Genetic Resources and Crop Evolution

, Volume 57, Issue 7, pp 1041–1052 | Cite as

The Frankincense tree (Boswellia sacra, Burseraceae) from Oman: ITS and ISSR analyses of genetic diversity and implications for conservation

  • Andrea Coppi
  • Lorenzo Cecchi
  • Federico Selvi
  • Mauro Raffaelli
Research Article


DNA sequences from the ITS region of the nuclear genome and Inter-Simple Sequence Repeat markers (ISSR) were used to estimate genetic diversity among and within populations of the Frankincense tree Boswellia sacra from Dhofar, Oman. This is a culturally and ecologically relevant species that is showing symptoms of decline due to anthropic factors and, possibly, global warming. ITS sequences were 511 bp long and showed low (6.4%) variation among geographically different populations. The four selected ISSR primers yielded 93 reproducible bands, of which 91 (97.9%) were polymorphic in the 97 individual profiles obtained. Total genetic diversity (H T) and average heterozygosity within populations (H S) resulted fairly low (0.22 and 0.136, respectively). The accession from Wadi Dowkah, an UNESCO world heritage site, showed the lowest level of genetic diversity (H E = 0.107), while the eastern populations from the Hasik area harboured a slightly greater amount of variation. Analysis of Molecular Variance showed that differentiation among populations was relatively high (38.1%), possibly due to the reduced gene flow between the largely isolated stands of Boswellia (N m = 0.39). Genetic distances and AMOVA suggested a clear differentiation between the eastern and western coastal populations, while those from the internal area did not form a consistent group. For conservation, the eastern sites should be given priority as core populations harbouring significant amounts of allelic diversity. Reasons for reinforcing the more depauperated stands, such as Wadi Dowkah, with local plant material only or, alternatively, with the introduction of germplasm from genetically distinct stands are discussed.


Boswellia sacra Dhofar Frankincense tree Genetic diversity ISSR markers ITS sequences Oman flora UNESCO sites 



The authors wish to acknowedge the Office for Cultural Affair of Sultanate of Oman for the logistic support in Dhofar. Dr. Stefano Mosti and sig. Marcello Tardelli helped with field sampling and Dr. Alessio Mengoni gave useful insights into the technical aspects of data analysis. An anonymous reviewer suggested useful comments on the first version of the manuscript. Research funds from M.I.U.R. 40% to M. Raffaelli are acknowledged.


