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Cited by 74Plant tissue culture and regenerationFlowering Plant Growth and CultivationGenetic diversity and population structure

Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant

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Rohela, Gulab Khan, Jogam, Phanikanth, Bylla, Prasad, Reuben, Christopher·2019·BioMed Research International
DOI 10.1155/2019/3698742PMC PMC6487127
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Abstract

Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.

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  • In studies using a combination of two or three types of markers (SCoT, ISSR, RAPD), research revealed genetic fidelity of regenerated plantlets, even for those that had passed through the callus stage.

    “There are a number of studies in which the somaclonal variation analysis is carried out using a combination of two or three types of markers (Table 2), such as SCot, ISSR, and RAPD, and, in all cases, the research revealed the genetic fidelity of the regenerated plantlets (even for those that had passed through the callus stage) [73,74,75,76,77,78].”

    From Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants (2023) · cites this paper as [77]

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  1. [2]Chopra A. Doiphode V. V. Ayurvedic medicine: core concept, therapeutic principles, and current relevance Medical Clinics of North America 2002 86 1 75 89 10.1016/S0025-7125(03)00073-7 2-s2.0-0036135782 11795092 · DOI
  2. [4]Sahu A. N. Hemalatha S. Sairam K. Phyto-Pharmacological review of Mesua ferrea Linn InternationalJournal of Phytology 2014 5 1 6 14
  3. [5]Lingaraj P. Medicinal Plants of India: With special reference to Odisha International Journal of Advanced Research In Ideas in Education 2016 2 5 121 135
  4. [8]Wiart C. Medicinal Plants of Asia and the Pacific 2006 Boca Raton, Fla, USA CRC Press, Taylor and Francis
  5. [9]Rajkarnikar K. M. Sainju H. K. Bhatta B. D. In vitro culture of Rauwolfia serpentina L. benth. ex. Kurz Proceedings of Nepal Japan Joint Symposium 2002 232 234
  6. [10]VAKIL R. J. A clinical trial of Rauwolfia serpentina in essential hypertension British Heart Journal 1949 9 4 350 355 10.1136/hrt.11.4.350 2-s2.0-0001147137 PMC503638 18140347 · DOI
  7. [11]Rohela G. K. Bylla P. Pendli S. Rajender K. Dharavath B. Thammidala C. R. ISSR marker based DNA profiling studies in Rauwolfia species Annals of Plant Sciences 2018 7 5 p. 2289 10.21746/aps.2018.7.5.13 · DOI
  8. [13]Singh P. Singh A. Shukla A. K. Singh L. Pande V. Nailwal T. K. Somatic embryogenesis and in vitro regeneration of an endangered medicinal plant sarpgandha (rauvolfia serpentina l.) Life Science Journal 2009 6 2 57 62 2-s2.0-71549120733
  9. [14]Faisal M. Ahmad N. Anis M. Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron Plant Cell, Tissue and Organ Culture 2005 80 2 187 190 2-s2.0-13244262851 10.1007/s11240-004-0567-x · DOI
  10. [15]Rohela G. K. Prasad B. Ravi C. H. In vitro clonal propagation of Rauwolfia tetraphylla, a relative of indian snake root plant Research Journal of Biotechnology 2015 10 23 31
  11. [17]Panda S. K. Debajoyti D. Bichitra N. Phyto-pharmacognostical studies and quantitative determination of reserpine in different parts of Rauwolfia(Spp) of Eastern Odisha by UV Spectrocsopy Method Asian Journal of Plant Sciences and Research 2012 2 2 151 162
  12. [19]Brahmachari G. Mandal L. C. Gorai D. Mondal A. Sarkar S. Majhi S. A new labdane diterpene from Rauvolfia tetraphylla Linn. (Apocynaceae) Journal of Chemical Research 2011 35 12 678 680 2-s2.0-84855568933 10.3184/174751911X13220462651507 · DOI
  13. [20]Ghosh K. C. Banerjee N. Influence of plant growth regulators on in vitro micropropagation of Rauwolfia tetraphylla L. Phytomorphology: An International Journal of Plant Morphology 2003 53 1 11 19 2-s2.0-0038541735
  14. [22]Mitra G. C. Studies on the formation of viable and non-viable seeds in Rauwolfia serpentinabenth Indian Journal of Experimental Biology 1976 14 1 54 56
  15. [23]Harisharanraj R. Suresh K. Saravanababu S. Rapid Clonal Propagation Rauvolfia tetraphylla L Academic Journal of Plant Sciences 2009 2 3 195 198
  16. [24]Sarma D. Sarma S. Baruah A. Micropropagation and in vitro flowering of Rauvolfia tetraphylla; a potent source of anti-hypertension drugs Planta Medica 1999 65 3 277 278 2-s2.0-0033046324 10.1055/s-2006-960778 10232082 · DOI
  17. [25]Rohela G. K. Bylla P. Kota S. Abbagani S. Chithakari R. Reuben T. C. In vitro plantlet regeneration from leaf and stem calluses of Rauwolfia tetraphylla (R.canescens) and confirmation of genetic fidelity of plantlets using the ISSR PCR method Journal of Herbs, Spices & Medicinal Plants 2013 19 1 66 75 10.1080/10496475.2012.741056 · DOI
