Acta Physiologiae Plantarum

, Volume 34, Issue 6, pp 2265–2273 | Cite as

Assessment of the role of meta-topolins on in vitro produced phenolics and acclimatization competence of micropropagated ‘Williams’ banana

  • Adeyemi O. Aremu
  • Michael W. Bairu
  • Lucie Szüčová
  • Karel Doležal
  • Jeffrey F. Finnie
  • Johannes Van Staden
Original Paper

Abstract

The effects of five topolins (meta-Topolin, mT; meta-Topolin riboside, mTR; meta-Methoxy topolin, MemT; meta-Methoxy topolin riboside, MemTR and meta-Methoxy topolin 9-tetrahydropyran-2-yl, MemTTHP) on the phenolic content and subsequent acclimatization potential of micropropagated ‘Williams’ bananas were compared to benzyladenine (BA). Sterile shoot-tip explants were cultured on modified Murashige and Skoog (MS) media containing 10, 20 or 30 μM of the above aromatic cytokinins (CKs) for 42 days. Phenolic contents were quantified spectrophotometrically. Growth parameters and photosynthetic pigments of the greenhouse-acclimatized plants were determined after 5 months. Total phenolic levels were highest in 10 μM mT-treated plantlets within the aerial parts and 30 μM MemTTHP for the underground parts. In the underground parts, 10 μM mT resulted in the production of the highest amount of proanthocyanidins which was approximately five-fold higher than in the control plants. Furthermore, 10 μM MemTTHP-treated plantlets had significantly higher total flavonoids (30.1 ± 0.24 mg CE/g DW) within the aerial parts. Plantlets regenerated using MemT, MemTR and MemTTHP had significantly longer roots and better shoot/root ratios compared to the control and BA-treated plants. In terms of root fresh weight, it was significantly higher in MemT-treated plantlets than in the control and BA treatments. Chlorophyll a/b ratio was significantly improved with the use of MemT, mTR and mT compared to control. Current findings indicate the potential of topolins in stimulating the accumulation of phenolic compounds in micropropagated plantlets. In view of the importance of plant secondary metabolites, their substantial accumulation probably enhanced the acclimatization and subsequent ex vitro survival of the micropropagated plantlets. Topolins, particularly, the new derivative MemTTHP could be an alternative CK for the micropropagation of plant species based on their stimulatory effect on ex vitro rooting that inevitably enhances acclimatization competence. Furthermore, topolins are demonstrated as potential elicitors in micropropagation.

