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In vitro shoot organogenesis and comparative analysis of endogenous phytohormones in kohlrabi (Brassica oleracea var. gongylodes): effects of genotype, explant type and applied cytokinins

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Abstract

Kohlrabi (Brassica oleracea var. gongylodes) cultivars Vienna Purple (VP) and Vienna White (VW) were tested for their ability of de novo organogenesis in vitro. Root, cotyledon, hypocotyl explants and intact seedlings were cultivated on Murashige and Skoog (MS) media supplemented with different cytokinins: benzyladenine (BA), thidiazuron (TDZ), trans- or cis-zeatin. All tested cytokinins, including cis-zeatin, induced shoot regeneration from hypocotyl explants and intact seedlings, with seedlings being most successful for regeneration efficiency and viability of regenerated shoots in both cultivars. The highest frequency of shoot regeneration was achieved on MS with BA (60 %) or TDZ (50 %) for VP; and with BA (50 %), TDZ (47.5 %) or transZ (37.5 %) for VW. Measurements of the endogenous cytokinin and indole-3-acetic acid (IAA) contents in both hypocotyl explants and seedlings with regenerated shoots (HRSs and SRSs) suggested that the observed differences in organogenic response between these two types of explants were related to their cytokinin and IAA contents. HRSs generally exhibited elevated amounts of total cytokinins, while SRSs displayed a higher IAA/bioactive cytokinins ratio. Shoots regenerated from seedlings were further successfully multiplicated on a medium supplemented with BA (0.5 mg L−1). The rooting potential of multiplicated shoots was tested on media supplemented with 2 or 4 mg L−1 indole-3-butyric acid (IBA), with the higher concentration of IBA leading to more efficient rooting. Rooted plantlets were successfully planted into soil and flow cytometric analysis did not reveal ploidy variations, indicating that the described protocol is fast and efficient for kohlrabi regeneration.

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Abbreviations

BA:

N 6-benzyladenine

cisZ:

cis-Zeatin

cisZR:

cis-Zeatin-9-riboside

cisZ7G:

cis-Zeatin-7-glucoside

cisZ9G:

cis-Zeatin-9-glucoside

cisZOG:

cis-Zeatin-O-glucoside

cisZROG:

cis-Zeatin-9-riboside-O-glucoside

cisZRMP:

cis-Zeatin-9-riboside-5′-monophosphate

CK:

Cytokinin

DNSO:

De novo shoot organogenesis

DHZ:

Dihydrozeatin

DHZR:

Dihydrozeatin-9-riboside

DHZ7G:

Dihydrozeatin-7-glucoside

DHZ9G:

Dihydrozeatin-9-glucoside

DHZOG:

Dihydrozeatin-O-glucoside

DHZROG:

Dihydrozeatin-9-riboside-O-glucoside

DHZRMP:

Dihydrozeatin-9-riboside-5′-monophosphate

HRS:

Hypocotyl with regenerated shoots

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

iP:

N 6-(∆2-isopentenyl)adenine

iPR:

N 6-(∆2-isopentenyl)adenosine

iP7G:

N 6-(∆2-isopentenyl)adenine-7-glucoside

iP9G:

N 6-(∆2-isopentenyl)adenine-9-glucoside

iPRMP:

N 6-(∆2-isopentenyl)adenosine -5′-monophosphate

PGR:

Plant growth regulator

SRS:

Seedling with regenerated shoots

TDZ:

Thidiazuron

transZ:

trans-Zeatin

transZR:

trans-Zeatin-9-riboside

transZ7G:

trans-Zeatin-7-glucoside

transZ9G:

trans-Zeatin-9-glucoside

transZOG:

trans-Zeatin-O-glucoside

transZROG:

trans-Zeatin-9-riboside-O-glucoside

transZRMP:

trans-Zeatin-9-riboside-5′-monophosphate

VP:

Vienna Purple cultivar

VW:

Vienna White cultivar

References

  • Abbasi BH, Khan M, Guo B, Bokhari SA, Khan MA (2011) Efficient regeneration and antioxidative enzyme activities in Brassica rapa var. turnip. Plant Cell Tissue Organ Cult 105:337–344