  18. [26]Rohela G. K. Jogam P. Shabnam A. A. Shukla P. Abbagani S. Ghosh M. K. In vitro regeneration and assessment of genetic fidelity of acclimated plantlets by using ISSR markers in PPR-1 (Morus sp.): An economically important plant Scientia Horticulturae 2018 241 313 321 2-s2.0-85049982861 10.1016/j.scienta.2018.07.012 · DOI
  19. [27]Murashige T. Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures Physiologia Plantarum 1962 15 3 473 497 10.1111/j.1399-3054.1962.tb08052.x 2-s2.0-84982358134 · DOI
  20. [29]Bylla P. Gulab K. R. Radha T. DNA profiling of commercial chilli pepper (Capsicum annuum L.) varieties using random amplified polymorphic DNA (RAPD) markers African Journal of Biotechnology 2013 12 30 4730 4735 10.5897/AJB2012.3017 · DOI
  21. [30]Rohela G. K. Bylla P. Korra R. Phytochemical screening and antimicrobial activity of leaf, stem, root and their callus extracts in Rauwolfia tetraphylla International Journal of Agriculture & Biology 2016 18 521 528
  22. [31]Anitha S. Ranjitha Kumari B. D. In vitro flowering in Rauvolfia tetraphylla L. Pakistan Journal of Biological Sciences 2006 9 3 422 424 2-s2.0-33646262602 10.3923/pjbs.2006.422.424 · DOI
  23. [32]Anitha S. Kumari D. R. In vitro callus culture in Rauvolfia tetraphylla L.: Indole alkaloid production Asian Journal of Plant Sciences 2012 12 1 1 4 10.3923/ajps.2013.28.33 2-s2.0-84879172581 · DOI
  24. [33]Anitha S. Ranjitha Kumari B. D. Reserplne accumulation in NaCI treated calli of Rauvolfia tetraphylla L ScienceAsia 2006 32 4 417 419 2-s2.0-33846358126 10.2306/scienceasia1513-1874.2006.32.417 · DOI
  25. [34]Huetteman C. A. Preece J. E. Thidiazuron: a potent cytokinin for woody plant tissue culture Plant Cell, Tissue and Organ Culture 1993 33 2 105 119 2-s2.0-0000814696 10.1007/BF01983223 · DOI
  26. [35]Pai S. R. Desai N. S. Ahmad. N. Faisal M. Effect of TDZ on various plant cultures Thidiazuron: From urea derivative to plant growth regulator 2018 Singapore Springer 10.1007/978-981-10-8004-3_25 · DOI
  27. [36]Ravi C. H. Avinash K. S. Gulab K. R. High frequency in vitro clonal propagation of Solanum surattense burm. F International Journal of Pharma and Biological Sciences 2012 3 147 151
  28. [37]Korra R. Bylla P. Rohela G. K. In vitro micro propagation and confirmation of genetic fidelity using RAPD marker in ethno medicinal plant Stachytarpheta jamaicensis L. Vahl International Journal of Advanced Research 2017 5 1494 1502
  29. [38]Azeez H. Ibrahim K. Pop R. Pamfil D. Hârţa M. Bobiș O. Changes induced by gamma ray irradiation on biomass production and secondary metabolites accumulation in Hypericum triquetrifolium Turra callus cultures Industrial Crops and Products 2017 108 183 189 2-s2.0-85021076284 10.1016/j.indcrop.2017.06.040 · DOI
  30. [39]Baskaran P. Van Staden J. Ultrastructure of somatic embryo development and plant propagation for Lachenalia montana South African Journal of Botany 2017 109 269 274 2-s2.0-85010993638 10.1016/j.sajb.2017.01.006 · DOI
  31. [40]Pai S. R. Upadhya V. Hedge H. V. Joshi R. K. Kholkute S. D. In vitro rapid multiplication and determination of triterpenoids in callus cultures of Achyranthes aspera Linn Indian Journal of Biochemistry and Biophysics 2018 17 1 151 159 2-s2.0-85051482870
  32. [42]Sujatha D. Ravi C. H. Raghuvardhanet L. In vitro plantlet regeneration and genetic transformation of sponge gourd (Luffa cylindrica L.) African Journal of Plant Sciences 2013 7 6 244 252
  33. [43]Mohd Din A. R. Iliyas Ahmad F. Wagiran A. Abd Samad A. Rahmat Z. Sarmidi M. R. Improvement of efficient in vitro regeneration potential of mature callus induced from Malaysian upland rice seed (Oryza sativa cv. Panderas) Saudi Journal of Biological Sciences 2016 23 1 S69 S77 10.1016/j.sjbs.2015.10.022 26858569 PMC4705290 · DOI
  34. [44]Khan Rohela G. Damera S. Bylla P. Korra R. Pendli S. Thammidala C. Somatic embryogenesis and indirect regeneration in Mirabilis jalapa Linn. Materials Today: Proceedings 2016 3 10 3882 3891 10.1016/j.matpr.2016.11.045 · DOI
  35. [45]Rohela G. K. Aftab A. S. Pawan S. In vitro clonal propagation of PPR-1, a superior temperate mulberry variety Indian Journal of Biotechnology 2018 17 619 625
  36. [46]Rohela G. K. Shabnam A. A. Shukla P. Kamili A. N. Ghosh M. K. Rapid one step protocol for the in vitro micro propagation of morus multicaulis var. goshoerami, an elite mulberry variety of temperate region Journal of Experimental Biology and Agricultural Sciences 2018 6 6 936 946 10.18006/2018.6(6).936.946 · DOI
