Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 133, Issue 1, pp 103–114 | Cite as

Pretreatment of seeds with thidiazuron delimits its negative effects on explants and promotes regeneration in chickpea (Cicer arietinum L.)

Original Article
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Abstract

Thidiazuron (TDZ) is an important cytokinin-like substance used in plant tissue culture. However, long exposure of plant tissues to TDZ results in abnormal regeneration. The present study reports an efficient regeneration protocol that avoids the TDZ overexposure to chickpea (Cicer arietinum L.) tissues. Two separate experiments were designed for seed pretreatment with TDZ, of which the first one involved short-term pretreatment with various concentrations of TDZ (15, 20 and 25 µM), whereas the second one lacked TDZ pretreatment. Axillary meristem explants prepared from TDZ-pretreated and TDZ-non-pretreated seeds were then analyzed in shoot induction medium (SIM) with or without 4 µM TDZ. Thus, four conditions were chosen to analyze the effect of TDZ: (i) TDZ pretreatment only without TDZ in SIM, (ii) TDZ pretreatment along with TDZ in SIM, (iii) no TDZ pretreatment but there was TDZ in SIM, and (iv) there was neither TDZ pretreatment nor TDZ supplied in SIM (control). The response in terms of percent regeneration (69%), number of shoots per explant (20.66 ± 0.5), minimum number of days taken for multiple shoot induction (7.3 ± 0.5) and effective rooting was the highest under the condition (i) with 20 μM TDZ pretreatment without subsequent TDZ supplementation into the SIM. In addition, shoot elongation medium (SEM-3) having 5 μM benzyladenine, 2 μM kinetin and 2 μM gibberellic acid showed the highest branching and maximum shoot length. In conclusion, we reported an easy and efficient regeneration method in chickpea using TDZ pretreatment only, which showed qualitative multiple shooting and effective rooting.

Keywords

Axillary meristem explants Chickpea Plant regeneration Shoot proliferation Thidiazuron 
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Notes

Acknowledgements

The authors are thankful to the Agriculture Research Station (ARS) Durgapura, Jaipur, India for the supply of different chickpea genotypes. We are thankful to Dr. Vinay K. Cheruvu, Department of Biostatistics, Kent State University, USA, for helping in statistical analysis.

Author contributions

PK, SY and SS conceived the idea. PK conducted the experiments with strategic experimental input from LSPT. PK and SY carried out data analysis with the input from LSPT. PK, SY and LSPT wrote the manuscript.

Compliance with ethical standards

Conflict of interest

No potential conflicts of interest were disclosed.

