Skip to main content
Log in

Production of chrysanthemum synthetic seeds under non-aseptic conditions for direct transfer to commercial greenhouses

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

A novel protocol for both production and sowing of chrysanthemum synthetic seeds in non-aseptic conditions for large-scale commercialization was successfully established. Effects of the availability of organic compounds, namely MS vitamins and sucrose, inside and outside gelling matrix on the microbial contaminations, plantlet formation and subsequent growth were investigated. Results showed that the presence of organic compounds, either in gelling matrix alone or in both gelling matrix and commercial substrate, caused microbial contaminations in all synthetic seeds and complete inhibition in regrowth. In contrast, the absence of organic compounds in both non-sterile gelling matrix and substrate resulted in 70 % plantlet formation after 6 weeks of sowing. However, organic compounds absent in gelling matrix but present in substrate induced lower plantlet formation frequency (34 %) after 6-week sowing. The removal of organic compounds from both gelling matrix and substrate, in comparison with the removal from only gelling matrix, also stimulated the formation of plantlets having more leaves, longer shoots and roots, and greater fresh and dry biomass accumulation, but equal leaf area, chlorophyll content, and number of nodes and roots. These findings suggest that the eradication of all organic compositions is a prerequisite for practical application of encapsulation technology to the mass production of chrysanthemum plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

ANOVA:

Analysis of variance

CaCl2·2H2O:

Dihydrate calcium chloride

DMRT:

Duncan’s multiple range test

MS:

Murashige and Skoog (1962)

Na-alginate:

Sodium-alginate

OCfree :

Both artificial endosperm and vermiculite substrate were free of sucrose and MS vitamins

OCseed :

Only artificial endosperm contained 3 % sucrose and MS vitamins

OCseed+soil :

Both artificial endosperm and vermiculite substrate contained 3 % sucrose and MS vitamins

OCsoil :

Only vermiculite substrate contained 3 % sucrose and MS vitamins

SPAD:

Special products analysis division

References

  • Ahmad N, Anis M (2010) Direct plant regeneration from encapsulated nodal segments of Vitex negundo. Biol Plant 54:748–752

    Article  Google Scholar 

  • Ahmed MR, Anis M, Al-Etta HA (2015) Encapsulation technology for short-term storage and germplasm exchange of Vitex trifolia L. Rend Fis Acc Lincei 26:133–139

  • Ara H, Jaiswal U, Jaiswal VS (2000) Synthetic seed: prospects and limitations. Curr Sci 78:1438–1444

    Google Scholar 

  • Banerjee S, Singh S, Pandey H, Pandey P, Rahman LU (2012) Conservation and storage of Curcuma amada Roxb. synseeds on Luffa sponge matrix and RAPD analysis of the converted plantlets. Ind Crop Prod 36:383–388

    Article  CAS  Google Scholar 

  • Bapat VA, Rao PS (1990) In vivo growth of encapsulated axillary buds of mulberry (Morus indica L.). Plant Cell Tissue Organ Cult 2:69–70

    Article  Google Scholar 

  • Bapat VA, Mhatre M, Rao PS (1987) Propagation of Morus indica L. (mulberry) by encapsulated shoot buds. Plant Cell Rep 6:393–395

    Article  CAS  PubMed  Google Scholar 

  • Caires ARL, Scherer MD, Santos TSB, Pontim BCA, Gavassoni WL, Oliveira SL (2010) Water stress response of conventional and transgenic soybean plants monitored by chlorophyll a fluorescence. J Fluoresc 20:645–649

    Article  CAS  PubMed  Google Scholar 

  • Chand S, Singh AK (2004) Plant regeneration from encapsulated nodal segments of Dalbergia sissoo Roxb.–a timber yielding leguminous tree. J Plant Physiol 161:237–243

    Article  CAS  PubMed  Google Scholar 

  • Duncan DB (1995) Multiple range and multiple F test. Biometrics 11:1–42

    Article  Google Scholar 

  • Faisal M, Anis M (2007) Regeneration of plants from alginate-encapsulated shoots of Tylophora indica (Burm. F.) Merrill., an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354

