Skip to main content
SpringerLink
Log in

Natural ventilation effectively reduces hyperhydricity in shoot cultures of Aloe polyphylla Schönland ex Pillans

  • Published:
Plant Growth Regulation Aims and scope Submit manuscript

Abstract

An investigation into the role of ventilation to reduce hyperhydricity in tissue cultures of Aloe polyphylla Schönland ex Pillans revealed that gaseous exchange between the in-vitro atmosphere and the outside environment is an essential prerequisite for controlling this disorder. In closed culture vessels, hyperhydricity affected as much as 84% of the newly-formed shoots on media gelled with gelrite. The leaves of hyperhydric shoots had a bright green colour, smooth epidermis and large, open stomata. Gaseous exchange was promoted by using modified lids with a hole covered with polyester or cotton mesh. In ventilated cultures, hyperhydricity was completely eliminated irrespective of the type of gelling agent used. Natural ventilation was further advantageous for the microplants in terms of leaf chlorophyll content as well as the deposition of epicuticular wax, indicating the onset of mechanisms that regulate water loss from the explants. Although culture ventilation was negatively correlated to the regeneration rate and shoot growth, it has the potential to control the appearance of abnormal phenotypes and can be easily adopted for routine A. polyphylla propagation in vitro.

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
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ANOVA:

Analysis of variance

CRD:

Completely randomized design

ESEM:

Environmental scanning electron microscope

FW:

Fresh weight

HS:

Hyperhydric shoots

IBA:

Indole-3-butyric acid

LSD:

Least significant difference

MS:

Murashige and Skoog (1962)

NS:

Normal shoots

References

  • Abrie AL, Van Staden J (2001) Micropropagation of the endangered Aloe polyphylla. Plant Growth Regul 33:19–23

    Article  CAS  Google Scholar 

  • Berger K, Schaffner W (1995) In vitro propagation of the leguminous tree Swartzia madagascariensis. Plant Cell Tissue Organ Cult 40:289–291

    Article  Google Scholar 

  • Böttcher I, Zoglauer K, Göring H (1988) Induction and reversion of vitrification of plants cultured in vitro. Physiol Plant 72:560–564

    Google Scholar 

  • Brainerd KE, Fuchigami LH (1981) Acclimatization of aseptically cultured apple plants to low relative humidity. J Am Soc Hortic Sci 106:515–518

    Google Scholar 

  • Casanova E, Moysett L, Trillas MI (2008) Effects of agar concentration and vessel closure on the organogenesis and hyperhydricity of adventitious carnation shoots. Biol Plant 52:1–8

    Article  Google Scholar 

  • Chanemougasoundharam A, Sarkar D, Pandey SK, Al-Biski F, Helali O, Minhas JS (2004) Culture tube closure-type affects potato plantlets growth and chlorophyll contents. Biol Plant 48:7–11

    Article  CAS  Google Scholar 

  • Crane J, Hughes H (1990) Medium overlays for improved hardening of micropropagated potatoes. HortScience 25:794–795

    Google Scholar 

  • Debergh PC (1983) Effects of agar brand and concentration on the tissue culture medium. Physiol Plant 59:270–276

    Article  CAS  Google Scholar 

  • Debergh P, Aitken-Christie J, Cohen D, Grout B, Von Arnold S, Zimmerman R, Ziv M (1992) Reconsideration of the term ‘vitrification’ as used in micropropagation. Plant Cell Tissue Organ Cult 30:135–140

    Article  Google Scholar 

  • Dillen W, Buysens S (1989) A simple technique to overcome vitrification in Gypsophila paniculata L. Plant Cell Tissue Organ Cult 19:181–188

    Article  Google Scholar 

  • Franck T, Kevers C, Gaspar T, Dommes J, Deby C, Greimers R, Serteyn D, Deby-Dupont G (2004) Hyperhydricity of Prunus avium shoots cultured on gelrite: a controlled stress response. Plant Physiol Biochem 42:519–527

    Article  PubMed  CAS  Google Scholar 

  • Ghashghaie J, Brenckmann F, Saugier B (1992) Water relations and growth of rose plants cultured in vitro under various relative humidities. Plant Cell Tissue Organ Cult 30:51–57

    Article  Google Scholar 

  • Grodzinski B, Boesel I, Horton K (1981) Effect of light and carbon dioxide on release of ethylene from leaves of Xanthium strumarium. Plant Physiol 67:272–273

    Google Scholar 

  • Hilton-Taylor C (1996) Red data list of Southern African plants. Strelitzia 4. National Botanical Institute, Pretoria

