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Investigating the Use of Fractional Replication and Taguchi Techniques in Cryopreservation: A Case Study Using Orthodox Seeds of a Tropical Rainforest Tree Species

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Abstract.

Cryopreservation is increasingly important for conserving endangered species including tropical rain forest germplasm where optimal cryopreservation protocols must be established rapidly sacrificing little germplasm. Currently, full factorial experiments analysed by ANOVA establish optimal conditions. However, these experiments can contain many treatment combinations whilst ANOVA identifies significant effects without guaranteeing to find robust optimal conditions. Taguchi optimization techniques efficiently identify robust conditions through fractional factorial experiments with an appropriate signal to noise ratio (SNR). This paper reports for the first time the use of Taguchi techniques in cryopreservation. An orthodox seed (Cassia siamea Lam.) was used to guarantee sufficient data to compare full and fractionally replicated experiments analysed using both ANOVA and SNR. For sprouting day (smaller is better), identical significant main effects were found for all experimental sizes for ANOVA and SNR. The 1/4 replicate did not allow investigation of interaction terms, but the significant main effects were the same for larger experiments. For shoot to root ratio (nominal is best), a significant main effect was found for all experimental sizes using ANOVA. This was also found using SNR, which identified additional significant main effect and interactions. No significant effects were found for dry weight (larger is better). We show smaller experiments are possible, provided important two level interactions are analysed. Differences determining the optimal treatment combination were found between ANOVA and SNR; with the Taguchi choice provides more robust solutions. Taguchi optimization techniques are recommended when germplasm is scarce and/or the experiment needs to be conducted rapidly.

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References

  1. I. Babiak E. Brzuska J. Perkowski (2000) ArticleTitleFractional factorial design of screening experiments on cryopreservation of fish sperm Aquacult. Res. 31 273–282 Occurrence Handle10.1046/j.1365-2109.2000.00383.x

    Article  Google Scholar 

  2. E.E. Benson (1999) Plant Conservation Biotechnology Taylor and Francis Ltd London

    Google Scholar 

  3. E.E. Benson B. Krishnapillay M. Marzalina (1996) The potential of biotechnology in the in vitro conservation of Malaysian forest germplasm: an integrated approach N. Hussein P.S. Bacon K.K. Choon (Eds) Forestry and Forest Products Research. Proceedings of the 3rd ConferenceForest Research Institute of Malaysia(FRIM) FRIM Publications Kepong, Malaysia 76–90

    Google Scholar 

  4. P. Berjak J.M. Farrant N.M. Pammenter (1989) The basis of recalcitrant seeds behaviour. Cell biology of the honoiohydrous seed condition R.B. Taylorson (Eds) Recent Advances in Development and Germination of Seeds Plenum Press New York 89–108

    Google Scholar 

  5. J.A. Callow B.V. Ford-Lloyd H.J. Newbury (1997) Biotechnology and Plant Genetic Resources Conservation and Use. Biotechnology in Agriculture. Series No 10 CAB International OxfordUK

    Google Scholar 

  6. Chai Y.M., Marzalina M. and Krishnapillay B. 1994. Effect of sucrose as a cryoprotective agent in the cryopreservation of some Dipterocarp species. Proceedings of Fifth National Biotechnology Symposium, Pulau Pinang, Malaysia.

  7. Draper N.R. and Smith I.H. 1998. Stepwise regression. In: Applied Regression Analysis. John Wiley and Sons, Inc, pp. 335–339.

  8. R.H. Ellis T.D. Hong E.H. Roberts (1990) ArticleTitleAn intermediate category of seed behaviour? 1. Coffee. J. Exp. Bot. 41 1167–1174

    Google Scholar 

  9. Engelmann F. 2000. In: Engelmann F. and Takagi H. (eds), Cryopreservation of Tropical Plant Germplasm. JIRCAS, IPGRI, Tsukuba, Japan, Rome, pp. 8–20.

  10. FAO 1993. Forest Resources Assessment 1990. Tropical Countries FAO Forest Paper 112. Rome.

  11. S.J.N. Gupta (1939) ArticleTitleDipterocurpus (gurja) forests in India and their regeneration Indian Forest Rec. 3 164

    Google Scholar 

  12. International Plant Names Index (IPNI). 1999. http://www.ipni.org/index/html.

  13. International Seed Testing Association. 1999. International rules for seed testing. Seed Sci. Technol.. 27 supplement 333pp.

  14. Kioko J., Berjak P., Pritchard H. and Matthews D. 2000. Seeds of the African pepper bark (Warburgia salutaris) can be cryopreseved after rapid dehydration in silica gel. Cryopreservation of tropical plant germplasm. Current Research Progress and Application. In: Engelmann F. and Takagi H. Japan International Research Centre for Agricultural Sciences, International Plant Genetic Resources InstituteTsukubaJapan, RomeItaly, 496pp.

  15. B. Kobmoo A.K. Hellum (1984) ArticleTitleHot water and acid improve the germination of Cassia siamea seeds The Embryon 1 IssueID1 27–33

    Google Scholar 

  16. Krishnapillay B. 1990. Towards the development of a protocol for cryopreservation of embryos a recalcitrant seed (Artocarpus heterophyllus Lam.). Ph.D. thesis. University of Agriculture MalaysiaSerdang.

  17. B. Krishnapillay M. Marzalina (1993) An Intermediate Category of Seeds Seed Centre News ASEAN-Canada Forest Tree Seed Centre Thailand

    Google Scholar 

  18. Krishnapillay B., Schroeder C. and Marzalina M. 1995. Towards the setting up of a Forest Tree Seed Center in Malaysia: possibilities and constraints. Proceedings of the IUFRO Symposium of the Project Group P.2.040.00 ȁ8Seed problemsȁ9 ArushaTanzania.