  37. [47]Lakshmanan V. Venkataramareddy S. R. Neelwarne B. Molecular analysis of genetic stability in long-term micropropagated shoots of banana using RAPD and ISSR markers Electronic Journal of Biotechnology 2007 10 1 p. 8 2-s2.0-33846437506
  38. [48]Thakur J. Dwivedi M. D. Sourabh P. Uniyal P. L. Pandey A. K. Genetic homogeneity revealed using SCoT, ISSR and RAPD markers in micropropagated Pittosporum eriocarpum Royle- an endemic and endangered medicinal plant PLoS ONE 2016 11 7 e0159050 2-s2.0-84979265944 PMC4951010 27434060
  39. [49]Collard B. C. Y. Mackill D. J. Start Codon Targeted (SCoT) polymorphism: a simple, novel dna marker technique for generating gene-targeted markers in plants Plant Molecular Biology Reporter 2009 27 1 86 93 10.1007/s11105-008-0060-5 2-s2.0-70349984257 · DOI
  40. [50]Gorji A. M. Poczai P. Polgar Z. Taller J. Efficiency of arbitrarily amplified dominant markers (SCOT, ISSR and RAPD) for diagnostic fingerprinting in tetraploid potato American Journal of Potato Research 2011 88 3 226 237 10.1007/s12230-011-9187-2 2-s2.0-79956357854 · DOI
  41. [51]Cabo S. Ferreira L. Carvalho A. Martins-Lopes P. Martín A. Lima-Brito J. E. Potential of Start Codon Targeted (SCoT) markers for DNA fingerprinting of newly synthesized tritordeums and their respective parents Journal of Applied Genetics 2014 55 3 307 312 2-s2.0-84904641607 10.1007/s13353-014-0211-3 24733248 · DOI
  42. [52]Fang-Yong C. Ji-Hong L. Germplasm genetic diversity of Myrica rubra in Zhejiang Province studied using inter-primer binding site and start codon-targeted polymorphism markers Scientia Horticulturae 2014 170 169 175 2-s2.0-84896956131 10.1016/j.scienta.2014.03.010 · DOI
  43. [53]Rathore N. S. Rai M. K. Phulwaria M. Rathore N. Shekhawat N. S. Genetic stability in micropropagated Cleome gynandra revealed by SCoT analysis Acta Physiologiae Plantarum 2014 36 2 555 559 2-s2.0-84893959684 10.1007/s11738-013-1429-0 · DOI
  44. [54]Rahmani M. Pijut P. M. Shabanian N. Nasri M. Genetic fidelity assessment of in vitro-regenerated plants of Albizia julibrissin using SCoT and IRAP fingerprinting In Vitro Cellular & Developmental Biology - Plant 2015 51 4 407 419 10.1007/s11627-015-9692-y · DOI
  45. [56]Sharma M. M. Verma R. N. Singh A. Batra A. Assessment of clonal fidelity of Tylophora indica (Burm. f.) Merrill “in vitro” plantlets by ISSR molecular markers Springer Plus 2014 3 1 9 10.1186/2193-1801-3-400 25170431 PMC4147081 · DOI
  46. [57]Kumar S. Mangal M. Dhawan A. K. Singh N. Assessment of genetic fidelity of micropropagated plants of Simmondsia chinensis (Link) Schneider using RAPD and ISSR markers Acta Physiologiae Plantarum 2011 33 6 2541 2545 2-s2.0-80054104772 10.1007/s11738-011-0767-z · DOI
  47. [58]Faisal M. Alatar A. A. Ahmad N. Anis M. Hegazy A. K. An efficient and reproducible method for in vitro clonal multiplication of Rauvolfia tetraphylla L. and evaluation of genetic stability using DNA-based markers Applied Biochemistry and Biotechnology 2012 168 7 1739 1752 10.1007/s12010-012-9893-3 2-s2.0-84874499458 22968591 · DOI
  48. [59]Razaq M. Heikrujam M. Chetri S. K. Agrawal V. In vitro clonal propagation and genetic fidelity of the regenerants of Spilanthes calva DC. using RAPD and ISSR marker Physiology and Molecular Biology of Plants 2013 19 2 251 260 2-s2.0-84877926290 10.1007/s12298-012-0152-4 24431493 PMC3656186 · DOI
  49. [60]Raji M. R. Lotfi M. Tohidfar M. Somatic embryogenesis of muskmelon (Cucumis melo L.) and genetic stability assessment of regenerants using flow cytometry and ISSR markers Protoplasma 2018 255 3 873 883 2-s2.0-85038128212 10.1007/s00709-017-1194-9 29248969 · DOI
  50. [61]Ilczuk A. Jacygrad E. In vitro propagation and assessment of genetic stability of acclimated plantlets of Cornus alba L. using RAPD and ISSR markers In Vitro Cellular & Developmental Biology - Plant 2016 52 4 379 390 2-s2.0-84983805341 10.1007/s11627-016-9781-6 27746667 PMC5042974 · DOI
  51. [62]Ramakrishnan M. Ceasar S. A. Duraipandiyan V. Ignacimuthu S. Efficient plant regeneration from shoot apex explants of maize (Zea mays) and analysis of genetic fidelity of regenerated plants by ISSR markers Plant Cell, Tissue and Organ Culture 2014 119 1 183 196 2-s2.0-84910135169 10.1007/s11240-014-0525-1 · DOI
  52. [63]Alatar A. Faisal M. Encapsulation of Rauvolfia tetraphylla microshoots as artificial seeds and evaluation of genetic fidelity using RAPD and ISSR markers Journal of Medicinal Plants Research 2012 6 7 1367 1374 10.5897/JMPR11.1632 · DOI
  53. [64]Faisal M. Alatar A. A. Ahmad K. H. Molecular and biochemical characterization in rauvolfia tetraphylla plantlets grown from synthetic seeds following in vitro cold storage Applied Biochemistry and Biotechnology 2012 10.1007/s12010-012-9977-0 23212849 · DOI