Keywords

Cytokinins Chlorophyll Phenolics Micropropagation Musa spp 

Abbreviations

BA

N6-Benzyladenine

CCE

Cyanidin chloride equivalents

CE

Catechin equivalents

CK

Cytokinin

DMRT

Duncan’s multiple range test

DW

Dry weight

Folin-C

Folin-Ciocalteu

FW

Fresh weight

GAE

Gallic acid equivalents

MemT

meta-Methoxy topolin

MemTR

meta-Methoxy topolin riboside

MemTTHP

meta- Methoxy topolin 9-tetrahydropyran-2-yl

MS

Murashige and Skoog basal medium

mT

meta-Topolin

mTR

meta-Topolin riboside

PPFD

Photosynthetic photon flux density

PTC

Plant tissue culture

References

  1. Adedipe NO, Hunt LA, Fletcher RA (1971) Effects of benzyladenine on photosynthesis, growth and senescence of the bean plant. Physiol Plant 25:151–153CrossRefGoogle Scholar
  2. Aremu AO, Bairu MW, Doležal K, Finnie JF, Van Staden J (2012) Topolins: a panacea to plant tissue culture challenges? Plant Cell Tissue Organ Cult 108:1–16CrossRefGoogle Scholar
  3. Bairu MW, Stirk WA, Doležal K, Van Staden J (2007) Optimizing the micropropagation protocol for the endangered Aloe polyphylla: can meta-topolin and its derivatives serve as replacement for benzyladenine and zeatin? Plant Cell Tissue Organ Cult 90:15–23CrossRefGoogle Scholar
  4. Bairu MW, Stirk WA, Doležal K, Van Staden J (2008) The role of topolins in micropropagation and somaclonal variation of banana cultivars ‘Williams’ and ‘Grand Naine’ (Musa spp. AAA). Plant Cell Tissue Organ Cult 95:373–379CrossRefGoogle Scholar
  5. Bairu MW, Aremu AO, Van Staden J (2011a) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173CrossRefGoogle Scholar
  6. Bairu MW, Novák O, Doležal K, Van Staden J (2011b) Changes in endogenous cytokinin profiles in micropropagated Harpagophytum procumbens in relation to shoot-tip necrosis and cytokinin treatments. Plant Growth Regul 63:105–114CrossRefGoogle Scholar
  7. Bari R, Jones JDG (2009) Role of plant hormones in plant defence responses. Plant Mol Biol 69:473–488PubMedCrossRefGoogle Scholar
  8. Baroja-Fernández E, Aguirreolea J, Martínková H, Hanus J, Strnad M (2002) Aromatic cytokinins in micropropagated potato plants. Plant Physiol Biochem 40:217–224CrossRefGoogle Scholar
  9. Bernier PY, Lamhamedi MS, Simpson DG (1995) Shoot:root ratio is of limited use in evaluating the quality of container conifer stock. Tree Planters Notes 46:102–106Google Scholar
  10. Binks RH, Greenham JR, Luis JG, Gowen SR (1997) A phytoalexin from roots of Musa acuminata var. Pisang sipulu. Phytochemistry 45:47–49CrossRefGoogle Scholar
  11. Buer CS, Imin N, Djordjevic MA (2010) Flavonoids: new roles for old molecules. J Integrat Plant Biol 52:98–111CrossRefGoogle Scholar
  12. Caponetti JD, Gray DJ, Trigiano RN (2005) History of plant tissue and cell culture. In: Trigiano RN, Gray DJ (eds) Plant development and biotechnology. CRC Press, Florida, pp 9–15Google Scholar
  13. Čatský J, Pospíšilová J, Kamínek M, Gaudinová A, Pulkrábek J, Zahradníček J (1996) Seasonal changes in sugar beet photosynthesis as affected by exogenous cytokinin N6-(m-hydroxybenzyl)adenosine. Biol Plant 38:511–518CrossRefGoogle Scholar
  14. Chow WS, Anderson JM, Melis A (1990) The photosystem stoichiometry in thylakoids of some Australian shade-adapted plant species. Aust J Plant Physiol 17:665–674CrossRefGoogle Scholar
  15. Collingborn FMB, Gowen SR, Mueller-Harvey I (2000) Investigations into the biochemical basis for nematode resistance in roots of three Musa cultivars in response to Radopholus similis infection. J Agric Food Chem 48:5297–5301PubMedCrossRefGoogle Scholar
  16. Criado M, Roberts I, Echeverria M, Barneix A (2007) Plant growth regulators and induction of leaf senescence in nitrogen-deprived wheat plants. J Plant Growth Regul 26:301–307CrossRefGoogle Scholar
