Absract Archive   February 2009 Research article Analysis of Microbial Pigment Productivety In Pseudomonas aerugionsa and Bacillus subtilis. Abstract Nature is rich in colour, and pigment producing microorganisms (Fungi, Yeast, and Bacteria) are quit common. Among the molecules produced are carotenoids, melanins, flavins, quinines, and more specifically monascins violancein or indigo. In the present study the microbial pigment production estimated on Pseudomonas aerugionsa and Bacillus subtilis. The test organism such as Pseudomonas aerugionsa and Bacillus subtilis were got from M.T.C.C Microbial Type Culture Collection Centre from Chandigar, Punjab. The Pseudomonas aerugionsa and Bacillus subtilis were inoculated into sterile nutrient broth and molasses. After the incubation pigments were analyzed from the fermentative broth. After fermentation microbial pigments were extracted and estimated. Maximum pigments production was noted in the Pseudomonas aerugionsa (0.18) compared the Bacillus subtilis (0.15). Optimize the pigments production by using various growth factors such as ph, temperature, carbon and nitrogen sources. Among the study of optimum products note Ph (7-7.5), temperature (400c) and the nutrient sources are lactose and ammonium chloride used as carbon and nitrogen respectively. Finally concluded the Pseudomonas aerugionsa and Bacillus subtilis used for large scale pigment production. The microbial pigment associated with quality and sensory properties of food. Key Words: Microbial pigments, Pseudomonas aeruginosa, Bacillus subtils, Introduction Colour is the most pleasing attribute of any article; red colour exudes warmth increases pulse rate and respiration, whereas, blue or green colour suggests cool and peaceful environment and encourages relaxation. The colour associated with quality and sensory properties of food. The first characteristic perceived by our senses is appearance or colour of food, which not only determines its appearance or colour of food, which not only determines its acceptance, its acceptance, but even helps in its recognition. Acceptability of colour in a given food, in turns, is influenced by cultural, geographical and sociological aspects of the population. Indeed, colour and other eating habits are viewed as a type of culinary anthropology and indigenous to specific region. However, certain food groups are acceptable only if they fall within certain range of colour gamuts and their acceptability is reinforced by their economic worth judged purely on the colour value. Authors:M. Muthuselvam,T. Sheeba,R. Rajasekaran.  Short communication QSAR Studies on the Action of an Algicide from a Cyanobacterium: Oscillatoria laetevirens, on Photosystem II Activity Abstract In this paper, we have discussed quantitative-structure-activity-relationship (QSAR) studies on the activity of an algicide from a cyanobacterium viz. Oscillatoria laetevirens, on photo system II activity.  For this purpose, we have used Ki values for inhibitors (mµ) for oscillatorin, atrazine, bentazon, diuron & metribuzing against Spinacia oleracea, Cassiatorra, Tephrosia purpuria, Alternathera Sessiles, Amaranthus viridis, and Parthenium hysterophorus and  modeled their photo system II activity using molecular descriptors usually called topological indices , Satisfactory results are obtained. KEYWORDS:  QSAR, Herbicide, Wiener Index Introduction An interesting achievement on QSAR studies was the effort to encode numerically molecule according to their structural features and to use these numbers in modeling the activity such as photo system II activity. The conversion of the structural formula into a numerical value, often called topological and graph-theoretical index, can be achieved in many ways1,2. It appears that among many topological indices that have been proposed so far, the Wiener index (W) is the first, oldest, and widely used topological index.  The same has been used3-5 in the present study for developing aforementioned quantitative structure-activity relationships (QSAR). Authors:Rahul Shrivastava Roma Sarkar,P. V. Khadikar,Aniruddha Chatterjee. Short communication Construction and analysis of cDNA library for tospovirus resistance genes Abstract In order to isolate a cDNA clone of tospovirus resistant R gene, a cDNA library was constructed in Uni-ZAP XL vector with poly(A) RNA purified from leaves of Lycopersicon peruvianum – EC 52071. The library showed around 40 fold more white plaques than blue plaques. To get the probe for screening the library, PCR of cDNA was conducted using the gene specific primers designed for a portion of the tospovirus resistance genes based on the already available sequence from the L. peruvianum Sw 5 gene.  