Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2011 | 33 | 1 |
Tytuł artykułu

RAPD based genetic stability analysis among micropropagated, synthetic seed derived and hardened plants of Bacopa monnieri (L.): a threatened Indian medicinal herb

Treść / Zawartość
Warianty tytułu
Języki publikacji
Bacopa monnieri (L.), a highly endangered miracle medicinal herb with global interest, is one of the popular ancient Indian ayurvedic plants. With ever increasing demand for Bacopa based formulations in pharmaceutical industries, there is a need to preserve the stocks of the plant through biotechnological approaches. Randomly amplified polymorphic DNA (RAPD) fingerprinting approach was applied to analyze the genetic stability of 19 different B.monnieri plants randomly selected after micropropagation, regrowth from alginate encapsulated uninodal cuttings (before and after storage at 4°C) and hardening with the mother plant (wild type). 16 arbitrary decamer primers amplified a total of 334 reproducible distinct DNA fragments ranging from 180 to 1,500 bp, of which 262 (78.4%) were monomorphic and the rest (21.5%) were polymorphic with an average of 20.8 bands per primer. The extent of polymorphism was low to moderate. Primers OPAK 14, OPM 15 and OPD 13 generated 69, 46 and 42% polymorphic patterns. Primers OPA 04, OPU 13 and OPD 08 generated 100% monomorphic pattern. Similarity matrix based on Jaccard’s coefficient revealed that pair wise values between the wild type and its analyzed plants ranged from 0.00 to 0.92 and among the micropropagated, synthetic seed derived and hardened plants, the range of genetic distance is from 0.67 to 0.92. Unweighted pair group method with arithmetic averages cluster analysis resulted in one loose group of the wild type with three subgroups. The present study paves the way for the identification and maintenance of genetically uniform B. monnieri plants micropropagated in the lab, plants regrown from synthetic seeds and hardened in the field.
Słowa kluczowe
Opis fizyczny
  • Alagappa University, Karaikudi, India
  • Alagappa University, Karaikudi, India
  • Alagappa University, Karaikudi, India
  • Alagappa University, Karaikudi, India
  • Anand A (2003) Studies on genetic stability of micropropagated plants and reintroduction in an endemic and endangered taxon: Syzgium travancorium Gamble. J Plant Biotechnol 5:201–207
  • Angel F, Barney BE, Tohme J, Roca WM (1996) Stability of Cassava plants at the DNA level after retrieval from 10 years of in vitro storage. Euphytica 90:307–313
  • Bindiya K, Kanwar K (2003) Random amplified polymorphic DNA (RAPDs) markers for genetic analysis in micropropagated plants of Robinia pseudoacacia. Euphytica 132:41–47
  • Brauner S, Crawford DJ, Stuessy TF (1992) Ribosomal DNA and RAPD variation in the rare plant family Lactoridaceae. Am J Bot 79:1436–1439
  • Ceasar SA, Maxwell SL, Prasad KB, Karthigan M, Ignacimuthu S (2009) Highly efficient shoot regeneration of Bacopa monnieri (L.) using a two-stage culture procedure and assessment of genetic integrity of micropropagated plants by RAPD. Acta physiol Plant. doi:10.1007/s11738-009-0419-8
  • Chandrika M, Thoyajaksha Ravishankar Rai V, Ramachandra Kini M (2008) Assessment of genetic stability of in vitro grown Dictyospermum ovalifolium. Biol Plant 52:735–739
  • Chowdhuri DK, Parmar D, Kakkar P, Shukla R, Seth PK, Srimal RC (2002) Antistress effects of bacosides of Bacopa monnieri: modulation of Hsp70 expression, superoxide dismutase and cytochrome p450 activity in rat brain. Phytother Res 16:639–645
  • Dahanukar SA, Thatte UM (1997) Current status of ayurveda. Phytomedicine 4:211–215
  • Darokar MP, Suman PSK, Shasany AK, Kumar Sushil (2001) Low levels of genetic diversity detected by RAPD analysis in geographically distinct accessions of Bacopa monnieri. Genet Resour Crop Evol 48:555–558
  • Devarumath R, Nandy S, Rani V, Marimuthu S, Muraleedharan N, Raina S (2002) RAPD, ISSR and AFLP fingerprint as useful markers to evaluate genetic integrity of micropropagated plants of three diploid and triploid elite tea clones representing Camellia sinensis (China type) and C. assamica ssp. assamica (Assam-India type). Plant Cell Rep 21:166–173
  • Elangovan V, Govindasamy S, Ramamoorthy N, Balasubramaniam K (1995) In vitro studies of the anti cancer activity of Bacopa monneri. Fitoterapia 66:211–215
  • Ganesh Ram S, Parthiban KT, Senthil Kumar R, Thiruvengadam V, Paramathma M (2008) Genetic diversity among Jatropha species as revealed by RAPD markers. Genet Resour Crop Evol 55:803–809
  • Gomes M, Kulothungan Mehetre SS, Eapen S (2004) Analysis of genetic relatedness in Gossypium species using RAPD. Ind J Biotechnol 3:41–46
  • Govindarajan R, Vijayakumar M, Pushpangadan P (2005) Antioxidant approach to disease management and the role of rasayana herbs of ayurveda. J Ethnopharmacol 19(2):165–178
  • Guo WL, Gong L, Ding ZF, Li YD, Li FX, Zhao SP, Liu B (2006) Genomic instability in phenotypically normal regenerants of medicinal plant Codonopsis lanceolata as revealed by ISSR and RAPD markers. Plant Cell Rep 25:896–906
  • Gupta R, Modgil M, Chakrabarthi SK (2009) Assessment of genetic fidelity of micropropagated apple rootstock plants, EMLA111, using RAPD markers. Indian J Exp Biol 47:925–928
  • Isabel N, Tremblay L, Michaud M, Tremblay FM, Bousquet J (1996) Occurrence of somoclonal among somatic embryo-derived white spruces (Picea glauca, Pinaceae). Am J Bot 83:1121–1130
  • Jaccard P (1908) Nouvelles recherches sur la distribution florale. Bull Soc Vaud Sci Nat 44:223–270
  • Jayanthi M, Mandal PK (2001) Plant regeneration through somatic embryogenesis and RAPD analysis of regenerated plants in Tylophora indica (Burm.F.Merrrii). In vitro Cell Dev Biol Plant 37:576–580
  • Kapoor LD (1990) CRC Handbook of ayurvedic medicinal plants. CRC Press, Boca Raton
  • Lakhanpaul S, Chada S, Bhat KV (2000) Random amplified polymorphic DNA (RAPD) analysis in Indian mung bean (Vigna radiata L.) Wilczek cultivars. Genetica 109:227–234
  • Lattoo SK, Bamotra S, Sapru DR, Khan S, Dhar AK (2006) Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceumm Baker—an endangered medicinal herb. Plant Cell Rep 25:499–506
  • Mehetre SS, Gomes M, Eapen S (2004) RAPD analysis of hybrid nature of the offspring of Gossypium hirsutum × G. raimondii. Curr Sci 87:25–28
  • Mohapatra HP, Rath SP (2005) In vitro studies of Bacopa monnieri— an important medicinal plant with reference to its biochemical variations. Indian J Exp Biol 43:373–376
  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497
  • National Medicinal Plants Board (2004) 32 Prioritized Medicinal Plants, National Informatics Centre, Ministry of Health and Family Welfare, Department of Ayush, Government of India Accessed 08 December 2009
  • National Medicinal Plants Board (2008) Agrotechniques of selected medicinal plants. National Medicinal Plants Board, Department of Ayush, Ministry of Health & Family Welfare, Govt of India. TERI Press, New Delhi, pp 33–38
  • National Medicinal Plants Board (NMPB) and Technology Information Forecasting and Assessment Council (TIFAC), Department of Science and Technology, Government of India (2007). Accessed 08 December 2009
  • Pei YL, Zou YP, Yin Z, Wang XQ, Zhang ZX, Hong DY (1995) Preliminary report of RAPD analysis in Paeonia suffruticosa subsp. Spontanea and P. rockii. Acta Phys Sin 33:350–356
  • Philip S, Banerjee NS, Das MR (1999) Genetic variation and micropropagation in three varieties of Piper longum L. Curr Sci 78:169–173
  • Potter R, Jones MGK (1991) An assessment of genetic stability of potato in vitro by molecular and phenotypic analysis. Plant Sci 76:239–248
  • Prasad R, Bagde US, Puspangadan P, Varma A (2008) Bacopa monniera L. pharmacological aspects and case studies involving Piriformospora indica. Int J Integr Biol 3:100–110
  • Praveen N, Naik PM, Manohar SH, Nayeem A, Murthy HN (2009) In vitro regeneration of brahmi shoots using semisolid and liquid cultures and quantitative analysis of bacoside A. Acta Physiol plant 31:723–728. doi:10.1007/s11738-009-0284-5
  • Pravina K, Ravindra KR, Goudar KS, Vinod DR, Joshua AJ, Wasim P, Venkateswarlu K, Saxena VS, Amit A (2007) Safety evaluation of BacoMind TM in healthy volunteers: a phase 1 study. Phytomedicine 12:301–308
  • Rai D, Bhatia G, Palit G, Pal R, Singh S, Singh HK (2003) Adaptogenic effect of Bacopa monniera (Brahmi). Pharmacol Biochem Behav 75:823–830
