The plant contains major compounds such as coumarins, psoralen, isopsoralen, angelicin and daidzein. The plant exhibits antitumour, antibacterial, antifungal and antioxidative activities⁴. It is used as laxative, aphrodisiac, anthelmintic, diuretic and diaphoretic in febrile conditions. It has been specially recommended in the treatment of leucoderma, leprosy, psoriasis and inflammatory diseases of the skin and prescribed both for oral administration and external application in the form of a paste or ointment⁵. The wild population of this medicinally important plant exhibits high-mortality of the seedlings and also declined very fast due to indiscriminate and illegal collections, and destruction of its habitats. Therefore this species was included in the endangered list⁴.

In pharmaceutical formulation it has great demand as sources of curative compounds to treat ailments. The highly demand of this plant in pharmaceutical industry, and in vitro micropropagation of Psoralea corylifolia has become a necessity in order to meet the pharmaceutical needs and also to prevent the plant from becoming endangered. Therefore the present review emphasizes on literature of Psoralea corylifolia covering the in vitro micropropagation and the occurrence of pharmacologically relevant compounds.⁵

Latha PG et al (1999) have reported the cytotoxic and antitumour properties of the hexane extract of the seeds of P. corylifolia. They investigated number of fatty acid glycerides by GC (Gas Chromatography) analysis from the hexane extract seed of P.corylifolia. GC analysis showed the presence of methyl esters of lauric acid (0.2%), myristic acid (0.13%), palmitic acid (22.07%), linoleic acid (67.7%), oleic acid (3.33%) and stearic acid (0.52%), besides 3 unidentified methyl esters. The extract of P. corylifolia seeds, depeting significant cytotoxic properties consisted of a number of fatty acid glycewrides⁶.

An efficient method for direct plantlet regeneration from nodal explants of P. corylifolia was investigated by Jeyakumar M et al (2000). Nodal explants cultured on Murashige and Skoog (MS) medium with different concentrations of BA (benzylaminopurine) and KIN (kinetin). The maximum number of multiple shoots was found on MS medium containing 0.5 mg BAP/l. Among the 3 auxin tested (IAA, indole-3-butyric acids (IBA) and naphthalene acitic acid (NAA), IAA at 1.0 mg/l was found to be best for rooting of shoots. Rooted plantlets were successfully established in the field by author⁷.

Rout GR et al (2001) studied on in vitro somatic embryogenesis of P. corylifolia. Somatic embryogenesis was achieved in callus derived from leaf and stem explants of P. corylifolia on modified Murashige and Skoog’s medium containing potassium nitrate (KNO₃) supplemented with kinetin, 2,4-dichlorophenoxyacetic acid (2,4-D) and sucrose. Somatic embryos proliferated rapidly by secondary somatic embryogenesis after transfer to MS medium supplemented with kinetin and 2,4-D in subsequent subcultures. A high percentage of cultures of somatic embryos developed from stem derived calluses as compared to leaf derived calli. Maturation and germination of somatic embryos were achieved on half strength MS basal medium supplemented with IBA and sucrose⁸.

Chand S et al (2002) developed an efficient plant regeneration protocol from root explants of P. corylifolia. Nodular embryogenic callus was cultured on Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid in combination with 6-benzylaminopurine, thiamine HCl, L-glutamine and sucrose. The maximum number of cotyledonary stage somatic embryos was obtained on MS medium containing 1.34 μM NAA and 13.2 μM benzylaminopurine (BA). Germination of somatic embryos occurred on MS medium without any growth regulators and also on MS medium enriched with BA, although the maximum germination frequency was obtained on 4.4 μM BA plus 1.45μM gibberellic acid (GA₃). The highest frequency of plant regeneration and mean number of plantlets were obtained on MS medium containing 1.34 μM NAA and 4.4 μM BA⁹.

Jeyakumar M et al (2002) has in vitro regenerated P. corylifolia from cotyledonary node. P. corylifolia gave rise to multiple shoots when cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of BAP and Kn (kinetin). The highest rate of shoot multiplication was obtained in MS containing 2.22 μM BAP. The regenerated shootlets were rooted on MS basal medium with different concentrations of indole-3-butyric acids (IBA). The maximum number of roots was produced on the medium containing 4.92 μM of IBA⁵.