The eluted 943 bp cDNA product obtained from RT-PCR using the above gene-specific primers were cloned in pTZ57R/T vector using TA cloning strategy and sequenced. The cDNA library was screened with the amplified fragment as a probe. Positive signals corresponding to around 150 plaques were seen in all 20 plates. All the 150 positive plaques were subjected to one more round of screening by replating and rehybridizing with the same probe. The presence of cDNA of interest in plaques selected at random were confirmed by PCR amplification. The cDNA library constructed using the ZAP Express® cDNA synthesis kit is a source for many genes in tomato. The library can also be utilized for isolation of other resistant genes and also of genes of agronomic importance.  Keywords: Tospovirus, cDNA library, L. peruvianum, resistance, RT-PCR. Introduction Tomato spotted wilt disease is one of the important virus diseases affecting tomato (Lycopersicon esculentum Mill). The causal agent, Tomato spotted wilt virus (TSWV) causes serious losses in the yield of groundnut and many other crops in Australia, India, Nepal, China, Thailand and USA (Reddy, 1985). Griep et al, 2000 estimated an annual crop loss due to TSWV at over one billion US dollars, which ranks TSWV among the top ten of the most damaging plant viruses in the world. Singh and Tripathi (1991) recorded a loss of 1.26 t in fruit yield per hectare due to infection by TSWV in tomato. The disease was first reported in Southern Australia and is now widespread in temperate and subtropical regions throughout the world (Brittlebank, 1919). Tospoviruses belong to the family Bunyaviridae (Van Regenmortel et al., 2000; Moyer, 1999). It is vectored by four species of thrips in a persistant manner (Francki and Halta, 1981). Attempts made to control TSWV by vector control have met with only little success (Carter, 1973). Since new infection often depends on thrips migration in tomato fields (Paterson, 1987), it was recognized that host resistance offered the best long term solution through development of resistant cultivars. Resistance to TSWV was reported to be available in some wild species of tomato. Many attempts have been made to breed commercial tomato varieties resistant to TSWV utilizing the wild species but met with limited success. The most effective way to manage tospoviruses is through genetic engineering. This is especially useful in crops where natural sources of resistance are not available or if available, are not amenable to transfer into agronomically superior cultivars by conventional breeding methods (Pappu, 1997). The wild species L. peruvianum has been reported to have broad resistance to different  isolates of TSWV (Smith, 1944). Gilbert and Tanaka (1971) released ‘Anahu’ as a TSWV resistant tomato cultivar that had L. peruvianum in its background. Authors:E. HEMAPRABHA ,R. BALASARASWATHI . Short communication Comparative study on efficiency  of genomic DNA Extraction methods in edible mushrooms Introduction White-rot fungi are a class of lignolytic basidiomycetes, capable of degrading lignin in wood. These fungi find their role in bioremediation, bioleaching and numerous other biotechnological applications (Bezalel et al.,1997). Pleurotus ostreatus is an edible basidiomycete of increasing biotechnological interest due to its ability to degrade both chemicals and wood related to lignin degradation products. Furthermore, this fungus produces secondary metabolites with pharmaceutical applications and some proteins of potential industrial use (Bobek et al.1993, Bobek and Ozdin 1994). Agaricus bisporus is also a commercially important edible mushroom. It is a secondary homothallic mushroom with intramictic breeding behavior and low level of genetic recombination (Raper, 1972, Fredricks, 2005). This cultivated species accounts for 90% of mushroom production in India and is a potential indoor crop for rural livelihood and health food. For molecular characterization of these economically important fungi, efficient genomic DNA extraction procedure is essential. Isolation of genomic DNA is an important step in any molecular biology technique. Even though many techniques are available for the isolation of genomic DNA, all have a common outline. The first step in the isolation of the genomic DNA is to disrupt the cellwall to release