Skip to main content

Direct shoot organogenesis from leaf explants of Populus deltoides and changes in selected enzymatic activities

Physiology and Molecular Biology of Plants volume 26, pages 399–407 (2020)Cite this article

Abstract

The direct shoot organogenesis was achieved from leaf explant of two commercially important clones of Populus deltoides on MS medium enriched with 15 mg/l adenine sulphate, 5 mg/l Ascorbic acid, 250 mg/l (NH4)2SO4 (referred to as PD1 medium) supplemented with 2.5 µM each of 6-benzylaminopurine and indole-3-acetic acid. Higher shoot organogenic potential was recorded from the explants of clone ‘G48’ as compared to clone ‘L34’. The age of leaf explant also affected the shoot organogenic potential, and maximum shoot organogenesis was recorded in case of 5th leaf from the top of microshoot. Histological studies revealed altered cell division resulting in the formation of meristematic pockets after 5 days of culture, these meristematic pockets grew into dome protuberances by 10th day. Organized shoots were visible after 15 days of culture. A clear three phases of shoot organogenesis viz induction (0–4 days), initiation and organization (4–10 days) and growth (11–16 days onwards) were observed. Marked variation in the activity of enzymes such as catalase, peroxidase, polyphenol oxidase and acid phosphatase was observed during these phases. The activity of these enzymes was found to increase in cultures grown on the medium resulting in shoot organogenesis during shoot development (after 7 days of culture).

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  • Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    CAS  PubMed  Google Scholar 

  • Aggarwal D, Kumar A, Reddy MS (2010) Shoot organogenesis in elite clones of Eucalyptus tereticornis. Plant Cell Tissue Organ Cult 102(1):45–52

    Google Scholar 

  • Aggarwal D, Kumar A, Reddy MS (2011) Agrobacterium tumefaciens mediated genetic transformation of selected elite clone (s) of Eucalyptus tereticornis. Acta Physiol Plant 33(5):1603–1611

    CAS  Google Scholar 

  • Aggarwal D, Jaiswal N, Kumar A, Reddy MS (2013) Factors affecting genetic transformation and shoot organogenesis of Bacopa monnieri (L.) Wettst. J Plant Biochem Biotechnol 22(4):382–391

    Google Scholar 

  • Brown DC, Thorpe TA (1986) Plant regeneration by organogenesis. Cell Cult Somat Cell Genet Plants 3:49–65

    Google Scholar 

  • Chaturvedi HC, Sharma AK, Agha BQ, Jain M, Sharma M (2004) Production of cloned trees of Populus deltoides through in vitro regeneration of shoots from leaf, stem and root explants and their field cultivation. Indian J Biotechnol 3(2):203–208

    Google Scholar 

  • Coleman GD, Ernst SG (1990) In vitro shoot regeneration of Populus deltoides: effect of cytokinin and genotype. Plant Cell Rep 8:459–462

    Google Scholar 

  • Cui HY, Lee HS, Oh CY, Han SH, Lee KJ, Lee HJ, Kang KS, Park SY (2014) High-frequency regeneration by stem disc culture in selected clones of Populus euramericana. J Plant Biotechnol 41:236–241

    Google Scholar 

  • De Klerk GJ, Arnholdt-Schmitt B, Lieberei R, Neumann KH (1997) Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biol Plant 39(1):53–66

    Google Scholar 

  • de Marco A, Roubelakis-Angelakis KA (1996) The complexity of enzymic control of hydrogen peroxide concentration may affect the regeneration potential of plant protoplasts. Plant Physiol 110(1):137–145

    PubMed  PubMed Central  Google Scholar 

  • Dougall DK (1962) On the fate of tyrosine in tobacco callus tissue II. The lack of incorporation into lignin. Aust J Biol Sci 15(4):619–622

    CAS  Google Scholar 

  • Giri CC, Shyamkumar B, Anjaneyulu C (2004) Progress in tissue culture, genetic transformation and applications of biotechnology to trees: an overview. Trees 18(2):115–135

    Google Scholar 

  • Goel A, Kaur A, Kumar A (2018) Biochemical and histological changes during in vitro rooting of microcuttings of Bacopa monnieri (L.) Wettst. Acta Physiol Plant 40(3):64

    Google Scholar 

  • Hiraga S, Sasaki K, Ito H, Ohashi Y, Matsui H (2001) A large family of class III plant peroxidases. Plant Cell Physiol 42(5):462–468