Ragendra NR et al (2004) investigated that the extracts obtained from seeds of Psoralea corylifolia showed several degrees of antifungal activity against Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum and Microsporum gypseum by the disc diffusion method on a Sabouraud dextrose agar (SDA) medium. Methanol extract of the seeds at 250 µg exhibited the maximum inhibition with a halo of 28 mm diameter. Six different bands were obtained when the methanol extract was subjected to Thin Layer Chromatography (TLC). ¹³C NMR and Mass spectra revealed that the active compound would be a flavonoid, 4'-methoxy flavone. Minimum inhibitory concentration (MIC) of the active compound along with standard miconazole was carried out using tube dilution technique¹⁰.

Jiangning G et al (2005) were tested extract of P. corylifolia by oxidative stability instrument (OSI) and were found to have strong antioxidant effects. Six compounds bakuchiol, psoralen, isopsoralen, corylin, corylifolin and psoralidin were isolated from the herb. Their antioxidant activities were investigated individually and compared with butylated hydroxytoluene (BHT) and α-tocopherol by OSI. The result showed that psoralen, isopsoralen had no antioxidant activities at 0.02% to 0.05% levels¹¹.

Sah P et al (2006) were isolated isomeric furocoumarins from the seeds of P. corylifolia. On the basis of physical and spectral parameters, i.e., UV, IR, NMR, mass and chemical reactions such as hydrolysis, alkali fusion and oxidation, they have been identified as 2H-furo[3',2'-g][1] benzopyran-2-one (1) and 2H-furo[2',3'-h] [1-] benzopyran-2-one¹².

Faisal M et al (2006) investigated on the effect of thidiazuron (TDZ) in vitro axillary shoot proliferation from nodal explant of P. corylifolia. Proliferation of shoots was achieved on MS medium supplemented with 0.5, 1, 2, 3, 4 and 5 μM TDZ. The maximum numbers of shoots per explant were obtained from nodal segment cultured on 2 μM TDZ for 4 weeks and this increased to 29.7 ± 2.1 on hormone free MS medium after 8 weeks. The in vitro proliferated and elongated shoots were transferred individually on a root induction medium containing 0.5 μM IBA and within 4 weeks 4.5 ± 0.5 roots per shoot were produced¹³.

Agarwal D et al (2006) were isolated and characterized isomeric furocoumarins from the seeds of P. corylifolia. On the basis of physical and spectral parameters, i.e., UV, IR, NMR, mass and chemical reactions such as hydrolysis, alkali fusion and oxidation, they have been identified as 2H-furo[1] benzopyran-2-one (1) and 2H-fu [1-] benzopyran-2-one¹⁴.

Chwan-Fwu Lin et al (2007) developed a HPLC method for simultaneously determining bakuchiol, psoralen and angelicin in Fructus Psoraleae (Buguzhi, the fruits of P. corylifolia and its commercial concentrated products. Extracted samples were analyzed by using a reverse-phase column and eluting with a gradient mobile phase consisting of 20% acetonitrile to acetonitrile at a flow rate of 1.0 mL/min with detection at a wavelength of 254 nm. They found that Bakuchiol, psoralen and angelicin contents were 36.2~71.0, 2.5~13.0 and 2.2~9.2 mg/g for ten raw material samples of Fructus Psoraleae, and 0.6~21.1, 0.6~5.2 and 0.6~5.3 mg/g for eight commercial concentrated products of Fructus Psoraleae, respectively¹⁵.

Baskaran P et al (2008) studied effect of growth regulators on rapid micropropagation and psoralen production in P. corylifolia. Multiple shoot buds were obtained in half-strength liquid Phillips–Collins medium supplemented with 5 μM BAand 5 μM TDZ from apical bud explants. The shoot buds were subcultured on enriched solid L2 medium supplemented with different concentrations and combinations of BA, kinetin 2-isopentenyladenine (2iP), TDZ, bavistin (BVN) and trimethoprim (TMP). Enriched solid L2 medium supplemented with 2 μM BA, 1 μM TDZ and 100 mg l−1 BVN were more effective in producing greater number of shoots per explant shoots/ after 4 weeks of culture. The regenerated shoots rooted and hardening upon transfer to 50 μM IBA for 15 min. The concentration of psoralen was evaluated in different tissues of ex vitro and in vivo grown plants by high-performance liquid chromatography (HPLC). Psoralen content was increased in leaves (2.97%), roots (2.38%), stems (5.40%) and seeds (1.63%) of ex vitro plants than the in vivo plants¹⁶.