  • Rajani M (2008) Bacopa monnieri, a nootropic drug. In: Ramawat KG, Merillon JM (eds) Bioactive molecules and medicinal plants. Springer, Berlin, pp 175–195
  • Ramesh M, Saravanakumar RM, Karutha Pandian S (2006) Benzyl amino purine and adenine sulphate induced multiple shoot and root induction from nodal explants of Brahmi, Bacopa monnieri (Linn.) Penn. Nat Prod Radiance 5:44–51
  • Ramesh M, Marx R, Mathan G, Karutha Pandian S (2009) Effect of bavistin on in vitro plant conversion from encapsulated uninodal microcuttings of micropropagated Bacopa monnieri (L.)—an ayurvedic herb. J Environ Biol 30:441–444
  • Ramser J, Peralta CL, Wetzel R, Weising K, Kahl G (1996) Genomic variation and relationships in aerial yam detected by RAPD. Genome 39:17–25
  • Rani V, Parida A, Raina SN (1995) Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant Cell Rep 14:459–462
  • Rastogi S, Pal R, Kulshreshtha DK (1994) Bacoside A3—a triterpenoid saponin from Bacopa monniera. Photochemistry 36:133–137
  • Rogers SO, Bendich AJ (1988) Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer Academic Publications, Dordrecht, pp 1–10
  • Rohlf FJ (2002) NTSYS-pc: numerical taxonomy system ver.2.1. Exeter Publishing Ltd., Setauket
  • Rout GR, Das G (2002) An assessment of genetic integrity of micropropagated plants of Plumbago zeylanica by RAPD markers. Biol plant 45:27–32
  • Rout GR, Das P, Goel S, Raina SN (1998) Determination of genetic stability of micropropagated plants of ginger using random amplified polymorphic DNA (RAPD) markers. Bot Bull Acad Sin 39:23–27
  • Salvi ND, George L, Eapen S (2002) Micropropagation and field evaluation of miccropropagated plants of turmeric. Plant Cell Tissue Organ Cult 68:143–151
  • Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
  • Shoyama Y, Zhu XX, Nakai R, Shiraishi S, Kohda H (1997) Micropropagation of Panax notoginseng by somatic embryogenesis and RAPD analysis of regenerated plantlets. Plant Cell Rep 16:450–453
  • Shrivastava N, Rajani M (1999) Multiple shoot regeneration and tissue culture studies on Bacopa monnieri (L.) Pennell. Plant Cell Rep 18:919–923
  • Singh RH, Singh L (1980) Studies on the anti anxiety effects of the medhya rasayana drug Bacopa monniera Wettst.). J Res Ayurveda and Siddha 1:33–148
  • Smith MK (1998) A review of factors influencing the genetic stability of micropropagated banana fruits. Fruits 43:219–223
  • Sneath PH, Sokal RR (1973) Numerical taxonomy. W.H. Freeman & Company. San Francisco
  • Su YJ, Wang T, Huang C (1999) RAPD analysis of different population of Dacydium pierrei. Acta Bot Sin 40:169–175
  • Tejavathi DH, Shailaja KS (1999) Regeneration of plants from the cultures of Bacopa monnieri (L.) Pennell. Phytomorphology 49:447–452
  • Tiwari V, Singh BD, Tewari KN (1998) Shoot regeneration and somatic embryogenesis from different explants of Brahmi (Bacopa monniera (L.) Wettst). Plant Cell Rep 17:538–543
  • Tiwari V, Tewari KN, Singh BD (2000) Suitability of liquid cultures or in vitro multiplication of Bacopa monnira (L.) Wettst. Phytomorphology 50:337–342
  • Tiwari V, Tewari KN, Singh BD (2001) Comparative studies of cytokinins on in vitro propagation of Bacopa monniera. Plant Cell Tissue Organ Cult 66:9–16
  • Tiwari V, Tewari KN, Singh BD (2006) Shoot bud regeneration from different explants of Bacopa monniera (L.) trimethoprim and bavistin. Plant Cell Rep 25:629–635
  • Tripathi YB, Chaurasia S, Tripathi E, Upadhyaya A, Dubey GP (1996) Bacopa monniera Linn. as an antioxidant: mechanism of action. Indian J Exp Biol 34:523–526
  • Turner S, Krauss SL, Bunn E, Senaratna T, Dixon K, Tan B, Touchell D (2001) Genetic fidelity and viability of Anigozanthos viridis following tissue culture, cold storage and cryo-preservation. Plant Sci 161:1099–1106
  • Wang XQ, Zou YP, Zhang DM, Hong DY (1996) RAPD analysis for genetic polymorphism in Cathaya argyrophylla. Science in China 26:437–441
  • Waugh R, Powell W (1992) Using RAPD markers for crop improvement. Trends Biotechnol 10:186–191
  • Williams JG, Kubelik AR, Livak KJ, Rafaleski JA, Tiongey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
Typ dokumentu
Identyfikator YADDA
JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.