  17. Curir P, VanSumere CF, Termini A, Barthe P, Marchesini A, Dolci M (1990) Flavonoid accumulation is correlated with adventitious roots formation in Eucalyptus gunnii Hook micropropagated through axillary bud stimulation. Plant Physiol 92:1148–1153PubMedCentralPubMedCrossRefGoogle Scholar
  18. De Klerk G-J, Guan H, Huisman P, Marinova S (2011) Effects of phenolic compounds on adventitious root formation and oxidative decarboxylation of applied indoleacetic acid in Malus ‘Jork 9’. Plant Growth Regul 63:175–185CrossRefGoogle Scholar
  19. Dixon RA, Paiva NL (1995) Stress-induced phenylpropanoid metabolism. Plant Cell 7:1085–1097PubMedCentralPubMedGoogle Scholar
  20. Escalona M, Cejas I, González-Olmedo J, Capote I, Roels S, Cañal MJ, Rodríguez R, Sandoval J, Debergh P (2003) The effect of meta-topolin on plantain propagation using a temporary immersion bioreactor. InfoMusa 12:28–30Google Scholar
  21. Genkov T, Tsoneva P, Ivanova I (1997) Effect of cytokinins on photosynthetic pigments and chlorophyllase activity in in vitro cultures of axillary buds of Dianthus caryophyllus L. J Plant Growth Regul 16:169–172CrossRefGoogle Scholar
  22. Jain SM (2004) Introduction: cellular biology and biotechnology including mutation techniques for creation of new useful banana genotypes. In: Jain SM, Swennen R (eds) Banana improvement: cellular, molecular biology, and induced mutations. Science Publishers, Inc., Enfield, pp xv–xviGoogle Scholar
  23. Lewinsohn E, Gijzen M (2009) Phytochemical diversity: the sounds of silent metabolism. Plant Sci 176:161–169CrossRefGoogle Scholar
  24. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. In: Douce R, Packer L (eds) Methods in enzymology, vol 148., Academic Press New York, USA, pp 350–382Google Scholar
  25. Liu X-N, Zhang X-Q, Sun J-S (2007) Effects of cytokinins and elicitors on the production of hypericins and hyperforin metabolites in Hypericum sampsonii and Hypericum perforatum. Plant Growth Regul 53:207–214CrossRefGoogle Scholar
  26. Makkar HPS, Sidhuraju P, Becker K (2007) Plant secondary metabolites. Humana Press Inc., New JerseyCrossRefGoogle Scholar
  27. Marinova D, Ribarova F, Atanassova M (2005) Total phenolics and total flavonoids in Bulgarian fruits and vegetables. J Univ Cheml Technol Metal 40:255–260Google Scholar
  28. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  29. Mutui TM, Mibus H, Serek M (2012) Effect of meta-topolin on leaf senescence and rooting in Pelargonium × hortorum cuttings. Postharvest Biol Technol 63:107–110CrossRefGoogle Scholar
  30. Namdeo AG (2007) Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev 1:69–79Google Scholar
  31. Palavan-Ünsal N, Çağ S, Çetin E (2004) The role of meta-topolin in senescence of wheat leaf segments. J Cell Mol Biol 3:23–31Google Scholar
  32. Peer WA, Murphy AS (2007) Flavonoids and auxin transport: modulators or regulators? Trends Plant Sci 12:556–563PubMedCrossRefGoogle Scholar
  33. Porter LJ, Hrstich LN, Chan BG (1985) The conversion of proanthocyanidins and prodelphinidins to cyanidin and delphinidin. Phytochemistry 25:223–230CrossRefGoogle Scholar
  34. Pospíšilová J, Synková H, Haisel D, Čatský J, Wilhelmová N, Šrámek F (1999) Effect of elevated CO2 concentration on acclimation of tobacco plantlets to ex vitro conditions. J Exp Bot 50:119–126CrossRefGoogle Scholar
  35. Pospíšilová J, Synková H, Haisel D, Semorádová S (2007) Acclimation of plantlets to ex vitro conditions: effects of air humidity, irradiance, CO2 concentration and abscisic acid (a review). Acta Hortic 748:29–38Google Scholar
  36. Quiala E, Cañal M-J, Meijón M, Rodríguez R, Chávez M, Valledor L, de Feria M, Barbón R (2012) Morphological and physiological responses of proliferating shoots of teak to temporary immersion and BA treatments. Plant Cell Tissue Organ Cult. 109:223–234CrossRefGoogle Scholar
  37. Ramachandra Rao S, Ravishankar GA (2002) Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20:101–153CrossRefGoogle Scholar