the cellular contents. Detergents like SDS or CTAB are used to denature the proteins. Other contaminants like  polysaccharides, fatty acids, cellwall materials are removed by precipitation followed by centrifugation. The DNA released in the supernatant is precipitated with ethanol or iso-propanol. The genomic DNA isolation methods used for plants are also employed to isolate DNA from  fungi with little modifications (Giardina et al. 1995). The efficiency of different methods used for the isolation  of DNA varies for different species of mushroom ( Zhang et al., 1996,Muller et al., 1998 and Melo et al., 1998) . The present study aims to find  a suitable method for the isolation of genomic DNA from edible mushrooms. 2. Materials and Method. Pleurotus ostreatus and Agaricus bisporus were obtained from commercially mushroom cultivation centers CTAB DNA extraction buffer (0.1 M Tris, 1% CTAB, 0.7 M NaCl, 1 mM EDTA and 1% 2-mercaptoethanol). SDS DNA extraction buffer (10 mM Tris Cl, 0.5 M NaCl, 1% SDS). 5 M Potassium acetate. Absolute ethanol TE buffer ( 10mM Tris-Cl, 1mM EDTA) Authors:Kabilan M, Arun, B. Saraswathy, N, Ramalingam  P.Rajasekaran. Short communication Effect of Azoreductase on the degradation of Carcinogenic Azo Dyes Abstract A total of four bacterial strains such as Bacillus, Klebsilla, Pseudomonas and Staphylococcus spp. were isolated from textile effluents to induce the secretion of extra cellular azoreductase for the biological degradation of azo dyes. Azoreductase was further purified and assayed which revealed 14000, 22000 and 36000 KDa protein molecules. Kinetic studies of azoreductase by Pseudomonas spp. to determine the suitable physical conditions during the enzyme production were performed under in vitro condition. The results showed that the optimum pH and temperature was found to be 8 and 40°C; respectively. The degradation of the azo dyes was evaluated by a UV-VIS spectrophotometer and HPLC system which revealed a prominent OD value at 519.57 nm was recorded in the samples treated with enzyme. Three minor peaks were noted with the retention time of 11.15 min, 16.44 min and 19.25 min in the samples treated with the same enzyme whereas only one peak was noted in the control samples with a retention time of 3.59 min. In conclusion, as a preliminary step in the development of textile effluent biotreatment using indigenous microbes, we have discovered some strains with potency to decolourize and/or remove COD and BOD. Introduction Azo dyes are synthetic organic colourants that are characterized by great structural variety. There are thousand of synthetic azo dyes being used in the textile, pharmaceutical, cosmetic and food industries. It has been reported that more than 500 azo dyes are available in the market which contain potentially carcinogenic aromatic amines (Wataru et al., 2006). Synthetic azo dyes are extensively used in the textile industries due to inefficiencies of the industrial dyeing process, as 10-15% of the dyes are lost in effluents of textile units, rendering them highly coloured. These discharge of wastewaters and industrial effluents containing recalcitrant residues into river and lakes lead to higher biological oxygen demand causing serious threat to native aquatic life(McMullan et al., 2001). Further, these compounds enter into the human body through ingestion of food, dermal contact and inhalation. They are metabolized via azoreductase to aromatic amines by intestinal microflora and in mammalian liver (Chung et al., 1992). The elimination of coloured effluents in wastewaters is based mainly on physical and chemical methods. They suffer from high cost, formation of hazardous by products and intensive energy requirements(Thomas et al., 1982). Biological degradation of azo dyes does not have similar problems, literature suggest that there is a great potential for developing microbiological decolourization system with total colour removal by reductive cleavage of the azo bonds with the help of NADH dependent azoreductase (Chen et al., 2005).To keep in mind, the present study was undertaken to isolate microbial consortium especially bacterial strains like Pseudomonas, Bacillus, Klebsilla and Staphylococcus spp. from textile effluents. The isolated strains were subjected to produce azoreductase for decolourization of azo dyes. Authors:K. Suresh Kumar, P. Ponmurugan, S. Murugesh. Short communication In vitro propagation of hybanthus enneaspermus linn., f. Muell. From leaf explant. Abstract Leaf explant of Hybanthus enneaspermus L. gave rise to multiple shoots when cultured on MS medium supplemented with different concentrations of BAP and NAA. The highest rate of shoot multiplication was obtained in MS medium containing 4.0mgL-1 BAP and 0.5mgL-1 NAA (14.42±1.322a).  