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Multiple shoot induction and plant regeneration of the endangered species Crepis novoana

  • Patricia Corral
  • Rubén Mallón
  • Juan Rodríguez-Oubiña
  • María Luz GonzálezEmail author
Original Paper

Abstract

In vitro culture is a useful tool in the ex situ conservation of rare, endemic, and threatened plant species. Crepis novoana (Compositae) is an endangered endemic in northwestern Spain. Use of in vitro culture tools is necessary due to the poor conservation status of populations of the species. The systems of in vitro propagation developed for this species in the present study were caulogenesis from leaf explants and growth of axillary buds from shoots. Explants were produced by placing fragments of leaves on Murashige and Skoog medium (MS) supplemented with 2.22 μM 6-benzyladenine (BA) and 2.69 μM naphthaleneacetic acid (NAA); caulogenesis was induced in 80% of explants, with development of a mean number of 2.48 shoots per explant. Axillary bud development from shoots was highest with MS supplemented with 4.44 μM BA and 0.54 μM NAA, resulting in production of a mean number of 49.77 shoots per explant. Immersion of the basal side of shoots in a solution of 5.37 mM NAA for 30 s yielded 90% success in the production of rooted shoots. Plantlets were well acclimatized, and almost 100% of plants transferred to soil recovered successfully.

Keywords

Endangered species Ex situ conservation Organogenesis 

Notes

Acknowledgments

The study was financed by the Xunta de Galicia, Spain, under projects PGIDIT 03RF020302PR and PGIDT/PGIDIT 07MDS009200PR.