References

  1. Ahmad M, Fautrier AG, McNeil DL, Hill GD, Burritt DJ (1997) In vitro propagation of Lens species and their F1 interspecific hybrids. Plant Cell Tissue Organ Cult 47:169–176CrossRefGoogle Scholar
  2. Ahmad N, Siddique I, Anis M (2006) Improved plant regeneration in Capsicum annuum L. from nodal segments. Biol Plantarum 50:701–704Google Scholar
  3. Akasaka Y, Daimon H, Mii M (2000) Improved plant regeneration from cultured leaf segments in peanut (Arachis hypogaea L.) by limited exposure to thidiazuron. Plant Sci 156:169–175CrossRefPubMedGoogle Scholar
  4. Akram M, Aftab F (2016) Establishment of embryogenic cultures and efficient plant regeneration system from explants of forced softwood shoots of Teak (Tectona grandis L.). Hortic Plant J 2:293–300CrossRefGoogle Scholar
  5. Alatar AA (2015) Thidiazuron induced efficient in vitro multiplication and ex vitro conservation of Rauvolfia serpentina-a potent antihypertensive drug producing plant. Biotechnol Biotechnol Equip 29:489–497CrossRefGoogle Scholar
  6. Anwar F, Sharmila P, Saradhi PP (2010) No more recalcitrant: chickpea regeneration and genetic transformation. Afr J Biotech 9:782–797CrossRefGoogle Scholar
  7. Bakshi S, Roy NK, Sahoo L (2012) Seedling preconditioning in thidiazuron enhances axillary shoot proliferation and recovery of transgenic cowpea plants. Plant Cell Tissue Organ Cult 110:77–91CrossRefGoogle Scholar
  8. Barna KS, Wakhlu AK (1993) Somatic embryogenesis and plant regeneration from callus culture of chickpea (Cicer arietinum L.). Plant Cell Rep 12:521–524CrossRefPubMedGoogle Scholar
  9. Bermejo C, Gatti I, Cointry E (2016) In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik). Plant Cell Tissue Organ Cult 127:585–590CrossRefGoogle Scholar
  10. Bhatnagar-Mathur P, Vincent V, Devi MJ, Lavanya M, Vani G, Sharma KK (2009) Genetic engineering of chickpea (Cicer arietinum L.) with the P5CSF129A gene for osmoregulation with implications on drought tolerance. Mol Breed 23:591–606CrossRefGoogle Scholar
  11. Biesaga-Kościelniak J, Kościelniak J, Janeczko A (2010) The impact of zearalenone and thidiazuron on indirect plant regeneration of oilseed rape and wheat. Acta Physiol Plant 32:1047–1053CrossRefGoogle Scholar
  12. Bohmer P, Meyer B, Jacobsen HJ (1995) Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast derived pea callus. Plant Cell Rep 15:26–29CrossRefPubMedGoogle Scholar
  13. Chakraborti D, Sarkar A, Das S (2006) Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.). Plant Cell Tissue Organ Cult 86:117–123CrossRefGoogle Scholar
  14. Chakraborti D, Sarkar A, Mondal HA, Das S (2009) Tissue specific expression of potent insecticidal, Allium sativum leaf agglutinin (ASAL) in important pulse crop, chickpea (Cicer arietinum L.) to resist the phloem feeding Aphis craccivora. Transgenic Res 18:529–544CrossRefPubMedGoogle Scholar
  15. Chauhan H, Desai SA, Khurana P (2007) Comparative analysis of the differential regeneration response of various genotypes of Triticum aestivum, Triticum durum and Triticum dicoccum. Plant Cell Tissue Organ Cult 91:191–199CrossRefGoogle Scholar
  16. Chhabra G, Chaudhary D, Varma M, Sainger M, Jaiwal PK (2008) TDZ-induced direct shoot organogenesis and somatic embryogenesis on cotyledonary node explants of lentil (Lens culinaris Medik.). Physiol Mol Biol Plant 14:347–353CrossRefGoogle Scholar
