Journal of Forestry Research

, Volume 28, Issue 6, pp 1169–1175 | Cite as

Plantlet regeneration of adult Pinus massoniana Lamb. trees using explants collected in March and thidiazuron in culture medium

Original Paper
First Online:
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Accepted:

Abstract

A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 μM TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 μM NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites.

Keywords

Thidiazuron Nodal segment Adventitious root Acclimatization NAA 
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Notes

Acknowledgements

This study was funded by the Science Research and Technology Development from the Department of Science and Technology of Guangxi (14125008-2-17, 1598006-5-7), the Natural Science Foundation of China (31360178), the Key Program of Guangxi Forestry Bureau ([2015]7), and as an independent project by the Key Laboratory of Gaungxi Fine Timber Forest Resources Cultivation (13A-01-01).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Ahmad N, Anis M (2007) Rapid plant regeneration protocol for cluster bean (Cyamopsis tetragonoloba L. Taub.). J Hortic Sci Biotechnol 82:585–589CrossRefGoogle Scholar
  2. Ahmad N, Siddique I, Anis M (2006) Improved plant regeneration in Capsicum annuum from nodal segment. Biol Plant 50:701–704CrossRefGoogle Scholar
  3. Alonso P, Moncaleán P, Fernández B, Rodríguez A, Centeno ML, Ordás R (2006) An improved micropropagation protocol for stone pine (Pinus pinea L.). Ann For Sci 63:879–885CrossRefGoogle Scholar
  4. Andersone U, Ievinsh G (2002) Changes of morphogenic competence in mature Pinus sylvestris L. buds in vitro. Ann Bot 90:293–298CrossRefPubMedPubMedCentralGoogle Scholar
  5. Campbell MM, Brunner AM, Jones HM, Strauss SH (2003) Forestry’s fertile crescent: the application of biotechnology to forest trees. Plant Biotechnol J 1:141–154CrossRefPubMedGoogle Scholar
  6. Capelle SC, Mok DWS, Kirchner SC, Mok MC (1983) Effects of thidiazuron on cytokinin autonomy and the metabolism of N6 (2-isopentenyl)[8-′4C] adenosine in callus tissue of Phaseolus lunatus L. Plant Physiol 73:796–802CrossRefPubMedPubMedCentralGoogle Scholar
  7. Chang YC, Lee N (1992) Tissue culture of Pleione formosana Hayata. J Chin Soc Hortsci 38:80–90Google Scholar
  8. Chen TY, Chen JT, Chang WC (2004) Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell Tissue Organ Cult 76:11–15CrossRefGoogle Scholar
  9. Davis JM, Becwar MR (2007) Examples of current implementation of cloning technology worldwide. In: Page A (ed) Developments in tree cloning. Pira International Ltd, Leatherhead, pp 32–33Google Scholar
  10. De Diego N, Montalbán IA, Fernández E, Moncaleán P (2008) In vitro regeneration of Pinus pinaster adult trees. Can J For Res 38:2607–2615CrossRefGoogle Scholar
  11. De Diego N, Montalbán IA, Moncaleán P (2010) In vitro regeneration of adult Pinus sylvestris L. trees. S Afr J Bot 76:158–162CrossRefGoogle Scholar
  12. Ding GJ, Zhou ZC, Wang ZR et al (2006) Cultivation and Utilization of Pulpwood Stand for Pinus massoniana[M]. China Forestry Press, Beijing, p 307Google Scholar
  13. Dumas E, Monteuuis O (1995) In vitro rooting of micropropagated shoots from juvenile and mature Pinus pinaster explants: influence of activated charcoal. Plant Cell Tissue Organ Cult 40:231–235CrossRefGoogle Scholar
  14. Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42CrossRefGoogle Scholar
  15. Ebert A, Taylor F, Blake J (1993) Changes of 6-benzylaminopurine and 2, 4 dichlorophenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal. Plant Cell Tissue Organ Cult 33:157–162CrossRefGoogle Scholar
  16. Evers PW, Donkers J, Prat A, Vermeer E (1988) Micropagation of forest trees through tissue culture. Pudoc Wageningen, Netherlands, p 84Google Scholar
  17. Faisal M, Ahmad N, Anis M (2005) Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron. Plant Cell Tissue Organ Cult 80:187–190CrossRefGoogle Scholar
  18. Hohtola A (1988) Seasonal changes in explant viability and contamination of tissue culture from mature Scots pine. Plant Cell Tissue Organ Cult 15:211–222CrossRefGoogle Scholar
  19. Hu DN, Shang-guan XC, Liu LY (2009) In vitro regeneration from stem segments of Cyclocarya paliurus (Batal.) Iljinskaja. Hubei Agric Sci 6:1300–1303Google Scholar
  20. Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33:105–119CrossRefGoogle Scholar
  21. Kanyand N, Dessai AP, Prakash SC (1994) Thidiazuron promotes high frequency regeneration of peanut (Arachis hypogea) plants in vitro. Plant Cell Rep 14:1–5CrossRefPubMedGoogle Scholar