    Google Scholar 

  • Inskeep WP, Bloom PR (1985) Extinction coefficients of chlorophyll a and b in N, N-dimethylformamide and 80% acetone. Plant Physiol 77:483–485

    Article  PubMed  CAS  Google Scholar 

  • Ivanova MV (2009) Regulation of hyperhydricity in Aloe polyphylla propagated in vitro. Dissertation, University of KwaZulu-Natal

  • Jakse M, Bohanec B, Ihan A (1996) Effect of media components on the gynogenic regeneration of onion (Allium cepa L.) cultivars and analysis of regenerants. Plant Cell Rep 15:934–938

    Article  CAS  Google Scholar 

  • Jones NB, Drennan PM, Van Staden J (1993) Leaf anatomy, chloroplast organization and photosynthetic rate of hyperhydric Eucalyptus saligna Sm. material. South African J Bot 59:551–555

    Google Scholar 

  • Kadota M, Imizu K, Hirano T (2001) Double-phase in vitro culture using sorbitol increases shoot proliferation and reduces hyperhydricity in Japanese pear. Sci Hort 89:207–215

    Article  CAS  Google Scholar 

  • Kevers C, Gaspar T (1985) Vitrification of carnation in vitro: changes in ethylene production, ACC level and capacity to convert ACC to ethylene. Plant Cell Tissue Organ Cult 4:215–223

    Article  CAS  Google Scholar 

  • Kevers C, Coumans M, Coumans-Gillès MF, Gaspar T (1984) Physiological and biochemical events leading to vitrification of plants cultured in vitro. Physiol Plant 61:69–74

    Article  CAS  Google Scholar 

  • Kozai T, Smith MAL (1995) Environmental control in plant tissue culture—general introduction and overview. In: Aitken-Christie J, Kozai T, Smith MAL (eds) Automation and environmental control in plant tissue culture. Kluwer Academic Publishers, Dordrecht, pp 301–318

    Google Scholar 

  • Kozai T, Fujiwara K, Kitaya Y (1996) Modelling, measurement and control in plant tissue culture. Acta Hort 393:63–73

    Google Scholar 

  • Lai C-C, Lin H-M, Nalawade SM, Fang W, Tsay H-S (2005) Hyperhydricity in shoot cultures of Scrophularia yoshimurae can be effectively reduced by ventilation of culture vessels. J Plant Physiol 162:355–361

    Article  PubMed  CAS  Google Scholar 

  • Maene L, Debergh P (1987) Optimisation of the transfer of tissue cultured shoots to in vivo conditions. Acta Hort 212:334–340

    Google Scholar 

  • Majada JP, Fal MA, Sánchez-Tamés R (1997) The effect of ventilation rate on proliferation and hyperhydricity of Dianthus caryophyllus L. In Vitro Cell Dev Biol Plant 33:62–69

    Google Scholar 

  • Makunga NP, Jäger AK, Van Staden J (2006) Improved in vitro rooting and hyperhydricity in regenerating tissues of Thapsia garganica L. Plant Cell Tissue Organ Cult 86:77–86

    Article  Google Scholar 

  • McCartan SA, Beckett RP, Ivanova MV, Van Staden J (2004) In vitro hardening—the role of ventilation on acclimation stress in Kniphofia leucocephala. Plant Growth Regul 43:49–55

    Article  CAS  Google Scholar 

  • Moran R, Porath D (1980) Chlorophyll determination in intact tissues using N, N-dimethylformamide. Plant Physiol 65:478–479

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nairn BJ, Furneaux RH, Stevenson TT (1995) Identification of an agar constituent responsible for hydric control in micropropagation of radiata pine. Plant Cell Tissue Organ Cult 43:1–11

    Article  CAS  Google Scholar 

  • Park SW, Jeon JH, Kim HS, Park YM, Aswath C, Joung H (2004) Effect of sealed and vented gaseous microenvironments on the hyperhydricity of potato shoots in vitro. Sci Hort 99:199–205

    Article  Google Scholar 

  • Pasqualetto P-L, Zimmerman RH, Fordham I (1988) The influence of cation and gelling agent concentrations on vitrification of apple cultivars in vitro. Plant Cell Tissue Organ Cult 14:31–40

    Article  CAS  Google Scholar 

  • Pérez-Tornero O, Egea J, Olmos E, Burgos L (2001) Control of hyperhydricity in micropropagated apricot cultivars. In Vitro Cell Dev Biol Plant 37:250–254

    Article  Google Scholar 

  • Phan CT, Letouzé R (1983) A comparative study of chlorophyll, phenolic and protein contents, and of hydroxycinnamate:CoA ligase activity of normal and ‘vitreous’ plants (Prunus avium L.) obtained in vitro. Plant Sci Lett 31:323–327