    Article  CAS  Google Scholar 

  • Ahmadabadi M, Bock R (2010) Development of a highly responsive leaf-based regeneration system for Peperomia species. Turk J Bot 34:329–334

    Google Scholar 

  • Akasaka-Kennedy Y, Yoshida H, Takahata Y (2005) Efficient plant regeneration from leaves of rape seed (Brassica napus L.): influence of AgNO3 and genotype. Plant Cell Rep 24:649–654

    Article  CAS  PubMed  Google Scholar 

  • Aremu AO, Plačková L, Bairu MW, Novák O, Plíhalová L, Doležal K, Finnie JF, Van Staden J (2014a) How does exogenously applied cytokinin type affect growth and endogenous cytokinins in micropropagated Merwilla plumbea? Plant Cell Tissue Organ Cult 118:245–256

    Article  CAS  Google Scholar 

  • Aremu AO, Plačková L, Bairu MW, Novák O, Szüčová L, Doležal K, Finnie JF, Van Staden J (2014b) Endogenous cytokinin profiles of tissue-cultured and acclimatized ‘Williams’ bananas subjected to different aromatic cytokinin treatments. Plant Sci 214:88–98

    Article  CAS  PubMed  Google Scholar 

  • Auer CA, Cohen JD, Laloue M, Cooke TJ (1992) Comparison of benzyl adenine metabolism in two Petunia hybrida lines differing in shoot organogenesis. Plant Physiol 98:1035–1041

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Bajguz A, Piotrowska A (2009) Conjugates of auxin and cytokinin. Phytochemistry 70:957–969

    Article  CAS  PubMed  Google Scholar 

  • Baroja-Fernández E, Aguirreolea J, Martínková H, Hanuš J, Strnad M (2002) Aromatic cytokinins in micropropagated potato plants. Plant Physiol Biochem 40:217–224

    Article  Google Scholar 

  • Bassil NV, Mok DWS, Mok MC (1993) Partial purification of a cis-trans-isomerase of zeatin from immature seed of Phaseolus vulgaris L. Plant Physiol 102:867–872

    PubMed Central  CAS  PubMed  Google Scholar 

  • Benková E, Michniewicz M, Sauer M, Teichmann T, Seifertová D, Jürgens G, Friml J (2003) Local, efflux-dependent auxin gradients as a common module for plant organ formation. Cell 115:591–602

    Article  PubMed  Google Scholar 

  • Bhalla PL, de Weerd N (1999) In vitro propagation of cauliflower, Brassica oleracea var. botrytis for hybrid seed production. Plant Cell Tissue Organ Cult 56:89–95

    Article  CAS  Google Scholar 

  • Bouza L, Jacques M, Sotta B, Miginiac E (1993) The differential effect of N6-benzyl-adenine and N6-(Δ2-isopentenyl)-adenine on in vitro propagation of Paeonia suffruticosa Andr. is correlated with different hormone contents. Plant Cell Rep 12:593–596

    Article  CAS  PubMed  Google Scholar 

  • Bretagne B, Chupeau MC, Chupeau Y, Fouilloux G (1994) Improved flax regeneration from hypocotyls using thidiazuron as a cytokinin source. Plant Cell Rep 14:120–124

    CAS  PubMed  Google Scholar 

  • Brugière N, Jiao S, Hantke S, Zinselmeier C, Roessler JA, Niu X, Jones RJ, Habben JE (2003) Cytokinin oxidase gene expression in maize is localized to the vasculature, and is induced by cytokinins, abscisic acid, and abiotic stress. Plant Physiol 132:1228–1240

    Article  PubMed Central  PubMed  Google Scholar 

  • Cardoza V, Stewart CN (2004) Brassica biotechnology: progress in cellular and molecular biology. In Vitro Cell Dev Biol Plant 40:542–551

    Article  CAS  Google Scholar 

  • Centeno ML, Rodríguez A, Feito I, Fernández B (1996) Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinia deliciosa tissue cultures. Plant Cell Rep 16:58–62

