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High-frequency in vitro shoot regeneration in Cucumis sativus by inhibition of endogenous auxin

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Abstract

Direct in vitro plantlet regeneration was studied in cucumber (Cucumis sativus), an important vegetable crop. The endogenous auxin concentration was quantified by HPLC in young cotyledon explants, and we observed that higher auxin content promotes callusing and inhibits direct shoot regeneration. Lowering the endogenous auxin concentration by the application of the auxin inhibitor 2,3,5-triiodobenzoic acid (TIBA), as confirmed by HPLC, increased the frequency of shoot regeneration. The auxin content was reduced by increasing the concentration of TIBA in cvs. Pant Kheera 1 and Pusa Uday. Regeneration efficiencies were dependent on explant age and cultivar. The 2-d-old explants gave the highest regeneration efficiencies, with Pusa Uday showing higher regeneration potential than PK-1. Shoot buds/small shoots transferred to Murashige and Skoog (MS) medium supplemented with 1 μM 6-benzylaminopurine grew vigorously and attained 1- to 2-cm length in 4 wk. These shoots rooted with 100% efficiency on MS basal medium or MS basal medium supplemented with 0.5 μM indole-3-acetic acid. The hardened plants showed normal growth and flowering under greenhouse conditions.

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References

  • Barnes WC (1961) Multiple disease resistant cucumbers. Proc Am Soc Hortic Sci 77:417–423

    Google Scholar 

  • Belaizi M, Paul H, Sangwan RS, Sangwan-Norreel BS (1991) Direct organogenesis from internodal segments of in vitro grown shoots of apple cv. Golden Delicious. Plant Cell Rep 9:471–474

    Article  CAS  PubMed  Google Scholar 

  • Bergervoet JHW, Van der Mark F, Custers JBM (1989) Organogenesis versus embryogenesis from long-term suspension cultures of cucumber (Cucumis sativus L.). Plant Cell Rep 8:116–119

    Article  CAS  PubMed  Google Scholar 

  • Bhau BS, Wakhlu AK (2001) Effect of genotype, explant type and growth regulators on organogenesis in Morus alba. Plant Cell Tissue Organ Cult 66:25–29

    Article  CAS  Google Scholar 

  • Bronsema FBF, van Oostveen WJF, van Lammeren AAM (2001) Influence of 2,4-D, TIBA and 3,5-D on the growth response of cultured maize embryos. Plant Cell Tissue Organ Cult 65:45–46

    Article  CAS  Google Scholar 

  • Burza W, Malepszy S, Rostek E (1995) An effect of simple and recurrent in vitro regeneration on cucumber inbred line under field cultivation. Hortic Sci 28:11–13

    Google Scholar 

  • Cade RM, Wehner TC, Blazich FA (1988) Embryogenesis from cotyledon-derived callus of Cucumis sativus L. Cucurbit Genet Coop Rep 11:3–4

    Google Scholar 

  • Cade RM, Wehner TC, Blazich FA (1990) Somatic embryos derived from cotyledons of cucumber. J Am Soc Hortic Sci 115:691–696

    CAS  Google Scholar 

  • Chee PP (1990) High frequency somatic embryogenesis and recovery of fertile cucumber plants. Sci Hortic 25:792–793

    Google Scholar 

  • Chee PP, Tricoli DM (1988) Somatic embryogenesis and plant regeneration from cell suspension cultures of Cucumis sativus L. Plant Cell Rep 7:274–277

    Article  CAS  PubMed  Google Scholar 

  • Christopher T, Rajam MV (1994) In vitro clonal propagation of Capsicum spp. Plant Cell Tissue Organ Cult 38:25–29

    Article  Google Scholar 

  • Deakin JR, Bohn GW, Whitaker TW (1971) Interspecific hybridization in Cucumis. Econ Bot 25:195–211

    Article  Google Scholar 

  • Detrez C, Sangwan RS, Sangwan-Norreel BS (1989) Phenotypic and karyotypic status of Beta vulgaris plants regenerated from direct organogenesis in petiole culture. Theor Appl Genet 77:462–468

    Article  CAS  PubMed  Google Scholar 

  • Esquinas-Alcazer JT, Gulick PJ (1983) Genetic resources of Cucurbitaceae. IBPGR, Rome

    Google Scholar 

  • Ezuka A, Komkada H (1974) Varietal difference in resistance of cucumber to downy mildew. Bull Tokai-Kinki Agric Exp Station 27:42–45

    Google Scholar 

  • Gambley RL, Dodd WA (1990) An in vitro technique for production of de novo of multiple shoots in cotyledon explants of cucumber (Cucumis sativus L.). Plant Cell Tissue Organ Cult 20:177–183

    CAS  Google Scholar 

  • Handley LW, Chambliss OL (1979) In vitro propagation of Cucumis sativus L. Sci Hortic 14:22–23

    CAS  Google Scholar 

  • Kim SG, Chang JR, Cha HC, Lee KW (1988) Callus growth and plant regeneration in diverse cultures of cucumber (Cucumis sativus L.). Plant Cell Tissue Organ Cult 12:67–74

    Article  Google Scholar 

  • Kim YJ, Oh YJ, Park WJ (2006) HPLC-based quantification of indole-3-acetic acid in the primary root tip of maize. J Nano Biotechnol 3:40–45