  1. Avanzini A (2000) Il porto di Sumharam nel Dhufâr. In: De Maigret A (ed) Yemen, nel paese della Regina di Saba. Skira, Milano, pp 231–233Google Scholar
  2. Becerra JX, Venable DL (1999) Nuclear ribosomal DNA phylogeny and its implications for evolutionary trends in Mexican Bursera (Burseraceae). Am J Bot 86:1047–1057CrossRefPubMedGoogle Scholar
  3. Bhat KV, Babrekar PP, Lakhanpaul S (1999) Study of genetic diversity in Indian and exotic sesame (Sesamum indicum L.) germplasm using random amplified polymorphic DNA (RAPD) markers. Euphytica 110:21–33CrossRefGoogle Scholar
  4. Borchsenius F (2007) FastGap 1.0.8. Software distributed by the author (
  5. Bornet B, Branchard M (2001) Nonanchored inter simple sequence repeat (ISSR) markers: reproducible and specific tools for genome fingerprinting. Plant Mol Biol Rep 19:209–215CrossRefGoogle Scholar
  6. Corpet F (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acid Res 16:10881–10890CrossRefPubMedGoogle Scholar
  7. El Quassani AS (1984) Dhofar the land of Frankincense. International Printing Press, RuwiGoogle Scholar
  8. Excoffier L, Smouse PE, Quattro M (1992) Analysis of molecular variance inferred from metric distances among DNA Haplotypes: application to human mitochondrial DNA restriction data. Genetics 131:479–491PubMedGoogle Scholar
  9. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  10. Feyissa T, Nybom H, Bartish IV, Welander M (2007) Analysis of genetic diversity in the endangered tropical tree species Hagenia abyssinica using ISSR markers. Genet Resour Crop Evol 54:947–958CrossRefGoogle Scholar
  11. Fleishman E, Launer AE, Switky KR, Yandell U, Heywood J, Murphy DD (2001) Rules and exceptions in conservation genetics: genetic assessment of the endangered plant Cordylanthus palmatus and its implications for management planning. Biol Conserv 98:45–53CrossRefGoogle Scholar
  12. Frankel OH, Brown AHD, Burdon JJ (1995) The conservation of plant biodiversity. Cambridge University Press, CambridgeGoogle Scholar
  13. Ge XJ, Yu Y, Yuan Y, Huang H, Yan C (2005a) Genetic diversity and geographic differentiation in endangered Ammopiptanthus (Leguminosae) populations in desert regions of northwest China as revealed by ISSR analysis. Ann Bot 95:843–851CrossRefPubMedGoogle Scholar
  14. Ge XJ, Zhang LB, Yuan YM, Hao G, Chiang TY (2005b) Strong genetic differentiation of the East-Himalayan Megacodon stylophorus (Gentianaceae) detected by inter-simple sequence repeats (ISSR). Biodivers Conserv 14:849–861CrossRefGoogle Scholar
  15. Ghazanfar SA (1994) Handbook of Arabian medicinal plants. CRC Press, Boca RatonGoogle Scholar
  16. Groom N (2000) I profumi d’Arabia. In: De Maigret A (ed) Yemen, nel paese della Regina di Saba. Skira, Milano, pp 87–89Google Scholar
  17. Hammer K, Gebauer J, Al Khanjari S, Buerkert A (2009) Oman at the cross-roads of inter-regional exchange of cultivated plants. Genet Resour Crop Evol 56:547–560CrossRefGoogle Scholar
  18. Hamrick JL, Godt MJW (1989) Allozyme diversity in plant species. In: Brown AHD, Clegg MT, Kahler AL, Weir BS (eds) Plant population genetics, breeding and genetic resources. Sinauer, Sunderland MA, pp 43–63Google Scholar
  19. Hatcher PE, Wilkinson MJ, Albani MC, Hebbern CA (2004) Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate. Biol Conserv 116:305–317CrossRefGoogle Scholar
  20. Hedrén M (2004) Species delimitation and the partitioning of genetic diversity – an example from the Carex flava complex (Cyperaceae). Biodivers Conserv 13:293–316CrossRefGoogle Scholar
  21. Holsinger KE, Gottlieb LK (1991) Conservation of rare and endangered plants: principle and prospect. In: Falk DA, Holsinger KE (eds) Genetics and conservation of rare plants. Oxford University Press, New York, pp 195–298Google Scholar
  22. Huenneke LF (1991) Ecological implications of genetic variations in plant populations. In: Falk DA, Holsinger KE (eds) Genetics and conservation of rare plants. Oxford University Press, New York, pp 31–44Google Scholar
  23. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163CrossRefPubMedGoogle Scholar
  24. Li J, Jin Z (2007) Genetic variation and differentiation in Torreya jackii Chun, an endangered plant endemic to China. Plant Sci 172:1048–1053CrossRefGoogle Scholar
  25. Lowe A, Harris S, Ashton P (2004) Ecological genetics. Design, analysis, and application. Blackwell, OxfordGoogle Scholar
  26. Luo X, Zhuang X, Yang Y (2007) Genetic diversity of Camellia changii Ye (Theaceae) using ISSR markers. J Trop Subtrop Bot 15(2):93–100Google Scholar
  27. Mantel N (1967) The detection of disease clustering and a generalized regression approach. Cancer Res 27:209–220PubMedGoogle Scholar
  28. Meloni M, Perini D, Filigheddu R, Binelli G (2006) Genetic variation in five mediterranean populations of Juniperus phoenicea as revealed by inter-simple sequence repeat (ISSR) markers. Ann Bot 97:299–304CrossRefPubMedGoogle Scholar
  29. Mengoni A, Selvi F, Cusimano N, Galardi F, Gonnelli C (2006) Genetic diversity inferred from AFLP fingerprinting in populations of Onosma echioides (Boraginaceae) from serpentine and calcareous soils. Plant Biosyst 140:211–782CrossRefGoogle Scholar
