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In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus

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Abstract

The present study develops a protocol for tetraploids of multiple genotypes induced from in vitro leaf explants of diploid full-sib progeny [(P. pseudo-simonii × P. nigra ‘Zheyin3#’) × (P. × beijingensis)] after a colchicine treatment. Leaf explants from a ten genotypes full-sib progeny were cultured in MS basal medium containing with 1.78 μM BA and 0.27 μM NAA for 4, 5, and 6 days and transferred to the same liquid MS medium containing different concentrations of colchicine (50, 75, or 100 μM) for 2, 3, and 4 days, respectively. The results indicate that preculture duration, colchicine concentration, and exposure time had significative impacts in tetraploid induction rate but no significant correlation with the genotype. The feasible protocol involves treating a leaf explants pre-culture for 5 days followed by their transfer to liquid MS with 75 μM colchicine for 3 days, for most genotypes of full-sib progeny which could regenerate from leaf explants in the shoot regeneration medium. Tetraploid plantlets are identified using flow cytometric analysis and further confirmed after chromosome counts. Size and frequency of leaf stomata was significantly distinguish the tetraploid from the diploid plants.

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Abbreviations

MS:

Murashige and Skoog (1962)

BA:

6-Benzyladenine

NAA:

1-Naphthaleneacetic acid

IBA:

Indole butyric acid

DAPI:

4,6-Diamidino-2-phenylindole

MOPS:

4-Morpholinepropane sulfonate

References

  1. Barchet GL, Dauwe R, Guy RD, Schroeder WR, Soolanayakanahally RY, Campbell MM, Mansfield SD (2014) Investigating the drought-stress response of hybrid poplar genotypes by metabolite profiling. Tree Physiol 34:1203–1219

  2. Bradshaw HD, Stettler RF (1995) Molecular genetics of growth and development in populus. IV. Mapping QTLs with large effects on growth, form, and phenology traits in a forest tree. Genetics 139:963–973

  3. Bradshaw HD, Ceulemans R, Davis J, Stettler R (2000) Emerging model systems in plant biology: poplar (Populus) as a model forest tree. J Plant Growth Regul 19:306–313

  4. Cai X, Kang XY (2011) In vitro tetraploid induction from leaf explants of Populus pseudo-simonii Kitag. Plant Cell Rep 30:1771–1778

  5. Chen ZH, Mao QS (2001) On the properties of triploid Chinese white poplar and its relation with pulp and paper making. China Pulp Pap Ind 21:17–20

  6. Cheng S, Zhu X, Liao T, Li Y, Yao P, Suo Y, Kang X (2015) Gene expression differences between high-growth Populus allotriploids and their diploid parents. Forests 6:839–857

  7. Comai L (2005) The advantages and disadvantages of being polyploid. Nat Rev Genet 6:836–846

  8. De Carvalho JFRP, De Carvalho CRDP, Otoni WC (2005) In vitro induction of polyploidy in annatto (Bixa orellana). Plant Cell, Tissue Organ Cult 80:69–75

  9. Dermen H, Henry FB (1944) A general cytohistological study of colchicine polyploidy in Cranberry. Am J Bot 31:451–463

  10. Dhooghe E, Laere KV, Eeckhaut T, Leus L, Huylenbroeck JV (2010) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell, Tissue Organ Cult 104:359–373

  11. Dong CB, Mao JF, Suo YJ, Shi L, Wang J, Zhang PD, Kang XY (2014) A strategy for characterization of persistent heteroduplex DNA in higher plants. Plant J 80:282–291

  12. Eckenwalder JE (1996) Systematics and evolution of Populus. Biol Populus Implic Manag Conserv 1:7–32

  13. Einspahr DW (1984) Production and utilization of triploid hybrid aspen. Iowa State J Res 58:401–409

  14. Ewald D, Ulrich K, Naujoks G, Schröder MB (2009) Induction of tetraploid poplar and black locust plants using colchicine: chloroplast number as an early marker for selecting polyploids in vitro. Plant Cell, Tissue Organ Cult 99:353–357

  15. FAO (2008) Synthesis of country progress reports. Food and Agriculture Organization of the United Nations: the 23rd session of the International Poplar Commission, Beijing

  16. FAO (2012) Synthesis of country progress reports. Food and Agriculture Organization of the United Nations: the 24rd session of the International Poplar Commission, Dehradun

  17. Galbraith DW, Harkins KR, Maddox JM, Ayres NM, Sharma DP, Firoozabady E (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220:1049–1051