References

  1. Bañares A, Blanca G, Güemes J, Moreno JC, Ortiz S (2003) Atlas y libro rojo de la flora vascular amenazada de España. Dirección General de Conservación de la Naturaleza, MadridGoogle Scholar
  2. Bunn E (2005) Development of in vitro methods for ex situ conservation of Eucaliptus impensa, an endangered malle from southwest Western Australia. Plant Cell Tiss Organ Cult 83:97–102. doi: 10.1071/BT06065 CrossRefGoogle Scholar
  3. Bunn E, Turner S, Panaia M, Dixon KW (2007) The contribution of in vitro technology and cryogenic storage to conservation of indigenous plants. Aust J Bot 55:345–355. doi: 10.1007/s11240-005-3275-2 CrossRefGoogle Scholar
  4. Cuenca S, Amo-Marco JB (2000) In vitro propagation of Centaures spachii from inflorescence stems. Plant Growth Regul 30:99–103. doi: 10.1023/A:1006356811180 CrossRefGoogle Scholar
  5. Cuenca S, Amo-Marco JB, Parra R (1999) Micropropagation from inflorescence stems of the Spanish endemic plant Centaurea paui Loscos ex Willk. (Compositae). Plant Cell Rep 18:674–679. doi: 10.1007/s002990050641 CrossRefGoogle Scholar
  6. Dhar U, Joshi M (2005) Efficient plant regeneration protocol though callus for Saussurea obvallata (DC.) Edgew. (Asteraceae): effect of explant type, age and plant growth regulators. Plant Cell Rep 24:195–200. doi:  10.1007/s00299-005-0932-1 Google Scholar
  7. Echeverriagaray S, Fracaro F, Andrade LB, Biasio S, Atti-Serafini L (2000) In vitro shoot regeneration from leaf explants of Roman Chamomile. Plant Cell Tiss Organ Cult 60:1–4. doi: 10.1023/A:1006368210670 CrossRefGoogle Scholar
  8. Evenor D, Reuveni M (2004) Micropropagation of Achillea filipendulina cv. ‘Parker’. Plant Cell Tiss Organ Cult 79:91–93. doi: 10.1023/B:TICU.0000049441.48404.70 CrossRefGoogle Scholar
  9. Fay MF (1992) Conservation of rare and endangered plants using in vitro methods. In Vitro Cell Dev Biol Plant 28:1–4. doi: 10.1007/BF02632183 Google Scholar
  10. Gonçalves S, Fernandes L, Romano A (2010) High-frequency in vitro propagation of the endangered species Tuberaria major. Plant Cell Tiss Organ Cult 101:359–363. doi: 10.1007/s11240-010-9683-y CrossRefGoogle Scholar
  11. Guo B, Gao M, Liu CZ (2007) In vitro propagation of an endangered medicinal plant Saussurea involucrata Kar et Kir. Plant Cell Rep 26:261–265. doi: 10.1007/s00299-006-0230-6 PubMedCrossRefGoogle Scholar
  12. Jayaram K, Prasad MNV (2007) Rapid in vitro multiplication of Drosera indica L.: a vulnerable, medicinally important insectivorous plant. Plant Biotechnol Rep 1:79–84. doi: 10.1007/s11816-007-0014-7 CrossRefGoogle Scholar
  13. Joshi M, Dhar U (2003) In vitro propagation of Saussurea obvallata (DC) Edgew.–an endangered ethnoreligious medicinal herb of Himalaya. Plant Cell Rep 21:933–939. doi: 10.1007/s00299-003-0601-1 PubMedCrossRefGoogle Scholar
  14. Koroch A, Juliani HR, Kapteyn J, Simon JE (2002) In vitro regeneration of Echinacea purpurea from leaf explants. Plant Cell Tiss Organ Cult 69:79–83. doi: 10.1023/A:1015042032091 CrossRefGoogle Scholar
  15. Ma G, Lü J, Teixeira da Silva JA, Zhang X, Zhao J (2010) Shoot organogenesis and somatic embryogenesis from leaf and shoot explants of Ochna integerrima (Lour). Plant Cell Tiss Organ Cult. doi:  10.1007/s11240-010-9812-7
  16. Maheshwari P, Kumar A (2006) Organogenesis, shoot regeneration, and flowering response of Vernonia cinerea to different auxin/citokynin combinations. In Vitro Cell Dev Biol Plant 42:589–595. doi: 10.1079/IVP2006825 CrossRefGoogle Scholar
  17. Mallón R, Bunn E, Turner SR, González ML (2008) Cryopreservation of Centaurea ultreiae (Compositae) a critically endangered species from Galicia (Spain). Cryo-Letters 29:363–370PubMedGoogle Scholar
  18. Mallón R, Rodríguez-Oubiña J, González ML (2010) In vitro propagation of the endangered plant Centaurea ultreiae: assessment of genetic stability by cytological studies, flow cytometry and RAPD analysis. Plant Cell Tiss Organ Cult 101:31–39. doi: 10.1007/s11240-009-9659-y CrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  20. Nikam TD, Shitole MG (1999) In vitro culture of Safflower L. cv Bhima: initiation, growth optimization and organogenesis. Plant Cell Tiss Organ Cult 55:15–22. doi: 10.1023/A:1026493616991 CrossRefGoogle Scholar
  21. Ortiz S, Soroña X, Rodriguez-Oubiña J (1997) Crepis novoana (Asteraceae), a new species restricted to coastal cliffs in northern Galicia (northwest Iberian Peninsula). Bot J Linn Soc 123:147–155. doi: 10.1111/j.1095-8339.1997.tb01409.x Google Scholar
  22. Ozel CA, Khawar KM, Mirici S, Ozcan S, Arslan O (2006) Factors affecting in vitro plant regeneration of the critically endangered Mediterranean knapweed (Centaurea tchihatcheffii Fisch et Mey). Naturwissenschaften 93:511–517. doi: 10.1007/s00114-006-0139-5 PubMedCrossRefGoogle Scholar
  23. Piovan A, Caniato R, Mariella Cappelletti E, Filippini R (2010) Organogenesis from shoot segments and via callus of endangered Kosteletzkya pentacarpos (L.) Ledeb. Plant Cell Tiss Organ Cult 100:309–315. doi: 10.1007/s11240-009-9652-5 CrossRefGoogle Scholar
  24. Radhika K, Sujhata M, Nageshwar Rao T (2006) Thidiazuron stimulates shoot regeneratiom in different explants. Biol Plant 50:174–179. doi: 10.1007/s10535-006-0003-7 CrossRefGoogle Scholar
  25. Raghu AV, Geetha SP, Martin G, Balachandram I, Ravindram PN (2006) Direct shoot organogenesis from leaf explants of Embelia ribes Burn f.: a vulnerable medicinal plant. J For Res 11:57–60CrossRefGoogle Scholar
  26. Sarasan V, Cripps R, Ramsay MM, Atherton C, McMichen M, Prendergast G, Rowntree JK (2006) Conservation in vitro of threatened plants—Progress in the past decade. In Vitro Cell Dev Biol Plant 42:206–214. doi: 10.1079/IVP2006769 CrossRefGoogle Scholar
  27. Sreedhar RV, Venkatachalam L, Thimmaraju M, Bhagyalakshmi N, Narayam NS, Ravishankar GA (2008) Direct organogenesis from leaf explants of Stevia rebaudiana and cultivation in bioreactor. Biol Plant 52:355–360. doi: 10.1007/s10535-008-0073-9 CrossRefGoogle Scholar
  28. Sujatha M, Dinesh Kumar V (2007) In vitro bud regeneration of Carthamus tinctorius and wild Carthamus species from leaf explants and axilary buds. Biol Plant 51:782–786. doi: 10.1007/s10535-007-0160-3 CrossRefGoogle Scholar
  29. Sujatha G, Ranjitha Kumari BD (2007) Effect of phytohormones on micropropagation of Artemisia vulgaris L. Acta Physiol Plant 29:189–195. doi: 10.1007/s11738-006-0023-0 CrossRefGoogle Scholar
  30. Vanegas PE, Cruz-Hernández A, Valverde ME, Paredes-López O (2002) Plant regeneration via organogenesis in marigold. Plant Cell Tiss Organ Cult 69:279–283. doi: 10.1023/A:1015610011374 CrossRefGoogle Scholar
  31. Wildi E, Schaffner W, Berger Büter K (1998) In vitro propagation of Petasites hybrid (Asteraceae) from leaf explants and from inflorescence buds. Plant Cell Rep 18:336–340. doi: 10.1007/s002990050582 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Patricia Corral
    • 1
  • Rubén Mallón
    • 3
  • Juan Rodríguez-Oubiña
    • 2
  • María Luz González
    • 1
    Email author
  1. 1.Departamento de Fisiología VegetalUniversidad de Santiago de CompostelaSantiago de CompostelaSpain
  2. 2.Departamento de BotánicaUniversidad de Santiago de CompostelaSantiago de CompostelaSpain
  3. 3.Instituto de Investigaciones Agrobiológicas de Galicia, CSICSantiago de CompostelaSpain

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