    Article  CAS  PubMed  Google Scholar 

  • Che P, Gingerich DJ, Lall S, Howell SH (2002) Global and hormone-induced gene expression changes during shoot development in Arabidopsis. Plant Cell 14:2771–2785

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Che P, Lall S, Nettleton D, Howell SH (2006) Gene expression programs during shoot, root, and callus development in Arabidopsis tissue culture. Plant Physiol 141:620–637

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Cheng ZJ, Wang L, Sun W, Zhang Y, Zhou C, Su YH, Li W, Sun TT, Zhao XY, Li XG, Cheng Y, Zhao Y, Xie Q, Zhang XS (2013) Pattern of auxin and cytokinin responses for shoot meristem induction results from the regulation of cytokinin biosynthesis by AUXIN RESPONSE FACTOR3. Plant Physiol 161:240–251

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chikkala VRN, Nugent G, Dix P, Stevenson T (2009) Regeneration from leaf explants and protoplasts of Brassica oleracea var. botrytis (cauliflower). Sci Hortic 119:330–334

    Article  CAS  Google Scholar 

  • Cogbill S, Faulcon T, Jones G, McDaniel M, Harmon G, Blackmon R, Young M (2010) Adventitious shoot regeneration from cotyledonary explants of rapid-cycling fast plants of Brassica rapa L. Plant Cell Tissue Organ Cult 101:127–133

    Article  Google Scholar 

  • Coleman GD, Ernst SG (1989) In vitro shoot regeneration of Populus deltoids: effect of cytokikin and genotype. Plant Cell Rep 8:459–462

    Article  CAS  PubMed  Google Scholar 

  • Ćosić T, Vinterhalter B, Vinterhalter D, Mitić N, Cingel A, Savić J, Bohanec B, Ninković S (2013) In vitro plant regeneration from immature zygotic embryos and repetitive somatic embryogenesis in kohlrabi (Brassica oleracea var. gongylodes). In Vitro Cell Dev Biol Plant 49:294–303

    Article  Google Scholar 

  • Cuesta C, Rodriguez A, Centeno ML, Ordas RJ, Fernandez B (2009) Caulogenic induction in cotyledons of tone pine (Pinus pinea): relationship between organogenic response and benzyladenine trends in elected families. J Plant Physiol 166:1162–1171

    Article  CAS  PubMed  Google Scholar 

  • Cuesta C, Novák O, Ordás RJ, Fernández B, Strnad M, Doležal K, Rodríguez A (2012) Endogenous cytokinin profiles and their relationships to between-family differences during adventitious caulogenesis in Pinus pinea cotyledons. J Plant Physiol 169:1830–1837

    Article  CAS  PubMed  Google Scholar 

  • Dai XG, Shi XP, Ye YM, Fu Q, Bao MZ (2009) High frequency plant regeneration from cotyledon and hypocotyl explants of ornamental kale. Biol Plant 53:769–773

    Article  Google Scholar 

  • Dobrev P, Kamínek M (2002) Fast and efficient separation of cytokinins from auxin and abscisic acid and their purification using mixed-mode solid-phase extraction. J Chromatogr A 950:21–29

    Article  PubMed  Google Scholar 

  • Dobrev P, Havlíček L, Vágner M, Malbeck J, Kamínek M (2005) Purification and determination of plant hormones auxin and abscisic acid using solid phase extraction and two-dimensional high performance liquid chromatograophy. J Chromatogr A 1075:159–166

    Article  CAS  PubMed  Google Scholar 

  • Duclercq J, Sangwan-Norreel B, Catterou M, Sangwan RS (2011) De novo shoot organogenesis: from art to science. Trends Plant Sci 16:597–606

    Article  CAS  PubMed  Google Scholar 

  • Dwivedi S, Vaňková R, Motyka V, Herrera C, Žižková E, Auer C (2010) Characterization of Arabidopsis thaliana mutant ror-1 (roscovitine-resistant) and its utilization in understanding of the role of cytokinin N-glucosylation pathway in plants. Plant Growth Regul 61:231–242

