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In vitro induction of tetraploid Ziziphus jujuba Mill. var. spinosa plants from leaf explants

Plant Cell, Tissue and Organ Culture (PCTOC) volume 131, pages 175–182 (2017)Cite this article

Abstract

Tetraploid Ziziphus jujuba Mill. var. spinosa plants were induced by combining the adventitious shoot organogenesis from leaf explants and polyploidization with colchicine. Leaf explants were cultured on WPM containing1.0 mg L−1 thidiazuron and 0.3 mg L−1 3-indolyl-butyric acid for 10 or 20 days. They were then transferred to the same liquid medium supplemented with 40, 60, or 80 mg L−1 colchicine and incubated for 24, 48, or 72 h. The highest tetraploid induction rate (6.67%) resulted from a 10-day pre-culture treatment followed by an incubation in 60 mg L−1 colchicine for 72 h. The ploidy levels of the regenerated shootlets were determined based on flow cytometry analysis and chromosome numbers. Additionally, there were significant morphological differences between the tetraploid and diploid plants regarding stomatal size and density as well as leaf and shoot characteristics. This variability represents an early screening to distinguish the shoot with a ploidy level alternation from the diploid ones. The initial field performance results showed a faster growth in tetraploids than in the diploids, as suggested by the basal diameter and plant height increases.

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References

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

    Article  CAS  PubMed  Google Scholar 

  • Campos JMS, Davide LC, Salgado CC et al (2009) In vitro inductionof hexaploid plants from triploid hybrids of Pennisetum purpureum and Pennisetum glaucum. Plant Breed 128:101–104

    Article  CAS  Google Scholar 

  • Chauvin JE, Label A, Kermarrec MP (2005) In vitro chromosome doubling in tulip (Tulipa gesneriana L.). J Hortic Sci Biotechnol 80:693–698

    Article  Google Scholar 

  • Chen J, Li Z, Maiwulanjiang M et al (2013) Chemical and biological assessment of Ziziphus jujuba fruits from China: different geographical sources and developmental stages. J Agric Food Chem 61:7315–7324

    Article  CAS  PubMed  Google Scholar 

  • Chung MY, Kim CY, Min JS et al (2014) In vitro induction of tetraploids in an interspecifichybrid of Calanthe (Calanthe discolor × Calanthe sieboldii) through colchicine and oryzalin treatments. Plant Biotechnol Rep 8:251–257

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Dhooghe E, Van Laere K, Eeckhaut T et al (2011) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tiss Organ Cult 104:359–373

    Article  Google Scholar 

  • Doležel J, Greilhuber J, Suda J (2007) Flow cytometry with plant cells. Wiley, Weinheim

    Book  Google Scholar 

  • Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11(1):1–42

  • Duren M, Morpurgo R, Dolezel J et al (1996) Induction and verification of autotetraploids in diploid banana (Musa acuminata) by in vitro techniques. Euphytica 88:25–34

    Article  Google Scholar 

  • Esen A (1972) Tetraploid progenies from 2X · 4X crosses of citrus and their origin. J Am Soc Hortic Sci 97:410–414

    Google Scholar 

  • Ewald D, Ulrich K, Naujoks G et al (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

    Article  Google Scholar 

  • Fasolo F, Zimmerman RH, Fordham I (1989) Adventitions shoot formation on excised leaves of in vitro grown shoots of apple cultivars. Plant Cell Tiss Organ Cult 16:75–87

    Article  CAS  Google Scholar 

  • Galbraith DH, Harkins KR, Maddox JM et al (1983) Rapid flow cytometric analysis of the cell cycle in intact plant tissue. Science 220:1049–1051

    Article  CAS  PubMed  Google Scholar 

  • Gao QH, Wu CS, Wang M (2013) The jujube (Ziziphus jujuba Mill.) fruit: a review of current knowledge of fruit composition and health benefits. J Agric Food Chem 61:3351–3363

    Article  CAS  PubMed  Google Scholar 

  • Glowacka K, Jezowski S, Kaczmarek Z (2010) In vitro induction of polyploidy by colchicine treatment of shoots and preliminary characterisation of induced polyploids in two Miscanthus species. Ind Crop Prod 32:88–96

    Article  CAS  Google Scholar 

  • Gu XF, Zhang JR (2005) An efficient adventitious shoot regeneration system for Zhanhua winter jujube (Zizyphus jujuba Mill.) using leaf explants. Plant Cell Rep 23:775–779