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Authors
Rohela, G. K., Jogam, P., Bylla, P., & Reuben, C.
Affiliations
  • Government of India (IN)
  • Kakatiya University (IN)
  • Department of Botany, Kakatiya University, Warangal 506 009, Telangana, India
Corresponding author
Rohela, Gulab Khan
ORCID
Venue
BioMed Research International
Publisher
Wiley
Published
2019-04-11
Volume
2019
Pages
1-14
ISSN
  • 1110-7243 (ISSN-L)
  • 2314-6133
  • 2314-6141
DOI
10.1155/2019/3698742
PMC
PMC6487127
OpenAlex
W2939785398
Cited by
74
References
0
Citations by year
2026: 7 · 2025: 3 · 2024: 7 · 2023: 10 · 2022: 8 · 2021: 19 · 2020: 16 · 2019: 4
FWCI
3.89
Type
journal article
Language
en
Open access
Licence
Locations
  • BioMed Research International (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
University Grants Commission (UGC), New Delhi
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Flowering Plant Growth and Cultivation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic diversity and population structure
Keywords
indirect regeneration, micropropagation, genetic fidelity, acclimatization, callus induction, shoot regeneration, plantlet rooting, SCoT markers, ISSR markers, RAPD markers, medicinal plant conservation
MeSH terms
Acclimatization, Culture Media, Genetic Markers, Indoleacetic Acids, Kinetin, Phenylurea Compounds, Plant Growth Regulators, Plants, Medicinal, Rauwolfia, Regeneration, Thiadiazoles, Genetic Variation, 2,4-Dichlorophenoxyacetic Acid, DNA Primers, Codon, Initiator, Plant Leaves, Plant Shoots, Plant Stems, DNA, Plant, Microsatellite Repeats, Cell Culture Techniques, Random Amplified Polymorphic DNA Technique

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