    Google Scholar 

  • Lall S, Mandegaran Z, Roberts AV (2005) Shoot multiplication in cultures of mature Alnus glutinosa. Plant Cell Tissue Organ Cult 83:347–350

    Article  CAS  Google Scholar 

  • Lomax TL, Muday GK, Rubery PH (1995) Auxin transport. In: Davies PJ (ed) Plant hormones—physiology, biochemistry and molecular biology. Kluwer, Dordrecht, pp 509–530

    Google Scholar 

  • Lou H, Kako S (1994) Somatic embryogenesis and plant regeneration in cucumber. Sci Hortic 29:906–909

    Google Scholar 

  • Malepszy S (1988) Cucumber (Cucumis sativus L.). In: Bajaj YPS (ed) Biotechnology in agriculture and forestry: crops II. Springer, Berlin, pp 276–293

    Google Scholar 

  • Malepszy S, Nadolska-Orczyk A (1983) In vitro culture of Cucumis sativus. I. Regeneration of plantlets from callus formed by leaf explants. Z Pflanzenphysiol 111:273–276

    Article  CAS  Google Scholar 

  • Malepszy S, Nadolska-Orczyk A (1989) In vitro culture of Cucumis sativus. VIII. Variation in the progeny of phenotypically not altered R1 plants. Plant Breed 102:66–72

    Article  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 

  • Rajasekaran K, Mulling MG, Nair Y (1983) Flower formation in vitro by hypocotyl explants of cucumber (Cucumis sativus L.). Ann Bot 52:417–420

    Google Scholar 

  • Selvaraj N, Vasudevan A, Manickavasagam M, Ganapathi A (2006) In vitro organogenesis and plant formation in cucumber. Biol Plant 50:123–126

    Article  CAS  Google Scholar 

  • Selvaraj N, Vasudevan A, Manickavasagam M, Kasthurirengan S, Ganapathi A (2007) High frequency shoot regeneration from cotyledon explants of cucumber via organogenesis. Sci Hortic 112:2–8

    Article  CAS  Google Scholar 

  • Shifriss OC, Myers CH, Chupp C (1942) Resistance to the mosaic virus in the cucumber. Phytopathology 32:773–784

    Google Scholar 

  • Singh SK, Syamal MM (2000) Anti-auxin enhance Rosa hybrida L. micropropagation. Biol Plant 43:279–281

    Article  CAS  Google Scholar 

  • Tabei Y (1997) Study on breeding of Cucurbitaceae using biotechnology. Bull Natl Inst Agrobiol Resour 11:1–107

    CAS  Google Scholar 

  • Tabei Y, Kanno T (1989) Effect of three kinds of auxins on the regeneration of cucumber (Cucumis sativus L.). Bull Natl Res Inst Veg Ornamental Plants Tea Ser 3:97–105

    Google Scholar 

  • Tetu T, Sangwan RS, Sangwan-Norreel BS (1987) Hormonal control of organogenesis and somatic embryogenesis in Beta vulgaris callus. J Exp Bot 38:506–517

    Article  CAS  Google Scholar 

  • Wehner TC, Locy RD (1981) Tissue culture propagation of field-grown cucumber selections. Cucurbit Genet Coop Rep 4:20–22

    Google Scholar 

  • Ziv M (1992) Micropropagation of Cucumis spp. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry: high-tech and micropropagation III. Springer, Berlin, pp 72–90

    Google Scholar 

  • Ziv M, Gadasi G (1986) Enhanced embryogenesis and plant regeneration from cucumber (Cucumis sativus L.) callus by activated charcoal in solid/liquid double-layer cultures. Plant Sci 47:115–122

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank Harshad Brahmbhatt for the assistance in HPLC analysis of the samples. The Analytical Discipline and Centralized Instrument Facilities are also duly acknowledged for providing the facilities. Financial assistance provided by a CSIR Empower project (OLP 0056) is gratefully acknowledged. P.S. Shukla is thankful to AcSIR for enrollment in the Ph.D. program.

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Authors and Affiliations

  1. Academy of Scientific and Innovative Research, CSIR-Central Salt and Marine Chemicals Research Institute, G. B. Marg, Bhavnagar, Gujarat, 364002, India

    Pushp Sheel Shukla, Bhavanath Jha & Pradeep K. Agarwal

  2. Analytical Discipline and Centralized Instrument Facility, CSIR-Central Salt and Marine Chemicals Research Institute, G. B. Marg, Bhavnagar, Gujarat, 364002, India

    Arun Kumar Das

  3. Discipline of Marine Biotechnology and Ecology, CSIR-Central Salt and Marine Chemicals Research Institute, G. B. Marg, Bhavnagar, Gujarat, 364002, India

    Bhavanath Jha

  4. Discipline of Wasteland Research, CSIR-Central Salt and Marine Chemicals Research Institute, G. B. Marg, Bhavnagar, Gujarat, 364002, India

    Pradeep K. Agarwal

Authors
  1. Pushp Sheel Shukla
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  2. Arun Kumar Das
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  3. Bhavanath Jha
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  4. Pradeep K. Agarwal
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Correspondence to Pradeep K. Agarwal.

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Editor: Susan Murch

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Shukla, P.S., Das, A.K., Jha, B. et al. High-frequency in vitro shoot regeneration in Cucumis sativus by inhibition of endogenous auxin. In Vitro Cell.Dev.Biol.-Plant 50, 729–737 (2014). https://doi.org/10.1007/s11627-014-9649-6

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