PALAMUR UENSE PU1 ^T AND METHODS THEREOF

TECHNICAL FIELD

[0001] The present invention generally relates to biofertihzers. More particularly, the present invention is related to biofertilizer compositions and methods thereof, for enhancing plant growth, wherein the composition comprises of a culture of Chryseomicrobium palamuruense PU1 ^T .

BACKGROUND

[0002] A fertilizer is any material, which can be of natural or synthetic origin that is applied to soils or to plant tissues for supplying plant nutrients essential for the growth of plants and for promoting plant growth. Broadly, fertilizers are classified as chemical fertilizers and organic fertilizers. Chemical fertilizers are composed of raw chemicals manufactured at a factory in both liquid and solid forms and specifically target plants' nutritional needs. In addition, a chemical fertilizer is fundamentally premeditated to mimic naturally occurring nutrients. Organic fertilizers are derived from natural materials through minimal processing and are normally applied in their original form. In recent years, with increase in emphasis about strengthening of environmental awareness and improving the quality of life, biological fertilizers have emerged as a class of efficient and green fertilizers since they do not pollute the environment.

[0003] A biofertilizer is a substance, which contains living microorganisms and application of the same to seeds, plant surfaces, or soil, colonizes the rhizosphere or the interior of the plant, and promotes growth by increasing the supply or availability of primary nutrients to the host plant. Biofertilizers supplement nutrients to the plants through the natural processes of nitrogen fixation, solubilizing phosphorus, and stimulating plant growth through the synthesis of growth- promoting substances. The microbial biofertilizers reinstate the soil's natural nutrient cycle and build soil organic matter. Based on several roles played by biofertilizers, scientific term coined for such beneficial bacteria is "plant-growth promoting rhizobacteria" (PGPR). Biofertilizers such as rhizobium, azotobacter, azospirilium and blue green algae (BGA) have been in use for a long time. Other types of bacteria, such as phosphate-solubilizing bacteria, are able to solubilize the insoluble phosphate from organic and inorganic phosphate sources.

[0004] Recently, there has been much debate concerning the benefits and disadvantages associated with chemical fertilizer in the fields, garden, or yard. Most of the chemical fertilizers are highly acidic in nature, which in turn increases the acidity of the soil, thereby reducing the population of beneficial organisms and exploiting plant growth. Long-term use of chemical fertilizers can eventually lead to ecological disturbance and create a chemical imbalance in the recipient plants. Chemical fertilizer need moderate application and increased quantities can easily "burn" the plants and sometimes even kill them. In addition, they contribute to groundwater pollution, and the process of manufacturing them is costly and they release greenhouse gases and other pollutants into the environment as well.

[0005] In the light of aforementioned discussion, there exists a need for a solution for imparting better soil nutrition in terms of development of natural, eco-friendly, and microbial based biofertilizer composition that can be employed as a bioinoculant for enhancing plant growth. The present invention discloses a biofertilizer composition comprising culture of Chryseomicrobium palamuruense PU1 along with an agriculturally acceptable carrier. The present invention further discloses methods of preparation of the biofertilizer composition for enhancing plant growth and the methods for application of the same to seed, seedling, plant, soil environment, and potting mixture. Chryseomicrobium palamuruense PU1 could be as used as a PGPR in bio formulation in poor soils that support least plant growth. Chryseomicrobium palamuruense PU1 possesses the potential to enhance commercial crops' growth with respect to physical, phytohormone production and biochemical characteristics.

BRIEF SUMMARY

[0006] The following presents a simplified summary of the disclosure in order to provide a basic understanding to the reader. This summary is not an extensive overview of the disclosure and it does not identify key/critical elements of the invention or delineate the scope of the invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later. [0007] Exemplary embodiments of the present disclosure are directed towards a biofertilizer composition comprising a culture of Chryseomicrobium palamuruense PU1 that enhances plant growth. The culture has a predetermined concentration of viable cells. The biofertilizer composition also comprises of an agriculturally acceptable carrier.

[0008] Yet another exemplary embodiment of the present disclosure is directed towards a method of preparing the biofertilizer composition for enhancing plant growth. The method comprises of growing the culture of Chryseomicrobium palamuruense PU1 under predetermined growth conditions. The culture has a predetermined concentration of viable cells. As an optional method, an agriculturally acceptable carrier can be added to the culture to get the biofertilizer composition, wherein the biofertilizer composition enhances plant growth.

[0009] Yet another exemplary embodiment of the present disclosure is directed towards a method of application of the biofertilizer composition. The method comprises of the following steps: firstly, providing the biofertilizer composition comprising of the culture of Chryseomicrobium palamuruense PU1 . The culture has a predetermined concentration of viable cells. Secondly, contacting the biofertilizer composition with a seed, a seedling, a plant, a soil environment, and a potting mixture, wherein the biofertilizer composition enhances plant growth.

[0010] It is an object of the present invention to disclose a biofertilizer composition comprising of the culture of Chryseomicrobium palamuruense PU1 along with an agriculturally acceptable carrier. Chryseomicrobium palamuruense PU1 could be as used as a PGPR in bioformulation in poor soils that support least plant growth and can be applied to a variety of crops, seeds, seedlings, plants, soil environment, and potting mixture. In the present invention, the biotechnological potential of Chryseomicrobium palamuruense PU1 as a biofertilizers, with respect to their plant growth promoting properties as efficient Phosphate solubilizing bacteria (PSB) has been explored by testing its potential on different crops.

BRIEF DESCRIPTION OF DRAWINGS

[0011] Other objects and advantages of the present invention will become apparent to those skilled in the art upon reading the following detailed description of the preferred embodiments, in conjunction with the accompanying drawings, wherein like reference numerals have been used to designate like elements, and wherein:

[0012] FIG. 1 is an image showing electron micrograph of negatively stained cells of Chryseomicrobium palamuruense PU1 (Bar, 0.5 μπι).

[0013] FIG. 2 is an image showing eight different cultivars of cotton inoculated with Chryseomicrobium palamuruense PU1 , in accordance with a non limiting exemplary embodiment of the present disclosure.

DETAILED DESCRIPTION

[0014] It is to be understood that the present disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.

[0015] The use of "including", "comprising" or "having" and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The terms "a" and "an" herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. Further, the use of terms "first", "second", and "third", and the like, herein do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.

[0016] According to different non limiting exemplary embodiments of the present disclosure, biofertilizer compositions that enhance plant growth are disclosed.

[0017] In accordance with different non limiting exemplary embodiments of the present disclosure, methods for preparing and applying biofertilizer compositions that enhance plant growth are disclosed. [0018] Referring to Figure 1, it is an image showing electron micrograph of negatively stained cells of Chryseomicrobium palamuruense PU1 (Bar, 0.5 μπι).

[0019] Referring to Figure 2, it is an image showing eight different cultivars of cotton inoculated with Chryseomicrobium palamuruense PU1 , in accordance with a non limiting exemplary embodiment of the present disclosure.

[0020] The species Chryseomicrobium palamuruense PU1 (accession No FN555708) is deposited with an identification reference of NBRC 106750 (Mar 5, 2010) at Culture Collection Division, Biological Resource Center, National Institute of Technology and Evaluation, Address:2-5-8 Kazusakamatari, Kisarazu-shi, Chiba, 292-0818 JAPAN which is identified as IDA under Budapest Treaty for the purpose of patent procedure. This species is also deposited at Culture Collection, University of Goteborg, Sweden with an identification reference of CCUG 59101 , at Japan Collection of Microorganism, Microbe Division / Japan Collection of Microorganisms, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan with an identification reference of JCM 16712 (18 Jan 2011), and at Korean Collection for Type Cultures, Biological Resource Center (BRC) Korea Research Institute of Bioscience and Biotechnology, KRIBB) with an identification reference of KCTC 13722 ^T (18 Jan 2010).

[0021] According to a non limiting exemplary embodiment of the present disclosure, a biofertilizer composition comprising a culture of Chryseomicrobium palamuruense PU1 . In a particular embodiment, the culture of Chryseomicrobium palamuruense PU1 with an accession no FN555708 is used in the biofertilizer composition. In the present invention, novel and efficient Chryseomicrobium palamuruense PU1 species was isolated from cotton rhizosphere soil nearby boy's hostel drainage in Palamuru University, Mahabubnagar district, Telangana, India. The species is described as Chryseomicrobium palamuruense (pa.la.mu.ren'se. N.L. masc. adj. palamuruense, of or belonging to Palamuru). Cells are gram-positive, motile with single mono polar flagellum, rod-shaped (0.6-0.7 μπι in width and 1.6-2 μπι in length) (FIGURE 1) and occur singly and multiply by binary fission. Colonies on nutrient agar are circular, 1-2 mm in diameter, smooth, cream in color, opaque, crateri form and entire. Cells grow from 18 to 40°C with an optimum temperature of 37 °C and tolerate up to 9.0 % NaCl (w/v). Growth occurs in a pH range of 7 to 10. Cells of the strain PU1 are positive for catalase, oxidase, phosphatase, lipase and urease and negative for gelatinase, amylase, protease, cellulase, lysine decarboxylase and ornithine decarboxylase. The fatty acids were dominated by saturated fatty acids (82.7 %), with a high abundance of iso-Cis _{: 0} (48.8 %), anteiso-Ci _{5 : 0} (7.3 %), iso-Ci _{6 :} o (11-9 %), C _{16 :} \ ic alcohol (11.9 %) and iso-C _{17 :} o (5.3 %). Chryseomicrobium palamuruense PU1 contains MK-8 as the major respiratory quinone and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine make up the phospholipid composition. The cell wall peptidoglycan contains meso-diaminopimelic acid as the diamino acid and cell wall sugars are D-glucose and D-galactose. The G + C content of DNA of the strain PU1 ^T is 48.5 mol%. The culture has a predetermined concentration of viable cells. The biofertilizer composition was found to enhance plant growth.

[0022] In accordance with a non limiting exemplary embodiment of the present disclosure, the biofertilizer composition comprises of a culture of Chryseomicrobium palamuruense PU1 and an agriculturally acceptable carrier.

[0023] According to a non limiting exemplary embodiment of the present disclosure, the culture of Chryseomicrobium palamuruense PU1 could be either a liquid suspension or a freeze dried powder.

[0024] In accordance with a non limiting exemplary embodiment of the present disclosure, the carrier could be either of the following: peat, clay, soil material, mineral, vermiculite, polymer, fertilizer, agriculturally compatible oil and mixtures thereof. Any other suitable carrier material known in the art that is agriculturally compatible and that would not reduce the viability of the cells can be used without limiting the scope of the present disclosure.

[0025] According to a non limiting exemplary embodiment of the present disclosure, the biofertilizer composition could be used in the form of powder, granules, liquid, semi-solid, suspension, emulsion, spray or any other suitable form that is known in the art without limiting the scope of the present disclosure.

[0026] In accordance with a non limiting exemplary embodiment of the present disclosure, the biofertilizer composition could be applied as a seed coating or applied to either soil environment or the potting mixture. [0027] In accordance with a non limiting exemplary embodiment of the present disclosure, a method of preparing a biofertilizer composition for enhancing plant growth is disclosed. The method involves growing a culture of Chryseomicrobium palamuruense PU1 under predetermined growth conditions, wherein the culture comprises of a specific concentration of viable cells. In a particular embodiment, the culture of Chryseomicrobium palamuruense PU1 with an accession no FN555708 is used for preparing the biofertilizer composition. This culture can be used on its own as a biofertilizer composition or can be mixed further with an agriculturally acceptable carrier to get the biofertilizer composition.

[0028] According to a non limiting exemplary embodiment of the present disclosure, the culture is further freeze dried as a powder before mixing with the carrier, and the powder has viable cells.

[0029] According to a non limiting exemplary embodiment of the present disclosure, a method of application of the biofertilizer composition comprising of the following steps is disclosed: a) Providing the biofertilizer composition, wherein the composition comprises of a culture of Chryseomicrobium palamuruense PU1 and the culture has a predetermined concentration of viable cells; and b) Contacting the biofertilizer composition with seed, seedling, plant, soil environment or the potting mixture. In a particular embodiment, the culture of Chryseomicrobium palamuruense PU1 with an accession no FN555708 is used in the biofertilizer composition.

[0030] In accordance with a non limiting exemplary embodiment of the present disclosure, the biofertilizer composition further comprises of an agriculturally acceptable carrier.

[0031] In accordance with a non limiting exemplary embodiment of the present disclosure, the biofertilizer composition can be used in the form of powder, granules, liquid, semi-solid, suspension, emulsion or spray. Examples Sample collection

[0032] According to a non-limiting exemplary embodiment of the present disclosure, all the experiments were carried out in the Department of Microbiology, Palamuru University, Mahabubnagar, Telangana, India. Cotton rhizosphere soil sample was collected nearby boy's hostel drainage of Palamuru University and screened for Chryseomicrobium palamuruense PU1 by serial dilution of the sample and dilution of 10 ^"4 was spread on Soya bean tripticase agar.

Preparation of standard inoculums of Chryseomicrobium palamuruense PU1 for the use as biofertilizer and seed preparation

[0033] In accordance with a non limiting exemplary embodiment of the present disclosure, inoculums of Chryseomicrobium palamuruense PU1 were prepared in the selective medium. Typically, 150 mL broth medium was inoculated in 500 mL conical flask and incubated at 28°C under shaking at 100-150 rpm for three days to give an optical density of 0.5. Broth culture of Chryseomicrobium palamuruense PU1 sp was inoculated to peat (100 mL per kg of peat) which was sterilized at 121 °C and 15 psi pressure for one hour. Peat based Chryseomicrobium palamuruense inoculums were incubated at 28°C by adding 10% sugar solution for 3-4 days to increase the population up to 10 8 CFU mL - ^" 1 . Chryseomicrobium palamuruense inoculations having @ 10 MPN bacterial cells per gram of peat were applied to Cajanus cajan, Sesbania grandiflora, Gliricidia maculata, Tomato, Arachis hypogea, Dolichas lablab, and Gossypium, as seed coating for a period of 90 days in sterilized soil.

[0034] Approximately 30-31 seeds of crops were surface sterilized in 5% NaOCl for 1 min and washed three times in SDW. Dry seeds were immersed in each bacterial suspension, and the preparation was stirred frequently for 5 min. The treated seeds were spread on a petri dish and air-dried overnight at room temperature. The number of bacterial cells per seed, determined via serial dilutions, was approximately 10 CFU/seed for a period of 90 days in sterilized soil. Preparation of sterilized soil

[0035] According to a non-limiting exemplary embodiment of the present disclosure, 1: 1 ratio of sand and red soil were mixed properly and sterilized in autoclave at 121°C, 15 lbs pressure. X

Production of IAA from Chryseomicrobium palamuruense PU1

[0036] According to a non-limiting exemplary embodiment of the present disclosure, indole acetic acid (IAA) production was detected as described by Brick et al. (1991). Chryseomicrobium palamuruense PU1 culture was grown for 48 h on the respective media at 36 °C. Well grown cultures were centrifuged at 3000 rpm for 30 min. The supernatant (2ml) was mixed with two drops of orthophosphoric acid and 4 ml of the Salkowski reagent (50ml, 35% of perchloric acid, 1 ml 0.5M FeC13 solution). Development of pink color indicates IAA production.

Siderophore production from Chryseomicrobium palamuruense PU1

[0037] In accordance with a non limiting exemplary embodiment of the present disclosure, siderophore production was detected based on method by Schwyn and Neilands (1987) using blue agar plates containing the dye chrom azurol S (CAS). An orange halo around the colony is indicative for siderophore production.

Phosphate solubilization of Chryseomicrobium palamuruense PU1

[0038] According to a non-limiting exemplary embodiment of the present disclosure, for phosphate solubilization assay, a medium containing 2 g yeast extract, 20 g glucose, 2 g tri calcium phosphate, 60 mg actidione, and 15 g agar mixed with 1000 ml water, adjusted to pH 7, was used. A loopful inoculum of strain Chryseomicrobium palamuruense PU1 was placed in the center of petri dishes containing the media and incubated at 28°C for 5 days. Bacterial colony forming clear zone was considered as phosphate solubilizer (Rosas et al. 2006).

Extraction and analysis of total protein

[0039] In accordance with a non limiting exemplary embodiment of the present disclosure, the total protein was extracted by homogenizing 0.5 g plant tissue in 10 ml of 0.2 M perchloric acid. The homogenate was centrifuged at 5000 g for 10 min at 24°C. Ethanol-ether-chloroform (2:2: 1; v/v/v) solvent mixture was used twice for the extraction of the pellet. To the residue, 0.2 M NaOH was added and left overnight. The supernatant was used for total protein estimation (Lowry et al. 1951). Estimation of sugars

[0040] According to a non-limiting exemplary embodiment of the present disclosure, total soluble sugar was analyzed by heating 1 g of the plant tissue with 0.2% anthrone reagent and reading the intensity at 625 nm using UV-VIS spectrophotometer (Spectronic D20) (Mahadevan and Shridhar 1986).

Estimation of proline

[0041] In accordance with a non limiting exemplary embodiment of the present disclosure, proline estimation was carried out as described by Bates et al. (1973). Fresh mass of 0.5 g leaf tissue was taken and homogenized in 5 ml of 3% (w/v) sulphosalicylic acid. The residue was removed by centrifugation at 5000 rpm for 10 min and the supernatant was filtered through Whatman # 2 filter paper. The filtrate was mixed with an equal volume of ninhydrin and glacial acetic acid and incubated at 95 °C for 1 h. The reaction was terminated by placing in an ice bath for about 30 min and then extracted with 4 ml toluene by mixing vigorously for 15 sec. The toluene phase containing the chromophore was aspirated, warmed to room temperature for 10 min and the proline content was determined colorimetrically and expressed in mg/g.

Estimation and extraction of chlorophyll

[0042] According to a non-limiting exemplary embodiment of the present disclosure, chlorophyll pigment was extracted from 1 g of cotton leaves inoculated with Chryseomicrobium palamuruense PU1 in 80% acetone and estimated as described by Harborne (1973). The extracts were filtered in dark. O.D values of filtrate were measured at 650 nm and 663 nm in UV-VIS spectrophotometer. The amount of total chlorophyll was calculated by Arnon's formula.

Estimation of IAA

[0043] In accordance with a non limiting exemplary embodiment of the present disclosure, lgram of leaf sample was crushed with 1ml of phosphate buffer. Samples were centrifuged and 2 drops of perchloric acid were added to the supernatant to make up the volume to 2 ml with Salkowaski reagent (i.e. 2% 0.5 M FeC13 in 35% perchloric acid). O.D values were taken after 25 min at 530 nm by UV- Visible spectrophotometer. Standard graph was prepared by plotting concentration of IAA in micrograms/ml Vs Optical Density at 530 nm. Collection of soil samples from different agricultural fields of cotton

[0044] According to a non limiting exemplary embodiment of the present disclosure, four different soil samples were collected from cotton fields of Mahabubnagar district viz., shallow black soil from Malleboinpally, red soil from Makthal, deep black soil from Kalwakurthy, and sandy soil from Narayanpet.

Physico-chemical characteristics of soil samples of agricultural fields of cotton

[0045] In accordance with a non limiting exemplary embodiment of the present disclosure, soil available nitrogen was estimated by alkaline potassium permanganate method (Subbiah and Asija 1956), available phosphorous was determined by Bray and Kurtz (1945) and potassium determined by flame photometrically (Jackson 1973).

Statistical Analysis

[0046] According to a non-limiting exemplary embodiment of the present disclosure, statistical analysis was performed using a statistical software SPSS for Windows version 17.0. ANOVA, t- test was applied on subjects to know the significance of multiple mean differences and mean difference respectively. P-values are significant at 5% level (or 95 confidence level).

Experiments on efficacy of the biofertilizer composition on different crops

[0047] According to a non limiting exemplary embodiment of the present disclosure, the efficacy of Chryseomicrobium palamuruense PU1 as a plant growth promoting rhizobacteria (PGPR) and bioformulant for few commercial crops including Cajanus cajan, Sesbania grandiflora, Gliricidia maculate, Tomato, Arachis hypogea, Dolichas lablab, and Gossypium was tested. Four soil samples that were known to be poor for plant growth were collected. These soil samples were inoculated with Chryseomicrobium palamuruense PU1 and uninoculated sample was taken as control. All the commercial crops as mentioned above were grown in these four different soils for a period of 90 days in order to see the efficiency of novel isolate in natural conditions. The growth of commercial crops inoculated with Chryseomicrobium palamuruense PU1 strain was much more beyond than the samples without inoculation (control) with the Chryseomicrobium palamuruense PU1 (physical growth parameters, phytohormone production and biochemical characteristics). Based on the data provided in Tables 1 and 2, it is apparent that Chryseomicrobium palamuruense PU1 could be as used as a PGPR in bio formulation in poor soils that support least plant growth. Table 1 reveals increase in height of the plant, plant fresh weight and plant dry weight with respect to shoot and root in 7 cultivars in the presence of Chryseomicrobium palamuruense PU1 as compared to control. Table 2 represents phytohormone and biochemical production of cultivars (60 days old) inoculated with Chryseomicrobium palamuruense PU1 . Table 3 represents physico-chemical characteristics, available Sulphur, and micronutrients content of four agricultural soils of Mahabubnagar district (NPK in kg/hec). Table 4 represents plant growth promoting characteristics of Chryseomicrobium palamuruense PU1 . It can be seen from the table that Chryseomicrobium palamuruense PU1 was tested as positive for Phosphate Solubilization test, IAA production, and Siderophore production.

[0048] In accordance with a non limiting exemplary embodiment of the present disclosure, the Chryseomicrobium palamuruense PU1 was tested for plant growth promoting characteristics in cotton (Gossypium) plant. Eight cotton cultivars showing maximum growth with Chryseomicrobium palamuruense PU1 (FIGURE 2) were analyzed for biochemical characteristics and phytohormone production. Among eight cultivars of cotton, Mahyco exhibited maximum levels of IAA, proteins sugars, chlorophyll, and minimum content of proline; whereas Sunny (NCS-108) showed minimum levels of IAA, proteins, sugars chlorophyll and high level of proline. Table 5 represents production of phytohormone and biochemical production of eight cotton cultivars (60 days old) inoculated with Chryseomicrobium palamuruense PU1 .

[0049] According to a non limiting exemplary embodiment of the present disclosure, efficacy of Chryseomicrobium palamuruense PU1 as a PGPR and bioformulant was tested in Mahyco cotton. Four different soil samples that were known to be poor for plant growth were collected and inoculated with Chryseomicrobium palamuruense PU1 and uninoculated sample was taken as control. Mahyco cultivar was grown in these four different soils for a period of 90 days in order to test the efficiency of Chryseomicrobium palamuruense PU1 in natural conditions. Mahyco growth (physical growth parameters, phytohormone production, and biochemical characteristics) was found to be good in all soil types, but maximum growth was recorded in deep black soil followed by shallow black soil and minimum growth was observed in sandy soil followed by red soil. Table 6 represents plant growth parameters, phytohormone production and biochemical characteristics of Mahyco cotton cultivar in four different agricultural soils of Mahabubnagar district. Table 7 also represents phytohormone production and biochemical characteristics of Mahyco cotton cultivar in four different agricultural soils of Mahabubnagar district.

Table 1: Plant growth parameters of cultivars inoculated with Chryseomicrobium palamuruense

p- Chryseomicrobium palamuruense PUl ; ^s - Shoot; ^R -Root; '-Significant at 0.05 Table 2: Phytohormone and biochemical production of cultivars (60 days old) inoculated with

Chryseomicrobium palamuruense PU1 ^T .

Highly Significant, * significant Table 3: Physico-chemical characteristics and available sulphur and micronutrients content of four agricultural soils of Mahabubnagar dist (NPK in kg/hec), (Sulphur and micronutrients in ppm).

Table 4: Plant growth promoting characteristics of Chryseomicrobium palamuruense

Table 5: Phytohormone and biochemical production of eight cotton cultivars (60 days old) inoculated with Chryseomicrobium palamuruense PU1 ^T .

Highly Significant, * significant

Table 6: Plant growth parameters of Mahyco cotton cultivar inoculated with Chryseomicrob palamuruense PU1 ^T in four different agricultural soils of Mahabubnagar district.

'-Mahyco cotton cultivar ^{B P} - Chryseomicrobium palamuruense PUl ^s - Shoot ^R -Root '-Significant at

0.05

Table 7: Phytohormone production and biochemical characteristics of Mahyco cotton cultivar in four different agricultural soils (with Mean +SE) of Mahabubnagar district.

Significant at 0.05

[0050] Although the present disclosure has been described in terms of certain preferred embodiments and illustrations thereof, other embodiments and modifications to preferred embodiments may be possible that are within the principles and spirit of the invention. The above descriptions and figures are therefore to be regarded as illustrative and not restrictive.

[0051] Thus the scope of the present disclosure is defined by the appended claims and includes both combinations and sub combinations of the various features described herein above as well as variations and modifications thereof, which would occur to persons skilled in the art upon reading the foregoing description.