    CAS  PubMed  Google Scholar 

  • Jehan H, Brown S, Marie D, Noin M, Prouteau M, Choursiqui D (1994) Ontogenesis and ploidy level of plantlets regenerated from Populus trichocarpa × deltoides cv. Hunnegem root, leaf and stem explants. J Plant Physiol 144(4–5):576–585

    CAS  Google Scholar 

  • Jiang C, Liu Z, Zheng Q (2015) Direct regeneration of plants derived form in vitro cultured shoots tips and leaves of poplar (Populus euramericana ‘Neva’). J Life Sci 9:366–372

    CAS  Google Scholar 

  • Kar M, Mishra D (1976) Catalase, peroxidase and polyphenol oxidase activities during rice leaf senescence. Plant Physiol 57:315–319

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kay LE, Basile D (1986) Specific peroxidase isozymes are correlated with organogenesis. Plant Physiol 84:99–105

    Google Scholar 

  • Kochba J, Lavee S, Spiegel-Roy P (1977) Differences in peroxidase activity and isoenzymes in embryogenic and non-embryogenic ‘Shamouti’ orange ovular callus lines. Plant Cell Physiol 18(2):463–467

    CAS  Google Scholar 

  • Kumar A (1996) Studies on in vitro propagation, biochemistry and field evaluation of two economically important plants: Rosa damascene Mill. and Gladiolus spp., Ph.D. Thesis. Kumaun University, Nainital

  • Kumar S, Chandra A (2009) Direct plant regeneration via multiple shoot induction in Stylosanthes seabrana. Cytologia 74(4):391–399

    CAS  Google Scholar 

  • Kumar A, Sood A, Palni LMS, Gupta AK (1999) In vitro propagation of Gladiolus hybridus Hort. Synergistic effect of heat shock and sucrose on morphogenesis. Plant Cell Tissue Organ Cult 57:105–112

    CAS  Google Scholar 

  • Kumar A, Aggarwal D, Gupta P, Reddy MS (2010) Factors affecting in vitro propagation and field establishment of Chlorophytum borivilianum. Biol Plant 54(4):601–606

    CAS  Google Scholar 

  • Kwon AR, Cui HY, Lee H, Shin H, Kang KS, Park SY (2015) Light quality affects shoot regeneration, cell division, and wood formation in elite clones of Populus euramericana. Acta Physiol Plant 37(3):65

    Google Scholar 

  • Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  PubMed Central  Google Scholar 

  • Martin KP, Joseph D, Madasser J, Philip VJ (2003) Direct shoot regeneration from lamina explants of two commercial cut flower cultivars of Anthurium andraeanum Hort. In Vitro Cell Dev Biol Plant 39(5):500–504

    Google Scholar 

  • McEwen CM Jr (1971) Monoamine oxidase. Methods Enzymol 17B:686–692

    Google Scholar 

  • Mhamdi A, Queval G, Chaouch S, Vanderauwera S, Van Breusegem F, Noctor G (2010) Catalase function in plants: a focus on Arabidopsis mutants as stress-mimic models. J Exp Bot 61(15):4197–4220

    CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497

    CAS  Google Scholar 

  • Noel N, Leple JC, Pilate G (2002) Optimization of in vitro micropropagation and regeneration for Populus × interamericana and Populus × euramericana hybrids (P. deltoides, P. trichocarpa, and P. nigra). Plant Cell Rep 20(12):1150–1155

    CAS  Google Scholar 

  • Panigrahi J, Behera M, Maharana S, Mishra RR (2007) Biomolecular changes during in vitro organogenesis of Asteracantha longifolia (L.) nees—a medicinal herb. Indian J Exp Biol 45:911–919

    CAS  PubMed  Google Scholar 

  • Patel KR, Thorpe TA (1984) Histochemical examination of shoot initiation in cultured cotyledon explants of radiata pine. Bot Gaz 145(3):312–322

    CAS  Google Scholar 

  • Rutledge CB, Douglas GC (1988) Culture of meristem tips and micropropagation of 12 commercial clones of poplar in vitro. Physiol Plant 72:367–373

    CAS  Google Scholar 

  • Sadasivam S, Manickam A (1992) Biochemical methods for agricultural sciences. Wiley Eastern Limited, New Delhi