  30. Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89:583–590PubMedGoogle Scholar
  31. Nybom H (2004) Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants. Mol Ecol 13:1143–1155CrossRefPubMedGoogle Scholar
  32. Nybom H, Bartish I (2000) Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants. Perspect Plant Ecol Evol Syst 3(2):93–114CrossRefGoogle Scholar
  33. Okun DO, Kenya EU, Oballa PO, Odee DW, Muluvi GM (2008) Analysis of genetic diversity in Eucalyptus grandis (Hill ex Maiden) seed sources using inter simple sequence repeats (ISSR) molecular markers. Afr J Biotechnol 7(13):2119–2123Google Scholar
  34. Pérez-Collazos E, Catalán P (2007) Genetic diversity analysis and conservation implications for the Iberian threatened populations of the irano-turanian relict Krascheninnikovia ceratoides (Chenopodiaceae). Biol J Linn Soc 92:419–429CrossRefGoogle Scholar
  35. Raffaelli M, Tardelli M (2007) L’incenso tra mito e realtà. Produzione, commercio e raccolta della resina, ieri e oggi. Le specie di Boswellia (Burseraceae) conosciute. Conservazione delle piante in natura. Centro Studi Erbario Tropicale, Università di Firenze. Pubbl. no. 108, FirenzeGoogle Scholar
  36. Raffaelli M, Mosti S, Tardelli M (2003a) The Frankincense Tree (Boswellia sacra Flueck., Burseraceae) in Dhofar, southern Oman: field-investigations on the natural populations. Webbia 58(1):133–149Google Scholar
  37. Raffaelli M, Tardelli M, Mosti S (2003b) The Wadi Doka Frankincense Park in Dhofar, Oman – first steps towards the safeguard and improvement of Boswellia sacra (Burseraceae). Pubbl. Erbario Tropicale di Firenze no. 93, FirenzeGoogle Scholar
  38. Raffaelli M, Mosti M, Tardelli M (2006) Boswellia sacra Flueck. (Burseraceae) in the Hasik area (eastern Dhofar, Oman) and a list of the surrounding flora. Webbia 61(2):245–251Google Scholar
  39. Raffaelli M, Tardelli M, Mosti S (2008) Preserving and restoring the frankincense tree (Boswellia sacra) at Wadi Doka: a work in progress. In Avanzini A. (Ed.), A port in Arabia between Rome and the Indian Ocean (3rd C. BC–5th C. AD), Khor Rori Report 2. “L’Erma” di Breitschneider. Roma, pp 715–723Google Scholar
  40. Rakoczy-Trojanowska M, Bolibok H (2004) Characteristics and comparison of three classes of microsatellite-based markers and their application in plants. Cell Mol Biol Lett 9:221–238PubMedGoogle Scholar
  41. Reddy MP, Sarla N, Siddiq EA (2002) Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128:9–17CrossRefGoogle Scholar
  42. Rees AR (1995) Frankincense and Myrrh. New Plantsman 2(1):55–59Google Scholar
  43. Richards CM (2000) Inbreeding depression and genetic rescue in a plant metapopulation. Am Nat 155:383–827CrossRefPubMedGoogle Scholar
  44. Rieseberg LH (1996) Homology among RAPD fragments in interspecific comparisons. Mol Ecol 5:99–105CrossRefGoogle Scholar
  45. Rohlf FJ (1990) NTSYS-pc. Numerical taxonomy and multivariate analysis system. Version 2.02. Exeter, New YorkGoogle Scholar
  46. Saitou N, Nei M (1987) The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  47. Schneider S, Rosselli D, Excoffier L (2000) Arlequin: a software for population genetics data analysis. Version 2.000. University of GenevaGoogle Scholar
  48. Sica M, Gamba G, Montieri S, Gaudio L, Aceto S (2005) ISSR markers show differentiation among Italian populations of Asparagus acutifolius L. BMC Genet 6:17CrossRefPubMedGoogle Scholar
  49. Simmons MP, Ochoterena H (2000) Gaps as characters in sequence-based phylogenetic analyses. Syst Biol 49:369–381CrossRefPubMedGoogle Scholar
  50. Slatkin M (1995) A measure of population subdivision based on microsatellite allele frequencies. Genetics 139:457–462PubMedGoogle Scholar
  51. Swofford DL (1998) PAUP*. Phylogenetic analysis using parsimony (and other methods) vers. 4.0. Sinauer, SunderlandGoogle Scholar
  52. Thulin M, Warfa AM (1987) The Frankincense trees (Boswellia ssp., Burseaceae) of Northern Somalia and Southern Arabia. Kew Bull 42(3):487–500CrossRefGoogle Scholar
  53. Weising K, Nybom H, Wolf K, Kahl G (2005) DNA fingerprinting in plants: principles, methods and applications. CRC Press, Boca RatonCrossRefGoogle Scholar
  54. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Shinsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322Google Scholar
  55. Xia T, Chen S, Chen S, Zhang D, Zhang D, Gao Q, Ge X (2007) ISSR analysis of genetic diversity of the Qinghai-Tibet Plateau endemic Rhodiola chrysanthemifolia (Crassulaceae). Biochem Syst Ecol 35:209–221CrossRefGoogle Scholar
  56. Xiao M, Li Q, Wang L, Guo L, Li J, Tang L, Chen F (2006) ISSR analysis of the genetic diversity of the endangered species Sinopodophyllum hexandrum (Royle) Ying from Western Sichuan Province, China. J Integr Plant Biol 48(10):1140–1146CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Andrea Coppi
    • 1
  • Lorenzo Cecchi
    • 1
  • Federico Selvi
    • 1
    • 2
  • Mauro Raffaelli
    • 1
    • 2
  1. 1.Department of Plant BiologyUniversity of FlorenceFlorenceItaly
  2. 2.Centro Studi Erbario TropicaleFlorenceItaly

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