  17. Croser JS, Ahmad JF, Clarke HJ, Siddique KHM (2003) Utilisation of wild Cicer in chickpea improvement-progress, constraints, and prospects. Aust J Agric Res 54:429–444CrossRefGoogle Scholar
  18. Cruz de Carvalho MH, Van LEB, Zuilyfodil Y, Phamthi AT, Tran Thanh Van K (2000) Efficient whole plant regeneration of common bean (Phaseolus vulgaris L.) using thin-cell-layer culture and silver nitrate. Plant Sci 159:223–232CrossRefPubMedGoogle Scholar
  19. Daffala HH, Abdellatef E, Elhadi EA, Khalafalla MM (2011) Effect of growth regulators on in vitro morphogenic response of Boscia senegalensis (Pers.) Lam. Poir. using mature zygotic embryos explants. Biotechnol Res Int 10:1–8CrossRefGoogle Scholar
  20. Debnath SC (2005) Strawberry sepal: another explant for thidiazuron-induced adventitious shoot regeneration. In Vitro Cell Dev Biol-Plant 41:671–676CrossRefGoogle Scholar
  21. Dewir YH, Chakrabarty D, Hahn EJ, Paek KY (2006) A simple method for mass propagation of Spathiphyllum cannifolium using an airlift bioreactor. In Vitro Cell Dev Biol-Plant 42:291–297CrossRefGoogle Scholar
  22. Din ARJM, Ahmad FI, Wagiran A, Samad AA, Rahmat Z, Sarmidi MR (2016) Improvement of efficient in vitro regeneration potential of mature callus induced from Malaysian upland rice seed (Oryza sativa cv. Panderas). Saudi J Biol Sci 23:S69–S77CrossRefGoogle Scholar
  23. Faisal M, Anis M (2006) Thidiazuron induced high frequency axillary shoot multiplication in Psoralea corylifolia. Biol Plant 50:437–444CrossRefGoogle Scholar
  24. Faisal M, Ahmad N, Anis M (2005) Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron. Plant Cell Tissue Organ Cult 80:187–190CrossRefGoogle Scholar
  25. Gatti I, Guindón F, Bermejo C, Espósito A, Cointry E (2016) In vitro tissue culture in breeding programs of leguminous pulses: use and current status. Plant Cell Tissue Organ Cult 127:543–559CrossRefGoogle Scholar
  26. Graner EM, Oberschelp GPJ, Brondani GE, Batagin-Piotto KD, Almeida CV, Almeida M (2013) TDZ pulsing evaluation on the in vitro morphogenesis of peach palm. Physiol Mol Biol Plant 19:283–288CrossRefGoogle Scholar
  27. Gubbuk H, Pekmezci M (2006) In vitro propagation of banana (Musa spp.) using thidiazuron and activated charcoal. Acta Agric Scand B Soil Plant Sci 56:65–69Google Scholar
  28. Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011) Thidiazuron: a multi-dimensional plant growth regulator. Afr J Biotech 10:8984–9000CrossRefGoogle Scholar
  29. Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant-tissue culture. Plant Cell Tissue Organ Cult 33:105–119CrossRefGoogle Scholar
  30. Jacob C, Carrasco B, Schwember AR (2016) Advances in breeding and biotechnology of legume crops. Plant Cell Tissue Organ Cult 127:561–584CrossRefGoogle Scholar
  31. Jahan AA, Anis M, Aref IM (2011) Preconditioning of axillary buds in thidiazuron-supplemented liquid media improves in vitro shoot multiplication in Nyctanthes arbor-tristis L. Appl Biochem Biotechnol 163:851–859CrossRefPubMedGoogle Scholar
  32. Jain M, Misra G, Patel RK, Priya P, Jhanwar S et al (2013) A draft genome sequence of the pulse crop chickpea (Cicer arietinum L.). Plant J 74:714–719CrossRefGoogle Scholar
  33. Jayanand B, Sudarsanam G, Sharma KK (2003) An efficient protocol for the regeneration of whole plants of chickpea (Cicer arietinum L.) by using axillary meristem explants derived from in vitro germinated seedlings. In Vitro Cell Dev Biol-Plant 39:171–179CrossRefGoogle Scholar
