Illa Patel

Biochemical Changes During In Vitro Organogenesis of Tylophora indica (Burm. F.) Merrill

Biochemical changes in respect to Peroxidase, Polyphenol oxidase, IAA oxidase, Total amino acid, Reduc- ing sugar and Protein content were studied during the process of shoot regeneration in the callus culture of Tylophora indica (Burm. F.) Merrill. The following stages: dedifferentiated callus, differentiating green callus, differentiating callus with shoot primordial, differentiating callus with well-developed multiple shoots were selected for present study. Mature leaf was used as control. From the present investigation, it was proved that the process of differentiation had high metabolic activity. High peroxidase and polyphenol oxidase and IAA oxidase activities were recorded in differentiating green callus. Protein content along with reducing sugar and total amino acid contents were decreased significantly during differentiation as compared to control. Increased metabolic rate during in vitro organogenesis help Tylophora for rapid growth and multiplication.

An effective validated method for HPTLC-fingerprinting of alkaloids and glycosides from multiple plant parts of three Terminalia spp.

Our study developed a HPTLC fingerprint profile of alkaloids and glycosides obtained from the methanol extracts of four different plant parts of Terminalia arjuna , T. bellerica and T. chebula , trees with cardio-protective values. The multiple qualitative phytochemical analyses of water, acetone, petroleum ether and methanol extracts from all the plant parts of Terminalia spp. detected the presence of alkaloids and glycosides, wherein the methanol extracts exhibited the presence of maximum alkaloids and glycosides. The chromatographic analysis of methanol extracts was carried out on silica gel 60F 254 HPTLC aluminium sheets with CAMAG Linomat 5 applicator. The plates were developed using ethyl acetate:toluene:formic acid (10:10:1; v / v / v ) mobile phase. Alkaloids and glycosides were detected at 254 nm, 366 nm and 540 nm (after derivatization). These developed fingerprints would eventually be of great benefit in identifying or differentiating the alkaloids and glycosides in the form of marker compounds in the three Terminalia spp. mentioned.

Study on Qualitative and Quantitative Estimation of Phytochemicals of Triphala Plants