    Article  CAS  Google Scholar 

  • Eklöf S, Åstot C, Blackwell J, Moritz T, Olsson O, Sandberg G (1997) Auxin-cytokinin interactions in wild-type and transgenic tobacco. Plant Cell Physiol 38:225–235

    Article  Google Scholar 

  • Emery RJN, Leport L, Barton JE, Turner NC, Atkins CA (1998) cis-Isomers of cytokinins predominate in chickpea seeds throughout their development. Plant Physiol 117:1515–1523

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Emery RJN, Ma Q, Atkins CA (2000) The forms and sources of cytokinins in developing white lupine seeds and fruits. Plant Physiol 123:1593–1604

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Escalona VH, Aguayo E, Artés E (2007) Extending the shelf life of kohlrabi stems by modified atmosphere packaging. J Food Sci 72:308–313

    Article  Google Scholar 

  • Gajdošová S, Spíchal L, Kamínek M, Hoyerová K, Novák O, Dobrev PI, Galuszka P, Klíma P, Gaudinová A, Žižková E, Hanuš J, Dančák M, Trávníček B, Pešek B, Krupička M, Vaňková R, Strnad M, Motyka V (2011) Distribution, biological activities, metabolism, and the conceivable function of cis-zeatin-type cytokinins in plants. J Exp Bot 62:2827–2840

    Article  PubMed  Google Scholar 

  • Ghnaya AB, Charles G, Branchard M (2008) Rapid shoot regeneration from thin cell layer explants excised from petioles and hypocotyls in four cultivars of Brassica napus L. Plant Cell Tissue Organ Cult 92:25–30

    Article  Google Scholar 

  • Glendening TM, Sjolund R (1988) In vitro propagation of kohlrabi from leaf explants. HortScience 23:772

    Google Scholar 

  • Gordon SP, Heisler MG, Reddy GV, Ohno C, Das P, Meyerowitz EM (2007) Pattern formation during de novo assembly of the Arabidopsis shoot meristem. Development 134:3539–3548

    Article  CAS  PubMed  Google Scholar 

  • Grosch R, Schneider JHM, Kofoet A (2004) Characterisation of Rhizoctonia solani anastomosis groups causing bottom rot in field-grown lettuce in Germany. Eur J Plant Pathol 110:53–62

    Article  Google Scholar 

  • Haddadi P, Moieni A, Karimzadeh G, Abdollahi MR (2008) Effects of gibberellin, abscisic acid and embryo desiccation on normal plantlet regeneration, secondary embryogenesis and callogenesis in microspore culture of Brassica napus L. cv. PF704. Int J Plant Prod 2:153–162

    CAS  Google Scholar 

  • Hansen LN, Ortiz R, Andersen SB (1999) Genetic analysis of protoplast regeneration ability in Brassica oleracea. Plant Cell Tissue Organ Cult 58:127–132

    Article  Google Scholar 

  • Jones B, Andersson Gunnerås S, Petersson SV, Tarkowski P, Graham N, May S, Dolezal K, Sandberg G, Ljung K (2010) Cytokinin regulation of auxin synthesis in Arabidopsis involves a homeostatic feedback loop regulated via auxin and cytokinin signal transduction. Plant Cell 22:2956–2969

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kamal GB, Illich KG, Asadollah A (2007) Effect of genotype, explant type and nutrient medium components on canola (Brassics napus L.) shoot in vitro organogenesis. Afr J Biotechnol 6:861–867

    CAS  Google Scholar 

  • Kamínek M, Pačes V, Corse J, Challice JS (1979) Effect of stereospecific hydroxylation of N6-(∆2-isopentenyl)adenosine on cytokinin activity. Planta 145:239–243

    Article  PubMed  Google Scholar 

  • Kamínek M, Motyka V, Vaňková R (1997) Regulation of cytokinin content in plant cells. Physiol Plant 101:689–700