  19. M. Marzalina B. Krishnapillay (1999) Recalcitrant seed biotechnology. Application to rain forest conservation (Chapter 17) E.E. Benson (Eds) Plant Conservation Biotechnology Taylor and Francis Ltd. London, UK 265–276

    Google Scholar 

  20. M. Marzalina M.N. Normah (2001) ArticleTitleDesiccation studies on Mahogany (Swietenia macrophylla) seeds J. Trop. Forest Sci. 13 IssueID3 405–414

    Google Scholar 

  21. Mascarenhas A.F. and Agarwal D.C. 1991. Prospects for in vitro conservation of tree germplasm. In Conservation of Plant Genetic Resources. Through In vitro Methods Zakri A.H., Normah M.N. and Abdul Karim A.G. (eds), FRIM/MNCPGR, pp. 231–250.

  22. G. Maury-Lechon A.M. Hassan D.R. Bravo (1981) ArticleTitleSeed Storage of Shorea parvifoliaDipterocarpus humeratus Malaysian Forester 44 267–280

    Google Scholar 

  23. H.T. Meryman R.J. Williams (1985) Basic principles of freezing injury to plant cells: natural tolerance and approaches to cryopreservation K. Kartha (Eds) Cryopreservation of Plant Cells and Organs CRC Press Boca Raton, FloridaUSA 13–48

    Google Scholar 

  24. D.C. Montgomery (1991) Design and Analysis of Experiments John Wiley New York 414–438

    Google Scholar 

  25. T. Murashige F. Skoog (1962) ArticleTitleA revised medium for rapid bioassays with tobacco tissue cultures Physiol. Plant. 15 473–497

    Google Scholar 

  26. M.N. Normah M.K. Narimah M.M. Clyde (1996) Techniques in in vitro conservation Proceeding of the International Workshop on In Vitro Conservation of Plant Genetic Resources, 4–6 July 1995, Kuala Lumpur University Kebangsaan Malaysia Publishers, Plant Biotechnology Laboratory Kuala LumpurMalaysia

    Google Scholar 

  27. H. Paz (2003) ArticleTitleRoot/shoot allocation and root architecture in seedlings: variation among forest sites, microhabitats, and ecological groups Biotropica 35 IssueID3 318–332

    Google Scholar 

  28. PROSEA [12.1.] 1999. In: de Padua L.S, Bunyapraphatatsava N. and Lemmens R.H.M.J. Medicinal and Poisonous Plant.

  29. Quinn G. and Keough M. 2002. Correlation and regression. In: Experimental Design and Data Analysis for Biologists. University Press, CambridgeUK. pp. 72–76.

  30. A.N. Rao (1988) In vitro culture studies on economically important tropical trees R.C. Umaly I. Umboh S. Halos N.M. Noor (Eds) The Application of Tissue Culture Techniques in Economically Important Tropical Trees Biotrop Special Publication No. 35, Seameo-Biotrop BogorIndonesia 9–30

    Google Scholar 

  31. E.H. Roberts (1973) ArticleTitlePredicting the storage life of seeds Seed Sci. Technol. 1 499–514

    Google Scholar 

  32. P.B. Tompsett (1985) ArticleTitleThe influence of moisture content and temperature on the viability of Shorea robustaS. roxburghii seeds Can. J. Forest Res. 15 1074–1079

    Google Scholar 

  33. R.C. Umaly R. Umboh S.S. Tjitrosomo N.M. Noor (Eds) (1992) Biotechnology for Forest Tree Improvement Biotrop Special Publication No. 49, SEAMEO-BIOTROP BogorIndonesia

    Google Scholar 

  34. L.A. Withers (1985) Cryopreservation of cultured plant cells and protoplasts K. Kartha (Eds) Cryopreservation of Plant Cells and Organs CRC Press Boca Raton, FloridaUSA 243–267

    Google Scholar 

  35. L.A. Withers P.J. King (1980) ArticleTitleA simple freezing unit and cryopreservation method for plant cell suspensions Cryo-letters 1 213–220

    Google Scholar 

  36. Wu J.C.F. and Hamada M. 2000. Fractional factorial experiments at two levels. In: Experiments: Planning, Analysis and Parameter Design Optimization. John Wiley & Sons, Inc, 630 pp.

  37. S.K. Yap (1981) ArticleTitleCollection, germination and storage of Dipterocarp seeds Malayan Forester 44 281–300

    Google Scholar 

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Authors and Affiliations

  1. SIMBIOS Centre, University of Abertay Dundee, Bell Street, DD1 1HG, Dundee, Scotland, UK

    JAYANTHI MUTHUSAMY & H.J. STAINES

  2. School of Contemporary Science, University of Abertay Dundee, Bell Street, DD1 1HG, Dundee, Scotland, UK

    E.E. BENSON

  3. Forest Research Institute of Malaysia (FRIM), 52109, Kuala Lumpur, Kepong, Malaysia

    JAYANTHI MUTHUSAMY, M. MANSOR & B. KRISHNAPILLAY

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  1. JAYANTHI MUTHUSAMY
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  2. H.J. STAINES
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  3. E.E. BENSON
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  4. M. MANSOR
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  5. B. KRISHNAPILLAY
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Correspondence to JAYANTHI MUTHUSAMY.

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MUTHUSAMY, J., STAINES, H., BENSON, E. et al. Investigating the Use of Fractional Replication and Taguchi Techniques in Cryopreservation: A Case Study Using Orthodox Seeds of a Tropical Rainforest Tree Species. Biodivers Conserv 14, 3169–3185 (2005). https://doi.org/10.1007/s10531-004-0385-9

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