reid DM (2006) Growth and physiological responses of canola (Brassica napus) to three components of global climate change: temperature, carbon dioxide and drought. Physiol Plant 128(4):710–721CrossRefGoogle Scholar
  31. Ran LP, Fang TT, Rong H, Jiang JJ, Fang YJ, Wang YP (2016) Analysis of cytosine methylation in early generations of resynthesized Brassica napus. J Integr Agric 15(6):1228–1238CrossRefGoogle Scholar
  32. Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B- and ozone-induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110:125–136CrossRefGoogle Scholar
  33. Rodriguez VM, Soengas P, Alonso-Villaverde V, Sotelo T, Cartea ME, Velasco P (2015) Effect of temperature stress on the early vegetative development of Brassica oleracea L. BMC Plant Biol 15:145–153CrossRefGoogle Scholar
  34. Seki M, Umezawa T, Urano K, Shinozaki K (2007) Regulatory metabolic networks in drought stress responses. Curr Opin Plant Biol 10:296–302CrossRefGoogle Scholar
  35. Shen Y, Zhang Y, Zou J, Meng J, Wang J (2015) Comparative proteomic study on Brassica hexaploid and its parents provides new insights into the effects of polyploidization. J Proteomics 112:274–284CrossRefGoogle Scholar
  36. Shi X, Zhang C, Ko DK, Chen ZJ (2015) Genome-wide dosage-dependent and—independent regulation contributes to gene expression and evolutionary in plant polyploids. Mol Biol Evol 32(9):2351–2366CrossRefGoogle Scholar
  37. Snyder J, Desborough S (1978) Rapid estimation of potato tuber total protein content with coomassie brilliant blue G-250. Theor Appl Genet 52:135–139CrossRefGoogle Scholar
  38. Soltis DE, Visger CJ, Marchant DB, Soltis PS (2016) Polyploidy: pitfalls and paths to a paradigm. Am J Bot 103:1146–1166CrossRefGoogle Scholar
  39. Song Q, Zhang T, Stelly DM, Chen ZJ (2017) Epigenomic and functional analysis reveal roles of epialleles in the loss of photoperiod sensitivity during domestication of allotetraploid cottons. Genome Biol 18:99–112CrossRefGoogle Scholar
  40. Urban MO, Vasek J, Klima M, Krtkova J, Kosova K, Prasil IT, Vitamvas P (2017) Proteomic and physiological approach reveals drought-induced changes in rapeseeds: water-saver and water-spender strategy. J Proteom 152:188–205CrossRefGoogle Scholar
  41. Valliyodan B, Nguyen HT (2006) Understanding regulatory networks and engineering for enhanced drought tolerance in plants. Curr Opin Plant Biol 9(2):189–195CrossRefGoogle Scholar
  42. Wang L, Jin X, Li Q, Wang X, Li Z, Wu X (2016) Comparative proteomics reveals that phosphorylation of β carbonic anhydrase 1 might be important for adaptation to drought stress in Brassica napus. Sci Rep 6:39024–39039CrossRefGoogle Scholar
  43. Wang N, Zhang W, Qin M, Li S, Qiao M, Liu Z, Xiang F (2017a) Drought tolerance conferred in soybean (Glycine max. L) by GmMYB84, a novel R2R3-MYB transcription factor. Plant Cell Physiol 58(10):1764–1776CrossRefGoogle Scholar
  44. Wang P, Yang C, Chen H, Song C, Zhang X, Wang D (2017b) Transcriptomic basis for drought-resistance in Brassica napus L. Sci Rep 7:40532–40551CrossRefGoogle Scholar
  45. Watkins J, Chapman JM, Muday GK (2017) Abscisic acid-induced reactive oxygen species are modulated by flavonols to control stomata aperture. Plant Physiol.  https://doi.org/10.1104/pp.17.01010 CrossRefPubMedPubMedCentralGoogle Scholar
  46. Wu L, Zhou H, Zhang Q, Zhang J, Ni F, Liu C, Qi Y (2010) DNA methylation mediated by a microRNA pathway. Mol Cell 38(3):465–475CrossRefGoogle Scholar
  47. Xia L, Yang L, Sun N, Li J, Fang Y, Wang Y (2016) Physiological and antioxidant enzyme gene expression analysis reveals the improved tolerance to drought stress of the somatic hybrid offspring of Brassica napus and Sinapis alba at vegetative stage. Acta Physiol Plant 38:88–97CrossRefGoogle Scholar
  48. Xiong LZ, Xu CG, Saghai MA, Zhang Q (1999) Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique. Mol Gen Genet 261:439–446CrossRefGoogle Scholar
  49. Yang C, Yang Z, Zhao L, Sun F, Liu B (2018) A newly formed hexaploid wheat exhibits immediate higher tolerance to nitrogen-deficiency than its parental lines. BMC Plant Biol 18:113–124CrossRefGoogle Scholar
  50. Zhao Q, Zou J, Meng J, Mei S, Wang J (2013) Tracing the transcriptomic changes in synthetic trigenomic allohexaploids of Brassica using an RNA-seq approach. PLoS One 8:e68883CrossRefGoogle Scholar
  51. Zhu JK (2016) Abiotic stress signalling and responses in plants. Cell 167:313–324CrossRefGoogle Scholar
  52. Zhu M, Simons B, Zhu N, Oppenheimer DG, Chen S (2010) Analysis of abscisic acid responsive proteins in Brassica napus guard cells by multiplexed isobaric tagging. J Proteomics 73:790–805CrossRefGoogle Scholar
  53. Zhu M, Monroe JG, Suhail Y, Villiers F, Mullen J, Pater D, Hauser F, Jeon BW, Bader JS, Kwak JM, Schroeder JI, McKay JK, Assmann SM (2016) Molecular and systems approaches towards drought-tolerant canola crops. New Phytol 210:1169–1189CrossRefGoogle Scholar
  54. Zhu X, Huang C, Zhang L, Liu H, Yu J, Hu Z, Hua W (2017) Systematic analysis of Hsf family genes in the Brassica napus genome reveals novel responses to heat, drought and high CO2 stresses. Front Plant Sci 8:1174–1188CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2019

Authors and Affiliations

  • Jinjin Jiang
    • 1
  • Yi Yuan
    • 1
  • Shuang Zhu
    • 1
  • Tingting Fang
    • 1
  • Liping Ran
    • 1
  • Jian Wu
    • 1
  • Youping Wang
    • 1
    Email author
  1. 1.Jiangsu Provincial Key Laboratory of Crop Genetics and PhysiologyYangzhou UniversityYangzhouChina

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