    Article  CAS  PubMed  Google Scholar 

  • Guo Y, Shan G (2010) The Chinese jujube. Shanghai Scientific and Technical Publishers, Shanghai

    Google Scholar 

  • Jaskani MJ, Kwon SW, Koh GC et al (2004) Induction and characterization of tetraploid watermelon. Hortic Environ Biotechnol 45:60–65

    CAS  Google Scholar 

  • Jiao Y, Wickett NJ, Ayyampalayam S et al (2011) Ancestral polyploidy in seed plants and angiosperms. Nature 473:97–100

    Article  CAS  PubMed  Google Scholar 

  • Kadota M, Niimi Y (2002) In vitro induction of tetraploid plants from a diploid Japanese pear cultivar (Pyrus pyrifolia N. cv. Hosui). Plant Cell Rep 21:282–286

    Article  CAS  Google Scholar 

  • Kaensaksiri T, Soontornchainaksaeng P, Soonthornchareonnon N et al (2011) In vitro clonal propagation of Centella asiatica (L.) urban using shoot tip explants pre-treated with thidiazuron. Med Plants 3:47–52

    Google Scholar 

  • Kermani MJ, Sarasan V, Roberts AV et al (2003) Oryzalin-induced chromosome doubling in Rosa and its effect on plant morphology and pollen viability. Theor Appl Genet 107:1195–1200

    Article  CAS  PubMed  Google Scholar 

  • Leblay C, Chevreau E, Raboin L M (1991) Adventitious shoot regeneration from in vitro leaves of several pear cultivars (Pyrus communis L.). Plant cell Tissue Organ Cult 25(2):99–105

    CAS  Google Scholar 

  • Leus L, Van Laere K, Dewitte A, Van Huylenbroeck J (2009) Flowcytometry for plant breeding. Acta Hortic 836:221–226

    Article  Google Scholar 

  • Li JW, Fan LP, Ding SD et al (2007) Nutritional composition of five cultivars of Chinese jujube. Food Chem 103:454–460

    Article  CAS  Google Scholar 

  • Li YH, Kang XY, Wang SD et al (2008) Triploid induction in Populus alba x P. glandulosa by chromosome doubling of female gametes. Silvae Genet 57:37

    Google Scholar 

  • Lyrene PM, Perry JL (1982) Production and selection of blueberry polyploids in vitro. J Hered 73:377–378

    Article  Google Scholar 

  • McCown BH, Lloyd G (1981) Woody plant medium (WPM)—a mineral nutrient formulation for microculture of woody plant-species. Hortic Sci 16(3):453–453

    Google Scholar 

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

    Google Scholar 

  • Miguel TP, Leonhardt KW (2011) In vitro polyploid induction of orchids using oryzalin. Sci Hortic 130:314–319

    CAS  Google Scholar 

  • Mishra BK, Pathak S, Sharma A, Trivedi PK, Shukla S (2010) Modulated gene expression in newly synthesized auto-tetraploid of Papaver somniferum L. S Afr J Bot 76:447–452

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nilanthi D, Chen XL, Zhao FC et al (2009) Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L. BioMed Res Int 1:1–7

  • Notsuka K, Tsuru T, Shiraishi M (2000) Induced polyploid grapes via in vitro chromosome doubling. J Jpn Soc Hortic Sci 69:543–551

    Article  CAS  Google Scholar 

  • Petersen KK, Hagberg P, Kristiansen K et al (2002) In vitro chromosome doubling of Miscanthus sinensis. Plant Breed 121:445–450

    Article  Google Scholar 

  • Petersen KK, Hagberg P, Kristiansen K (2003) Colchicine and oryzalin mediated chromosome doubling in different genotypes of Miscanthus sinensis. Plant Cell Tissue Organ Cult 73:137–146

    Article  Google Scholar 

  • Roy A, Leggett G, Koutoulis A (2001) In vitro tetraploid induction and generation of tetraploids from mixoploids in hop (Humulus lupulus L.). Plant Cell Rep 20:489–495

    Article  CAS  Google Scholar 

  • Rugini E, Pannelli G, Ceccarelli M, Muganu M (1996) Isolation of triploid and tetraploid olive (Olea europaea L.) plants from mixoploid cv. ‘frantoio’ and ‘leccino’ mutants by in vivo and in vitro selection. Plant Breeding 115:23–27