  18. Gilbert SK, Patterson FL (1965) Colchicine induced mutants in Decatur barley. Crop Sci 5:44–47

  19. Gu XF, Yang AF, Meng H, Zhang JR (2005) In vitro induction of tetraploid plants from diploid Zizyphus jujube Mill. Cv. Zhanhua. Plant Cell Rep 24:671–676

  20. Huang Z, Xu C, Li Y, Wang P, Li Y, Kang X (2015) Induction of somatic embryogenesis by anther-derived callus culture and plantlet ploidy determination in poplar (Populus × beijingensis). Plant Cell, Tissue Organ Cult 120:949–959

  21. Jiao Y, Wickett NJ, Ayyampalayam S, Chanderbali AS, Landherr L, Ralph PE, Leebens-Mack J (2011) Ancestral polyploidy in seed plants and angiosperms. Nature 473:97–100

  22. Johnsson H (1945) The triploid progeny of the cross diploid × tetraploid Populus tremula. Hereditas 31:411–440

  23. Kang X, Zhu Z, Zhang Z (1999) Suitable period of high temperature treatment of 2n pollen of Populus tomentosa × P. bolleana. J Beijing For Univ 22:1–4

  24. Li YH, Kang XY, Wang SD, Zhang ZH, Chen HW (2008) Triploid induction in Populus alba × P. glandulosaby chromosome doubling of female gametes. Silvae Genetica 57:37–40

  25. Li Y, Zhen H, Wang PQ, Li Y, Kang XY (2014) Establishment of leaf regeneration system for hybrids of Populus cathayana Rehd. J Nucl Agric Sci 28:0978–0984

  26. Liu G, Li Z, Bao M (2007) Colchicine-induced chromosome doubling in Platanus acerifolia and its effect on plant morphology. Euphytica 157:145–154

  27. Lu M, Zhang P, Wang J, Kang X, Wu J, Wang X, Chen Y (2014) Induction of tetraploidy using high temperature exposure during the first zygote division in Populus adenopoda Maxim. Plant Growth Regul 72:279–287

  28. Luckett D (1989) Colchicine mutagenesis is associated with substantial heritable variation in cotton. Euphytica 42:177–182

  29. Mashkina OS, Burdaeva LM, Belozerova MM, Vyunova LN (1989a) Ethod of obtaining diploid pollen of woody species. Lesoved 1:19–25

  30. Mashkina OS, Burdaeva LM, Vyunova LN (1989b) Experimental mutagenesis and polyploidy in breeding forest trees. Lesnaya genetika, selektsiya i fiziologiya drevesnykh rasteniĭ, Voronezh, 25-30 sentyabrya, 1989: Materialy Mezhdunarodnogo simpoziuma, pp 136–137. http://www.cabdirect.org/abstracts/19901696883.html

  31. Mergen F, Lester DT (1971) Colchicine induced polyploidy in Abies. For Sci 7:314–319

  32. Mohrdiek O, Melchior GH (1976) Combined hybrid and polyploid breeding as a promising method for poplars of the section Leuce: comparison of diploid and triploid grey poplars [Populus canescens] of the same progenies. Holzzucht 30:7–10

  33. Mu JJ (2013) Transcriptomic Analysis of Growth and Reproductive Changes in Birch (Betula platyphylla) Autotetraploids. Dissertation, Northeast Forestry University

  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

  35. Nilanthi D, Chen XL, Zhao FC, Yang YS, Wu H (2009) Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L. BioMed Res Int. doi:10.1155/2009/343485

  36. Nilsson-Ehle H (1936) Note regarding the gigas from of Populus tremula found in nature. Hereditas 21:372–382

  37. Seitz FW (1954) The occurrence of triploids after self-pollination of anomalous androgynous flowers of a grey poplar. Zeitschrift für Forstgenetik and Forstpflanzenzüchtung 3:1–6

  38. Sharman BC (1945) Leaf and bud initiation in the Gramineae. Bot Gaz 106:269–289

  39. Tang ZQ, Chen DL, Song ZJ, He YC, Cai DT (2010) In vitro induction and identification of tetraploid plants of Paulownia tomentosa. Plant Cell, Tissue Organ Cult 102:213–220

  40. Tavan M, Mirjalili MH, Karimzadeh G (2015) In vitro polyploidy induction: changes in morphological, anatomical and phytochemical characteristics of Thymus persicus (Lamiaceae). Plant Cell, Tissue Organ Cult 122:573–583

  41. Taylor GAIL (2002) Populus: Arabidopsis for forestry. Do we need a model tree. Ann Bot 90:681–689

  42. Thao NTP, Ureshino K, Miyajima I, Ozaki Y, Okubo H (2003) Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell, Tissue Organ Cult 72:19–25