Shinde AN et al (2009) induced high frequency shoot regeneration and enhanced isoflavones production in Psoralea corylifolia. In vitro regenerated shoots were induced from germinated seedling on MS medium supplemented with thidiazuron and N₆–benzylaminopurine. The results revealed that optimum concentrations of thidiazuron into the medium increased shoot regeneration frequency.High concentrations of Indole-3-aceticacid (IAA) into the rooting medium resulted in slow growth. Regenerated shoots and roots enhanced isoflavones production compared to field grown plants. A reverse phase high performance liquid chromatography analysis revealed that in vitro regenerated shoots accumulated 0.85% dry wt of daidzein and 0.06% dry wt of genistein. Maximum daidzein (1.23% dry wt) and genistein (0.38% dry wt) were accumulated by roots which obtained from regenerated shoots, which is 6.3-fold more daidzein and 77-fold more genistein respectively than field grown plants. The regeneration protocol developed successfully in this study showed the possibility for rapid propagation of P. corylifolia and enhanced isoflavones production¹⁷.

Baskaran P et al (2009) was developed a protocol for in vitro regeneration of P. corylifolia from hypocotyl-derived callus. nodular calli were induced from 3 day-old hypocotyl explants on Phillips and Collins medium containing 25 g l/1 sucrose, 7 g l/1 agar and supplemented with 10mM NAA and 2m M TDZ. Higher shoot regeneration (89.5±1.18) was achieved in enriched L2 medium supplemented with 2mM BA, 4m M TDZ and 50 mg l/1 BVN. This system would be useful for mass propagation and germplasm conservation of P. corylifolia¹⁸.

Ghosh P et al (2009) was studied effect of ethanolic extract of P. corylifolia on H. pylori infection in laboratory rats. Naproxen was administered orally to produce ulcers and then rats were orally inoculated with H. pylori. The extent of neutrophil infiltration was estimated by myeloperoxidase assay. Clarithromycin treated animal’s demonstrated absence of infection at 4^th, 7^th and 10^th week whereas the control group of animals showed the presence of infection throughout the treatment regimen. Ethanolic extract of Psoralea eradicated the H. pylori infection in a dose and time dependent manner. The level of myeloperoxidase was elevated in the control group whereas significantly reduced in the clarithromycin and extract treated groups when compared with the vehicle treated group of animals but decreased in the clarithromycin and ethanol extract treated groups. It was concluded that ethanol extract showed anti H. pylori activity¹⁹.

Khushboo PS et al (2009) developed and validated an accurate and precise high-performance thin-layer chromatographic method for quantification of psoralen from Psoralea corylifolia. The method employed TLC aluminium plates precoated with silica gel 60F- 254 as the stationary phase. Linear regression analysis of the calibration plots showed good linear relationship between peak area and peak height (r² = 0.99828 and 0.99649) in the concentration range 10–100 ng/ spot. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits. The average recovery of the method was 99.73 %. The amount of psoralen in seed powder extract was found to be 0.928%. This method can be used as quality control method for checking the purity of P. corylifolia seed powder, extract and its formulation²⁰.

Somasundaram T et al (2010) screened aqueous and alcoholic extracts from P. corylifolia leaves for the presence of chemically active compounds by standard methods and evaluated for their antimicrobial activity in vitro by disc diffusion method. The results revealed the presence of saponins, tannins, flavonoids, glycosides, carbohydrates, tannins and phenolic compounds, gums and mucilages, fixed oils and fats. Alkaloids were not detected from any of the leaves extract under study. Aqueous and alcoholic extracts exhibited broad-spectrum antibacterial and antifungal activity against Eschericia coli, Pseudomonas aeruginosa, Staphylococcus pyogenes and Candida albicans. Alcoholic extract is better than that of aqueous extract of P. corylifolia leaves in respect to their antimicrobial activity and the broad spectrum of activity makes it a promising indigenous drug²¹.

Three chalcones and two flavanones displaying tyrosinase inhibitory activity were isolated from the seed of P. corylifolia by Jun Young Kim et al (2010) The isolated compounds were elucidated as known isovabachalcone, 4'-O-methylbavachalcone, isobavachromene, corylifolin, and bavachinin. The inhibitory potencies of the polyphenols toward monophenolase activity of mushroom tyrosinase were investigated. The inhibitory concentration (IC₅₀) values of compounds 1-5 for monophenolase activity were determined to be 12.3, 48.8, 15.8, 23.6, and 143.9 μM, respectively. The potent inhibitors 1-4 exhibited the competitive inhibition characteristics in analysis of Lineweaver-Burks and Dixon-plot²².