    Article  Google Scholar 

  • Kamínek M, Březinová A, Gaudinová A, Motyka V, Vaňková R, Zažímalová E (2003) Purine cytokinins: a proposal of abbreviations. Plant Growth Regul 32:253–256

    Article  Google Scholar 

  • Kasahara H, Takei K, Ueda N, Hishiyama S, Yamaya T, Kamiya Y, Yamaguchi S, Sakakibara H (2004) Distinct isoprenoid origins of cis- and trans-zeatin biosyntheses in Arabidopsis. J Biol Chem 279:14049–14054

    Article  CAS  PubMed  Google Scholar 

  • Khan MR, Rashid H, Ansar M, Chaudry Z (2003) High frequency shoot regeneration and Agrobacterium-mediated DNA transfer in Canola (Brassica napus). Plant Cell Tissue Organ Cult 75:223–231

    Article  Google Scholar 

  • Klemš M, Slámová Z, Motyka V, Malbeck J, Trávníčková A, Macháčková I, Holík J, Procházka S (2011) Changes in cytokinin levels and metabolism in tobacco (Nicotiana tabacum L.) explants during in vitro shoot organogenesis induced by trans-zeatin and dihydrozeatin. Plant Growth Regul 65:427–437

    Article  Google Scholar 

  • Klíma M, Vyvadilová M, Kučera V (2004) Production and utilization of doubled haploids in Brassica oleracea vegetables. Hortic Sci Prague 31:119–123

    Google Scholar 

  • Koh WL, Loh CS (2000) Direct somatic embryogenesis, plant regeneration and in vitro flowering in rapid-cycling Brassica napus. Plant Cell Rep 19:1177–1183

    Article  CAS  Google Scholar 

  • Lexa M, Genkov T, Malbeck J, Macháčková I, Brzobohatý B (2003) Dynamics of endogenous cytokinin pools in tobacco seedlings: a modelling approach. Ann Bot 91:585–597

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Lillo C, Shanin EA (1986) Rapid regeneration of plants from hypocotyl protoplasts and root segments of cabbage. HortScience 21:315–317

    CAS  Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127

    Article  CAS  Google Scholar 

  • Liu J, Mehdi S, Topping J, Tarkowski P, Lindsey K (2010) Modelling and experimental analysis of hormonal crosstalk in Arabidopsis. Mol Syst Biol 6: Article number 373

  • LoSchiavo F, Pitto L, Giuliano G, Torti G, Nuti-Ronchi V, Marazziti D, Vergara R, Orselli S, Terzi M (1989) DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs. Theor Appl Genet 77:325–331

    Article  CAS  PubMed  Google Scholar 

  • Martin RC, Mok MC, Habben JE, Mok DWS (2001) A cytokinin gene from maize encoding an O-glucosyltransferase specific to cis-zeatin. Proc Natl Acad Sci USA 98:5922–5926

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Miyawaki K, Matsumoto-Kitano M, Kakimoto T (2004) Expression of cytokinin biosynthetic isopentenyltransferase genes in Arabidopsis: tissue specificity and regulation by auxin, cytokinin, and nitrate. Plant J 37:128–138

    Article  CAS  PubMed  Google Scholar 

  • Moghaieb R, El-Awady M, El Mergawy R, Youssef S, El-Sharkawy A (2006) A reproducible protocol for regeneration and transformation in canola (Brassica napus L.). Afr J Biotechnol 5:143–148

    CAS  Google Scholar 

  • Mollika SR, Sarker RH, Hoque MI (2011) In vitro plant regeneration in Brassica spp. Plant Tissue Cult Biotechnol 21:127–134

    Article  Google Scholar 

  • Moncaleán P, Rodríguez A, Fernández B (2003) Effect of different benzyladenine time pulses on the endogenous levels of cytokinins, indole-3-acetic acid and abscisic acid in micropropagated explants of Actinidia deliciosa. Plant Physiol Biochem 41:149–155

    Article  Google Scholar 

  • Montalbán IA, Novák O, Rolčik J, Strnad M, Moncaleán P (2013) Endogenous cytokinin and auxin profiles during in vitro organogenesis from vegetative buds of Pinus radiata adult trees. Physiol Plant 148:214–231