  43. Tosca A, Pandolfi R, Citterio S, Fasoli A, Sgorbati S (1995) Determination of flow cytometry of the chromosome doubling capacity of oryzalin and colchicine in gynogenetic haploids of gerbera. Plant Cell Rep 14:455–458

  44. Ulrich K, Ewald D (2014) Breeding triploid aspen and poplar clones for biomass production. Silvae Genet 63:47–58

  45. Van Tuyl JM, Meijer B, Van Diën MP (1992) The use of oryzalin as an alternative for colchicine in in vitro chromosome doubling of Lilium and Nerine. In VI international symposium on flower bulbs, vol 325, pp 625–630

  46. Viehmannová I, Cusimamani EF, Bechyne M, Vyvadilová M, Greplová M (2009) In vitro induction of polyploidy in yacon (Smallanthus sonchifolius). Plant Cell, Tissue Organ Cult 97:21–25

  47. Wahid A, Gelani S, Ashraf M, Foolad MR (2007) Heat tolerance in plants: an overview. Environ Exp Bot 61:199–223

  48. Wan Y, Petolin JF, Widholm JM (1989) Efficient production of doubled haploid plants through colchicine treatment of antherderived maize callus. Theor Appl Genet 77:889–892

  49. Wang XJ, Wu JY, Cheng Y, Liao DZ (2006) Progress on research of improved variety propagation of Populus adenopoda Maxim. Hunan For Sci Technol 5:28

  50. Wang J, Kang XY, Shi L, Lu M, Yang J (2010) Tetraploid induction of Populus hybrid in section Tacamahaca through zygotic chromosome doubling with physical and chemical treatments. J Beijing For Univ 32:63–66

  51. Weisgerber H (1980) 25 years of forest tree breeding in Hesse. Allgemeine Forstzeitschrift 26:665–712

  52. Xie X, Agüero CB, Wang Y, Walker MA (2015) In vitro induction of tetraploids in Vitis × Muscadinia hybrids. Plant Cell, Tissue Organ Cult 122:675–683

  53. Yang X, Cao Z, An L, Wang Y, Fang XW (2006) In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.). Euphytica 152:217–224

  54. Yao C, Pu J (1998) Timber characteristics and pulp properties of the triploid of populus tomentosa. J Beijing For Univ 5:18–21

  55. Zhang S, Qi L, Chen C, Li X, Song W, Chen R et al (2004) A report of triploid Populus of the section Aigeiros. Silvae Genet 53:69–75

  56. Zhang Z, Kang X, Zhang P, Li Y, Wang J (2007) Incidence and molecular markers of 2n pollen in Populus tomentosa Carr. Euphytica 154:145–152. doi:10.1007/s10681-006-9280-7

  57. Zhang Z, Dai H, Xiao M, Liu X (2008) In vitro induction of tetraploids in Phlox subulata L. Euphytica 159:59–65

  58. Zhang J, Wei Z, Li D, Li B (2009) Using SSR markers to study the mechanism of 2n pollen formation in Populus × euramericana (Dode) Guinier and P. popularis. Ann For Sci 66:53–62

  59. Zhang Q, Luo F, Liu L, Guo F (2010) In vitro induction of tetraploids in crape myrtle (Lagerstroemia indica L.). Plant Cell, Tissue Organ Cult 101:41–47

  60. Zhu Z (2006) Genetic improvement of Populus tomentosa. Chinese Forestry Press, Beijing

  61. Zhu Z, Lin HB, Kang X (1995) Studies on allotriploid breeding of Populus tomentosa B301 clones. Sci Silvae Sin 31:499–505

  62. Zhu Z, Kang X, Zhang Z (1998) Studies on selection of natural triploid of Populus tomentosa. Sci Silvae Sin 34:22–31

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Acknowledgments

This research was supported by a Grant from National Natural Science Foundation of China for Molecular basis of the vegetative growth advantage in allotriploid poplar (31530012), Supported by a Grant from the Special Fund for Forest Scientific Research in the Public Welfare (201404113) and Supported by Program for Changjiang Scholars and Innovative Research Team in University (IRT13047).

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Correspondence to Xiangyang Kang.

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Xu, C., Huang, Z., Liao, T. et al. In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus . Plant Cell Tiss Organ Cult 125, 1–9 (2016). https://doi.org/10.1007/s11240-015-0922-0

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Keywords

  • Tetraploid
  • Colchicine
  • Chromosome doubling
  • Genotypes
  • Populus