World is endowed with a rich wealth of medicinal plants and Natural products of plants offer vast resource of newer medicinal agents with potential in clinical use. Extraction methods and solvent selections are important for the collection of the phytochemicals because all compounds are not dissolving in all extracts and those compounds quantity rates are also different in different solvents. For that which solvent obtain maximum phytochemicals is most important things and the present study was an attempt to analyze the phytochemical analysis of Triphala plants (Terminalia chebula Retz., Terminalia belerica Retz., Phyllanthus emblica L). Introduction The natural resources how so ever large are bound to diminish hence need effective strategy is needed for sustainable utilization. Cultivation of medicinal and aromatic plants is constrained due to lack of suitable technology, which led to low yield and poor quality. Consequently, medicinal herbs are predominantly harvested in sufficient quantities from the wild in an unregulated manner (Shabbir et al., 2003). There are many astonishing things to learn from the collector of these herbs. Triphala is the combination of three medicinal plants exteensively used in ayurveda from ancient time. In trifala plants, one is Terminalia chebula Retz. (Harde), second one is Terminalia bellerica Retz. (Behada) and third one is Phyllanthus emblica L. (Ambla). T. chebula Retz , T. bellerica both the plant belong to family combretaceae and P.emblica L.belong to family Euphorbiaceae. T. chebula Retz. is used in many diseases laxative, diuretic, cardiotonic (Jagtap and Karkera, 1999) and also reported to be an antimicrobial (Sato et al., 1997; Ahmad and Aqil ,2007), hepatoprotective (Tasaduq et al., 2003; Tasaduq et al., 2006), antiinflammatory (Srikumar et al., 2005), antioxidant (Lee et al., 2007) and adaptogenic (Rege et al., 1999). P. emblica has been used for the treatment of several disorders and the important constituent of plant leaves extracts also posses several pharmacological properties like anti-viral (HIV, AIDS, HERPES VIRUS, CMV) antimutegenic, anti-allergic, anti-bacterial activities (Khopde et al., 2000).P. emblicaLcontains different class of secondary metabolites (Calixto et al, 1998). T.bellerica is used in the treatment of fever, cough, asthma, urinary diseases, piles, chronic diarrhea, dysentery, flatulence, vomiting, colic and enlarged spleen and liver (Cheng et al., 2003). The present study was carried out with an aim to estimate the amount of phenolic acids and flavonoids which are the constituents that may be responsible for the various pharmacological activities exhibited by the plan. Methodology Collection and preparation of Plant Extracts Materials The Sample of triphala plants were collected from Botanical garden of Hemchandracharya North Gujara University. The leaf and stem of the plant samples were collected from botanical garden. Here extraction was prepared in maceration method. Collected leaves and stems are washed under tap water than cut in to small pieces and again washed in distill water Then leaves and stems were left for drying under shade and after air drying they were grinding. 1 gms of each sample was soaked in conical flask containing 20ml water, methanol, Petroleum ether, chloroform, ethyl acetate and acetone for 24hour. After filtered of extracts the supernatants were collected, covered, labeled, stored at 20° C and used for the screening of various phytochemical and quantative estimation (Prashant et al., 2011). Qualitative and Quantitative estimation of phytochemical Primary qualitative analysis was done by Talari et al., 2012 and Rajeshvari et al., 2011 methods. Total phenol was determined by Folin Ciocalteu reagent (McDonald et al., 2001) and Aluminum chloride colorimetric method was used for flavonoids determination (Chang et al., 2002). Quantitative estimation was done by regression method. For Total Phenol gallic Acid was used as standard (Fig:1) and For Total Flavonoids quercetin was used as standard (Fig:2). Result and Discussion Qualitative Analysis The result of qulitative analysis was shown in Table No; 1.Phytochemicals screening of Terminalia chebula Retz. alkaloids was detected in water, methanol, petroleum ether and acetone of leaves but for stem alkaloids was detected in methanol and in acetone extracts. In Terminalia bellerica Retz. that was showed positive result in water, methanol, ethyl acetate and acetone extracts of leaves and same result was observed in Phyllanthus emblica L but only for leaves. For stem extracts showed positive result in water and methanol extracts of T.bellerica Retz. and in Phyllanthus emblica L, showed same result like T.chebula Retz. Alkaloids test was obtained positive in water, methanol and acetone extracts for leaves of all the plants and same things observed positive in water and methanol but for stem. Among three plants, in two plants tannins were detected in water, methanol and acetone but in T.bellerica Retz. Fig 1: Standard of Gallic acid for Total phenol. Fig 2: Standard of Quercetin for Total Flavonoids. Table 1; Qualitive analisys of phytochemical in various solvents

Direct shoot organogenesis from bulbs explants of Polianthes tuberosa cultivars (Prajwal and Shringar)

