Plumbago zeylanica is an of import medicinal works and has been used in Ayurveda from ancient times. In the present survey we have observed the consequence of variable concentration of auxin and cytokinin on the regeneration potency. P. zeylanica has shown the better micropropagation consequence when nodal bud was used as explants. The budding portion respond best in the medium C holding IAA and BAP concentrations 4 mg/l & A ; 5mg/l severally in MS media, ensuing in the development of shoot regeneration. After three hebdomads shoot regeneration additions without farther development in the roots. At the same clip it has been observed that explants has shown regeneration potency in media D and G, ( BAP-3.5mg/l, NAA-2.5 mg/l ) but the growing rate was slower. Phytochemical analysis of rough infusion of this works indicates the presence of photochemical i.e. alkaloids, glycosides and phenolic compound etc.
Introduction
Plumbago zeylanica ( known vernacularly as Chitraka, Chitramulamu, Tellachitramulamu, Agnichela, Agnimaala or by its trade or popular names of “ Lead wort-white flowered ” and “ Ceylon Lead wort ” ) of the Plumbaginaceae, is distributed as a weed in throughout the tropical and semitropical states of the world.A This works is a spreading or subscandent, herbaceous, suffrutescent works, 1 – 2 m in length. The root contains medicinally of import compound known as ‘Plumbagin ‘ . Plumbagin is present in all the assortment of graphite to a upper limit of approximately 0.91 % . This active chemical is vesicatory, has belongingss of vitamin K, and is antibiotic on several human pathogens ( Burkill, 1985 ) . The works is continuously collected from the wild and over exploited from ancient times. In position of this, there is an demand for alternate agencies of clonal extension with a position to forestalling the works from extinction.Biotechnological tools such as In-Vitro micropropagation can play of import function in the preservation of endangered medicative workss every bit good as for rapid generation and quality betterment of valuable medicinally of import works species. In-vitro regeneration of Plumbago zeylanica, workss via callosity civilizations ( Rout et al. , 1999 ) and through direct organogenic distinction have been worked out in recent old ages ( Selvakumar et al.,2001 ; Verma et al. , 2002 ) . Clonal generation of Plumbago rosea ( Kumar and Bhavanandan,1988 ) and Plumbago indica ( Chetia and Handique,2000 ) had besides been reported earlier. In-vitro production of “ plumbagin ” – the coveted secondary metabolite of P.zeylanica, from A.rhizogenes mediated haired root civilizations had besides been published presently ( Verma et al. , 2002 ) .
Methodology
The experiment was performed in the tissue civilization research lab of IMS Engineering college, Ghaziabad in the twelvemonth 2008-09 Fresh and healthy works stuffs of P. zeylanica collected from the local baby’s room and nodal bud and foliage were used as explants for the present survey. The standard MS media were used during full survey with variable concentration of Auxin and Cytokinin The pH of each medium was adjusted to 5.8 prior to the add-on of 8gm/litere of agar ( Quligens ) . Media and instruments were sterilized by autoclaving for 15 – 30 min at 121A°C ( 1 standard pressure ) . Aseptic transportations were performed in a laminar flow goon ( S.M Scientific ) . The explants were washed exhaustively with liquid detergents and followed by under running tap H2O for 30 proceedingss and concluding sterilisation was carried out with 0.1 % HgCl2 and rinsed decently with unfertile distilled autoclaved H2O for 4-5 times. After the surface sterilisation of explants it was aseptically transferred into the pre- sterilised media boottles ( Conical flask ) . Cultures were maintained 240C in the civilization suites and exposed to 8-10 hours visible radiation. Contaminated civilizations were treated with 0.1 % HgCl2 and bomber cultured in fresh media.
The petroleum infusion of Plumbago zelanica was extracted through soxlet setup taking foliage and flower parts as works stuff utilizing ethyl alcohol and hexane as dissolver. Solvent is evaporated utilizing vacuity rotatory evaporator. Crude infusion was analyzed through chemical reaction for the presence of different photochemical
Result & A ; Analysis
The full experiment were performed in 3 rhythms which gives the undermentioned consequences in variable concentration of auxin and cytokinin
Cycle I
Observations
IBA-4
BAP-5
IBA-4
BAP-4
IAA-4
BAP-4
IAA-4
BAP-5
Survival
3
2
3
4
Growth
2
1
1
1
Contamination
0
2
2
0
Dead
1
1
0
1
CYCLE II:
Observations
IBA-3mg/ml
NAA-2.5mg/ml
IBA-2.5mg/ml
NAA-2mg/ml
IBA-3.5mg/ml
NAA-3.0mg/ml
Growth
2
3
5
Contamination
4
1
2
Dead
0
0
0
Conformation analysis of different phytochemical nowadays in the Plumbago ( foliages ) works stuff
Trial
Class
Consequence
Alkaloid
Dragandroff ‘s reagent
positive
Carbohydrates
Molish trial
positive
Glycosides
Balijet trial
negative
Phenolic resin
Lead ethanoate
positive
Flavanoids
Ammonia trial
Negative
Protein & A ; amino Acid
Xanthoprotein
Positive
Conformation analysis of different phytochemicals nowadays in the Plumbago ( flower ) works stuff
Trial
Class
Consequence
Alkaloid
Dragandroff ‘s
reagent
Positive
Carbohydrates
Molish trial
Positive
Glycosides
Balijet trial
Negative
Phenolic resin
Lead ethanoate
positive
Ferric chloride
negative
Flavanoids
Ammonia trial
negative
Protein & A ; amino acid
Xanthoprotein
positive
Saponins
Foam
negative
P. zeylanica has shown the better micropropagation consequence when nodal bud was used as explants. The budding portion respond best in the medium C holding IAA and BAP concentrations 4 mg/l & A ; 5mg/l severally in MS media, ensuing in the development of shoot regeneration. After three hebdomads shoot regeneration additions without farther development in the roots. At the same clip it has been observed that explants has shown regeneration potency in media D and G, ( BAP-3.5mg/l, NAA-2.5 mg/l ) but the growing rate was slower. Phytochemical analysis of rough infusion of this works indicates the presence of photochemical i.e. alkaloids, glycosides and phenolic compound etc.