    CAS  Google Scholar 

  • Felek W, Mekibib F, Admassu B (2015) Optimization of explants surface sterilization condition for field grown peach (Prunus persica L. Batsch. cv. Garnem) intended for in vitro culture. Afr J Biotechnol 14:657–660

    Article  Google Scholar 

  • George EF (1993) Plant propagation by tissue culture. Part 1. The technology. Exegetics Ltd., Edington

    Google Scholar 

  • Germanà MA, Micheli M, Pulcini L, Standardi A (2007) Perspective of the encapsulation technology in the nursery activity of Citrus. Caryo 60:192–195

    Article  Google Scholar 

  • Hung CD, Trueman SJ (2010) Nutrient responses differ between node and organogenic cultures of Corymbia torelliana × C. citriodora (Myrtaceae). Aust J Bot 58:410–419

    Article  Google Scholar 

  • Hung CD, Trueman SJ (2011a) Encapsulation technology for short-term preservation and germplasm distribution of the African mahogany Khaya senegalensis. Plant Cell Tissue Organ Cult 107:397–405

    Article  CAS  Google Scholar 

  • Hung CD, Trueman SJ (2011b) In vitro propagation of the African mahogany Khaya senegalensis. New For 42:117–130

    Article  Google Scholar 

  • Hung CD, Trueman SJ (2012a) Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora. Acta Physiol Plant 34:117–128

    Article  CAS  Google Scholar 

  • Hung CD, Trueman SJ (2012b) Preservation of encapsulated shoot tips and nodes of the tropical hardwoods Corymbia torelliana × C. citriodora and Khaya senegalensis. Plant Cell Tissue Organ Cult 109:341–352

    Article  Google Scholar 

  • Hung CD, Johnson K, Torpy F (2006) Liquid culture for efficient micropropagation of Wasabia japonica (Miq.) Matsumura. In Vitro Cell Dev Biol Plant 42:548–552

    Article  CAS  Google Scholar 

  • Kamada H (1985) Artificial seeds. In: Tanaka (ed) Practical technology on the mass production of clonal plants. CMC Publisher, Tokyo, p 48

  • Kinoshita I, Saito A (1992) Regeneration of Japanese white birch plants from encapsulated axillary buds. In: Kuwahara M, Shimada M (eds) Proceedings of the 5th International Conference on Biotechnology. Pulp and Paper Industry, Kyoto, pp 27–30

    Google Scholar 

  • Kulus D, Zalewska M (2014) In vitro plant recovery from alginate encapsulated Chrysanthemum × grandiflorum (Ramat.) Kitam. shoot tips. Prop Orn Plants 14:3–12

    Google Scholar 

  • Le Roux JJ, Van Staden J (1991) Micropropagation and tissue culture of Eucalyptus–a review. Tree Physiol 9:435–477

    Article  PubMed  Google Scholar 

  • Lee BC, Lee SK, Kim TS, Lee JS, Kim YW (1990) Encapsulation of in vitro shoot buds with alginate in Betula davurica. Res Rep Inst For Genet Korea 26:69–74

    Google Scholar 

  • Leifert C, Morris CE, Waites WM (1994) Ecology of microbial saprophytes and pathogens in tissue culture and field-grown plants: reasons for contamination problems in vitro. Crit Rev Plant Sci 13:139–183

    Article  Google Scholar 

  • Ma XM, Wu CF, Wang GR (2011) Application of artificial seeds in rapid multiplication of Pseudostellaria heterophylla. Afr J Biotechnol 10:15744–15748

    Article  CAS  Google Scholar 

  • Maruyama E, Kinoshita I, Ishii K, Shigenaga H, Ohba K, Saito A (1997) Alginate-encapsulation technology for the propagation of the tropical forest trees: cedrela odorata L., Guazuma crinite Mart., and Jacaranda mimosaefolia D. Don. Silvae Genet 46:17–23

    CAS  Google Scholar 

  • Micheli M, Pellegrino S, Piccioni E, Standardi A (2002) Effects of double encapsulation and coating on synthetic seed conversion in M.26 apple rootstock. J Microencapsul 19:347–356