    Article  PubMed  Google Scholar 

  • Motte H, Vereecke D, Geelen D, Werbrouck S (2014) The molecular path to in vitro shoot regeneration. Biotechnol Adv 32:107–121

    Article  CAS  PubMed  Google Scholar 

  • Motyka V, Vaňková R, Čapková V, Petrášek J, Kamínek M, Schmülling T (2003) Cytokinin-induced upregulation of cytokinin oxidase activity in tobacco includes changes in enzyme glycosylation and secretion. Physiol Plant 117:11–21

    Article  CAS  Google Scholar 

  • Msikita W, Skirvin RM (1989) In vitro regeneration from hypocotyl and seedling cotyledons of tronchuda (Brassica oleracea var. tronchuda Bailey). Plant Cell Tissue Organ Cult 19:159–165

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • National Institute for Health and Welfare, Nutrition Unit. Fineli (2011) Finnish Food Composition Database Release 14, Helsinki 2011. http://www.fineli.fi/food.php?foodid=329&lang=en. Accessed 8 June 2012

  • Neelakandan AK, Wang K (2012) Recent progress in the understanding of tissue culture-induced genome level changes in plants and potential applications. Plant Cell Rep 31:597–620

    Article  CAS  PubMed  Google Scholar 

  • Nikolić R, Mitić N, Miletić R, Nešković M (2006) Effects of cytokinins on in vitro seed germination and early seedling morphogenesis in Lotus corniculatus L. J Plant Growth Regul 25:187–194

    Article  Google Scholar 

  • Nordström A, Tarkowski P, Tarkowska D, Norbaek R, Åstot C, Dolezal K, Sandberg G (2004) Auxin regulation of cytokinin biosynthesis in Arabidopsis thaliana: a factor of potential importance for auxin-cytokinin-regulated development. Proc Natl Acad Sci USA 101:8039–8044

    Article  PubMed Central  PubMed  Google Scholar 

  • Okubo H, Wada K, Uemoto S (1991) In vitro morphogenetic response and distribution of endogenous plant hormones in hypocotyl segment of snapdragon (Antirrhinum majus L.). Plant Cell Rep 10:501–504

    Article  CAS  PubMed  Google Scholar 

  • Ono Y, Takahata Y, Kaizuma N (1994) Effect of genotype on shoot regeneration from cotyledonary explants of rapeseed (Brassica napus L.). Plant Cell Rep 14:13–17

    Article  CAS  PubMed  Google Scholar 

  • Otto F (1988) High resolution DNA—flow cytometry using DAPI, Protocol 1, Partec Arlesheim, Münster

  • Ovesná J, Ptacek L, Opatrny Z (1993) Factors influencing the regeneration capacity of oilseed rape and cauliflower in transformation experiments. Biol Plant 35:107–112

    Article  Google Scholar 

  • Parker CW, Entsch B, Letham DS (1986) Inhibitors of two enzymes which metabolize cytokinins. Phytochemistry 25:303–310

    Article  CAS  Google Scholar 

  • Paul S, Sikdar SR (2005) Regeneration of plants from root explant of two Indian cultivars of Brassica campestris L. through somatic embryogenesis. Curr Sci 89:1323–1326

    Google Scholar 

  • Pavlović S, Vinterhalter B, Mitić N, Adžić S, Pavlović N, Zdravković M, Vinterhalter D (2010) In vitro shoot regeneration from seedling explants in Brassica vegetables: red cabbage, broccoli, Savoy cabbage and cauliflower. Arch Biol Sci 62:337–345

    Article  Google Scholar 

  • Pellegrineschi A (1997) In vitro plant regeneration via organogenesis of cowpea [Vigna unguiculata (L.) Walp.]. Plant Cell Rep 17:89–95

