Skip to main content
SpringerLink
Log in

Effect of liquid pulses with 6-benzyladenine on the induction of somatic embryogenesis from coffee (Coffea arabica L.) callus cultures

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

Abstract

We investigated the effect of the physical state of the nutrient medium on the induction of somatic embryogenesis on cell cultures derived from coffee (Coffea arabica L.). Non-embryogenic callus tissues were pulsed initially with 50 μM 6-benzyladenine (BA) for 6, 24 or 48 h in half-strength liquid Murashige and Skoog (MS) medium. After pretreatment, calli were transferred to agar-solidified half-strength MS medium supplemented with 50 μM BA (‘standard induction medium’). Control callus tissues were incubated directly on the solid standard induction medium. Callus growth was promoted by longer pretreatment periods. Formation of globular somatic embryos was observed on callus tissues pretreated with BA for 24 or 48 h, which developed fully to cotyledonary-stage within only 2 weeks after transfer to agar-solidified medium supplemented with BA. No embryo formation occurred in control cultures. Pretreatment with BA in liquid medium was associated with changes in the redox status of cultured cells, such as alterations of the ascorbate–glutathione redox systems and the accumulation of free radicals and oxidized lipids, as well as the possible reduction of cytochrome c-mediated apoptotic pathways. In particular, the induction of somatic embryogenesis was highly positively correlated (r 2 = 0.822) with the accumulation of protein carbonyls. The physiological role of BA as an inducer of both embryonic differentiation and cellular death is discussed.

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
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

AOX:

Ascorbate oxidase

APX:

Ascorbate peroxidase

Asc:

Ascorbate acid

BA:

6-Benzyladenine

CDNB:

1-Chloro-2,4-dinitrobenzene

DHAR:

Deydroascorbate reductase

DTT:

1-4-Dithiothreitol

GR:

Glutathione reductase

GSSG:

Glutathione oxidised

GSH:

Glutathione reduced

HCl:

Hydrochloric acid

HEPES:

N-2-Hydroxyethylpiperazine-N′-2-ethanesulfonic acid

H2O2 :

Hydrogen peroxide

H2DCF-DA:

2′,7′-Dichlorofluorescein diacetate

MDA:

Malondialdehyde

MS:

Murashige and Skoog basal medium

NaOH:

Sodium hydroxide

NADPH:

Nicotinamide adenosine dinucleotide phosphate

PGR:

Plant growth regulator

PPFD:

Photosynthetic photon flux density

TBA:

Thiobarbituric acid

TCA:

Trichloroacetic acid

Tris/EDTA:

Ethylenediamine tetraacetic acid buffer solution

References

  • Akutsu M, Sato H (2002) Induction of proembryos in liquid culture increases the efficiency of plant regeneration from Alstroemeria calli. Plant Sci 163:475–479

    Article  CAS  Google Scholar 

  • Ali MB, Hahn EJ, Paek K-Y (2006) Antioxidative responses of Echinacea angustifolia cultured roots to different levels of CO2 in bioreactor liquid cultures. Enzyme Microb Tech 39:982–990

    Article  CAS  Google Scholar 

  • Bearden JC (1978) Quantitation of submicrogram quantities of protein by an improved protein dye binding assay. Biochim Biophys Acta 533:525–529

    PubMed  CAS  Google Scholar 

  • Belmonte MF, Macey J, Yeung EC, Stasolla C (2005) The effect of osmoticum on ascorbate and glutathione metabolism during white spruce (Picea glauca) somatic embryo development. Plant Physiol Bioch 43:337–346

    CAS  Google Scholar 

  • Berros B (1996) Ontogenia y caracterizacion molecular e histologica de la organogenesis-embriogenesis en avellano. Dissertation, University of Oviedo, Spain

  • Braeckman BP, Hoothoofd K, Νemie De Vreese A, Vanfleteren JR (2002) Assaying metabolic activity in ageing Caenorhabditis elegans. Mech Ageing Dev 123:105–119

    Article  PubMed  CAS  Google Scholar 

  • Canal MJ, Fernandez H, Fernandez P, Centeno ML, Fernandez B (2000) Uptake and metabolism of 6-benzyladenine in ventilated cultures of kiwifruit plants. Plant Growth Reg 30:209–214

