14 International Journal of Research in Pure and Applied Microbiology 2013; 3(1): 14-16 ISSN 2277–3843 Original Article PHYSIOLOGICAL AND GROWTH PROMOTING CHARACTERISTICS OF PSEUDOMONAS PUTIDA ISOLATED FROM FOREST LITTER OF TIRUMALA HILL FOREST Prasada Babu Gundala*, JayaKumar, K and Paramageetham Chinthala *Department of Botany, Sri Venkateswara University, Tirupati - 517 502. Andhra Pradesh, India Department of Microbiology, Sri Venkateswara University, Tirupati - 517 502 Andhra Pradesh, India Corresponding Author E-mail: prag.babu@gmail.com Received 09 February 2013; accepted 25 February 2013 Abstract Pseudomonas putida was isolated from forest litter of Tirumala hills which have dense deciduous forest and a part in Seshachalam hill range. Based on phenotypic and physiological characteristics it was identified as P.putida. The isolate is able to promote plant growth either directly or indirectly by secreting enzymes and by producing growth promoting substances such as IAA , siderophore etc., © 2013 Universal Research Publications. All rights reserved KEY WORDS : P.putida, siderophore, Phosphate solubilizing ability, IAA,HCN production 1. INTRODUCTION The genus Pseudomonas has been heterogeneous since Migula first named it in 1894. Pseudomonads are gram – negative, strictly aerobic, polarly flagellated rods. Pseudomonads are ubiquitous soil microorganisms and common inhabitants of rhizosphere. Stainer et al.,(1966) (1) conducted a fundamental study on the Pseudomonas that result in an extensive phenotypic characterization in which the genus was subdivided into species and species groups. They are directly or indirectly involved in plant growth and productivity .The direct way of promotion of plant growth is by secreting plant growth promoting substances or facilitation of uptake of certain nutrients from the soil (2). In indirect way they also prevent the proliferation of phytopathogens and thereby support the plant growth (3). Among the Pseudomonads P.putida and P.florescence were identified as organisms with ability of effective disease suppression and plant growth promotion. Keeping the importance of the P.putida a study was conducted to isolate the P.putida from forest litter and characterized its physiological and plant growth promoting traits. 2. MATERIALS AND METHODS Soil samples were collected from forest litter of sacred Tirumala hills, which is a dense forest area of Andhra Pradesh state, India. Randomized block design was employed to collect the samples. Collected soils were sealed in sterile polyethylene bags. One gram soil from different sampling sites was placed in 9 ml of saline solution and incubated for 2 hours in an orbital shaking incubator at 180 rpm. Later a loop of the resulting bacterial suspension was spread plated on to King’s B Agar medium (4) and incubated at 37 0 C. After 2 days the colonies were screened for fluorescence under UV light (366nm). In order to identify the species Phenotypic characters like gram’s reaction, levan production, optimum growth temperature, fluorescence, Gelatin hydrolysis, Citrate utilization test, Oxidase, β- galactosidase activity, Catalase test, Indole production were conducted for the bacterial isolates. Substrate utilization profiles were tested using Hi carbohydrates (Hi media, Mumbai, India). Cell suspension was established in sterile saline using 24 hours grown culture. The density of the suspension was made to 0.5 O.D at 620 nm. An aliquot of 50µl of this suspension was inoculated at 30 0 C for 48 hrs and on to the substrates such as lactose, xylose, fructose, galactose, glycerol, trehalose, mannitol, sucrose, ribose, glucose and incubated at 30 0 C for 48 hrs. Functional properties of the isolates such as ability to secrete enzymes such as catalase, protease, gelatinase, cellulase and pectinase and plant growth promoting traits like phosphate solubilization , Siderophore production and HCN properties were also studied (5). Phosphate solubilizing enzyme production was assessed by using Available online at http://www.urpjournals.com International Journal of Research in Pure and Applied Microbiology Universal Research Publications. All rights reserved