Mishra A et al (2010) investigated in vivo hypoglycemic and antidiabetic potential of methanolic extract of seeds of P. corylifolia in glucose loaded animals and alloxan induced diabetic animals. In both the models P. corylifolia reduced the blood glucose level when compared to diabetic control group and exert a significant hypoglycemic and antidiabetic activity. P. corylifolia methanolic extract also reversed the body weight in normal and alloxan induced diabetic animals. The results of this study revealed the presence of a significant antidiabetic potential of methanolic extract of P. corylifolia in alloxan induced diabetic rats²³.

Baskaran P et al (2010) developed a protocol for in vitro regeneration of P. corylifolia through adventitious shoots regeneration from hypocotyl explants cultured on MS medium with various combination and concentration of plant growth regulators. The highest response was obtained on MS medium containing 3.0µM BA, 1.0µM NAA, 5.0µM ascorbic acid (AA) and 100mg /L casein hydrolysate (CH). Shoots were rooted on half basal media suplemented with 3.0µM IBA²⁴.

Gidwani B et al (2010) were determining anti-inflammatory and antimicrobial activity of hexane extract of seeds of P. corylifolia by carrageenan induced rat paw edema assay. The seed of plant was extracted in soxhlet apparatus using hexane as solvent. The evaluation was done by Hind paw method and compared with standard and control groups. Further, the antimicrobial activity of P. corylifolia was studied by Disc diffusion method using different gram positive and gram negative bacterial and fungal strains, which showed that it has prominent activity compared with the standard used. The hexane extract of P. corylifolia showed significant anti-inflammatory activity (44%) when compared with the standard (55%). For antimicrobial activity, zone of inhibition were found to be in concentration of 5μg/ml for bacterial strains and 4μg/ml for fungal strains²⁵.

Wang et al (2011) were screen and identified bioactive compounds with anticancer activity from the seeds of Psoralea corylifolia. One volatile fraction (fraction I) and three other fractions (fraction II, III, IV) from methanol extraction of P. corylifolia were obtained. Bioactivities of these fractions were evaluated by the cytotoxicity on KB, KBv200, K562, K562/ADM cancer cells with MTT assay. Major components in the active fraction were identified by HPLC/MSn. Fraction IV significantly inhibits the growth of cancer cells in a dose-dependent manner²⁶.

Anwar M et al (2011) were standardized the crude extract of P. corylifolia and determine its pharmacological activities by high performance liquid chromatography (HPLC), Fourier transform infra red (FTIR), Fourier transform near infra red (FTNIR) and ultra violet (UV) spectrophotometric methods. Pharmacological study and its fractions were carried out on smooth muscle of rabbit intestine. The results revealed quick decrease in the normal intestinal movement followed by a gradual dose dependant increase in the rhythmic activity of intestine. The antispasmodic response of the crude extract was found most significant at 20 mg/ml. The chloroform fraction of the crude extract exhibited maximum antispasmodic response at 10 mg/ml. n-butanol and aqueous fractions produced 39.29 and 13.79% antispasmodic effect, respectively, whereas, ethyl acetate fraction produced spasmogenic effect. The crude extract of P. corylifolia exhibited positive antifungal activity against Candida albicans and positive antibacterial activity against Staphylococcus aureus. Significant analgesic effect was also observed with 300 and 500 mg/kg dose at p < 0.05. The analgesic effect of the plant extract was found to be higher than that of diclofenac sodium²⁷.

Characterization, isolation and purification of Acid phosphatase from the cotyledons of Psoarlea corylifolia was carried out by Singh S et al (2011). The characterization of acid phosphatase includes the optimization of kinetic parameters over the range of substrate concentrations, influence of different divalent metal ions, modulators, various inorganic and organic phosphates on the acid phosphatase mediated hydrolysis of p-Nitrophenyl phosphates. The purification processes included the ammonium sulphate precipitation, DEAE cellulose separation and gel filtration of enzyme through sepharose column. They found that the Partially purified acid phosphatase protein (226.08 fold) obtained by ammonium sulphate precipitation and gel filtration exhibited very high specific activity (264211.157 μmole min-1 mg-1). Acid phosphatase showed spots at ~28 kDa and ~30 kDa in two dimensional gel electrophoresis followed by western blot analysis²⁸.