    Article  CAS  Google Scholar 

  • Carimi F, Zottini M, Formentin E, Terzi M, Lo Schiavo F (2003) Cytokinins: new apoptotic inducers in plants. Planta 216:413–421

    PubMed  CAS  Google Scholar 

  • Carimi F, Terzi M, de Michele R, Zottini M, Lo Schiavo F (2004) High levels of the cytokinin BAP induce PCD by accelarating senescente. Plant Sci 166:963–969

    Article  CAS  Google Scholar 

  • Carol RJ, Takeda S, Linstead P, Durrant MC, Kakesova H, Derbyshire P, Drea S, Zarsky V, Dolan L (2005) A RhoGDP dissociation inhibitor spatially regulates growth in root hair cells. Nature 438:1013–1016

    Article  PubMed  CAS  Google Scholar 

  • Debergh P, De Riek J, Matthys D (1994) Nutrient supply and growth of plants in culture. In: Lumsden PJ, Nicholas JR, Davies WJ (eds) Physiology, growth and development of plants in culture. Kluwer Academic Publishers, Dordrecht, pp 58–68

    Google Scholar 

  • De Feria M, Jimenez E, Barbon R, Capote A, Chavez M, Quiala E (2003) Effect of dissolved oxygen concentration on differentiation of somatic embryos of Coffea arabica cv. Catimor 9722. Plant Cell Tiss Org 72:1–6

    Article  Google Scholar 

  • De Gara L, De Pinto MC, Moliterni VMC, D’Egidio MG (2003) Redox regulation and storage processes during maturation in kernels of Triticum durum. J Exp Bot 54:249–258

    Article  PubMed  Google Scholar 

  • Desamero NV, Adelberg JW, Hale A, Young RE, Rhodes BB (1993) Nutrient utilization in liquid/membrane system for watermelon micropropagation. Plant Cell Tiss Org 33:265–271

    Article  CAS  Google Scholar 

  • D’Onofrio C, Morini S (2006) Somatic embryo, adventitious root and shoot regeneration in in vitro grown quince leaves as influenced by treatments of different length with growth regulators. Sci Hortic Amsterdam 107:194–199

    Article  CAS  Google Scholar 

  • Etienne-Barry D, Bertrand B, Schlönvoigt A, Etienne H (2002) The morphological variability within a population of coffee somatic embryos produced in a bioreactor affects the regeneration and the development of plants in the nursery. Plant Cell Tiss Org 68:153–162

    Article  Google Scholar 

  • Feito I, Gonzalez A, Centeno ML, Fernandez B, Rodriguez A (2001) Transport and distribution of benzyladenine in Actinidia deliciosa explants cultured in liquid and solid media. Plant Physiol Bioch 39:909–916

    Article  CAS  Google Scholar 

  • Foreman J, Demidchik V, Bothwell JHF, Mylona P, Miedema H, Torresk MA, Linstead P, Costa S, Brownlee C, Jonesk JDG, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446

    Article  PubMed  CAS  Google Scholar 

  • Goldfarb B, Howe GT, Bailey LM, Strauss SH, Zaerr JB (1991) A liquid cytokinin pulse induces adventitious shoot formation from Douglas-fir cotyledons. Plant Cell Rep 10:156–160

    CAS  Google Scholar 

  • Habig WH, Pabst MJ, Jacoby WB (1974) Glutathione S-transferase. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    PubMed  CAS  Google Scholar 

  • Jablonski PP, Anderson WJ (1978) Light-dependent reduction of oxidized glutathione by ruptured chloroplasts. Plant Physiol 61:221–225

    PubMed  CAS  Google Scholar 

  • Jiménez A, Hernández JA, del Río LA, Sevilla F (1997) Evidence for the presence of the ascorbate–glutathione cycle in mitochondria and peroxisomes of pea leaves. Plant Physiol 114:275–284

    PubMed  Google Scholar 

  • Kadota M, Niimi Y (2004) Improvement of micropropagation of Japanese yam using liquid and gelled medium culture. Sci Hortic Amsterdam 102:461–466

