VEGETOS: An International Journal of Plant Research & Biotechnology
(Society For Plant Research)

Research Articles

A SOCIETY FOR PLANT RESEARCH PUBLICATION


Volume: 33, Issue: 3, September 2020


Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Page Visits: 47

Doi: 10.1007/s42535-020-00124-7
Doi Link: https://doi.org/10.1007/s42535-020-00124-7
First Page: 420
Last Page: 429
Published: 10 May, 2020

Shoot organogenesis and assessment of clonal fidelity of regenerated plants of Ocimum tenuiflorum L.: Queen of Herbs


Abstract:

An efficient and reliable shoot organogenesis protocol was developed from leaf explants taken from microshoots of Ocimum tenuiflorum L. Cultures were established using nodal explants taken from field grown mature plant on MS medium supplemented with 6-benzyladenine (BA; 2.5 μM). The effect of different cytokinins namely BA, kinetin (KIN) and thidiazuron (TDZ) in combination with 0.5 μM α-naphthalene acetic acid (NAA) was examined on shoot proliferation and elongation. Maximum number of shoots per culture vessel (38) was recorded on MS medium supplemented with 5.0 μM BA, whereas the maximum number of elongated shoots per culture vessel (15) along with maximum shoot length (4.80 cm) was observed on medium supplemented with 2.5 μM BA. Shoot organogenesis was attempted from the segments of fully expanded leafs and 34.45% explants regenerated shoots on MS medium supplemented with 5.0 µM each of BA and NAA. Incorporation of cefotaxime (300 mg/l) was found to be beneficial for shoot organogenesis and further increased shoot regeneration frequency to 36% explants. On the other hand, addition of carbenicillin into the medium was found to inhibit shoot organogenesis. For root induction in microshoots, among various auxins used, IBA was found to be the best. A pulse treatment of 50.0 μM IBA for 36 h improved rooting of microshoots and a maximum of 89% microshoots rooted. Rooted microshoots were successfully acclimatized under polyhouse conditions with a survival rate of 80%. Later acclimatized plants were established in their natural conditions. Random Amplified polymorphic DNA and Inter Simple Sequence Repeat markers established a clonal fidelity of micropropagated plants with that of mother plant.

Vegetos

Keywords:


ISSR, RAPD, Tulsi, Medicinal plants, Pulse treatment, Antibiotic


References:


  1. Aggarwal D, Barna KS (2004) Tissue culture propagation of elite plants of Aloe vera Mill. J Plant Biochem Biotech 13:77–79

  2. Aggarwal D, Kumar A, Reddy MS (2010) Shoot organogenesis from elite plants of Eucalyptus tereticornis. Plant Cell Tiss Org Cult 102:45–52

  3. Aggarwal D, Kumar A, Reddy MS (2011) Agrobacterium tumefaciens mediated genetic transformation of selected elite clones of Eucalyptus tereticornis. Acta Physiol Plant 33:1603–1611

  4. Aggarwal D, Kumar A, Sharma J, Reddy MS (2012) Factors effecting micropropagation and acclimatization of an elite clone of Eucalyptus tereticornis. In Vitro Cell Dev Biol Plant 48:521–529

  5. Asghari F, Hossieni B, Hassani A, Shirzad H (2012) Effect of explants source and different hormonal combinations on direct regeneration of Ocimum basilicum. Aust J Agr Engg 3:12–17

  6. Bansal M, Kumar A, Reddy MS (2014) Diversity amongst wild accessions of Bacopa monnieri (L.) Wettst. and their morphogenetic potential. Acta Physiol Plant 36:1177–1186

  7. Bast F, Rani P, Meena D (2014) Chloroplast DNA phylogeography of holy basil (Ocimum tenuiflorum) in Indian subcontinent. Sci World J 70(3):277–285

  8. Bhau BS, Wakhlu AK (2001) Effect of some antibiotics on the in vitro morphogenetic response from callus cultures of Coryphantha elephantidens. Biol Plant 44:19–24

  9. Bodeker G, Ong CK, Grundy C, Burford G, Shein K (2005) WHO global atlas of traditional, complementary and alternative medicine: text volume. WHO Centre Health Development xiv:216

