Biotechnology

Phosphorus and nitrogen are very much crucial for growth of Plant. Nitrogen fixing bacteria
and phosphate solubilizing bacteria (PSB) promote plant growth and enhance various cellular
processes like root elongation, proliferation and changes of root architecture, seed development
and normal crop maturity. In present study the inoculation effect of phosphate solubilizing and
Nitrogen fixing bacteria, isolated from rhizosphere and non-rhizosphere soil of North Gujarat,
India are investigated. The isolated bacteria were tested for indole acetic acid (IAA), ammonia
production and phosphate solubilization in vitro. Various parameters like root and shoot height,
number of root hair, length, width and weight of leaves and chlorophyll content under greenhouse
condition was measured. The study shows highest IAA production 0.1822 µg mL-1 by isolate N8 and
highest phosphate production 0.029 µg mL-1 by isolate P6. The DNA of effective bacterial isolates
was amplified using 16S r-DNA primers. Amplified PCR product were purified and sequenced to
identify the isolates up to species level using BLAST. The isolates were identified as Azotobacter
vinelandii and Bacillus niabensis, respectively (Accession number: JX564632 and KF535156).

India ranks among the largest producer of cotton worldwide with high economic value. During
the time of cultivation cotton plant normally suffers from the bacterial blight diseases, leading
to colour change of affected leaves into light brown exhibiting a blighted appearance. While
retrieving the causative agent on nutrient agar plate, yellow pigmented bacteria were consistently
recovered which was thought to be caused by Xanthomonas oryzae pv. oryzae, the cotton bacterial
blight pathogen. However, physiological and molecular analysis of isolated causative agent was
of the bacterium  Pantoea  sp. eighteen strains of active antimicrobial metabolites producing
Streptomyces  sp. were isolated from the soil samples of agricultural field of Mehsana district,
North Gujarat, India. All the isolates were assessed for antagonistic activity against Pantoea sp.
A causative agent of bacterial blight disease of cotton plant. Among the strains tested, isolate
Gray-1 showed strong antimicrobial activity. This isolate was identified as a Streptomyces rochei
through genetic analysis. The antimicrobial compounds obtained from ethyl acetate extract of
Streptomyces rochei were investigated using FTIR and Gas Chromatography-Mass Spectrometry
(GC-MS). GC-MS analysis showed the presence 20 bioactive compounds. The most active
compounds are 3-Hydroxy-2-Methylbenzaldehyde, 2-Methyl-3-Beta-Furyl propenal, 5-Endo-5-Exo-(Epoxymethano)-6-Methylidene-7 oxabicycloheptane, 4-Hydroxy-Methylbenzaldehyde, 3Hydroxy 4
Methylbenzaldehyde and 3,4 (Methylenedioxy) Toluene, 2-Hydroxy 5 Methylisophthalaldehyde

Twenty eight actinomycetes strains were isolated from cotton plant rhizosphere soil of North Indian region of India. The isolates
were identified as actinomycetes by morphological studies. Among them 11 strain are selected on basis of cellulolytic activities
by primary screening. Cellulolytic activities were exhibited by strain A1, A2, A3, A8, A11, A13, A16, A18, Grey 72, AB1, AB4
and  it  was  confirmed  by  formation  of  clear  zone  of  hydrolysis  around  the  colonies.  Among  the  isolates  maximum  cellulolytic
activity were exhibited by strains AB4 and it was determined by Enzyme assay.

The free living bacteria having the ability to fix molecular nitrogen can be distinguished into
aerobic, facultative aerobic and anaerobic organisms. A total of 8 selected strains of free living nitrogen fixing
bacteria were isolated from different rhizospheric soils of North Gujarat. All the isolates show Acetylene
reduction and few isolates produce IAA. In the present study active culture of N2-fixing bacteria were
applied on Cotton plant in pot. Statistical data of plant growth promotion shows that applications of these
cultures significantly increased plant height, number of leaves, and number of branches per plant, high
chlorophyll and protein content, when compared with control. The results of this study support that the
Am5 strain is very much effective for plant growth promotion and it is closely match with uncultured
bacterial sp. Further investigation may prove this strain as a novel species

Twenty eight actinomycetes strains were isolated from cotton  plant rhizosphere soil  of North  Indian region  of  India.  The
isolates  were  identified  as  actinomycetes  by  morphological  studies.  Among  them  11  strain  are  selected  on  basis  of
cellulolytic  activities  by  primary  screening.  Cellulolytic  activities  were  exhibited  by  strain  A1,  A2,  A3,  A8,  A11,  A13,
A16, A18, Grey 72, AB1, AB4 and it was confirmed by formation of clear zone of hydrolysis around the colonies. Among
the isolates maximum cellulolytic activity were exhibited by strains AB4 and it was determined by Enzyme assay.