    Article  CAS  PubMed  Google Scholar 

  • Murashige T (1977) Plant cell and organ culture as horticultural practices. Acta Hort 78:17–30

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nemoto J, Horikawa M, Ohnuki K, Shibata T, Ueno H, Hoshino M, Kaneko M (2007) Biophotofuel cell (BPFC) generating electrical power directly from aqueous solutions of biomass and its related compounds while photodecomposing and cleaning. J Appl Electrochem 37:1039–1046

    Article  CAS  Google Scholar 

  • Nieves N, Zambrano Y, Tapia R, Cid M, Pina D, Castillo R (2003) Field performance of artificial seed-derived sugarcane plants. Plant Cell Tissue Organ Cult 75:279–282

    Article  Google Scholar 

  • Onishi N, Sakamoto Y, Hirosawa T (1994) Synthetic seeds as an application of mass production of somatic embryos. Plant Cell Tissue Organ Cult 39:137–145

    Article  Google Scholar 

  • Palanyandy SR, Gantait S, Suranthran P, Sinniah UR, Subramaniam S (2015) Storage of encapsulated oil palm polyembryoids: influence of temperature and duration. In Vitro Cell Dev Biol Plant 51:118–124

    Article  Google Scholar 

  • Pattnaik SK, Sahoo Y, Chand PK (1995) Efficient plant retrieval from alginate encapsulated vegetative buds of mature mulberry trees. Sci Hortic 61:227–239

    Article  Google Scholar 

  • Pérez FJ, Meza P, Berti M, Pinto M (2000) Effect of carbon source and sucrose concentration on growth and hexose accumulation of grape berries cultured in vitro. Plant Cell Tissue Organ Cult 61:37–40

    Article  Google Scholar 

  • Perveen S, Anis M (2014) Encapsulation of internode regenerated adventitious shoot buds of Indian Siris in alginate beads for temporary storage and twofold clonal plant production. Acta Physiol Plant 36:2067–2077

    Article  CAS  Google Scholar 

  • Piccioni E, Standardi A (1995) Encapsulation of micropropagated buds of six woody species. Plant Cell Tissue Organ Cult 42:221–226

    Article  Google Scholar 

  • Pinker I, Abdel-Rahman SSA (2005) Artificial seeds for propagation of Dendranthema × grandiflora (Ramat.). Prop Orn Plant 5:186–191

    Google Scholar 

  • Rai MK, Asthana P, Singh SK, Jaiswal VS, Jaiswal U (2009) The encapsulation technology in fruit plants–a review. Biotechnol Adv 27:671–679

    Article  PubMed  Google Scholar 

  • Ravi D, Anand P (2012) Production and applications of artificial seeds: a review. Int Res J Biol Sci 1:74–78

    Google Scholar 

  • Reddy MC, Murthy KSR, Pullaiah T (2012) Synthetic seeds: a review in agriculture and forestry. Afr J Biotechnol 11:14254–14275

    Google Scholar 

  • Redenbaugh K, Nichol JW, Kossler ME, Paasch BD (1984) Encapsulation of somatic embryos for artificial seed production. In Vitro Cell Dev Biol Plant 20:256–257

    Google Scholar 

  • Redenbaugh K, Paasch BD, Nichol JW, Kossler ME, Viss PR, Walker KA (1986) Somatic seeds: encapsulation of asexual plant embryos. Nat Biotechnol 4:797–801

    Article  Google Scholar 

  • Rout GR, Das P (1997) Recent trends in the biotechnology of Chrysanthemum: a critical review. Sci Hort 69:239–257

    Article  Google Scholar 

  • Sato N (2012) Scientific élan vital: entropy deficit or inhomogeneity as a unified concept of driving forces of life in hierarchical biosphere driven by photosynthesis. Entropy 14:233–251

    Article  CAS  Google Scholar 

  • Shaheen A, Shahzad A (2015) Nutrient encapsulation of nodal segments of an endangered white cedar for studies of regrowth, short term conservation and ethylene inhibitors influenced ex vitro rooting. Ind Crop Prod 69:204–211