  10. Bueren ETL, Jones SS, Tamm L, Murphy KM, Myers JR, Leifert C, Messmer MM (2011) The need to breed crop varieties suitable for organic farming, using wheat, tomato and broccoli as examples: a review. NJAS Wageningen J Life Sci 58:193–205

  11. Cortizo M, Cuesta C, Ml C, Rodríguez A, Fernández B, Ordás R (2009) Benzyladenine metabolism and temporal competence of Pinus pinea cotyledons to form buds in vitro. J Plant Physiol 166(10):1069–1076

  12. Debergh PC, Read PE (1991) Micropropagation. In: Debergh PC, Zimmerman RH (eds) Micropropagation technology and application. Kluwer Academic Publishers, Dordrecht, pp 1–14

  13. Dode B, Bobrowski LV, Braga BJE, Seixas KF, Schuch WM (2003) In vitro propagation of Ocimum basilicum L. Acta Sci Biol Sci 25:435–437

  14. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:39–40

  15. Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42

  16. Ekor M (2013) The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol 4:177

  17. Faisal M, Alatar AA, El-Sheikh MA, Abdal-Salam EM, Qahtan AA (2018) Thidiazuron induced in vitro morphogenesis for sustainable supply of genetically true quality plantlets of Brahmi. Ind Crop Prod 118:173–179

  18. Gaddaguti V, Talluri VR, Rao AP (2015) In vitro propagation of Ocimum tenuiflorum var. CIM-AYU from nodal explants. J Appl Bio Sci Res 6:1–7

  19. Gang DR, Wang J, Dudareva N, Nam KH, Simon JE, Lewinsohn E, Pichersky E (2001) An investigation of the storage and biosynthesis of phenylpropenes in sweet basil. Plant Physiol 125:539–555

  20. Gantait S, Sinniah UR, Mandal N, Das PK (2012) Direct induction of protocorm-like bodies from shoot tips, plantlet formation and clonal fidelity analysis in Anthurium andreanum cv. CanCan Plant Growth Regul 67:257–270

  21. Goel A, Kaur A, Kumar A (2018) Biochemical and histological changes during in vitro rooting of microcuttings of Bacopa monnieri (L.) Wettst. Acta Physiol Plant 40:64

  22. Ho CK, Chang SH, Tsay JY, Tsai CJ, Chiang VL, Chen ZZ (1998) Agrobacterium tumefaciens mediated transformation of Eucalyptus camaldulensis and production of transgenic plants. Plant Cell Rep 17:675–680

  23. Husain MK, Anis M (2006) Rapid in vitro propagation of Eclipta alba (L.) Hassk. through high frequency axillary shoot proliferation. Acta Physiol Plant 28:325–330

  24. Ilczuk A, Jacygrad E (2016) In vitro propagation and assessment of genetic stability of acclimated plantlets of Cornus alba L. using RAPD and ISSR markers. Vitro Cell Dev Biol Plant 52:379–390

  25. Jawla S, Gupta AK, Singla R, Gupta V (2009) General awareness and relative popularity of allopathic, ayurvedic and homeopathic systems. J Chem Pharm Res 1(1):105–112

  26. Joseph B, Nair MV (2013) Ethanopharmacological and phytochemical aspects of Ocimum sanctum L.—the Elixir of Life. Br J Pharm Res 3:273–292

  27. Kaur A, Reddy MS, Kumar A (2017) Efficient, one step and cultivar independent shoot organogenesis of potato. Physiol Mol Biol Plants 23:461–469

  28. Khan S, Fahim N, Singh P, Rahman LU (2015) Agrobacterium tumefaciens mediated genetic transformation of Ocimum gratissimum: a medicinally important crop. Ind Crop Prod 71:138–146

  29. Kumar A (1996) Studies on in vitro propagation, biochemistry and field evaluation of two economically important plants: Rosa damascene Mill. and Gladiolus spp., Ph.D. Thesis. Kumaun University, Nainital

  30. Kumar A, Sood A, Palni LMS, Palni UT, Gupta AK (2001) Micropropagation of Rosa damascene from selected bushes. J Hortic Sci Biotechnol 76:30–34

