Integrated disease direction is a non-separable portion of all the eco-friendly stable agricultural plans. Plant infusions have played important function in the suppression of pathogens and in the betterment of harvest production. Cuscuta reflexa Roxb. is a parasitic works. Agriculturists consider Cuscuta spp is merely a destructive weed and effort to eliminate it. The chief intent of this survey was to measure its significance in developing works based preparations for fungous disease direction. In vitro, fungicidal potency of C. reflexa ( parasitizing Bougainvillea glabra L. ) infusion was evaluated against five different pathogenic Fungi viz. , Alternaria surrogate, Aspergillus Niger, Fusarium solani, Fusarium oxysporium and Macrophomina phaseolina. Different aqueous concentrations ( 0, 10, 20, and 30 % ) of C. reflexa were prepared. Aqueous concentrations of C. reflexa were evaluated against fungous isolate by good diffusion method. The fungitoxicity of infusion in footings of suppression zone diameter was calculated. Consequences indicated that fungous growing suppression was straight relative to the concentration of C. reflexa infusion. Cuscuta infusion exhibited important fungicidal activity against all trial fungal isolates. However, infusion was extremely effectual against F. solani, F. oxysporium and M. phaseolina, and was least effectual against A. Niger. It was besides found that 30 % ( w/v ) concentration was significantly effectual in cut downing the mycelial growing of fungous isolates after 6 yearss of incubation. Further probes nevertheless are required to analyse nature of fungicidal compounds in C. reflexa and their stableness.
Cardinal words: Cuscuta reflexa, parasitic workss, fungicidal infusion, infective Fungi, parasitic weeds
Cuscuta reflexa Roxb. is an angiospermic hustorial progress, obligate, hemiparasite belonging to household cuscutaceae. It parasitizes wild and cultivated workss, and is particularly destructive to such commercially valuable harvests as flax, lucerne, beans, and murphies. It besides grows on common cosmetic workss though the programs of Pakistan ( Bhattacharya, 1976 ; Malik et al. , 1980 ) . Agriculturists consider Cuscuta species a destructive weed and effort to eliminate it. However on biological facets, strong fungicide and antibacterial compounds have been extracted from C. reflexa ( Loffler et al. , 1997 ; Da-Nian Qin et al. , 2000 ) . Its curative belongingss such as anticancer, antidiabetic, antiviral, and anti-inflammatory are besides good documented ( Awasthi, 1981 ; Poudel, 2002 ) . Surveies have besides proved that the presences of some stable phytochemicals in Cuscuta species are irrespective of host workss and locations ( Loffler et al. , 1997 ) . Plant metabolites and plant-based pesticides appear to be one of the better options as they are known to hold minimum environmental impact and in the betterment of harvest production ( Varma and Dubey, 1999 ; Nwachukwu and Umechuruba, 2001 ) . Conventional genteelness of disease-resistant cultivars and works protection based on extended usage of agrochemicals represent still common schemes in the competition between human attacks to battle microbic nutrient rivals and the evolutionary adaptability of bacterial and fungous phytopathogens. These are responsible for tremendous harvest losingss worldwide and hence threaten human nutrition ( Osusky et al. , 2000 ) . Integrated disease direction is a non-separable portion of all the eco-friendly stable agricultural plans. Biological control of works diseases and works pathogens is of great significance in forestry and agribusiness ( Inderjit, 2006 ) . Therefore, new schemes to contend phytopathogens have to be explored ( Moffat, 2001 ) . Keeping in position the strong phytochemical potency of C. reflexa, this survey has been carried out to measure fungicidal activity against of import infective fungous species with the ultimate purpose of developing works based preparations for works disease direction.
