Skip to main content
SpringerLink
Log in

High throughput in vitro micropropagation of sugarcane (Saccharum officinarum L.) from spindle leaf roll segments: Cost analysis for agri-business industry

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

Abstract

High throughput micropropagation (HTM) protocol in sugarcane through direct shoot regeneration comprising five stages, was developed for agri-business industry. The distinction of the protocol lies in direct adventitious shoot regeneration without an intervening callus phase and liberty over the number of subculture passages leading to high rates of synchronous plant production. Stage 0 dealt with selection and maintenance of field-grown stock plants; Stage I marked the initiation of in vitro propagation on culture initiation medium using spindle leaf roll segments from apical tops; Stage II involved five passages (of 14 days each) on shoot multiplication medium followed by one additional passage (of 10 days) on shoot multiplication-cum-elongation medium. The shoot multiplication rates ranged from 4 to 25-fold in CoPb 91 and CoJ 83 cultivars, respectively; Stage III involved in vitro rooting on root induction medium and hardening for 14 days at each step; Stage IV comprised separation of clumps into single plantlets and transfer to soil. The complete plants were produced in 157 days with 97 percent survival rate. The fidelity of the protocol for agri-business industry was tested by producing approximately 0.1 million saleable HTM sugarcane plants in a small scale (150 m2) tissue culture unit. The recurring (energy, labour, media, culture container, transplanting material) and non-recurring (laboratory infrastructure, glasshouse, equipment) parameters accounted for 75.56 and 24.44 percent production costs, respectively. The low-cost options for media, culture containers and transplanting material were incorporated and cost per plant was calculated to be US$ 0.13. The HTM protocol was adopted by two sugar mills, and the HTM plants produced in our tissue culture unit were sold for seed multiplication to eight sugar mills and 430 farmers throughout North India.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

Abbreviations

BAP:

6-Benzyl amino purine

CIM:

Culture initiation medium

GA3 :

Gibberellic acid

HTM:

High throughput micropropagation

IAA:

Indole acetic acid

IBA:

Indole-3-butyric acid

ISSR:

Inter simple sequence repeat

Kin:

Kinetin

MS:

Murashige and Skoog (1962)

NAA:

Naphthalene acetic acid

RAPD:

Random amplified polymorphic DNA

RIM:

Root induction medium

SMEM:

Shoot multiplication-cum-elongation medium

SMM:

Shoot multiplication medium

SSR:

Simple sequence repeat

TDS:

Total dissolved solids

References

  • Altman A, Loberant B (1998) Micropropagation: clonal plant propagation in vitro. In: Altman A (ed) Agricultural biotechnology. Marcel Dekker Inc, New York, pp 19–42

    Google Scholar 

  • Amâncio S, Rebordão JP, Chaves MM (1999) Improvement of acclimatization of micropropagated grapevine: photosynthetic competence and carbon allocation. Plant Cell Tiss Organ Cult 58:31–37

    Article  Google Scholar 

  • Anonymous (2002) Indian tissue culture industry, Part-I. Dalal Consultants and Engineers Limited. Floriculture Today (June), India

  • Anonymous (2005) Summary report on market survey on tissue cultured plants. Biotech Consortium India Limited for Department of Biotechnology and SFAC, New Delhi

  • Babu CN (1990) Sugar cane. Allied Publishers Limited, Bombay

    Google Scholar 

  • Barathi N (2002) Plant TC listserv. www.agro.agri.umn.edu/plant-tc/listserv/

  • Baucum LE, Rice RW, Schueneman TJ (2002) An overview of Florida sugarcane, SS-AGR-232. Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida, USA

    Google Scholar 

  • Bhatia R, Singh KP, Jhang T, Sharma TR (2009) Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers. Sci Hortic 119:208–211

    Article  CAS  Google Scholar 

  • Bhojwani SS, Dantu PK (2013) Plant tissue culture: an introductory text. Springer Science & Business Media, New York

    Book  Google Scholar 

  • Boxus P (1978) The production of fruit and vegetable plants by in vitro culture: actual possibilities and perspectives. In: Hughes KW, Henker R, Constantin M (eds) Propagation of higher plants through tissue culture. Technical Information Centre, US Dept Energy, Springfield, pp 44–58

    Google Scholar 

  • Brink JA, Woodward BR, DaSilva EJ (1998) Biotechnology: a tool for development in Africa. Electron J Biotechnol 1: http://ejb.ucv.cl/content/vol1/issue1/full/1/index.html

  • Conrath U, Pieterse CMJ, Mauch-Mani B (2002) Priming in plant-pathogen interactions. Trends Plant Sci 7:210–216

