Physiology and Molecular Biology of Plants

, Volume 24, Issue 6, pp 1285–1294 | Cite as

In vitro propagation of some old Persian cypress accessions (Cupressus sempervirens L.) by embryo culture

  • Hamid Reza KarimiEmail author
  • Kobra Janghorban
  • Mahmoud Raqamy
  • Homayoun Farahmand
Research Article


In order to study in vitro propagation of some old Persian cypress genotypes (Cupressus sempervirens L.), embryos of twenty old cypress accessions were cultured on MS and SH media containing 100 and 200 mg L−1 myo-inositol. Germination percentage and growth parameters of produced plantlets and their hardening off were evaluated. Results showed that the highest germination percentage and germination rate was obtained with MS medium containing 100 mg L−1 myo-inositol, although no significant difference was observed with MS containing 200 mg L−1 myo-inositol and SH containing 100 mg L−1 myo-inositol. Furthermore, based on the results, the highest root length was gained with SH medium containing 200 mg L−1 myo-inositol. Germination percentage of isolated embryos and shoot length of produced plantlets were affected by genotype, so that the highest germination percentage and shoot length was obtained with KB and KT genotypes, respectively. The oldest genotype which was 4000 years old (Abarkuh cypress) showed no significant difference with other genotypes in terms of shoot and root length. In hardening-off step, first all genotypes were initially established but after 2 weeks, KJ2, KK, KSHN, KD1, KB genotypes died.