  22. Kartsonas E, Papafotiou M (2007) Mother plant age and seasonal influence on in vitro propagation of Quercus euboica Pap., an endemic, rare and endangered oak species of Greece. Plant Cell Tissue Organ Cult 90:111–116CrossRefGoogle Scholar
  23. Khan MI, Ahmad N, Anis M (2011) The role of cytokinins on in vitro shoot production in Salix tetrasperma Roxb.: a tree of ecological importance. Trees 25:577–584CrossRefGoogle Scholar
  24. Kumar R, Sharma K, Agrawal V (2005) In vitro clonal propagation of Holarrhena antidysenterica (L.) Wall. through nodal explants from mature trees. In Vitro Cell Dev Biol Plant 41:137–144CrossRefGoogle Scholar
  25. Laukkanen H, Rautiainen L, Taulavuori E, Hohtola A (2000) Changes of cellular structures and enzymatic activities during browning of Scots pine callus derived from mature buds. Tree Physiol 20:467–475CrossRefPubMedGoogle Scholar
  26. Lee N, Teng WL (1987) Endogenous growth promoter and inhibitor levels in Pleione formosana corms. Acta Hortic 205:225–231CrossRefGoogle Scholar
  27. Lelu-Walter MA, Bernier-Cardou MB, Klimaszewska K (2008) Clonal plant production from self- and cross-pollinated seed families of Pinus sylvestris (L.) through somatic embryogenesis. Plant Cell Tissue Organ Cult 92:31–45CrossRefGoogle Scholar
  28. Lu CY (1993) The use of thidiazuron in tissue culture. In vitro Cell Dev Biol Plant 29:92–96CrossRefGoogle Scholar
  29. Merkle SA, Dean JF (2000) Forest tree biotechnology. Curr Opin Biotechnol 11:298–302CrossRefPubMedGoogle Scholar
  30. Mok MC, Mok DWS, Armstrong DJ, Shudo K, Isogai Y, Okamoto T (1982) Cytokinin activity of N-phenyl-N′-1,2,3-thidiazol-5-ylurea (thidiazuron). Phytochemistry 21:1509–1511CrossRefGoogle Scholar
  31. Moncaleán P, Alonso P, Centeno ML, Cortizo M, Rodríguez A, Fernández B, Ordás R (2005) Organogenic response of Pinus pinea cotyledons to hormonal treatments: BA metabolism and cytokinin content. Tree Physiol 25:1–9CrossRefPubMedGoogle Scholar
  32. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  33. Murch SJ, Saxena PK (2001) Molecular fate of thidiazuron and its effects on auxin transport in hypocotyls tissues of Pelargonium x hortorum Bailey. Plant Growth Regul 35:269–275CrossRefGoogle Scholar
  34. Pan MJ, Van Staden J (1998) The use of charcoal in in vitro culture—a review. Plant Growth Regul 26:155–163CrossRefGoogle Scholar
  35. Pirttilä AM, Laukkanen H, Hohtola A (2002) Chitinase production in pine callus (Pinus sylvestris L.): a defense reaction against endophytes? Planta 214:848–852CrossRefPubMedGoogle Scholar
  36. Saklani K, Singh S, Purohit VK, Prasad P, Nautiyal AR (2015) In vitro propagation of Rudraksha (Elaeocarpus sphaericus (Gaertn.) K. Schum): a biotechnological approach for conservation. Physiol Mol Biol Plants 21(4):611–615CrossRefPubMedPubMedCentralGoogle Scholar
  37. Shekhawat MS, Manokari M (2016) In vitro propagation, micromorphological studies and ex vitro rooting of cannon ball tree (Couroupita guianensis aubl.): a multipurpose threatened species. Physiol Mol Biol Plants 22(1):131–142CrossRefPubMedPubMedCentralGoogle Scholar
  38. Siddique I, Anis M (2007) In vitro shoot multiplication and plantlet regeneration from nodal explants of Cassia angustifolia: a medicinal plant. Acta Physiol Plant 29:333–338CrossRefGoogle Scholar
  39. Stojicic D, Budimir S (2002) Cytokinin-mediated axillary shoot formation in Pinus heldreichii. Biol Plant 48:477–479CrossRefGoogle Scholar
  40. Victor JMR, Murthy BNS, Murch SJ, Krihnaraj S, Saxena PK (1999) Role of endogenous purine metabolism in thidiazuron-induced somatic embryogenesis of peanut (Arachis hypogea). Plant Growth Regul 28:41–47CrossRefGoogle Scholar
  41. Von Aderkas P, Bonga JM (2000) Influencing micropropagation and somatic embryogenesis in mature trees by manipulation of phase change, stress and culture environment. Tree Physiol 20:921–928CrossRefGoogle Scholar
  42. Yao RL, Wang Y, Wu YM (2016) Key factors affecting rooting of Pinus massoniana by tissue culture. Guihaia 36(11):1288–1294Google Scholar
  43. Zhu LH, Wu XQ, Qu HY, Ji J, Ye JR (2010) Micropropagation of Pinus massoniana and mycorrhiza formation in vitro. Plant Cell Tissue Organ Cult 102(1):121–128CrossRefGoogle Scholar

Copyright information

© Northeast Forestry University and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Guangxi Key Laboratory of Superior Timber Trees Resource CultivationGuangxi Forestry Research InstituteNanningPeople’s Republic of China

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