    Article  CAS  PubMed  Google Scholar 

  • Debergh PC, Read PE (1991) Micropropagation. In: Debergh PC, Zimmerman RH (eds) Micropropagation technology and applications. Kluwer Acad Pub, Dordrecht, pp 1–13

    Chapter  Google Scholar 

  • Estrada-Luna AA, Davies FT Jr, Egill JN (2000) Mycorrhizal fungi enhancement of growth and gas exchange of micropropagated guava plantlets (Psidium guajava L.) during ex vitro acclimatization and plant establishment. Mycorrhiza 10:1–8

    Article  CAS  Google Scholar 

  • Ganapathi TR, Mohan JS, Suprasanna P, Bapat VA, Rao PS (1995) A low-cost strategy for in vitro propagation of banana. Curr Sci 68:646–665

    Google Scholar 

  • Geijskes RJ, LiFang W, Lakshmanan P, McKeon MG, Berding N, Swain RS, Elliott, Grof CPL, Jackson JA, Smith GR (2003) SmartSett ™ seedlings: Tissue cultured seed plants for the Australian sugar industry. Sugarcane Int. May/June:13-17

  • Gerdemann JW (1968) Vesicular-arbuscular mycorrhiza and plant growth. Annu Rev Phytopathol 6:397–418

    Article  Google Scholar 

  • Gill R, Malhotra PK, Gosal SS (2006) Direct plant regeneration from cultured young leaf segments of sugarcane. Plant Cell Tiss Organ Cult 84:227–231

    Article  CAS  Google Scholar 

  • Gosal SS, Thind KS, Dhaliwal HS (1998) Micropropagation of sugarcane-An efficient protocol for commercial plant production. Crop Improv 25:167–171

    Google Scholar 

  • Hendre RR, Mascarenhas AF, Nadgir AL, Pathak M, Jagannathan V (1975) Growth of mosaic virus free sugarcane plants from apical meristems. Indian Phytopathol 18:175–178

    Google Scholar 

  • Jalaja NC, Neelamathi D, Sreenivasan TV (2008) Micropropagation for quality seed production in sugarcane in Asia and the Pacific. Food and Agriculture Organization of the United Nations, Asia-Pacific Consortium on Agricultural Biotechnology, New Delhi

    Google Scholar 

  • Jeong BR, Fujiwara K, Kozai T (1995) Environmental control and photoautotrophic micropropagation. Horticult Rev 17:123–170

    Google Scholar 

  • Joshi P, Dhawan V (2007) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51:22–26

    Article  CAS  Google Scholar 

  • Kodym A, Zapata AFJ (2001) Low-cost alternatives for the micropropagation of banana. Plant Cell Tiss Organ Cult 66:67–71

    Article  CAS  Google Scholar 

  • Kozai T, Kubota C, Jeong BR (1997) Environmental control for the large-scale production of plants through in vitro techniques. Plant Cell Tiss Organ Cult 51:49–56

    Article  Google Scholar 

  • Lakshmanan P, Geijskes RJ, Elliott AR, Wang LF, McKeon MG, Swain RS, Borg Z, Berding N, Grof CPL, Smith GR (2002) A genotype-independent in vitro method for rapid, high frequency direct regeneration of sugarcane (Saccharum spp.). Proc Aust Soc for Biochemistry and Molecular Biology Conf, Sydney, Australia Pos-Wed-128

  • Lee TSG (1987) Micropropagation of sugarcane (Saccharum spp). Plant Cell Tiss Org Cult 10:47–55

    Article  Google Scholar 

  • Leva AR, Petruccelli R, Rinaldi LMR (2012) Somaclonal variation in tissue culture: a case study with olive. In: Leva A, Rinaldi LMR (eds) Recent advances in plant in vitro culture. InTech Pub, pp 123–150 (from:http://www.intechopen.com/books/)

  • Levin R, Alper Y, Stav R, Watad AA (1997) Methods and apparatus for liquid media and semi automated micropropagation. Acta Horticult 447:659–663

    Google Scholar 

  • Liu F, Huang LL, Li YL, Reinhoud P, Jongsma MA, Wang CY (2011) Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers. Plant Cell Tiss Organ Cult 104:111–117

    Article  CAS  Google Scholar 

  • Nowak J (1998) Benefits of in vitro “biotization” of plant tissue cultures with microbial inoculants. In Vitro Cell Dev Biol Plant 34:122–130

    Article  Google Scholar 

  • Pandey RN, Singh SP, Rastogi J, Sharma ML, Singh RK (2012) Early assessment of genetic fidelity in sugarcane (Saccharum officinarum) plantlets regenerated through direct organogenesis with RAPD and SSR markers. Aus J Crop Sci 6:618–624