    Article  CAS  Google Scholar 

  • Pernisová M, Klíma P, Horák J, Válková M, Malbeck J, Souček P, Reichman P, Hoyerová K, Dubová J, Friml J, Zažímalová E, Hejátko J (2009) Cytokinins modulate auxin-induced organogenesis in plants via regulation of the auxin efflux. Proc Natl Acad Sci USA 106:3609–3614

    Article  PubMed Central  PubMed  Google Scholar 

  • Raspor M, Motyka V, Žižková E, Dobrev PI, Trávníčková A, Zdravković-Korać S, Simonović A, Ninković S, Dragićević IC (2012) Cytokinin profiles of AtCKX2-overexpressing potato plants and the impact of altered cytokinin homeostasis on tuberization in vitro. J Plant Growth Regul 31:460–470

    Article  CAS  Google Scholar 

  • Ravanfar SA, Aziz MA, Kadir MA, Rashid AA, Haddadi F (2011) In vitro adventitious shoot regeneration and acclimatisation of Brassica oleracea subsp. italica cv Green Marvel. Afr J Biotechnol 10:5614–5619

    CAS  Google Scholar 

  • Ravanfar SA, Aziz MA, Rashid AA, Salim S (2014) In vitro adventitious shoot regeneration from cotyledon explant of Brassica oleracea subsp. italica and Brassica oleracea subsp. capitata using TDZ and NAA. Pak J Bot 46:329–335

    CAS  Google Scholar 

  • Schmitz RY, Skoog F, Playtis AJ, Leonard NJ (1972) Cytokinins: synthesis and biological activity of geometric and position isomers of zeatin. Plant Physiol 50:702–705

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Selman LW, Kulasegaram S (1966) Development of the stem tuber in kohlrabi. J Exp Bot 18:471–490

    Article  Google Scholar 

  • Sharma KK, Thorpe TA (1989) In vitro regeneration of shoot buds and plantlets from seedling root segments of Brassica napus L. Plant Cell Tissue Organ Cult 18:129–141

    Article  Google Scholar 

  • Souza BM, Kraus JE, Enders L, Mercier H (2003) Relationships between endogenous hormonal levels and axillary bud development of Ananas comosus nodal segments. Plant Physiol Biochem 41:733–739

    Article  CAS  Google Scholar 

  • Sparrow PAC, Townsend TM, Morgan CL, Dale PJ, Arthur AE, Irwin JA (2004) Genetic analysis of in vitro shoot regeneration from cotyledonary petioles of Brassica oleracea. Theor Appl Genet 108:1249–1255

    Article  CAS  PubMed  Google Scholar 

  • Spíchal L, Rakova NY, Riefler M, Mizuno T, Romanov GA, Strnad M, Schmülling T (2004) Two cytokinin receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, differ in their ligand specificity in a bacterial assay. Plant Cell Physiol 45:1299–1305

    Article  PubMed  Google Scholar 

  • Sretenović-Rajičić T, Ninković S, Vinterhalter B, Miljuš-Đukić J, Vinterhalter D (2004) Introduction of resistance to herbicide Basta® in Savoy cabagge. Biol Plant 48:431–436

    Article  Google Scholar 

  • Sretenović-Rajičić T, Ninković S, Miljuš-Đukić J, Vinterhalter B, Vinterhalter D (2006) Agrobacterium rhizogenes-mediated transformation of Brassica oleracea var. sabauda and B. oleracea var. capitata. Biol Plant 50:525–530

    Article  Google Scholar 

  • Stirk WA, Novák O, Václavíková K, Tarkowski P, Strnad M, van Staden J (2008) Spatial and temporal changes in endogenous cytokinins in developing pea roots. Planta 227:1279–1289

    Article  CAS  PubMed  Google Scholar 

  • Stirk WA, Václavíková K, Novák O, Gajdošová S, Kotland O, Motyka V, Strnad M, van Staden J (2012) Involvement of cis-zeatin, dihydrozeatin, and aromatic cytokinins in germination and seedling establishment of maize, oats, and lucerne. J Plant Growth Regul 31:392–405