Differentiated shoot buds elongated to 4–5 cm within 4 weeks. The regenerated shootlets were rooted on half strength MS basal medium with different concentrations of IBA and NAA. The maximum number of roots was achieved on the medium containing 2.0mgL-1 NAA. In vitro regenerated plantlets were transferred on to plastic pots containing coco peat as potting mix and thereafter successfully established under ex vitro conditions. The survival percentage of transplanted plantlets was 60. Key words: Hybanthus enneaspermus, Medicinal plant, Regeneration, Plantlets. Abbreviations: MS-Murashige Skoog medium, NAA-Napthalene Acetic Acid, BAP-Benzyl Amino Purine, IBA-Indole Butric Acid.  Introduction Mass propagation of plant species through in vitro culture is one of the best and most successful examples of commercial application of plant tissue culture technology. Recently, there has been much progress in this technology for some medicinal plants. Tissue culture propagation and its importance in conservation of genetic resources and clonal improvement have been described by many workers; Barz et al., (1977), Datta and Datta (1985), Kukreja et al., (1989) and Jusekutty et al., (1993). Hybanthus enneaspermus (L.) F. Muell. (Violaceae) (Singh, 1988) known as Lakshmisheshta, Padmavati, Padmacharini or Purusharathna in Sanskrit, is an important plant in the Indian system of medicine. It is a small suffrutescent perennial herb and grows 15-30 cm in height with many diffuse or ascending branches and is pubescent in nature (Kirtikar and Basu, 1991). Traditionally the plant is used as an aphrodisiac, demulcent, tonic, diuretic, in urinary infections, diarrhea, leucorrhoea, dysuria, and sterility (Yoganarasimhan, 2000). The plant is also attributed to its antimicrobial and antiplasmodial action (Rajakaruna et al., 2002). Various phytoconstituents viz. dipeptide alkaloids, aurantiamide acetate, isoarborinol, and β-sitosterol have been isolated from different parts of this plant (Majumder et al., 1979; Prakash et al., 1999; Yoganarasimhan, 2000; Retnam and De Britto, 2003). Moreover, the plant is reported, in ancient ayurvedic literature, to cure conditions of “kapha” and “pitta”, urinary calculi, strangury, painful dysentery, vomiting, burning sensation, wandering of the mind, urethral discharges, blood troubles, asthma, epilepsy, cough, and to give tone to the breasts (Kirtikar and Basu, 1991). Authors:M. JOTHI BASU, N. YOGANANTH. Short communication Studies on Influence of  Sugar Mill waste water on growth of Rhizobium Abstract An investigation was carried out to study the influence of sugar mill effluent on growth of rhizobium.  For this study Azorhizobium was isolated from the sugar mill effluent polluted soil root nodules of Subanioa rostrata study sites.A pot trail was conducted to know the impact of sugar mill effluent on growth and nodulation  of rhizobium.  Nodule number, dry weight, total proten amino acid lipids and protease content s were reduced in effluent treated pots then untreated one. Key wards: Sugar mill effluent, Azorhizobium, Subanioa rostrata Introduction The sugar industry in India is playing an important role in the economic development of our country. More than 60% of the world’s production is from sugarcane. (World Bank,1995). The effluents are discharged during the production of sugar. They contain high polluted organic and inorganic wastes. Sugar industries and distilleries have been playing an important role in our social economy but their effluents have been guite obnoxious, creating a serious problem of water pollution. These effluents caused severe damage to the flora and fauna of the water bodies, if mixed with them. Sesbania is an aquatic species, which nodulate with Rhizobium. Sesbania produce nodule on both root and stem. Water loging was highly favorable for Sesbina root nodule development (Ramani et al., 1990). Azorhizobium is a unique genus havinga remarkable characteristic of producing photosynthetic green stem nodules, in addition to the root nodules in the host plant called Sesbania rostrata.  Which is present in waste water loging condition  also. Hence, a comprehensive study was undertaken to understand the impact of sugar mill wastewater on growth and nodulation on rhizobium. Author:.V.Mahesh.