    Article  CAS  Google Scholar 

  • Saher S, Piqueras A, Hellín E, Olmos E (2004) Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress. Physiol Plant 120:152–161

    Article  PubMed  CAS  Google Scholar 

  • Smith MAL, Spomer LA (1995) Vessels, gels, liquid media, and support systems. In: Aitken-Christie J, Kozai T, Smith MAL (eds) Automation and environmental control in plant tissue culture. Kluwer, Dordrecht, pp 371–404

    Google Scholar 

  • Smith EF, Roberts AV, Mottley J (1990) The preparation in vitro of Chrysanthemum for transplantation to soil. 3. Improved resistance to desiccation conferred by reduced humidity. Plant Cell Tissue Organ Cult 21:141–145

    Article  Google Scholar 

  • Tanaka K, Fujiwara K, Kozai T (1992) Effects of relative humidity in culture vessel on the transpiration and net photosynthetic rates of potato plantlets in vitro. Acta Hort 319:59–64

    Google Scholar 

  • Thomas P, Mythili JB, Shivashankara KS (2000) Explant, medium and vessel aeration affect the incidence of hyperhydricity and recovery of normal plantlets in triploid watermelon. J Hort Sci Biotechnol 75:19–25

    Google Scholar 

  • Tsay H-S (1998) Effects of medium composition at different recultures on vitrification of carnation (Dianthus caryophyllus) in vitro shoot proliferation. Acta Hort 461:243–249

    Google Scholar 

  • Tsay H-S, Lee C-Y, Agrawal DC, Basker S (2006) Influence of ventilation closure, gelling agent and explant type on shoot bud proliferation and hyperhydricity in Scrophularia yoshimurae—a medicinal plant. In Vitro Cell Dev Biol Plant 42:445–449

    Article  CAS  Google Scholar 

  • Williams RR, Taji AM (1991) Effect of temperature, gel concentration and cytokinins on vitrification of Olearia microdisca (J.M. Black) in vitro shoot cultures. Plant Cell Tissue Organ Cult 26:1–6

    Article  CAS  Google Scholar 

  • Ziv M (1986) In vitro hardening and acclimatization of tissue culture plants. In: Withers LA, Alderson PG (eds) Plant tissue culture and its agricultural applications. Butterworths, London, pp 187–196

    Google Scholar 

  • Ziv M (1991) Vitrification: morphological and physiological disorders of in vitro plants. In: Debergh PC, Zimmerman RH (eds) Micropropagation: technology and application. Kluwer, Dordrecht, pp 45–69

    Google Scholar 

  • Ziv M, Ariel T (1992) On the relationship between vitrification and stomatal cell wall deformity in carnation leaves in vitro. Acta Hort 314:121–129

    Google Scholar 

  • Ziv M, Ariel T (1994) Vitrification in relation to stomatal deformation and malfunction in carnation leaves in vitro. In: Lumsden PJ, Nicholas J, Davies WJ (eds) Physiology, growth and development of micropropagated plants. Kluwer, Dordrecht, pp 143–154

    Google Scholar 

  • Ziv M, Schwartz A, Fleminger D (1987) Malfunctioning stomata in vitreous leaves of carnation (Dianthus caryophyllus) plants propagated in vitro; implications for hardening. Plant Sci 52:127–134

    Article  CAS  Google Scholar 

  • Zobayed SMA, Armstrong J, Armstrong W (1999) Evaluation of a closed system, diffusive and humidity-induced convective throughflow ventilation on the growth and physiology of cauliflower in vitro. Plant Cell Tissue Organ Cult 59:113–123

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The National Research Foundation, Pretoria is thanked for financial support. Mrs B. White and Mr V. Bandu (Centre for Electron Microscopy, University of KwaZulu-Natal) are gratefully acknowledged for their assistance with the electron microscopy work.

Author information

Authors and Affiliations

  1. Research Centre for Plant Growth and Development, School of Biological and Conservation Sciences, University of KwaZulu-Natal Pietermaritzburg, Private Bag X01, Scottsville, 3209, South Africa

    Mariyana Ivanova & Johannes Van Staden

Authors
  1. Mariyana Ivanova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Johannes Van Staden
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Johannes Van Staden.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ivanova, M., Van Staden, J. Natural ventilation effectively reduces hyperhydricity in shoot cultures of Aloe polyphylla Schönland ex Pillans. Plant Growth Regul 60, 143–150 (2010). https://doi.org/10.1007/s10725-009-9430-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-009-9430-8

Keywords

Search

Navigation