    Article  CAS  Google Scholar 

  • Kahia JW (1999) In vitro propagation of the new disease resistant Coffea arabica L. Hybrid Ruiru 11. PhD Thesis, University of London, UK

  • Kintzios S, Drossopoulos J, Shortsianitis E, Peppes D (2000) Induction of somatic embryogenesis from young, fully expanded leaves of chilli pepper (Capsicum annuum L.): effect of leaf position, illumination and explant pretreatment with high cytokinin concentrations. Sci Hortic Amsterdam 85:137–144

    Article  CAS  Google Scholar 

  • Kintzios S, Seretti E, Blouhos P, Drossopoulos JB, Kitsaki CK, Liopa-Tsakalidis A (2002) Growth regulator pre-treatment improves somatic embryogenesis from leaves of squash and melon. Plant Cell Rep 21:1–8

    Article  CAS  Google Scholar 

  • Konstas J, Kintzios S (2003) Developing a scale-up system for the micropropagation of cucumber (Cucumis sativus L.): the effect of growth retardants, liquid culture and vessel size. Plant Cell Rep 21:538–548

    PubMed  CAS  Google Scholar 

  • Kuijpers AM, Bouman H, De Klerk GJ (1996) Increase of embryogenic callus formation in cucumber by initial culture on high concentration of 2,4-dichlorophenoxyacetic acid. Plant Cell Tiss Org 46:81–83

    Article  CAS  Google Scholar 

  • Kumar Tewari R, Kumar P, Tewari N, Srivastava S, Nand Sarma P (2004) Macronutrient deficiencies and differential antioxidant responses—influence on the activity and expression of superoxide dismutase in maize. Plant Sci 166:687–694

    Article  CAS  Google Scholar 

  • Labrou NE, Mello LV, Clonis YD (2001) Functional and structural roles of the glutathione-binding residues in maize (Zea mays) glutathione S-transferase I. Biochem J 358:101–110

    Article  PubMed  CAS  Google Scholar 

  • LeBel CP, Ischiropoulos I, Bondy SC (1992) Evaluation of the probe 2′,7′-dichlorofluorescein as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol 5:227–231

    Article  PubMed  CAS  Google Scholar 

  • Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER (1990) Determination of carbonyl conent in oxidatively modified proteins. Method Enzymol 186:464–478

    Article  CAS  Google Scholar 

  • Madhulatha P, Anbalagan M, Jayachandran S, Sakthivel N (2004) Influence of liquid pulse treatment with growth regulators on in vitro propagation of banana (Musa spp. AAA). Plant Cell Tiss Org 76:189–191

    Article  CAS  Google Scholar 

  • Maene L, Debergh P (1985) Liquid medium additions to established tissue cultures to improve elongation and rooting in vitro. Plant Cell Tiss Org 5:23–33

    Article  CAS  Google Scholar 

  • Mlejnek P, Dolezel P, Procházka S (2005) Intracellular conversion of cytokinin bases into corresponding mononucleotides is related to cell death induction in tobacco BY-2 cells. Plant Sci 168:389–395

    Article  CAS  Google Scholar 

  • Molina DM, Aponte ME, Cortina H, Moreno G (2002) The effect of genotype and explant age on somatic embryogenesis of coffee. Plant Cell Tiss Org 71:117–123

    Article  CAS  Google Scholar 

  • Moncaleán P, Rodriguez A, Fernandez B (2001) In vitro response of Actinidia deliciosa explants to different BA incubation periods. Plant Cell Tiss Org 67:257–266

    Article  Google Scholar 

  • Moschopoulou G, Papanastasiou I, Makri O, Lambrou N, Economou G, Soukouli K, Kintzios S (2007) Cellular redox-status is associated with regulation of frond division in Spirodela polyrrhiza. Plant Cell Rep 26:2063–2069

    Google Scholar 

  • Mozer TJ, Tiemeier DC, Jaworski EG (1983) Purification and characterization of corn glutathione S-transferase. Biochem US 22:1068–1072

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised method for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:472–497

    Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • Nhut DT, Hanh NTM, Tuan PQ, Nquyet LTM, Tram NTH, Chinh NC, Nguyen NH, Vinh DN (2006) Liquid culture as a positive condition to induce and enhance quality and quantity of somatic embryogenesis of Lilium longiflorum. Sci Hortic Amsterdam 110:93–97

    Article  Google Scholar 

  • Nielsen TH, Ulvskov P (1992) Cytokinins and leaf development in sweet pepper (Capsicum annuum L.) II. Sink metabolism in relation to cytokinin-promoted leaf expansion. Planta 188:78–84

    Article  CAS  Google Scholar 

  • Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Phys 49:249–279

    Article  CAS  Google Scholar 

  • Ralph RK, Wojcik SJ (1982) Cytokinin effects on mitochondria and calcium metabolism. Plant Sci Lett 25:321–328

    Article  CAS  Google Scholar 

  • Ravishankar GA, Giridhar P, Indu EP, Chandrasekar A (2004) Influence of triacontanol on somatic embryogenesis in Coffea arabica L. and Coffea canephora P. ex Fr. In Vitro Cell Dev Biol Plant 40:200–203

    Article  Google Scholar 

  • Rodriguez AA, Grunberg KA, Taleisnik EL (2002) Reactive oxygen species in the elongation zone of maize leaves are necessary for leaf elongation. Plant Physiol 129:1627–1632

    Article  PubMed  CAS  Google Scholar 

  • Smirnoff N (1996) The function and metabolism of ascorbic acid in plants. Ann Bot Lond 78:661–669

    Article  CAS  Google Scholar 

  • Staritsky G (1970) Embryoid formation in callus tissues of coffee. Acta Bot Neerl 19:509–514

    Google Scholar 

  • Stark GR, Dawson CR (1963) Ascorbic acid oxidase. In: Boyer PD, Lardy H, Myrbäck K (eds) The enzymes, 2nd ed, vol 8. Academic Press, New York, pp 297–311

    Google Scholar 

  • Stasolla C, Loukanina N, Ashihara H, Yeung EC, Thorpe TA (2001) Ascorbic acid changes the pattern of purine metabolism during germination of white spruce somatic embryos. Tree Physiol 21:359–367

    Google Scholar 

  • Tommasi F, Paciolla C, de Pinto C, De Gara L (2001) A comparative study of glutathione and ascorbate metabolism during germination of Pinus pinea L. seeds. J Exp Bot 362:1647–1654

    Article  Google Scholar 

  • Van Boxtel J, Berthouly M (1996) High frequency somatic embryogenesis from coffee leaves. Factors influencing embryogenesis and subsequent proliferation and regeneration in liquid medium. Plant Cell Tiss Org 44:7–17

    Article  Google Scholar 

  • Very AA, Davies JM (2000) Hyperpolarization-activated calcium channels at the tips of Arabidopsis root hairs. P Natl Acad Sci USA 97:9801–9806

    Article  CAS  Google Scholar 

  • Washko PW, Welch RW, Dhariwal KR, Wang Y, Levine M (1992) Ascorbic acid and dehydroascorbic acid analyses in biological samples. Anal Biochem 204:1–14

    Article  PubMed  CAS  Google Scholar 

  • Yasuda T, Fujii Y, Yamaguchi T (1985) Embryogenic callus induction from Coffea arabica leaf explants by benzyladenine. Plant Cell Physiol 26:595–597

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Plant Physiology, Faculty of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, Athens, 11855, Greece

    Iosif Papanastasiou, Katerina Soukouli, Georgia Moschopoulou & Spiridon Kintzios

  2. International Coffee Research Institute, Ruiru, Kenya

    Jane Kahia

Authors
  1. Iosif Papanastasiou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Katerina Soukouli
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Georgia Moschopoulou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jane Kahia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Spiridon Kintzios
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Spiridon Kintzios.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Papanastasiou, I., Soukouli, K., Moschopoulou, G. et al. Effect of liquid pulses with 6-benzyladenine on the induction of somatic embryogenesis from coffee (Coffea arabica L.) callus cultures. Plant Cell Tiss Organ Cult 92, 215–225 (2008). https://doi.org/10.1007/s11240-007-9326-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-007-9326-0

Keywords

Search

Navigation