Ornamental plant Polianthes tuberosa (Amaryllidaceae) contains flowers which are used in wedding ceremonies, garlands, decoration and various traditional rituals. The present investigation given emphasis on: 1. In vitro propagation of two hybrid variety of Polianthes tuberosa namely Prajwal and Sringar from bulb as explant. 2. Effect of different combinations of BAP and Kinetin for its shoot proliferation, and to standardize the optimum concentration of growth regulators for proper growth of these hybrid varieties of Polianthes tuberosa. The rate of shoot proliferation frequency was gradually raised and reached maximum at 2.0mg/l concentration of both BAP and Kn in Prajwal as compared to 0.2mg/l in Shringar. The plantlets were incubated in a solution of IBA (0.5 mg l–1) and IAA (2.0 mg l–1) and IBA (0.5mgl–1) and IAA (2.5 mgl–1) at 50C to induce rooting of Prajwal and Shringar varieties respectively. INTRODUCTION The tuberose (Polianthes tuberosa) is an herbaceous, perennial plant belongs to Amaryllidacea family. Tuberose is a popular flower in floral arrangements and their scent is used to produce perfumes the world over (Sangavai, 2008). The Tuberose is a night blooming plant thought to be native to Mexico and is mostly grown in southern hemisphere but can do nicely in the north if planted in a protected sunny location (Singh, 1995; Trueblood, 1973). The long spikes of flowers are excellent for cut flower and people like their sweet fragrance (Moazz Hassanpour Asil, 2011). Tuberose inflorescences (spikes) bear 10 to 20 pairs of florets which open. Unopened flower buds scarcely open after harvest, and thus display quality of Tuberose spikes is limited (Michael Reid, 1996) .Its bloom time is August to September. The Tuberose exists only in cultivation. Flowers single, or double with twice or three times the number of petals. They are slow growers and one will need to have patience while waiting for them to pop out of the ground but these tropical beauties are well worth of time. It prefers to be kept on the dry side and needs rich well-drained, somewhat sandy, soil. The flowers are used in wedding ceremonies, garlands, decoration and various traditional rituals.AgNO3, CaCl22H2O and Tri-Miltox Forte treatments delayed flower opening and also extended vase-life. (Muhammad Akbar et al., 2001). Moreover Tri-Miltox Forte treatments maintained the fragrance of flowers for a longer period (Muhammad Akbar et al., 2001). In tuberose, various explants have been tried to produce regenerable cultures via in vitro morphogenesis are shoot tips (Hutchinson et al., 2004), bulb scale (Muralidhar and Mehta, 1982; Bose et al., 1987; Khan et al., 2000; Rajeshkaran et al., 2000; Nazneen et al., 2003) rhizome (Sangavai and Chellapandi, 2008). The traditional method of propagation through bulbs is rather slow to meet the growing demand and, therefore, direct shoot organogenesis through tissue culture may be utilized for rapid and large-scale multiplication. MATERIALS & METHODS Explant material & Inoculation Two cultivars of Polianthes tuberosa were selected for the study. Two cultivars viz: Prajwal and Shringar were procured from Navasari Agriculture University, Navasari. (Navasari -Surat). The explants were placed on solid Murashige and Skoog’s (MS) basal medium (Murashige, T. and Skoog, F., 1962) containing 0.8% (w/v) agar, and various concentrations of BAP (6-benzylaminopurine), Kn (kinetin) and IAA (indol-3-acetic acid) in combination used for shoot proliferation and root proliferation. These media were supplemented with growth regulators of different concentration (0.2 mg/l, 0.5 mg/l, 1.0 mg/l, 1.5 mg/l, 2.0 mg/l, and 2.5 mg/l) and adjusted to pH 5.8 prior to autoclaving at 120°C for 20 minutes. Culture media (25ml) were dispensed into 50 ml test tubes and plugged with cotton. These were maintained under long day conditions (16 hours light/8 hours dark cycle at an intensity of 2000-lux luminance at 25±2 oC) and 60% RH (Gajbhiye et al, 2011) .Shoot proliferation was observed after 4 to 5 days of inoculation. The effect of hormones on shoot proliferation and root proliferation was studied and effort was made determine the appropriate hormone combinations for optimal shoot proliferation and root proliferation. RESULT Hybrid Variety Prajwal The research conducted to optimize the medium for the multiplication of Prajwal of Polianthes tuberosa L. have been depicted in Tables 1 and 2. Effect of BAP and Kn The shoot initiation of hybrid variety Prajwal of Polianthes tuberosa L. from explant (Bulb) with different concentration of growth regulators BAP (0.2-2.5mg/l) and Kn (0.2 – 2.5mg/l) and the results are presented in Table 1 and shoot growth is depicted in Plate1(A). Effect of IBA and IAA The effects of the different concentration of growth regulators IBA (0.2 mg/l) and IAA (0.2 – 2.5mg/l) on root induction of hybrid variety Prajwal of Polianthes tuberosa L. from explant (Bulb) were presented in Table 2.The data showed that in Prajwal the combination of IBA (0.5mg/l) and BAP (2.0mg/l) gave best result for initiation of root at 50C. Hybrid Variety Shringar Effect of BAP and Kn The shoot initiation of hybrid variety Shringar of Polianthes tuberosa L. from explant (Bulb) with different concentration of growth regulators BAP (0.2-2.5mg/l) and Kn (0.2 – 2.5mg/l) and the results are presented in Table 1 and shoot growth is depicted in Plate1(B).