  38. Szücová L, Spíchal L, Dolezal K, Zatloukal M, Greplová J, Galuszka P, Krystof V, Voller J, Popa I, Massino FJ, Jørgensen J-E, Strnad M (2009) Synthesis, characterization and biological activity of ring-substituted 6-benzylamino-9-tetrahydropyran-2-yl and 9-tetrahydrofuran-2-ylpurine derivatives. Bioorg Med Chem 17:1938–1947PubMedCrossRefGoogle Scholar
  39. Valero-Aracama C, Kane M, Wilson S, Vu J, Anderson J, Philman N (2006) Photosynthetic and carbohydrate status of easy-and difficult-to-acclimatize sea oats (Uniola paniculata L.) genotypes during in vitro culture and ex vitro acclimatization. In Vitro Cell Dev Biol Plant 42:572–583CrossRefGoogle Scholar
  40. Valero-Aracama C, Kane M, Wilson S, Philman N (2010) Substitution of benzyladenine with meta-topolin during shoot multiplication increases acclimatization of difficult- and easy-to-acclimatize sea oats (Uniola paniculata L.) genotypes. Plant Growth Regul 60:43–49CrossRefGoogle Scholar
  41. Van Staden J, Cook EL, Noodén LD (1988) Cytokinins and senescence. In: Noodén LD, Leopold AC (eds) Senescence and aging in plants. Academic Press, Inc., San Diego, pp 281–328Google Scholar
  42. Vogt T (2010) Phenylpropanoid biosynthesis. Mol Plant 3:2–20PubMedCrossRefGoogle Scholar
  43. Vuylsteke DR (1998) Shoot-tip culture for the propagation, conservation, and distribution of Musa germplasm. International Institute of Tropical Agriculture, IbadanGoogle Scholar
  44. Werbrouck SPO, Van der Jeugt B, Dewitte W, Prinsen E, Van Onckelen HA, Debergh PC (1995) The metabolism of benzyladenine in Spathiphyllum floribundum ‘Schott Petite’ in relation to acclimatisation problems. Plant Cell Rep 14:662–665PubMedCrossRefGoogle Scholar
  45. Werbrouck SPO, Strnad M, Van Onckelen HA, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture? Physiol Plant 98:291–297CrossRefGoogle Scholar
  46. Werner T, Motyka V, Strnad M, Schmülling T (2001) Regulation of plant growth by cytokinin. Proc Natl Acad Sci 98:10487–10492PubMedCrossRefGoogle Scholar
  47. Wu HC, du Toit ES, Reinhardt CF, Rimando AM, van der Kooy F, Meyer JJM (2007) The phenolic, 3,4-dihydroxybenzoic acid, is an endogenous regulator of rooting in Protea cynaroides. Plant Growth Regul 52:207–215CrossRefGoogle Scholar
  48. Wuyts N, Lognay G, Sági L, De Waele D, Swennen R (2005) Secondary metabolites in roots and implications for nematode resistance in banana (Musa spp.). In: Turner DW, Rosales FE (eds) Banana root system: towards a better understanding for its productive management. Proceedings of an international symposium. The International Network for the Improvement of Banana and Plantain, Montpellier, pp 238–246Google Scholar
  49. Wuyts N, Lognay G, Verscheure M, Marlier M, De Waele D, Swennen R (2007) Potential physical and chemical barriers to infection by the burrowing nematode Radopholus similis in roots of susceptible and resistant banana (Musa spp.). Plant Pathol 56:878–890CrossRefGoogle Scholar
  50. Yokoyama M, Naito K, Suzuki H (1980) Effects of benzyladenine on chlorophyll, DNA, RNA and protein content of attached young bean (Phaseolus vulgaris L.) leaves. Ann Bot 45:649–653Google Scholar
  51. Zhishen J, Mengcheng T, Jianming W (1999) The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 64:555–559CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2012

Authors and Affiliations

  • Adeyemi O. Aremu
    • 1
  • Michael W. Bairu
    • 1
  • Lucie Szüčová
    • 2
    • 3
  • Karel Doležal
    • 2
    • 3
  • Jeffrey F. Finnie
    • 1
  • Johannes Van Staden
    • 1
  1. 1.Research Centre for Plant Growth and Development, School of Life SciencesUniversity of KwaZulu-Natal PietermaritzburgScottsvilleSouth Africa
  2. 2.Laboratory of Growth RegulatorsPalacký University and Institute of Experimental Botany AS CROlomoucCzech Republic
  3. 3.Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of SciencePalacký UniversityOlomoucCzech Republic

Personalised recommendations