    Google Scholar 

  • Sarkar S, Jha S (2017) Morpho-histological characterization and direct shoot organogenesis in two types of explants from Bacopa monnieri on unsupplemented basal medium. Plant Cell Tissue Organ Cult 130(2):435–441

    CAS  Google Scholar 

  • Sellmer JC, McCown BH, Haissig BE (1989) Shoot culture dynamics of six Populus clones. Tree Physiol 5(2):219–227

    CAS  PubMed  Google Scholar 

  • Sharma S, Millam S (2004) Somatic embryogenesis in Solanum tuberosum L.: a histological examination of key developmental stages. Plant Cell Rep 23:115–119

    CAS  PubMed  Google Scholar 

  • Son SH, Hall RB (1990) Multiple shoot regeneration from root organ cultures of Populus alba × P. grandidentata. Plant Cell Tissue Organ Cult 20:53–57

    CAS  Google Scholar 

  • Sujatha M, Sivaraj N, Prasad MS (2000) Biochemical and histological changes during in vitro organogenesis in Jatropha integerrima. Biol Plant 43(2):167–171

    CAS  Google Scholar 

  • Syono K (1965) Physiological and biochemical changes of carrot root callus during successive cultures. Plant Cell Physiol 6(3):371–392

    CAS  Google Scholar 

  • Thakur AK, Sharma S, Srivastava DK (2005) Plant regeneration and genetic transformation studies in petiole tissue of Himalayan poplar (Populus ciliata Wall.). Curr Sci 89(4):664

    CAS  Google Scholar 

  • Thakur AK, Saraswat A, Srivastava DK (2012) In vitro plant regeneration through direct organogenesis in Populus deltoides clone G48 from petiole explants. J Plant Biochem Biotechnol 21(1):23–29

    CAS  Google Scholar 

  • Thorpe TA, Gaspar T (1978) Changes in isoperoxidases during shoot formation in tobacco callus. Vitro 14(6):522–526

    CAS  Google Scholar 

  • Van Staden J, Zazimalova E, George EF (2008) Plant growth regulators II: cytokinins, their analogues and antagonists. In: George EF, Hall MA, Klerk GJD (eds) Plant propagation by tissue culture. Springer, Dordrecht, pp 205–226

    Google Scholar 

  • Vatankhah E, Niknam V, Ebrahimzadeh H (2010) Activity of antioxidant enzyme during in vitro organogenesis in Crocus sativus. Biol Plant 54(3):509–514

    CAS  Google Scholar 

  • Vinoth A, Ravindhoursan R (2013) In vitro propagation—a potential method for plant conservation. Int J Comput Algorithm 2:268–272

    Google Scholar 

  • Winton LL (1968) Plantlets from aspen tissue cultures. Science 160:1234–1235

    CAS  PubMed  Google Scholar 

  • Wolter KL (1968) Root and shoot initiation in aspen callus cultures. Nature 219:509–510

    CAS  PubMed  Google Scholar 

  • Yadav R, Arora P, Kumar D, Katyal D, Dilbaghi N, Chaudhury A (2009) High frequency direct plant regeneration from leaf, internode, and root segents of Eastern Cottonwood (Populus deltoides). Plant Biotechnol Rep 3(3):175–182

    Google Scholar 

  • Yepes LM, Aldwinekle HS (1994) Factors that effect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis. Plant Cell Tissue Organ Cult 37(3):257–269

    CAS  Google Scholar 

Download references

Acknowledgements

Funding was provided by Council of Scientific and Industrial Research, India (Grant No. (38(1326)/12/EMR-II)).

Author information

Affiliations

  1. TIFAC-Centre of Relevance and Excellence (CORE) in Agro and Industrial Biotechnology, Department of Biotechnology, Thapar Institute of Engineering and Technology, Patiala, 147004, India

    Saloni Sharma, M. S. Reddy & Anil Kumar

Authors
  1. Saloni Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. S. Reddy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anil Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anil Kumar.

Additional information

Publisher's Note

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

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Sharma, S., Reddy, M.S. & Kumar, A. Direct shoot organogenesis from leaf explants of Populus deltoides and changes in selected enzymatic activities. Physiol Mol Biol Plants 26, 399–407 (2020). https://doi.org/10.1007/s12298-019-00755-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12298-019-00755-4

Keywords

  • Enzyme activity
  • Explant maturity
  • Micropropagation
  • Organogenesis
  • Plant growth regulators