  34. Jones MPA, Yi Z, Murch SJ, Saxena PK (2007) Thidiazuron-induced regeneration of Echinacea purpurea L.: micropropagation in solid and liquid culture systems. Plant Cell Rep 26:13–19CrossRefPubMedGoogle Scholar
  35. Kaur H, Gupta AK, Kaur N, Sandhu JS (2012) High acid invertase activity for a prolonged period in developing seeds/podwall of wild chickpea is detrimental to seed filling. Indian J Exp Biol 50:735–743PubMedGoogle Scholar
  36. Ket NV, Hahn EJ, Park SY, Chakarbarty D, Paek KY (2004) Micropropagation of an endangered orchid Anoectochilus formosanus. Biol Plant 48:339–344CrossRefGoogle Scholar
  37. Malik KA, Saxena PK (1992) Thidiazuron induces high frequency shoot regeneration in intact seedlings of pea (Pisum sativum), chickpea (Cicer arietinum) and lentil (Lens culinaris). Aust J Plant Physiol 19:731–740CrossRefGoogle Scholar
  38. Malik S, Sharma S, Sharma M, Ahuja PS (2010) Direct shoot regeneration from intact leaves of Arnebia euchroma (Royle) Johnston using thidiazuron. Cell Biol Int 34:537–542CrossRefPubMedGoogle Scholar
  39. Mehta UJ, Barretto SM, Hazra S (2004) Effect of thidiazuron on germinating tamarind seedlings. In Vitro Cell Dev Biol-Plant 40:279–283CrossRefGoogle Scholar
  40. Mekala GK, Juturu VN, Mallikarjuna G, Kirti PB, Yadav SK (2016) Optimization of Agrobacterium-mediated genetic transformation of shoot tip explants of green gram (Vigna radiata (L.) Wilczek). Plant Cell Tissue Organ Cult 127:651–663CrossRefGoogle Scholar
  41. Mihaljevic S, Vrsek I (2009) In vitro shoot regeneration from immature seeds of Epimedium alpinum induced by thidiazuron and CPPU. Sci Hortic 120:406–410CrossRefGoogle Scholar
  42. Mukhtar S, Ahmad N, Khan MI, Anis M, Ibrahim MA (2012) Influencing micropropagation in Clitoria ternatea L. through the manipulation of TDZ levels and use of different explant types. Physiol Mol Biol Plant 18:381–386CrossRefGoogle Scholar
  43. Murthy BNS, Victor J, Singh RP, Fletcher RA, Saxena PK (1996) In vitro regeneration of chickpea (Cicer arietinum L.) stimulation of direct organogenesis and somatic embryogenesis by thidiazuron. Plant Growth Regul 19:233–240CrossRefGoogle Scholar
  44. Murthy BNS, Murch SJ, Saxena PK (1998) Thidiazuron: a potent regulator of in vitro plant morphogenesis. In Vitro Cell Dev Biol-Plant 34:267–275CrossRefGoogle Scholar
  45. Nguyen AH, Hodgson LM, Erskine W, Barker SJ (2016) An approach to overcoming regeneration recalcitrance in genetic transformation of lupins and other legumes. Plant Cell Tissue Organ Cult 127:623–635CrossRefGoogle Scholar
  46. Ochatt S, Conreux C, Smýkalová I, Smýkal P, Mikić A (2016) Developing biotechnology tools for ‘beautiful’ vavilovia (Vavilovia formosa), a legume crop wild relative with taxonomic and agronomic potential. Plant Cell Tissue Organ Cult 127:637–648CrossRefGoogle Scholar
  47. Ouyang Y, Chen Y, Lü J, Jaime A, da Silva T, Zhang X, Mab G (2016) Somatic embryogenesis and enhanced shoot organogenesis in Metabriggsia ovalifolia W.T. Wang. Sci Rep 6:24662CrossRefPubMedPubMedCentralGoogle Scholar
  48. Podwyszynska M, Novák O, Doležal K, Strnad M (2014) Endogenous cytokinin dynamics in micropropagated tulips during bulb formation process influenced by TDZ and iP pre-treatment. Plant Cell Tissue Organ Cult 119:331–346CrossRefGoogle Scholar
  49. Polowick PL, Baliski DS, Mahon JD (2004) Agrobacterium tumefaciens mediated transformation of chickpea (Cicer arietinum L.): gene integration, expression and inheritance. Plant Cell Rep 23:485–491CrossRefPubMedGoogle Scholar
