Analysis of nuclear DNA content, genetic stability, Bacoside A quantity and antioxidant potential of long term in vitro grown germplasm ... View Full Text


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Article Info

DATE

2014-09-04

AUTHORS

Muthiah Joe Virgin Largia, Jayabalan Shilpha, Govindan Pothiraj, Manikandan Ramesh

ABSTRACT

The true-to-type nature of shoot cultures of Bacopa monnieri maintained in vitro for 5 years has been confirmed by means of flow cytometry (FCM), inter simple sequence repeat (ISSR), directed amplification of mini satellite region DNA (DAMD) analysis and chemical profiling. The nuclear DNA content of B. monnieri has been estimated as 1.86 pg/2C based on the FCM analysis of nuclei from young leaves. It has also detected no significant variation in the nuclear DNA contents between long term maintained germplasm lines and field grown plants. ISSR and DAMD analysis produced about 93 amplification products and all of them were found to be monomorphic among the field grown and seven germplasm lines. Even though morphological growth parameters varied slightly between the lines, the bioactive compound production capacity has been retained. The mean amount of Bacoside A accumulated by the germplasm lines has been quantified as 25.02 mg g−1 dry weight (DW) using high performance liquid chromatography analysis which is comparable with the quantity of Bacoside A accumulated by the field grown plants (26 mg g−1 DW). The total phenolics (folin–ciocalteau method; expressed in milligram Gallic acid equivalents per gram dry weight of the sample) and the flavonoids content (aluminium chloride method; expressed in milligram Quercetin equivalents per gram dry weight of the sample) of the germplasm lines (53.50–56.62 mg GAE/g DW; 273.51–277.16 mg QRE/g DW) were found to be slightly higher than the field grown plants (46.66 mg/g GAE; 256.65 mg/g QRE) respectively. Further, the antioxidant potential of the methanolic whole plant extracts determined by means of (1,1-diphenyl-2-picrylhydrazyl) DPPH assay revealed that the germplasm lines showed little higher antioxidant potential with IC50 values ranging from 39.51 to 43.71 µg g−1 DW when compared to the IC50 value of field grown (45.68 µg g−1 DW). Hence the protocol devised using axillary bud culturing can be utilized effectively for the long term maintenance of aseptic clones with high genetic and chemical fidelity. The approach tackles the increasing market demand through the continuous availability of high quality plant materials all round the year besides preserving the lines under monitored in vitro conditions. More... »

PAGES

399-406

References to SciGraph publications

  • 2013-03-02. Ploidy stability in embryogenic cultures and regenerated plantlets of tamarillo in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2011-04-22. Genetic variation among highly endangered Bacopa monnieri (L.) Pennell from Southern India as detected using RAPD analysis in GENETIC RESOURCES AND CROP EVOLUTION
  • 2009-03-21. Determination of genetic stability of long-term micropropagated plantlets of Platanus acerifolia using ISSR markers in BIOLOGIA PLANTARUM
  • 2013-03-21. Phenolic acid and DNA contents of micropropagated Eryngium planum L. in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2008-01-22. Minisatellites as DNA markers to classify bermudagrasses (Cynodon spp.): confirmation of minisatellite in amplified products in JOURNAL OF GENETICS
  • 2007-09-06. Estimation of nuclear DNA content in plants using flow cytometry in NATURE PROTOCOLS
  • 2014-06-11. Efficient plant regeneration from shoot apex explants of maize (Zea mays) and analysis of genetic fidelity of regenerated plants by ISSR markers in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2007-03-22. Assessment of genetic stability of the germplasm lines of medicinal plant Scutellaria baicalensis Georgi (Huang-qin) in long-term, in vitro maintained cultures in PLANT CELL REPORTS
  • 2014-01-10. Improved in vitro propagation, solasodine accumulation and assessment of clonal fidelity in regenerants of Solanum trilobatum L. by flow cytometry and SPAR methods in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2001-09. Assessment of Age-Related Polyploidy in Quercus Robur L. Somatic Embryos and Regenerated Plants Using DNA Flow Cytometry in BIOLOGIA PLANTARUM
  • 2001-02. Oxidative stress and physiological, epigenetic and genetic variability in plant tissue culture: implications for micropropagators and genetic engineers in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
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