    Article  CAS  Google Scholar 

  • Suri SS, Saini ARK, Ramawat KG (2005) High frequency regeneration and Agrobacterium tumefaciens-mediated transformation of broccoli (Brassica oleracea var. italica). Eur J Hortic Sci 70:71–78

    CAS  Google Scholar 

  • Takei K, Yamaya T, Sakakibara H (2004) Arabidopsis CYP735A1 and CYP735A2 encode cytokinin hydroxylases that catalyze the biosynthesis of trans-zeatin. J Biol Chem 279:41866–41872

    Article  CAS  PubMed  Google Scholar 

  • Teo W, Lakshmanan P, Kumar P, Goh CJ, Swarup S (1997) Direct shoot formation and plant regeneration from cotyledon explants of rapid-cycling Brassica rapa. In Vitro Cell Dev Biol Plant 33:288–292

    Article  Google Scholar 

  • Valdés AE, Ordas RJ, Fernandez B, Centeno ML (2001) Relationships between hormonal contents and the organogenic response in Pinus pinea cotyledons. Plant Physiol Biochem 39:377–384

    Article  Google Scholar 

  • Veach YK, Martin RC, Mok DWS, Malbeck J, Vaňková R, Mok MC (2003) O-Glucosylation of cis-zeatin in maize. Characterization of genes, enzymes, and endogenous cytokinins. Plant Physiol 131:1374–1380

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Verma SK, Yücesan BB, Şahin G, Gürel S, Gürel E (2011) Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turk J Bot 35:689–695

    Google Scholar 

  • Vinterhalter D, Sretenović-Rajičić T, Vinterhalter B, Ninković S (2007) Genetic transformation of Brassica oleracea vegetables. Transgenic Plant J 1:340–355

    Google Scholar 

  • Wang Y, Tong Y, Li Y, Zhang Y, Zhang J, Feng J, Feng H (2011) High frequency plant regeneration from microspore-derived embryos of ornamental kale (Brassica oleracea L. var. acephala). Sci Hortic 130:296–302

    Article  CAS  Google Scholar 

  • Wong KW, Loh CS (1988) In vitro regeneration of plantlets from root segments of Brassica alboglabra Bailey. Sci Hortic Amst 35:7–14

    Article  Google Scholar 

  • Yonekura-Sakakibara K, Kojima M, Yamaya T, Sakakibara H (2004) Molecular characterization of cytokinin-responsive histidine kinases in maize. Differential ligand preferences and response to cis-zeatin. Plant Physiol 134:1654–1661

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Zhang Y, Xu J, Han L, Wei W, Guan Z, Cong L, Chai T (2006) Efficient regeneration and Agrobacterium-mediated transformation of Brassica juncea. Plant Mol Biol Rep 24:255a–255i

    Article  Google Scholar 

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Acknowledgments

This work was supported by the Ministry of Education, Science and Technological Development of Serbia (No. 173015) and the Czech Science Foundation (P506/11/0774).

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    Authors and Affiliations

    1. Institute for Biological Research “Siniša Stanković”, University of Belgrade, Bulevar despota Stefana 142, 11060, Belgrade, Serbia

      Tatjana Ċosiċ, Martin Raspor, Jelena Savić, Aleksandar Cingel, Branka Vinterhalter, Dragan Vinterhalter & Slavica Ninković

    2. Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, 16502, Prague 6, Czech Republic

      Václav Motyka, Alena Trávníčková & Petre I. Dobrev

    3. Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia

      Borut Bohanec

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    1. Tatjana Ċosiċ
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    Correspondence to Tatjana Ċosiċ.

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    Tatjana Ċosiċ and Václav Motyka have contributed equally to this work.

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    Ċosiċ, T., Motyka, V., Raspor, M. et al. In vitro shoot organogenesis and comparative analysis of endogenous phytohormones in kohlrabi (Brassica oleracea var. gongylodes): effects of genotype, explant type and applied cytokinins. Plant Cell Tiss Organ Cult 121, 741–760 (2015). https://doi.org/10.1007/s11240-015-0742-2

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    • DOI: https://doi.org/10.1007/s11240-015-0742-2

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