Cypress Hardening Embryo culture 


  1. Afzal-Rafii Z, Dodd RS (1994) Biometrical variability of foliage and cone characters in Cupressus bakeri (Cupressaceae). Plant Syst Evol 192:151–163CrossRefGoogle Scholar
  2. Bridgen MP (1994) A review of plant embryo culture. HortSci 29:1243–1246Google Scholar
  3. Capuana M, Gannini R (1997) Micropropagation of young and adult plants of cypress (Cupressus sempervirens L.). J Hortic Sci 72:453–460CrossRefGoogle Scholar
  4. Capuana M, Lambardi ML (1995) Cutting propagation of common cypress (Cupressus sempervirens L.). New Forest 9:111–122CrossRefGoogle Scholar
  5. Chee PP (1994) In vitro culture of zygotic embryos of Taxus species. HortScience 29:695–697Google Scholar
  6. Danti R, Rocca GD, Panconesi A (2013) Cypress canker. In: Gonthier P, Nocolotti G (eds) Infectoos forest diseases. CAB International. Plant Protection Institute, National Research Council, FlorenceGoogle Scholar
  7. Darikvand R, Zolfaghari R (2014) Effects of some ecological factors on seed and germination characteristics of Cupressus sempervirens L. var. horizontalis: a case study in Tange Soulak Forest Reserve, Kuhgiluyeh and Boyer- Ahmad province, Iran. Iran J Appl Ecol 2:65–75Google Scholar
  8. Farahmand H (2015) Ornamental trees and shrubs (Gymnosperms). Entesharat-e-Jahad Daneshgahi Mashhad (In Persian) Google Scholar
  9. Farahmand H, Karimi HR (2016) Old Persian cypress accessions, a rich and unique gene pool for cypress (Cupressus sempervirens L.) in the world. In: International symposium on role of plant genetic resources on reclaimed lands and environment deteriorated by human and natural actions. Shiraz University, Shiraz, IranGoogle Scholar
  10. Farahmand H, Nazari F (2015) Environmental and anthropogenic pressures on geophytes of Iran and the possible protection strategies: a review. Int J Hortic Sci Technol 2:111–132Google Scholar
  11. Farahmand H, Hashemipour M, Karimi HR, Mohammadi-Nejad G, Bagheri V (2015) Characterization and evaluation of genetic diversity of some old cypress genotypes (Cupressus sempervirens L.) in Iran using leaf mineral concentration, biochemical characteristics and SSR markers. Plant Syst Evol 301:761–772CrossRefGoogle Scholar
  12. Fossi D, Lipucci Di Paola M, Tognoni F (1981) Induzione in vitro di gemme ascellari della specie Cupressus sempervirens L. Rivista di Ortoflorofrutticoltura Italiana 65:293–299Google Scholar
  13. Ghorpade RP, Chopra A, Nikam TD (2010) In vitro zygotic embryo germination and propagation of an endangered Boswellia serrata Roxb. As a source of boswellic acid. Physiol Mol Biol Plants 16:160–165CrossRefGoogle Scholar
  14. Giannini R, Capuana M, Giovannelli A (1999) Produzione di piante. In: Teissier du Cros E (ed) Cipresso-Manuale Tecnico II. Studio Leonardo, Firenze, pp 44–53Google Scholar
  15. Giovanelli A, De Carlo A (2007) Micropropagation of mediterranean cypress (Cupressus sempervirens L.). In: Jain SM, Haggman H (eds) Protocols for micropropagation of woody trees and fruits. Springer, Dordrecht, Finland, pp 93–105CrossRefGoogle Scholar
  16. Gianinazzi S, Gianinazzi-Pearson V (1988) Mycorrhiza, a plants health insurance. Chiamica Oggi, October, France, pp 56–58Google Scholar
  17. Hao B, Li W, Linchun M, Li Y, Rui Z, Mingxia T, Weikai B (2006) A study of conservation genetics in Cupressus chengiana, an endangered endemic of China, using ISSSR markers. Biochem Genet 44:29–43CrossRefGoogle Scholar
  18. Hartman HT, Kester DE, Davis FT, Geneve RL (2011) Hartman and Kester’s plant propagation, principles and practices. Prentice Hall, Upper Saddle RiverGoogle Scholar
  19. Karimi HR, Farahmand H, Hashemipour M (2013) Morphological diversity of some old genotypes of Cupressus Sempervirens L. in Iran. Plant Syst Evol 299:1379–1386CrossRefGoogle Scholar
  20. Kaur R, Sharma N, Kumar K, Sharma DR, Sharma SD (2006) In vitro germination of walnut (Juglans regia L.) embryos. Sci Hortic 109:385–388CrossRefGoogle Scholar
  21. Kauth P (2005) In vitro seed germination and seedling development of Calopogon tuberosus and Sacoila lanceolata var. lanceolata: two Florida native terrestrial orchids. MS.c. Thesis, University of FloridaGoogle Scholar
  22. Kauth PJ, Vendrame EA, Kane ME (2006) In vitro seed culture and seedling development of Calopogon tuberosus. Plant Cell Tissue Organ Cult 85:91–102CrossRefGoogle Scholar
  23. Lambardi M (2000) Somatic embryogenesis in cypress (Cupressus sempervirens L.). In: Jain SM, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants. Kluwer Academic Publisher, Dordrecht, pp 553–567CrossRefGoogle Scholar
  24. Lambardi M, Harry IS, Menabeni D, Thorpe TA (1995) Organogenesis and somatic embryogenesis in Cupressus sempervirens. Plant Cell Tissue Organ Cult 40:179–182CrossRefGoogle Scholar