  50. Radhakrishnan R, Ramachandran A, RanjithaKumari BD (2009) Rooting and shooting: dual function of thidiazuron in in vitro regeneration of soybean (Glycine max. L). Acta Physiol Plant 31:1213–1217CrossRefGoogle Scholar
  51. Rizvi SMH, Singh RP (2000) In-vitro plant regeneration from immature leaflet-derived callus cultures of Cicer arietinum L. via organogenesis. Plant Cell Biotechnol Mol Biol 1:109–114Google Scholar
  52. Shaik NM, Arha M, Nookaraju A, Gupta SK, Srivastava S, Yadav AK, Kulkarni PS, Abhilash OU, Vishwakarma RK, Singh S (2009) Improved method of in vitro regeneration in Leucaena leucocephala—a leguminous pulpwood tree species. Physiol Mol Biol Plant 15:311–318CrossRefGoogle Scholar
  53. Sharma R, Shahzad A (2008) Thidiazuron (TDZ) Induced regeneration from cotyledonary node explant of Abelmoschus moschatus Medik. L., (a valuable medicinal plant). World J Agr Sci 4:449–452Google Scholar
  54. Sharma VK, Hansch R, Mendel RR, Schulze J (2005) Influence of picloram and thidiazuron on high frequency plant regeneration in elite cultivars of wheat with long-term retention of morphogenecity using meristemic shoot segments. Plant Breed 124:242–246CrossRefGoogle Scholar
  55. Siddique I, Anis M (2007) In vitro shoot multiplication and plantlet regeneration from nodal explants of Cassia angustifolia (Vahl.): a medicinal plant. Acta Physiol Plant 29:233–238CrossRefGoogle Scholar
  56. Singh V, Chauhan NS, Singh M, Idris A, Madanala R, Pande V, Mohanty CS (2014) Establishment of an efficient and rapid method of multiple shoot regeneration and a comparative phenolics profile in in vitro and greenhouse-grown plants of Psophocarpus tetragonolobus (L.) DC. Plant Signal Behav 9:e970443CrossRefPubMedPubMedCentralGoogle Scholar
  57. Thomas TD (2003) Thidiazuron induced multiple shoot induction and plant regeneration from cotyledonary explants of mulberry. Biol Plant 46:529–533CrossRefGoogle Scholar
  58. Tulac S, Leljak-Levanic D, Krsnik-Rasol M, Jelaska S (2002) Effect of BAP, TDZ and CPPU on multiple shoot formation in pea (Pisum sativum L.) in culture in vitro. Acta Biol Cracovien 44:161–168Google Scholar
  59. Upreti J, Dhar U (1996) Micropropagation of Bauhinia vahlii Wight & Arnott—a leguminous liana. Plant Cell Rep 16:250–254PubMedGoogle Scholar
  60. Varshney RK, Close TJ, Singh NK, Hoisington DA, Cook DR (2009) Orphan legume crops enter the genomics era! Curr Opin Plant Biol 12:202–210CrossRefPubMedGoogle Scholar
  61. Zatloukal M, Gemrotov M, Dolezal K, Havlıcek L, Spıchal L, Strnad M (2008) Novel potent inhibitors of A. thaliana cytokinin oxidase/dehydrogenase. Bioorg Med Chem 16:9268–9275CrossRefPubMedGoogle Scholar
  62. Zayed EMM, Abdelbar OH (2015) Morphogenesis of immature female inflorescences of date palm in vitro. Ann Agric Sci 60:113–120Google Scholar
  63. Zhihui S, Tzitzikas M, Raemakers K, Zhengqiang M, Visser R (2009) Effect of TDZ on plant regeneration from mature seeds in pea (Pisum sativum). In Vitro Cell Dev Biol-Plant 45:776–782CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Life ScienceSinghania UniversityJhunjhunuIndia
  2. 2.Department of BiotechnologyHemvati Nandan Bahuguna Garhwal (Central) UniversitySrinagar GarhwalIndia
  3. 3.Institute of Research and DevelopmentDuy Tan UniversityDa NangVietnam
  4. 4.Signaling Pathway Research UnitRIKEN Center for Sustainable Resource ScienceYokohamaJapan

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