Material and Methods
Plant stuff and extraction
Fresh works stuff of C. reflexa ( parasitizing Bougainvillea glabra L. ) , turning in the premises of University of the Punjab, was collected. Plant stuff was washed exhaustively under running tap H2O, dried with blotting paper and cut into little pieces. A 50 % ( w/v ) aqueous infusion was prepared by homogenising works stuff ( 50 g ) in a liquidizer with 100ml sterile distilled H2O for 5 min and allowed to stand for 24 hours. The mixture was filtered through muslin fabric and so centrifuged at 4000 g for 30 min ; the supernatant was filtered through Whatman No.1 filter paper to take the dust. Three concentrations ( 10, 20 and 30 % ) were prepared by dilution of stock solution. Each concentration was simmered at 40o C for 15 proceedingss, cooled and stored at 4 & A ; Acirc ; & A ; deg ; C. To avoid taint and prospective chemical alteration, the infusions were used within 2-3 yearss.
Pathogenic fungal isolates viz. , Alternaria surrogate, Aspergillus Niger, Fusarium solani, Fusarium oxysporium and Macrophomina phaseolina were obtained from the First Fungal Culture Bank of Pakistan, University of Punjab, Lahore, Pakistan. Fungal civilizations were maintained on 2 % Malt infusion agar ( MEA ) medium at 25 & A ; Acirc ; ±2o C for seven yearss.
Antifungal Assay- Well Diffusion Technique
Three concentrations ( 10, 20 and 30 % ) were evaluated against five different fungal pathogens. Antifungal activity of C. reflexa infusion was assayed by good diffusion method ( Rani et al. , 2008 ) with some alteration. For fungicidal check, inoculant phonograph record ( 10mm ) was prepared aseptically from seven yearss old civilization. Wells ( 10mm in diameter ) were made on 2 % MEA plates utilizing a unfertile glass tubing. Each home base contained three Wellss, equally distributed around the inoculant phonograph record of trial Fungi that was placed aseptically at the centre. For each trial concentration, 100 & A ; Acirc ; µL infusion was poured into each well. Each intervention was replicated thrice. For control, sterile distilled H2O was filled in Wellss. The home bases were incubated at 25 & A ; Acirc ; ±2o C for 6 yearss. The clear zone environing each good indicated suppression activity of C. reflexa. The existent diameter of zone of suppression was got after deducting the diameter of the well. All the information was analyzed statistically.
Consequences and Discussions
Antifungal activity of C. reflexa was analyzed by good diffusion check against five infective Fungi. The consequences are shown in Fig1 and Fig 2. The size of the zone of suppression is normally related to the degree of antimicrobic activity nowadays in the sample or merchandise – a larger zone of suppression normally means that the disinfectant is more powerful. Consequences showed that C. reflexa infusion was significantly effectual against all trial Fungi. The C. reflexa infusion exhibited variable grade of suppression with regard to each trial Fungi. C. reflexa infusion was more active against M. phaseolina, F. solani and F. oxysporium and was least effectual A. Niger ( Fig 1 ) . It had been observed that at 10 % ( w/v ) , C. reflexa infusion showed least anifungal activity. C. reflexa infusion at 20 % and 30 % , showed important suppression potency against M. phaseolina, F. solani and F. oxysporium but least effectual against A. surrogate and A. niger. Results besides indicated that size of the zone of suppression was straight relative to the concentration of C. reflexa infusion. Surveies have reported turning involvement in antimicrobic potency of C. reflexa ( Loffler et al. , 1997 ; Da-Nian Qin et al. , 2000 ) . Antifungal ( Mohammad et al. , 1984 ) and insecticidal ( Chavan et al. , 1982 ) consequence of C. reflexa has been studied. The methanol infusion of C. reflexa and Corchorus olitorius showed a wide spectrum of antibacterial activity ( Pal et al. , 2006 ) . However, in instance of F. moniliforme, C. reflexa extract inhibit fungous growing merely up to 25 % ( Yasmin et al. , 2008 ) . This survey showed that C. reflexa contains strong fungicidal potency, exercising strong suppression against the growing of some infective Fungis. It is concluded that C. reflexa infusion can be helpful in developing a strong plant-based preparations for works disease direction. The certification of the fungicidal value of this ill-famed and quickly turning works brings a batch of alleviation to the works preservation community, which is concerned about its consequence on other biodiversity. In consequence, this works has potency of being exploited for its fungicidal potency and hence commanding its land screen in the universe at big.