  25. Lambardi M, Lachance D, Seguin A, Charest PJ (1997) Evaluation of micro projectile mediated DNA delivery and reporter genes for genetic transformation of the Mediterranean cypress (Cupressus sempervirens L.). Plant Cell Rep 18:198–202CrossRefGoogle Scholar
  26. Maher JT, Alqudan AM, Arabiat YS (2010) Comparative study between Cupressus semprvirens and Cupressus arizanica in seed germination and seedling vigour. Crop Res 40:174–181Google Scholar
  27. Manescu C, Hamamouch N, Maios C, Harfouche A, Doulis AG, Aravanopoulos FA (2011) Linkage mapping of the Mediterranean cypress, Cupressus sempervirens, based on molecular and morphological markers. Genet Mol Res 10:1891–1909CrossRefGoogle Scholar
  28. Mohammadi H, Kazemi Sh, Farahmand H (2014) Phaeoacremonium and Botryosphaeriacea species associated with cypress (Cupressus sempervirens L.) decline in Kerman province (Iran). Phytopathol Mediterr 53:27–39Google Scholar
  29. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497CrossRefGoogle Scholar
  30. Paudel M, Prandham S, Pant B (2012) In vitro seed germination and seedling development of Esmeralda clarkia Rchb. F. (Orchidaceae). Plant Tissue Cult Biotechnol 22:107–111Google Scholar
  31. Rai MK (2001) Current advances in mycrorrhization in micropropagation. In Vitro Cell Dev Biol Plant 37:158–167CrossRefGoogle Scholar
  32. Razi M, Jalili Marandi R, Doulati Baneh H, Hosseini B, Darvishzadeh R (2013) Effect of paternal genotypes sprays with BA and IAA concentration on embryo rescue of F1 progenies from ‘Askari’ (Vitis vinifera L.) cultivar. J Agric Sci Technol 15:1023–1032Google Scholar
  33. Romano A, Noronha C, Martins-Loução MA (1995) Role of carbohydrates in micro propagation of cork oak. Plant Cell Tissue Organ Cult 40:159–167CrossRefGoogle Scholar
  34. San SB, Yidirim AN (2009) Seed and in vitro embryo germination in an aged almond. Seed Sci Techol 37:365–371CrossRefGoogle Scholar
  35. Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50(1):199–204CrossRefGoogle Scholar
  36. Shadang R, Dwivedi P, Hegde SN, Ahmed A (2007) Effects of different culture media on seed germination and subsequent development of protocorms of Hygrochilus parishii (Veith & Rchb.f.) Pfitz (Orchidaceea). Indian J Biotechnol 6:256–261Google Scholar
  37. Shi G, Zhou Z, Xie Z (2011) Cupressus foliage shoots and associated seed cones from he oligocene ningming formation of Guanxi, South China. Rev Paleo Palyn 166:325–334CrossRefGoogle Scholar
  38. Smith JA (2010) Seiridium canker of leyland cypress. School of Forest Resource and Conservation, Florida Cooperation Extention Service, Institute of Food and Agricultural Sciences, University of Florida FOR279.
  39. Song LL, Zhang HN, Zhao HQ, Jiang YL, Hou MF (2014) In vitro germination and seedling development of Taxus chinensis var. mairei embryo culture. J Agric Sci Technol 16:1355–1363Google Scholar
  40. Spanos KA, Pirri A, Woodward S (1997) Micropropagation of Cupressus sempervirens L. and Chamaeycyparis lawsoniana (A. Murr.) Par. Silva Gene 46:291–295Google Scholar
  41. Sparapano L, Bruno G (2004) Cupressus callus and cell suspension cultures: effect of seiridins on their growth and sensitivity. In vitro Cell Dev Biol Plant 40:617–625CrossRefGoogle Scholar
  42. Stankova T, Panetsos K (1997) Vegetative propagation of Cupressus sempervirens L. of Cretan origin by softwood stems cuttings. Silva Genet 46:137–144Google Scholar
  43. Teissier du Cros E (1999) Introduction. In: Teissier du Cros E (ed) Cypress: a practical handbook. Studio Leonardo, Firenze, pp 7–9Google Scholar
  44. Xenopoulos S, Tsopelas P (2000) Sphaeropsis canker, a new disease of cypress in Greece. Forest Pathol 30:121–126CrossRefGoogle Scholar
  45. Yang Z-Y, Ran J-H, Wang X-Q (2012) Three genome-based phylogeny of Cupressaceae s.l.: Further evidence for the evolution of gymnosperms and Southern Hemisphere biogeography. Mol Phylogenet Evol 64(3):452–470CrossRefGoogle Scholar
  46. Zhiri A, Jaziri M, Homes J, Vanhaelen M, Shimomura K (1994) Factors affecting the In vitro rapid germination of Taxus embryos and the evaluation of taxol content in the plantlets. Plant Cell Tissue Organ Cult 39:261–263CrossRefGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  • Hamid Reza Karimi
    • 1
    Email author
  • Kobra Janghorban
    • 1
  • Mahmoud Raqamy
    • 1
  • Homayoun Farahmand
    • 2
  1. 1.Department of Horticultural Science, Faculty of AgricultureVale-e- Asr University of RafsanjanRafsanjanIran
  2. 2.Department of Horticultural Science, Faculty of AgricultureShahid Bahonar University of KermanKermanIran

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