    Article  Google Scholar 

  • Rugini E, Cristofori V, Silvestri C (2016a) Genetic improvement of olive (Olea europaea L.) by conventional and in vitro biotechnology methods. Biotechnol Adv 34:687–696

    Article  CAS  PubMed  Google Scholar 

  • Rugini E, Silvestri C, Ceccarelli M, Muleo R, Cristofori V (2016b) Mutagenesis and biotechnology techniques as tools for selecting new stable diploid and tetraploid olive genotypes and their dwarfing agronomical characterization. Hortic Sci 51:799–804

    Google Scholar 

  • Sari N, Abak K, Pitrat M (1999) Comparison of ploidy level screening methods in watermelon: Citrullus lanatus (Thunb.) Matsum and Nakai. Sci Hort 82:265–277

    Article  Google Scholar 

  • Shi Y, Wang Q, Zhou G et al (1990) Genome engineering breeding of apple in vitro. Fruit Breed Genet 317:13–22

    Google Scholar 

  • Shi QH, Liu P, Liu MJ et al (2015) A novel method for rapid in vivo induction of homogeneous polyploids via calluses in a woody fruit tree (Ziziphus jujuba Mill.). Plant Cell Tissue Organ Cult 121:423–433

    Article  CAS  Google Scholar 

  • Sun Q, Sun H, Li L et al (2009) In vitro colchicine-induced polyploid plantlet production and regeneration from leaf explants of the diploid pear (Pyrus communis L.) cultivar ‘Fertility’. J Hortic Sci Biotechnol 84:548–552

    Article  CAS  Google Scholar 

  • Tang ZQ, Chen DL, Song ZJ et al (2010) In vitro induction and identification of tetraploid plants of Paulownia tomentosa. Plant Cell Tissue Organ Cult 102:213–220

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Urwin NAR, Horsnell J, Moon T (2007) Generation and characterisation of colchicine-induced autotetraploid Lavandula angustifolia. Euphytica 156:257–266

    Article  Google Scholar 

  • Väïnölä A (2000) Polyploidization and early screening of Rhododendron hybrids. Euphytica 112:239–244

    Article  Google Scholar 

  • Van Laere K, França SC, Vansteenkiste H et al (2011) Influence of ploidy level on morphology, growth and drought susceptibility in Spathiphyllum wallisii. Acta Physiol Plant 33:1149–1156

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Yang X, Wang C (2005) Studies on induction and characteristics of grape polyploid. J Gansu Agric Univ 6:008

    Google Scholar 

  • Ye YM, Tong J, Shi XP et al (2010) Morphological and cytological studies of diploid and colchicine-induced tetraploid lines of crape myrtle (Lagerstroemia indica L.). Sci Hortic 124:95–101

    Article  Google Scholar 

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

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

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Acknowledgements

This research was financially supported by the Fundamental Research Funds for the Central Universities (2016ZCQ05) and the National Science Foundation of China (31400578).

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    Authors and Affiliations

    1. National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Beijing, China

      Yanhong Cui, Lu Hou, Xiang Li, Feiyi Huang, Xiaoming Pang & Yingyue Li

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    1. Yanhong Cui
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    2. Lu Hou
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    3. Xiang Li
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    4. Feiyi Huang
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    5. Xiaoming Pang
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    6. Yingyue Li
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    Contributions

    YL conceived and designed research; YC, LH and FH conducted experiments; YC and LH collected and analyzed the data; YC wrote the manuscript; YL and LH provided valuable suggestions on the manuscript; XP and YL revised the manuscript; YL obtained funding and is responsible for this article. All authors read and approved the manuscript.

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    Correspondence to Yingyue Li.

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    The authors declare that they have no conflict of interest.

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    Communicated by Maria Antonietta Germanà.

    Yanhong Cui and Lu Hou have contributed equally to this work.

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    Cui, Y., Hou, L., Li, X. et al. In vitro induction of tetraploid Ziziphus jujuba Mill. var. spinosa plants from leaf explants. Plant Cell Tiss Organ Cult 131, 175–182 (2017). https://doi.org/10.1007/s11240-017-1274-8

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    • DOI: https://doi.org/10.1007/s11240-017-1274-8

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