    CAS  Google Scholar 

  • Pierik RLM (1987) In vitro culture of higher plants. Martinus Nijhoff Pub, Dordrecht

    Book  Google Scholar 

  • Prakash S (1993) Production of ginger and turmeric through tissue culture methods and investigations into making tissue culture propagation less expensive. Dissertation, University of Bangalore

  • Purohit SD (2012) Introduction to plant cell tissue and organ culture. PHI Learning Pvt Ltd, New Delhi

    Google Scholar 

  • Razdan MK (2008) Introduction to plant tissue culture. Oxford and IBH Publishing Company Pvt Ltd, New Delhi

    Google Scholar 

  • Sandhu SK, Gosal SS, Thind KS, Uppal SK, Sharma B, Meeta M, Singh K, Cheema GS (2009) Field performance of micropropagated plants and potential of seed cane for stalk yield and quality in sugarcane. Sugar Tech 11:34–38

    Article  Google Scholar 

  • Savangikar VA, Savangikar C, Daga RS, Pathak S (2002) Reduction in cost in micropropagation: Achievements and further prospects. Ist International Symp. Liquid system for in vitro mass propagation of plants, 29. Agricultural University, Norway

    Google Scholar 

  • Sharma TR, Singh BM (1995) Simple and cost-effective medium for propagation of ginger (Zingiber officinale). Indian J Agric Sci 65:506–508

    Google Scholar 

  • Singh G, Shetty S (2011) Impact of tissue culture on agriculture in India. Biotechnol Bioinf Bioeng 1:279–288

    Google Scholar 

  • Singh B, Yadav GC, Lal M (2001) An efficient protocol for micropropagation sugarcane using shoot tip explants. Sugar Tech 3:113–116

    Article  Google Scholar 

  • Sinha RK, Majumdar K, Sinha S (2000) In vitro differentiation and plant regeneration of Albizia chinensis (osb.) Merr. In Vitro Cell Dev Biol Plant 36:370–373

    Article  CAS  Google Scholar 

  • Sivanesan I, Jeong BR (2007) Direct shoot regeneration from nodal explants of Sida cordifolia Linn. In Vitro Cell Dev Biol Plant 43:436–441

    Article  CAS  Google Scholar 

  • Sood N, Gupta PK, Srivastava RK, Gosal SS (2006) Comparative studies on field performance of micropropagated and conventionally propagated sugarcane plants. Plant Tissue Cult Biotech 16:25–29

    Google Scholar 

  • Standaert D, Metsenaere REA (1991) Economic considerations. In: Debergh PC, Zimmerman RH (eds) Micropropagation. Kluwer Acad Pub, Dordrecht, pp 123–140

    Google Scholar 

  • Thiyagarajan M, Venkatachalam P (2012) Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: pharmaceutically important and antidiabetic medicinal herb. Ind Crops Prod 37:111–117

    Article  CAS  Google Scholar 

  • Thorpe T (2007) History of plant tissue culture. J Mol Microbiol Biotechnol 37:169–180

    CAS  Google Scholar 

  • Walker KA (1986) Automation of in vitro plant propagation: Has its time finally arrived. Genetic Engineering News July-August, pp 52

  • Yin ZF, Zhao B, Bi WL, Chen L, Wang QC (2013) Direct shoot regeneration from basal leaf segments of Lilium and assessment of genetic stability in regenerants by ISSR and AFLP markers. In Vitro Cell Dev Biol Plant 49:333–342

    Article  CAS  Google Scholar 

  • Ziv M (1986) Enhanced shoot and cormlet proliferation in liquid cultured gladiolus buds by growth retardants. Plant Cell Tiss Organ Cult 17:101–110

    Article  Google Scholar 

  • Ziv M (1991) Vitrification. In: Debergh PC, Zimmerman RH (eds) Micropropagation technology and application. Kluwer Acad Pub, Dordrecht, pp 45–69

    Chapter  Google Scholar 

Download references

Acknowledgments

The authors thankfully acknowledge the Punjab Rural Development Board, Chandigarh for providing financial support for conducting research on production of super elite planting material of sugarcane through micropropagation.

Author information

Authors and Affiliations

  1. School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana, 141 004, India

    Ajinder Kaur & Jagdeep Singh Sandhu

Authors
  1. Ajinder Kaur
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jagdeep Singh Sandhu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jagdeep Singh Sandhu.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kaur, A., Sandhu, J.S. High throughput in vitro micropropagation of sugarcane (Saccharum officinarum L.) from spindle leaf roll segments: Cost analysis for agri-business industry. Plant Cell Tiss Organ Cult 120, 339–350 (2015). https://doi.org/10.1007/s11240-014-0610-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-014-0610-5

Keywords

Search

Navigation