  31. Kumar A, Aggarwal D, Gupta P, Reddy MS (2010) Factors effecting in vitro propagation and field establishment of Chlorophytum borivillianum. Biol Plant 54(4):601–606

  32. Li X, Krasnyanski SF, Korban SS (2002) Optimization of the uidA gene transfer into somatic embryos of rose via Agrobacterium tumefaciens. Plant Physiol Biochem 40:453–459

  33. Ling HQ, Kriseleit D, Ganal MW (1998) Effect of ticarcillin/potassium clavulanate on callus growth and shoot regeneration in Agrobacterium-mediated transformation of tomato (Lycopersicon esculentum Mill.). Plant Cell Rep 17:843–847

  34. Morales MR, Simon JE (1997) ‘Sweet Dani’: a new culinary and ornamental lemon basil. HortScience 32:148–149

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

  36. Naing AH, Ai TN, Jeon SM, Park KI, Lim SH, Lim KB, Kim CK (2016) Novel antibiotics enhance regeneration and genetic transformation with RsMYB1 gene of recalcitrant chrysanthemum cv. Shinma Plant Biosyst 151:98–107

  37. Oliveira MLP, Costa MGC, Silva CV, Otoni WC (2010) Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars. Pesq Agropec Bras Brasília 45(7):654–660

  38. Overvoorde P, Fukaki H, Beeckmans T (2010) Auxin control of root development. Cold Spring Harb Perspect Biol 2:a001537

  39. Phippen WB, Simon JE (1998) Anthocyanins in basil. J Agric Food Chem 46:1734–1738

  40. Rai V, Vajpayee P, Singh SN, Mehrotra S (2004) Effect of chromium accumulation on photosynthetic pigments, oxidative stress defense system, nitrate reduction, proline level and eugenol content of Ocimum tenuiflorum L. Plant Sci 167:1159–1169

  41. Saini S, Sharma I, Kaur N, Pati PK (2013) Auxin: a master regulator in plant root development. Plant Cell Rep 32(6):741–757

  42. Saxena C, Samantaray S, Rout G, Das P (2000) Effect of Auxins on in vitro rooting of Plumbago zeylanica: peroxidase activity as a marker for root induction. Biol Plant 43:121–124

  43. Siddique I, Anis M (2008) An improved plant regeneration system and ex vitro acclimatization of Ocimum basilicum L. Acta Physiol Plant 30:493–499

  44. Simon JE, Quin J, Murray RG (1990) Basil: a source of essential oils. In: Janik J, Simon JE (eds) Advances in new crops. Timber Press, Portland, pp 484–489

  45. Supe UJ (2007) In vitro regeneration of Aloe barbadensis. Biotech 6(4):601–603

  46. Tambarussi EV, Rogalski M, Nogueira FTS, Brondani GE, De Martin VF, Carrer H (2015) Influence of antibiotics on indirect organogenesis of Teak. Ann For Res 58(1):177–183

  47. Veira RF, Grayer RJ, Paton A, Simon JE (2001) Genetic diversity of Ocimum gratissimum L. based on volatile oil constituents, flavonoids and RAPD markers. Biochem Syst Ecol 29(3):287–304

  48. Wakchaure R, Ganguly S, Praveen PK (2017) Ocimum sanctum (Tulsi), the Queen of Herbs: a review. In: Abbas AM, Sharma YK, Abid M, Khan MMAA (eds) Biochemistry and therapeutic uses of medicinal plants. Discovery Publishing House Pvt. Ltd., New Delhi, pp 166–173

  49. Yu Y, Wei ZM (2008) Influences of cefotaxime and carbenicillin on plant regeneration from wheat mature embryos. Biol Plant 52:553–556


  50.  


Acknowledgements :



The authors are thankful to Maharishi Markandeshwar (Deemed to be University), Mullana-Ambala and TIFAC-CORE, Thapar Institute of Engineering and Technology, Patiala for providing necessary facilities required to carry out the work.


Author Information:



Diwakar Aggarwal
Department of Biotechnology, TIFAC-Center of Relevance & Excellence in Agro & Industrial Biotechnology (TIFAC-CORE), Thapar Institute of Engineering and Technology, Patiala, India
diwakaraggarwal@yahoo.co.in




Pdf Download
Share: