utopia of global education€¦ · forest research institute, dehradun, u.k., india 3 department of...

S.R.S.D. Memorial Shiksha Shodh Sansthan, Agra, India

Keywords: Dhamani, Energy filed, Electromagnetic field, Prana, Pranic healing.

.

Key words: Pseudomonas fluorescence, Dalbergia sissoo, Solanum tuberosum, soil, bio-

fertilizer, Plant growth promoting rhizobecteria, plant, soil

Introduction

The soil acts as a reservoir for millions of microorganisms, out of which more than 85%

are beneficial for plant life. In particular, the varied genetic and functional activities of the

extensive microbial populations have a critical impact on soil functions, based on the fact that

microorganisms are driving forces for fundamental metabolic process involving specific enzyme

activities (Nannipieri, et al. 2003). Many microbial interactions, which are regulated by specific

molecules/ signals, are responsible for key environmental accesses such as the biogeochemical

cycling of nutrients and organic matter and the maintenance of plant health and soil quality

(Pace, 1997; Barea et al. 2004)

UTOPIA OF GLOBAL EDUCATION

A Peer Reviewed Refereed International Research Journal

ONLINE ISSN-2454-7387 Volume III, Issue I, June 2017

www.srsshodhsansthan.org

Studies on the effect of biofertilizer comprising of the plant growth promoting rhizobacterial Pseudomonas fluorescence strains on

leguminous and non leguminous plants height at different soil pH

Kalpana Sharma1, S.B. Singh

2, T.C. Pokhriyal

3

1 & 2 Department of Forest Soil and Land Reclamation Division

Forest Research institute, Dehradun, U.K., India 3 Department of Plant Physiology Division

Forest Research institute, Dehradun, U.K., India

Abstract

Soil fertility improvement is an important area which needs attention since most of the hazardous inputs

added into the forestry system are in the form of chemicals. Since plant growth and soil normal flora are

affected by input of chemical fertilizer. Bio-fertilizer microorganisms make the nutrients available to

plants. The present study was concerning to isolate of Pseudomonas fluorescence species that possess a

promising properties which make it a better bio-fertilizer bacteria. Pseudomonas fluorescence species

was isolated from rhizospheric soil of Dalbergia sissoo and Solanum tuberosum tested for soil fertility

and plant growth .Pseudomonas fluorescence directly inoculated with the soil pH and showed the

increased plant height. Pseudomonas fluorescence enhances the growth and role of the plant growth

promoting rhizobacteria. The concluded that plant growth promoting rhizobacteria (PGPR) can be used

as a good tool in the enhancement of plant height and alkaloid contents in medicinal plants and other

plants. As result, all tested isolates of Pseudomonas fluorescence have given strength to soil and

increased plant height. So it could be summarized that all Pseudomonas fluorescence isolated strains

can be used as bio-fertilizers for soil fertility improvement.

Vol. III, Issue I, June 2017 ONLINE ISSN-2454-7387

Studies on the effect of biofertilizer comprising …. 12 | P a g e

Many conditions of the effect soil micro-organism. Among the most important are the

supplies of oxygen, moisture, temperature, nature of the soil, organic matter, pH and the amount

of exchangeable calcium present. Brady (1996) has described the conditions in detail which

effect the growth of soil microorganism.

Soil pH

Soil pH generally reduces nodulation, acid soil may also be low in levels of available

calcium, magnesium, phosphorus, molybdenum (Munns, 1977) and all of these factors influence

nitrogen fixation. Nodule activity with respect to salinity has been reviewed extensively by

Zaharan and Sprent, (1986), who observed a decline in the colonization of root at both 50 and

100 mm of NaCl tested. They also concluded that early infection and nodulation makes them

better able to withstand stress. On the other hand 0.1% solutions of Na2CO3 or NaHCO3 also

caused significant reduction in nodulation.

Plant Growth promoting Rhizobacteria (PGPR)

The groups of organisms that colonize roots or rhizosphere in the crop roots are referred

to as plant growth promoting rhizobateria (PGPR). These microorganisms are beneficial to crops.

These PGPR are referred to as bio-stimulants and the phytohormones. They produce indole

acetic acid, cytokinins, gibberellines and inhibitors of ethylene production. During late 1980‟s

Kloepperwas the first to recognize plant growth promoting rhizobateria who told that these are

major player in crop production. Thus, science of PGPR is relatively young in comparison to

nitrogen fixing bacteria. In Kloepper, et al. (1989) reported that PGPR are beneficial to plant

growth.

Activities of PGPR

Pseudomonas, a PGPR enhances the plant growth productivity by production of

metabolites such as siderophores, HCN, chitin and -1, 3-gluconase, antibiotic production which

control the pathogenic microorganism in rhizosphere. Pseudomonades produceindole acetic acid

(IAA), Phosphate solubilates and vitamins by which plant root elongates and rhizosphere

bacterial population is also improved. The association of PGPR with the crop makes

beneficial effects on the roots zone. Major microbial form includes Azotobacter, Azospirillum,

Pseudomonas, Acetobacter, Burkholderiaand Bacilli (Kloepper, et al. 1989). PGPR influence

plant growth in two ways: i.e., (1) indirect and (2) direct. The direct growth promotion comprises

of providing the plants with a compound that is synthesized by the bacterium which facilitate the

uptake of certain nutrients from the environment.

The mechanism of PGPR to increase crop performance is not well understood. There are

several PGPR inoculants currently commercialized that seem to promote growth through at least

one mechanism; suppression of plant disease (termed bio-protectants), improved nutrient

acquisition (termed bio-fertilizers), or phytohormone production (termed bio-stimulants).



Development of inoculants (organisms capable of killing other organisms pathogenic or

disease causing to lower production of crops) has been most successful to deliver biological

agents of plant disease.

Effect of plant growth promoting rhizobateria

PGPR in agriculture

Two of the most important types of agriculture crops are cereals and legumes. Nitrogen

fixation contributes directly through symbiotic relationship to productivity of forage and grain

legumes. Less direct contribution may be made to the growth of rice by bacteria, cynobacteria,

azolla and the other graminae both grain and forage species by associative bacteria, free living

bacteria and cynobacteria. Legume inoculants as commercial products were in vogue in USA and

UK even 100 years ago and a wealth of understanding on biological nitrogen fixation process has

been developed as indicated by review (Becking, 1970).

PGPR in forestry

The use of PGPRs on tree species is gaining interest. Since the usual pathogens in

agricultural crops are also common to tree nurseries inoculation of PGPRs in forest tree nurseries

has proved to be crucial in enhancing survival of young seedlings when transplanted to the field.

Plant selected most bacteria that are more beneficial for their health by realizing organic

compounds through exudates (Lynch, 1990), creating a very selective environment where

diversity is low (Marilley and Aragno, 1999). Therefore, the rhizosphere of wild plants species

seems to be the best source to isolate plant growth promoting rhizobacteria (Lucus Garcia, et al.,

2001). Consequent rhizosphere of wild populations of Cistus ladanifer would be a good source

for putative PGPRs.

Pseudomonas

All species and strains of Pseudomonas are Gram negative rods. Fluorescent

Pseudomonas hasbeen implicated in suppressive soils by many researchers (Cook and Rovira,

1976; Thomashow and Weller, 1988; Baker, et al. 1990; Defago and Hass, 1990). Pseudomonas

are good candidates for biological control for the following reasons: (i) Many of the secondary

metabolites from Pseudomonas inhibit other microorganisms (ii) A higher production of

Pseudomonasare inhibitory to other microorganisms than is found with other soil bacteria

(Rovira and Ridge, 1973); (iii). Pseudomonas when introduced via seed coating, they can

dominate the rhizosphere population (iv) There can be extensive colonization of the

endorhizosphere by Pseudomonas (Van Peer, et al., 1990) Certain members of the Pseudomonas

genus have been applied directly to soils as a way of preventing the growth or establishment of

crop pathogens. This practice is known as biocontrol. The biocontrol properties of Pseudomonas

fluorescence strains CHAO or Pf-5 are currently best understood, although it is not clear exactly

how the plant growth promoting properties of Pseudomonas fluorescence are activated. Theories



include that the bacteria might induce systemic resistance in the host plant. So it can better resist

attack by the pathogens, the bacteria might out compete other (pathogenic) soil microbes, e.g., by

siderophores giving a competitive advantage at scavenging for iron; the bacteria might produce

compounds antagonistic to other soil microbes, such as phenazine type antibiotics or HCN.

There is an experimental evidence to support all of these theories, in certain conditions, a good

review of the topic is written by Haas and Defago, 2005.

Tracking of Pseudomonas in the rhizosphere, plant residues and soil

The use of mutants resistant to the antibiotics, rifampin and nalidixic acid, has

proven a powerful tool to study the survival, growth and spread of biocontrol Pseudomonas in

soil and on plants. Useful immuno-fluorescence colony- staining technique developed by

Leeman, et al., (1991). Uses polyclonal antiserum to a fluorescent Pseudomonas that detects as

few as colony forming units per gram of soil.

Variability in biological control with Pseudomonas

There is often considerable variability in the effectiveness of biological control, especially when

measured by yields rather than by disease (Cook and Baker, 1983). An example of the need to

understand the influence of soil is in the research of Stutz, et al. (1989) which showed that soils

actually suppressive to black root rot of tobacco were based on vermiculate clays, while

conductive soils were manly based on little clays. While conductive soils were mainly based on

little clays. They found that biological control with a Pseudomonas .However, biological,

chemical and physical means of control, including crop rotations or time showing, will not result

in growth or yield response unless condition favor the disease, e.g., cold and wet phythium root

rot, water stress for Fusarium, early showing and wet conditions for take all. These factors

should be taken in to account when analyzing results from field experiments on biological

control of root diseases.

Material and Method

Present study basically planned to assess the “Studies on the effect of biofertilizer

comprising of the plant growth promoting rhizobacterial Pseudomonas fluorescence strains

on leguminous and non leguminous plants height at different soil pH” required mainly

following materials:

1. Bacterial strain

2. Soils

3. Plant species

Bacterial strain

Two strains of bacteria isolated from the rhizospheric soils of Dalbergia sissoo(Ds) and

Solanum tuberosum (St) were used in present study. Those strains were isolated as below:



(i) Dalbergia sissoo bacterial strain

Pseudomonas fluorescence from the rhizospheric soil of Dalbergia sisssoo (Shisham)

was isolated in following manner: For isolation of Pseudomonas fluorescence from shisham, soil

from adjoining site of Forest Soil and Land Reclamation (FS&LR) Division, F.R.I. was

conducted and packed in polythene bag. Seeds of Dalbergia sissoo collected from Gorakhpur

were taken from seed testing laboratory, Silviculture Division F.R.I. and were shown on wet

filter papers in plastic tray for germination. After germination and attaining about one and half

centimeter height, seedlings were transplanted in collected soil and irrigated as per requirement.

After two months when plants established well, plants were uprooted, roots and shoots were

separated and kept in sterile polythene bags. 5-6 roots with rhizospheric soil were cut with the

help of scissor into the petri dish. 10 g of rhizosphericsoil was mixed with 100ml sterile water

and well shaked in a magnetic shaker for 15 minutes. One ml of above rhizospheric soil as

transferred in test tube containing 9 ml of distilled water in each test tube. 106 strength of

therhizospheric solution as prepared by serial dilution. Then one drop of diluted bacterial

suspension was spread on pre-sterilized Petri dish seeded with King‟s B medium under the

Laminar Air Flow. Plates were incubated at 28oC for 24 hours. Bacterial colonies appeared on

the surface of King‟s B medium. The fluorescent pigment produced by the bacteria was observed

under UV light. Pseudomonas from same source of Dalbergia sissoo was also acquired from

Plant Pathology Division, Forest Research Institute and compared with isolated in FS & LR

Division.

(ii) Solanumtuberosum bacterial strain

Pseudomonas fluorescence from the rhizospheric soil of Solanum tuberosum (potato)

was isolated in manner described below: For isolation of Pseudomonas florescence from potato,

soil from adjoining site of Forest Soil and Land Reclamation (FS&LR) Division, F.R.I. as

collected and packed in polythene bag. Solanum tuberosum (potato) was purchased from local

market. Potato seeds (Kufri Himsona) were shown in soil and irrigated as per requirement. After

one month when plant established well, potato plants were uprooted, roots and shoots were

separated, properly kept in sterile polythene bags. From rhizospheric soil 5-6 roots were cut with

the help of scissor, separated and kept in sterile petri dish. 10 g of rhizospheric soil was mixed

with 100 ml sterile water and well shaked in a magnetic shaker for 15 minutes. One ml of above

rhizospheric soilwas transferred in test tube containing 9 ml of distilled water in each test tube.

106

strength of rhizospheric solution was prepared by serial dilution. Then one drop of diluted

bacterial suspension was spread on pre sterilized petri dish seeded with King‟s B medium

(King‟s B medium‟ was prepared by dissolving following reagents in one litter of distilled water

as described by Dubey and Maheshwari, 2004.

Protease Peptone 20.0 g

Glycerine 15.0 ml

K2HPO4 2.5 g



MgSO4.7H2O 6.0 g

Agar 20.0 g

The medium was autoclaved at 120oC (15 lb) for 20 minutes. When the temperature of the

medium came down to about 60oC, 15 ml of the medium was dispensed into petri plate (10 cm

diameter) and allowed to solidify. Petri plates with medium were kept in the refrigerator for

future use) under the Laminar Air flow. Plates were incubated at 28oC for 4 hours. Then bacterial

colonies appeared on the surface of the Pseudomonas was also acquired from plant pathology,

Forest Research Institute and compared with present prepared strain.

Isolated strains were preserved in freezer for further use. Biochemical characterization of

isolated Pseudomonas fluorescence strains were done (Aneja, 2003). The both strains performed

equally in the tests. Results shown in Table- 3.

Soils

Following Soils from 0-17 cm depth was collected for the experimental purpose:

(i) Forest soil

Forest soil was collected from the Pinus roxbergii (Pine) plantation of Champion Block,

Forest Research Institute (FRI), Dehradun. Plantation is of about 70-years old that consists about

95% canopy cover and 6-8 cm organic residue on the surface. About 08 hectares of pine

plantation was covered for sample collection. Soil samples were collected from five different

places and mixed. Composite sample was brought to laboratory for experimental purpose.

(ii) Agriculture Soil

Agriculture soil was collected from the agriculture land of Bungalow No. 04, Canning

Road, Forest Research Institute. Paddy and wheat crop rotation is common on the site where

urea, single super phosphate and muriate of potash are generally applied in the soil to supplement

nitrogen, phosphorus and potash. Scattered use of Farm Yard Manure (FYM) is also practiced. In

the area of about of five hectares, soil samples were collected from four different places, mixed

and composites soil was brought to laboratory for the experiment.

(iii)Upland Soil

Upland site located in village Kaulagarh, Dehradun was selected to collect soil. Such site

spread over in about 15 ha in this village is occasionally used for agriculture under rain fed

condition. Bajara is commonly grown without application of fertilizer and manure. Four places

were chosen to collect soil sample. Soil collected from four different places was collected, mixed

and composite samples as brought to laboratory.



(iv) Grassland Soil

Soil was collected from where no crop is grown. Grasses, generally in rainy season, are

commonly seen in the area that dry during summer. Villagers use this site as pasture. Soils were

collected from three locations, mixed and composite sample was brought to laboratory. Farm

Yard Manure (FYM), collected from cow owner‟s farm, was mixed with each soil in 1:5 ratios

(by volume). Clods of soils and FYM were broken by wooden roller and mixed with fingers.

After well mixing, sample of each soil was analyzed for their physical and chemical properties

and sterilized at 15 lb (120oC) for one hour in laboratory of Forest Entomology Division, Forest

Research Institute (FRI), Dehradun and tightly packed.

Plant species

Keeping the suitability of plants for plantation in agro-forestry and wastelands, following four

plant species were selected for study:

(i) Acacia nilotica (ii) Albizia lebbeck

(ii) Terminalia arjuna (iv)Eucalyptus camaldulensis

(i)Acacia nilotica

Acacia nilotica is a leguminous plant species that fixes atmospheric nitrogen in the soil.

This species is often planted around the farmer‟s field. This species is suitable for plantation on

degraded soil also and require comparatively less soil moisture to survive. The leaves of this

species are used as fodder and bark in disease like cough, leprosy, dysentery and diarrhea. Root

of the plant is useful in tuberculosis. Wood is used for tool of agriculture, doors and Window of

houses and lumber of boats.

(ii)Albizia lebbeck

This species, suitable for alkaline soils, is leguminous that fixes atmospheric nitrogen in

the soil. The tree is used as an astringent, to treat boils, to treat the eye, flue, gingivitis and lungs

problems, as a tonic and to treat abdominal tumors. Having density of 0.55-0.66 g/cm3 or higher

the tree is also used as timber for making house doors, window.

(iii)Terminalia arjuna

Terminalia arjuna, suitable for waterlogged soils is a medicinal plant of the genus

Terminalia, widely praised & used by Ayurvedic physician for its curative properties in

organic/functional heart problems like angina, hypertension, deposits in arteries etc.

(iv)Eucalyptus camaldulensis

Eucalyptus camaldulensis suitable for saline and sodic soil is somewhat brittle and is

often cross-grained. Traditionally used in rot resistant applications like stumps, fence posts and



sleepers, more recently it has been recognized in craft furniture for its spectacular deep red

colour and typical fiddle back figure. It is quite hard, dense (about 900 kg/ m3), can take a fine

polish and carves well. It is a popular timber for wood turners, particularly if old and well-

seasoned. It is also popular for use as firewood. The wood makes fine charcoal and is

successfully used in Brazil for iron and steel production. In addition, this plant is use for

beekeeping in Brazil and Australia.

Raising of seedlings

Healthy seeds of selected plant species (Acacia nilotica, Albizia lebbeck, Terminalia arjuna and

Eucalypyus camaldulensis) were collected from Seed Testing Laboratory Forest Research

Institute (FRI), Dehradun and scattered on wet filter papers, placed in plastic tray. After seedling,

seeds were partially covered with sterilized sand mass and kept continuously wet for uniform and

healthy germination. Seedlings were allowed to grow on blotting paper for the period of 20 days.

After 20 days, seedlings growing on filter papers were separately transplanted in polythene bags

filled with 3.00 kg of sterilized forest, agriculture, upland and grassland soils collected as

described earlier.

Details of experiment and application of treatments

After establishment of transplanted seedlings in polythene bags, experiment as laid-out to

assess the effect of three treatments of pseudomonas fluorescence strains on four plant species in

four types of soil (Table-1).Experiment as conducted in Factorial Design with three replications

in polythene bags under green house condition.

Table-1: Details of treatments with their symbol are given below:

Factors Treatments Symbol

Psudomonas strain control C

Dalbergia sissoo Ds

Solanum tuberosum St

Plant species Acacia nilotica An

Albizia lebbeck Al

Eucalyptus camaldulensis Ec

Terminalia arjuna Ta

Soils Forest soil F

Agriculture soil A

Upland soil U

Grassland soil G

Note: 50 ml pseudomonas strains were applied in soil, in control treatment. Treated and non

treated pots applied with sterilized water two times daily.



The study was conducted at Forest Research Institute (F.R.I., Dehradun, located between

30 20‟40” N latitude and 77 52‟12” elongitute)in the sylvan surrounding to DoonVally at 640.08

m above mean sea level. Metrological data (mean monthly, maximum and minimum

temperature, rain fall and humidity are given in Table-2).

Table-2:Rainfall, temperature and relative humidity in Forest Research Institute Estate

Month Rainfall

(mm)

Rainy

days

Average temperature (oC) Relative humadity(%)

Minimum Maximum Mean 07.19am 2.19pm

January 37.0 3.1 3.9 19.8 11.4 96.6 48.0

February 52.5 3.1 6.6 22.4 14.2 95.7 46.0

March 52.4 3.6 10.0 26.8 18.3 89.4 39.1

April 19.5 1.9 14.0 33.0 23.5 72.5 28.4

May 57.7 4.3 18.2 34.8 25.8 70.3 37.3

June 168.9 9.3 21.0 33.8 26.6 78.2 54.0

July 502.5 18.7 22.9 30.9 26.3 91.6 74.0

August 456.2 19.7 20.5 30.5 25.8 93.4 75.5

September 214.7 12.1 20.2 30.1 24.6 92.8 68.9

October 39.0 2.0 13.7 29.1 21.0 93.9 53.3

November 0.9 0.1 8.1 25.8 16.5 96.9 46.2

December 6.5 1.1 4.6 22.2 12.9 97.3 45.9

Total 1607.9 79.0

Mean 13.8 28.3 20.6 89.1 51.4

Observations

Observation on soil and plants were recorded at 3 (July, 2007), 6 (October, 2007), 9

(January, 2008) and 12 months (April, 2008) after application of treatments for the following

attributes:

Plants Height

Plant height was recorded in centimeters (cm) by measuring scale of 100 cm and rounded

up to second point of dismal in average.

Soil pH

Soil pH was measured in 1:2.5 soil-water suspensions by potentiometer using digital pH

meter with glass electrode as described by Jackson (1967).

Results and Discussion

An endeavor has been made to express the effects of various treatments on the soil

properties and growth & nutrients expression of the experimental plant species.



As detailed in material and method, present investigation was carried out to assess the

effects of three treatments of pseudomonas strain (i-control, soil treated with strain of ii-

Solanum tuberosum and iii-Dalbergia sissoo) and four types of soils (Forest, agriculture, upland

and grassland soil) on Four plant species (Acacia niloticca, Albizia lebbeck, Eucalyptus

camaldulensis and Terminalia arjuna). Observations were taken at 3 (July, 2007), 6 (October,

2007), 9 (January, 2008) and 12 months (April, 2008) months after treatments.

Effect of main and combined treatments of soil pH and plant height with their CD value

at 5% is tabulated. In each diagram „a‟ show the effect of strain treatment, „b‟ the effect of strain

in soil, „c‟ the effect of strain in species and„d‟ the effect of strain at different time of

observation. Similarly, effect of soil, effect of soil on different species and at different time of

observations represented in fig „e‟ „f‟ and „g‟ respectively where as figure „h‟ shows he response

of two plant species at different time of observation. Here in the proceeding paragraphs, only

statistically significant results are highlighted in detail in order to provide quick understanding of

the trend exhibited by treatments.

Microbial activity tests

Several tests for microbial activities of the one isolates of Pseudomonas (Solanum tuberosum

and Dalbergia sissoo) were performed in four types of soil planted with different plant species.

The results of the various tests are summarized in Table-3.

Table-3: Microbial activities in Solanum tuberosum and Delbergia sissoo

Test/Treatments Solanum tuberosum Delbegia sissoo

Gram staining -ve -ve

Catalase Activity + +

Growth on GPA + +

Ability to produce 3-ketolactose + +

Carbohydrate fermentation + +

Amylase production - -

Urease test - -

Growth Hofer’s + +

Antibiotic Sensitivity test R R

Tests were carried out for microbial activities in all the soils at initial stage after getting

the soil sterilized. All soil samples gave negative results for microbial activity. Test in microbial

activity in all soils showed the absence of Pseudomonas strain. After 3 months (July, 2007) of

inoculation and plantations, all soils were again tested for presence of Pseudomonas treatments.

Soils of control where no strain was given, strain was found absent again.



Soil pH

It is clear from Table-4 (Fig.-1,a)that Pseudomonas strains of Solanum tuberosum and

Dalbergia sissoo resulted lower soil pH (7.03-7.02) than control(7.05).There was no change in

pH of forest soil whereas soil pH n agriculture soil significantly decreased from 7.78 in control

to 7.76 in Solanum tuberosum and to 7.74 in strain Dalbergia sissoo inoculated soils,

respectively. In upland soil also, pH significantly decreased from 6.99 in control to 6.97 in soil

treated with Solanum tuberosum and to 6.96 in soil treated with Dalbergia sissoo strain.

Difference in pH of upland soil with Solanum tubersoum inoculation and inoculation of

Dalbergia sissoo was statistically equal (6.97 and 6.96) but lower than soil of control. Grassland

soil showed equal pH in control and Solanum tuberosum strain treated soil showed equal pH in

control and Solanum tuberosum strain treated soil (6.90) which was significantly lower than the

soil treated with strain of Dalbergia sissoo(Fig.-1,b).

Soil pH under different species did not change significantly due to strain application which

was 7.03 with strain of Solanum tuberosum and 7.02 in soil treated with Dalbergia sissoo strain

over control 7.04 in soil of Acacia nilotica. Soil of Albizia lebbeck under control treatment

showed pH of 7.05 which was statistically equal to soil of Solanum tuberosum treated (7.04) and

Dalbergia sissoo treated strain (7.03).

In case of Eucalyptus camaldulensis, control soil showed the ph of 7.05 which was equal to

pH of Solanum tuberosum treated strain soil (7.05 which was equal to pH of Solanum tuberosum

treated strain soil (7.03) but higher (CD = 0.03) than soils treated with Dalbergia sissoo(7.02).

Same soil pH was observed in case of Tarminalia arjuna under different strains treatments (Fig.-

1, c). Though there was no significant change in soil pH (7.05 o 7.01) during different months of

observation but it decreased with increase of plant age under all pseudomonas treatments.

Different soils collected for experiment were significantly different in soil pH. Forest soil

was lowest in pH (6.15) and agriculture the highest (7.76). Forest soil under different plantations

showed no significant change in pH that was minimum in Albizia lebbeck (6.50) followed by

Acacia nilotica, Eucalyptus camaldulensis(6.51 and 6.51) and Tarminalia arjuna(6.52). In

agriculture, soil pH under plantation of all species remained same (7.76). In upland soil,

minimum soil pH was seen under Acacia nilotica(6.95) followed by Tarminalia arjuna(6.97),

Eucalyptus camaldulensis(6.96) and Albizia lebbeck(6.99).

Grassland soil recorded higher pH in Acacia nilotica(6.90) than the soils of rest species i.e.

Albizia lebbeck, Eucalyptus camaldulensis and Tarminalia arjuna was equal (6.89). Though soil

pH under all plantations decreased with the age of plans but agriculture soil showed maximum

and decrease in soil pH with increase of plant age was seen in different soil also.



Table-4:Effect of different treatments on soil pH.

Strains Soil Species Months

F A U G Mean An Al Ec Ta Mean 3 6 9 12 Mean

C 6.51 7.78 6.99 6.90 7.05 7.04 7.05 7.05 7.05 7.05 7.07 7.05 7.04 7.03 7.05

St 6.51 7.76 6.97 6.90 7.03 7.03 7.04 7.03 7.03 7.03 7.05 7.04 7.03 7.01 7.03

Ds 6.51 7.74 6.96 6.88 7.02 7.02 7.03 7.02 7.02 7.02 7.04 7.04 7.01 7.00 7.02

Mean 6.51 7.76 6.97 6.89 7.03 7.04 7.04 7.04 7.05 7.04 7.02 7.01

Factors Species Months Factors Months

An Al Ec Ta 3 6 9 12 Mean 3 6 9 12 Mean

F 6.51 6.50 6.57 6.52 6.54 6.53 6.05 6.48 6.51 An 7.05 7.04 7.02 7.00 7.03

A 7.76 7.76 7.76 7.76 7.78 7.76 7.75 7.74 7.76 Al 7.06 7.05 7.03 7.02 7.04

U 6.95 6.99 6.98 6.97 6.99 6.98 6.97 6.95 6.97 Ec 7.06 7.05 7.03 7.02 7.04

G 6.90 6.89 6.89 6.89 6.91 6.91 6.89 6.88 6.89 Ta 7.05 7.04 7.03 7.02 7.04

Mean 7.03 7.04 7.04 7.04 7.05 7.04 7.02 7.01 7.05 7.04 7.02 7.01

C = Control, St = Solanum tuberosum,Ds =Dalbergia sissoo strain

An = Acacia nilotica, Al = Albizia lebbeck, Ec = Eucalyptus camaldulensis , Ta = Tarminalia arjuna

F = Forest soil, A = Agriculture soil, U =Upland soil, G = Grassland soil

3=July, 6=October, 9=January, 12=April

Note CD (5%)

Strains = 0.020 Soil X Species =NS Soil X Month=NS

Species=NS Species X Month=NS Soil X Strain=NS

Soil =0.020 Species X Strain=NS Strain X Month=NS

Month=NS

b: Effect of strains in different soil c: Effect of strains in differentspecies d: Effect of strains in different months A: Effect of Pseudomonas strains

7

7.01

7.02

7.03

7.04

7.05

7.06

C St Ds

C St

5.5

6

6.5

7

7.5

8

F A U G

C St Ds

7.0057.01

7.0157.02

7.0257.03

7.0357.04

7.0457.05

7.055

An Al Ec Ta

C St Ds

6.96

6.98

7

7.02

7.04

7.06

7.08

July. Oct. Jan. Apl.

C St Ds



Figure: 1: Effect of Pseudomonas strains and soil types on soil pH.

Plant Height

Application of Pseudomonas strains increased plant height significantly (Table- 5). Plant

height noted in control treatment was recorded to 39.26 cm which significantly increased to

42.42 cm in Solanum tuberosum and to 42.22 cm in Dalbergia sissoo strain treatments.

Effect of strain inoculation in different soil was different. Both, Solanum tuberosum and

Dalbergia sissoo strains resulted in statistically equal plant height in forest and grassland

soils that was higher than height noted in control treatment (Fig.-2, b). In agriculture soil, soil

treated with strain of Solanum tuberosum showed higher plant height (47.88 cm) than treated

with strain of Dalbergia sissoo(47.49 cm). In upland soil, plant shoed height in order of

control (35.01 cm) <Solanum tuberosum (38.46 cm) and <Dalbergia sissoo (39.47 cm).

Though strain treatments did not show significant response to plant height of individual

species but Eucalyptus camaldulensis and Tarminalia arjuna responded to comparatively

lower under Solanum tuberosum inoculation (54.93 and 35.55 cm). In case of Acacia nilotica

and Albizia lebbeck response of strain treatments for plant height was in increasing order of

control (35.38 and 37.18 cm), Solanum tuberosum (38.24 and 41.39 cm) and Dalbergia

sissoo(38.04 and 41.14 cm) strains. All strain treatments resulted increasing plant height with

increase of inoculation time.

Different soils showed significant difference in plant height (CD = 0.57). Plant height

noted in forest soil was maximum (49.29 cm) and in grassland the minimum (30.86 cm).

Tarminalia arjuna showed minimum plant height and Eucalyptus camaldulensis the

maximum in all soil types, (Fig.-2, c). All species undertaken in present study showed

increasing plant height with their age in different soils.

e: Mean effect of different soils f: Effect of soils on different species g: Effect of soils in different months h:Effect of species in different months

5.86

6.26.46.66.8

77.27.47.67.8

8

F A U G

F A U G

5.86

6.26.46.66.8

77.27.47.67.8

8

F A U G

An Al Ec Ta

0123456789

July Oct. Jan. Apl.

F A U G

6.976.986.99

77.017.027.037.047.057.067.07

July Oct. Jan. Apl.

An Al Ec TA



Table- 5:Effect of different treatments on plant Height (cm).

Strains Soil Species Months

F A U G Mean An Al Ec Ta Mean 3 6 9 12 Mean

C 48.44 44.85 35.01 28.75 39.26 35.30 37.18 51.59 32.89 39.26 27.95 34.88 43.07 51.14 39.26

St 49.67 47.88 39.47 31.86 42.22 38.24 41.39 54.49 34.76 42.22 30.78 37.79 46.07 54.24 42.22

Ds 49.76 49.49 38.46 31.96 42.42 38.04 41.14 54.3 35.55 42.42 30.81 38.29 45.75 54.81 42.42

Mean 49.29 47.41 37.64 30.86 37.22 39.90 5.67 34.40 29.85 36.99 44.96 53.40

Fact

ors

Species Months Fa

cto

rs

Months

An Al Ec Ta 3 6 9 12 Mean 3 6 9 12 Mean

F 44.29 46.36 65.56 40.95 34.91 45.49 53.62 63.13 49.29 An 27.12 33.48 41.48 46.84 37.22

A 43.46 45.66 59.98 40.53 33.36 42.84 52.70 60.73 47.41 Al 28.76 35.90 43.72 51.23 39.90

U 33.50 37.84 49.18 30.05 26.63 31.03 40.55 52.37 37.64 Ec 39.73 47.63 58.48 68.83 53.67

G 27.64 29.74 39.95 26.09 24.50 28.58 32.98 37.36 30.86 Ta 23.78 30.93 36.20 46.70 34.40

Mea

n

37.22 39.90 53.67 34.41 29.85 36.99 44.96 53.4 29.85 36.99 44.96 53.40

C = Control, St = Solanum tuberosum,Ds = Dalbergia sissoo strain

An = Acacia nilotica, Al = Albizialebbeck, Ec = Eucalyptus camaldulensis , Ta =

Tarminaliaarjuna

F = Forest soil, A = Agriculture soil, U =Upland soil, G = Grassland soil

3=July, 6=October, 9=January, 12=April

Note CD (5%)

Strains = 0.492 Soil X Species =1.136 Soil X Month=1.136

Species=0.568 Species X Month=NS Soil X Strain=0.980

Soil =0.568 Species X Strain=NS Strain X Month=NS

Month=0.568

a: Mean of effect of Pseudomonas

strains

b: Effect of strains in different

soils

c: Effect of strains in different species d: Effect of strains in different

months

37

38

39

40

41

42

43

C St Ds

C St Ds

0

10

20

30

40

50

60

F A U G

C St Ds

0

10

20

30

40

50

60

An Al Ec Ta

C St Ds

0

10

20

30

40

50

60

July Oct. Jan. Apl.

C St Ds



Figure : 2: Effect of different treatments on height (cm) in plants.

Conclusion

Strains of Solanum tuberosum and Dalbergia sisssoo reduced soil pH. There was no

effect of forest and grassland soils. Strains reduced pH in agriculture and upland soils. Reduction

in pH was, however, more in soil treated with strain of Dalbergia sisssoo. Though decrease in

soil pH due to strain was insignifica-nt but it was more in soils treated with strain of Dalbergia

sisssoo.

Inoculation of strains insignificantly increased plant height also. Application of Solanum

Tuberosum strain, however recorded significantly higher height than soil treated with Dalbergia

sissoo. In forest and grassland soils, plant height was in order of Solanum Tuberosum =

Dalbergia sissoo > control. In agriculture soil, effect of strains was in order of control <

Dalbergia sissoo and Solanum Tuberosum whereas in upland soil it was in order of control <

Solanum Tuberosum and Dalbergia sisso.

Height of Acacia nilotica, Albizia lebbeck and Terminalia arjuna in different soil was in

order of grassland < upland < agriculture = forest whereas Eucalyptus camaldulensis recorded

plant eight in order of grassland < upland, agriculture and , forest soils.

In the present study that applications of different Pseudomonas strains have given

strength to soil and increased plant growth. Further, different types of soils have response of

different to different species. In the light of above results, there is need to test the response of

different strains and different soils on growth of different plants in field conditions. Therefore,

must be conducted for selection of Pseudomonas strains as well as to demonstrate efficacy of

different soils on plant species.

0

10

20

30

40

50

60

F A U G

F A U G

e: Mean effect of different soils

0

10

20

30

40

50

60

70

An Al Ec Ta

F A U G

f: Effect of soils on different species

0

10

20

30

40

50

60

70

July Oct. Jan. Apl.

F A U G

g: Effect of soils in different

months

0

20

40

60

80

July Oct. Jan. Apl.

An Al Ec Ta

h: Effect of species in different

months



Stasistical Analysis

The data generated were subjected for analysis of variance through Genstate 3.2 (vsni

Laboratories, USA) and Microsoft excel version 2003 (Microsoft technology USA). For better

insight of differences between all the sources of variation, critical difference (CD) value was

calculated at 5% probability level.

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