    Article  CAS  Google Scholar 

  • Sharma TR, Singh BM, Chauhan RS (1994) Production of disease free encapsulated buds of Zingiber officinale Rosc. Plant Cell Rep 13:300–302

    Article  CAS  PubMed  Google Scholar 

  • Sharma S, Shahzad A, Sahai A (2009) Artificial seeds for propagation and preservation of Spilanthes acmella (L.) Murr., a threatened pesticidal plant species. Int J Plant Dev Biol 3:62–64

    Google Scholar 

  • Sharma S, Shahzad A, Teixeira da Silva JA (2013) Synseed technology–a complete synthesis. Biotechnol Adv 31:186–207

    Article  CAS  PubMed  Google Scholar 

  • Sharma S, Shahzad A, Kumar J, Anis M (2014) In vitro propagation and synseed production of scarlet salvia (Salvia splendens). Rend Fis Acc Lincei 25:359–368

    Article  Google Scholar 

  • Singh AK, Sharma M, Varshney R, Agarwal SS, Bansal KC (2006) Plant regeneration from alginate to encapsulated shoot tips of Phyllanthus amarus Schum and Thonn, a medicinally important plant species. In Vitro Cell Dev Biol Plant 42:109–113

    Article  CAS  Google Scholar 

  • Singh SK, Rai MK, Asthana P, Sahoo L (2010) Alginate-encapsulation of nodal segments for propagation, short-term conservation and germplasm exchange and distribution of Eclipta alba (L.). Acta Physiol Plant 32:607–610

    Article  Google Scholar 

  • Sudipta KM, Kumara-Swamy M, Anuradha M (2013) Influence of various carbon sources and organic additives on in vitro growth and morphogenesis of Leptadenia reticulata (Wight & Arn), a valuable medicinal plant of India. Int J Pharm Sci Rev Res 21:174–179

    CAS  Google Scholar 

  • Tan TK, Loon WS, Khor E, Loh CS (1998) Infection of Spathoglottis plicata (Orchidaceae) seeds by mycorrhizal fungus. Plant Cell Rep 18:14–19

    Article  CAS  Google Scholar 

  • Teixeira da Silva JA (2003) Chrysanthemum: advances in tissue culture, cryopreservation, postharvest technology, genetics and transgenic biotechnology. Biotechnol Adv 21:715–766

    Article  CAS  PubMed  Google Scholar 

  • Teixeira da Silva JA (2004) Ornamental chrysanthemums: improvement by biotechnology. Plant Cell Tissue Organ Cult 79:1–18

    Article  Google Scholar 

  • Teixeira da Silva JA, Kulus D (2014) Chrysanthemum biotechnology: discoveries from recent literature. Folia Hortic 25:133–140

    Google Scholar 

  • Teixeira da Silva JA, Kim H, Engelmann F (2015) Chrysanthemum low-temperature storage and cryopreservation: a review. Plant Cell Tissue Organ Cult 120:423–440

    Article  CAS  Google Scholar 

  • Tomaszewska-Sowa M, Figas A (2011) Optimization of the processes of sterilization and micropropagation of cup plant (Silphium perfoliatum L.) from apical explants of seedlings in in vitro cultures. Acta Agrobot 64:3–10

    Article  Google Scholar 

  • Yaseen M, Ahmad T, Sablok G, Standardi A, Hafiz IA (2013) Review: role of carbon sources for in vitro plant growth and development. Mol Biol Rep 40:2837–2849

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was funded by the Korea Ministry of Trade, Industry and Energy under the Industrial Technology Research Program (No. N0000004), and by Cao Dinh Chay and Cao Thi Yen through CDH’s postdoctoral research project conducted at the LED Agri-Bio Fusion Technology Research Center of the Chonbuk National University, Iksan, South Korea.

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Cao Dinh Hung.

Additional information

Cao Dinh Hung and Cao Dinh Dung have contributed equally to this article.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hung, C.D., Dung, C.D. Production of chrysanthemum synthetic seeds under non-aseptic conditions for direct transfer to commercial greenhouses. Plant Cell Tiss Organ Cult 122, 639–648 (2015). https://doi.org/10.1007/s11240-015-0797-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-015-0797-0

Keywords

Navigation