Kiran B et al (2011) were tested antifungal activity of aqueous and solvent extract of seeds of P. corylifolia against five seed borne fungi of maize. In aqueous extract, maximum inhibition was observed in A. alternata and recorded 95.4% inhibition at 50% concentration followed by C. lunata (86.0%), Rhizopus sp. (82.3%), D. halodes (68.0%) and C. cladosporioides (57.7%). Significant activity was also observed in 10, 20, 30 and 40% concentration. In solvent extracts tested at 250,500, 750 and 1000μl concentration, maximum inhibition was observed in petroleum ether extract and moderate activity was observed in methanol extract²⁹.

Chand S et al (2011) the antibacterial activity of aqueous and methanolic extracts of the plants was evaluated against 5 microorganisms by agar well diffusion method. Amongst the 13 plants screened, P. corylifolia showed best antibacterial activity. The seed and aerial parts of P. corylifolia were extracted successively using a series of various organic solvents. The antibacterial activity of these extracts was done against 5 microorganisms by agar disc diffusion method. All the extracts of seed and aerial parts were active against S. epidermidis and P. morganii. Maximum antibacterial activity was shown by dioxan extract of the seed. The present findings suggest that the dioxan extract of seed of P. corylifolia can be used as a promising novel antibacterial agent in the near future³⁰.

Purkayastha S et al (2012) evaluated phytochemical constituents, antibacterial activity and TLC bioautography assay of P. corylifolia. They demonstrated the presence of flavonoids, tannins, steroids, terpenoids, glycosides and saponins. Antibacterial activity of P. corylifolia was assessed on eight multi-drug resistant (MDR) clinical isolates from both Gram-positive and Gram-negative bacteria and two standard strains. It showed broad antibacterial activity against both Gram-positive bacteria and Gram-negative bacteria. The highest in vitro inhibitory activity was observed for MDR Enterococcus sp. with wide inhibition zone diameters (21±0.16 mm) followed by standard S. aureus ATCC 25923 (16±0.11 mm) and K. pneumoniae (15±0.056 mm). Thin layer chromatography and bioautography assay demonstrated well-defined growth inhibition zones against Enterococcus species in correspondence with tannins observed at R_fvalues of 0.10-0.15 and 0.70-0.83. This established a good support to the use of this essential oil in herbal medicine³¹.

Sangeetha S et al (2012) were tested Antifungal activity of petroleum ether extract of P. corylifolia seed, against Fusarium species was evaluated by agar well diffusion assay. The chromatographic fractionation yielded a new phenyl derivative of pyranocoumarin (PDP). The PDP had a potent antifungal activity with a minimum inhibitory concentration. Molecular docking using Grid-Based Ligand Docking with Energetics (GLIDE, Schrodinger) was carried out with the Tri101, trichothecene 3-O-acetyltransferase, as target protein to propose a mechanism for the antifungal activity. The ligand PDP showed bifurcated hydrogen bond interaction with active site residues at TYR 413 and a single hydrogen bond interaction at ARG 402. This indicated a strong binding of the ligand with the trichothecene 3-O-acetyltransferase, preventing as a result the acetylation of the trichothecene mycotoxin and destruction of the “self-defense mechanism” of the Fusarium species³².

CONCLUSION:

The prime importance of in vitro propagation of rare, critically endangered, endangered and vulnerable plants would be to generate a large number of planting materials from a single explant without destroying the mother plant and subsequently their restoration in the natural habitat, thus conserving the biodiversity. Psoralea corylifolia specially recommended in the treatment of leucoderma, leprosy, psoriasis and inflammatory diseases of the skin and prescribed both for oral administration and external application in the form of a paste or ointment. Pharmaceutical companies largely depend upon materials procured from naturally occurring stands causing rapid depletion of this important source of medicinal herb. Hence, it has become imperative to establish a suitable protocol to generate enough materials to ensure its supply for pharmaceutical industries without further depopulating this species. Rapid multiplication of elite clones, production of healthy and disease-free plants and faster introduction of novel cultivars with desirable traits are of urgent need in Psoralea corylifolia improvement programme. In this regard, in vitro propagation techniques are likely to play a vital role. The significance of an efficient in vitro protocol would be to obtain maximum number of plantlets in minimum period of time with proper rooting along with acclimatization in the field.

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Received on 04.11.2012

Modified on 12.11.2012

Accepted on 20.11.2012

Research J. Science and Tech. 4(6): November –December, 2012: 237-242