Research on single cell proteins to cope with the problem of worldwide food scarcity has been progressing fast for a couple
of years. New technologies are being developed to reduce the cost and to enhance the quality of the product. Present study
focuses  on  the  comparative  analysis  and  includes  isolation  of  yeast,  inoculums  development,  collection  of  waste,  and
biomass and protein estimation. Turbidity due to growth of yeast cells was checked in media and found the highest in the
GSFH. This media shows better results than others in the study. Wet and dry masses were recorded as 29.8 g/l and 12.6 g/l
respectively.  Protein  concentration  was  obtained  0.6  g/g  of  dry  biomass  in  500  ml  of  media.  So  in  all experiments  our
media gave the best results and can be used for the production of single cell proteins in the cost effective manner.

There  are  various  enzymes  used  in  different  industries  including  paper,  textile  and  food.  For  this  propose  Halophilic
Bacterial strain isolated from  India’s  1st Marine National Park at Jamnagar. Total nine isolates were selected for amylase
production on solid agar media containing 1.5% of starch as a substrate. Selected isolates were subjected to morphological,
colonial  and  biochemical  characterization.  The  specific  activity  of  selected  one  isolate Halobaterium  sp. GB24  is  33.62
U/mg with purification fold 2.26 and Yield-38.53% by Ammonium sulphate precipitation method and further purification
by Gel Filtration Chromatography by using Sephadex G-25 shows specific activity 30.24 U/mg, Purification Fold-2.04 and
24.19% Yield. The enzyme acted optimally at pH-6, temperature-40 ºC and 20% NaCl respectively. The 16S r-RNA gene
sequence analysis shows 99% similarity with Halobaterium sp. and the GenBank accession number is KY606985.

The  North  Gujarat  region  is  geographically  having  varied  terrestrial  ecosystem  which  is  comprised  of
semiarid to arid and highly arid saline soil types. It also has tropical dry deciduous Forest patches. Thus,
considering  these  ecoclimatic  conditions,  it  is  thought  to  have  different  diazotrophic  populations  which
might be explored for potential biofertilizer to improve crop productivity in the region. In this context, the
long-lasting  challenges  in  the  development  of  effective  methods  to know  the  types  of  microorganisms
present  in  soils,  and  to  determine  functions  which  the  microbes  perform  in  situ.  Total  68  free  living
nitrogen fixing bacteria having ability to grow on N2 free media were categorized in different group and
identified at species level by ARDRA analysis and 16S r-RNA sequences. One isolate is closely related to
uncultured  bacteria  based  on  complete  sequence  analysis.  Our  result  indicated  that  higher  diversity  of
free-living nitrogen-fixers was mainly affected by the physico-chemical properties of the soils and plant
species, especially the changes of nutrient.

P-Nitrophenol  is  the  potent  contaminant  in  the  environment. We  have  isolated  and  characterize  the  Nocardiodies  nitrophenolicus  strain  use  for
degradation of P-Nitrophenol. Greatest degradation of PNP was observed at 35°C and under slightly alkaline pH (pH 79) conditions. Effective
degradation rates slowed as the concentration of PNP was increased. Addition of glucose from 0.1% to 0.5% generally enhanced the degradation of
PNP at high concentration (325 mg/l) although acidification as a result of glucose metabolism had a negative effect on PNP depletion. On the basis of
16S rRNA gene sequencing the bacterium is a species of Nocardiodies, closely related to Nocardiodies nitrophenolicus.

With  an  objective  of  identifying  the  free  living  Nitrogen  fixing  bacterial  diversity  from  the  soils  of  North  Gujarat
area, a total 35 free living nitrogen fixing bacterial isolates from different regions of North Gujarat were isolated.
Pure  cultures  were  obtained  on  selective  media.  One  individual  isolate  was  selected  for  the  molecular
characterization and  phylogenetic relatedness.  The biochemical  and  molecular  characterization  of the  isolate  was
carried  out.  The  bacterial  isolate  was identified  as  Azotobacter  salinestris.  The  phylogenetic tree  was  constructed
and it showed strong homology of the isolate with Azotobacter salinestris.

It is well known that certain strains of each rhizobial group that are effective plant growth
promoters by  producing plant growth hormones, in addition to their ability in N2 fixation.
Such strains can be useful not only for their host, but also for other plants. Six Rhizobium
strains were isolated from root nodules of Abrus precatorius collected from different regions
of  North  Gujarat.  Four  of  the  Rizobium strains  produced  indole  acetic  acid  (IAA),  but
maximum  amount  was  produced  by  only  two  strains  in  Yeast  Extract  Mannitol  (YEM)
medium supplemented with L-tryptophan. The strains were found to elaborate maximum IAA
when  fed  with  5  mg/ml  L-tryptophan.  The  strains  were  examined  for  production  of
Exopolysaccharide  (EPS)  and  Indole  acetic  acid  (IAA)  by  utilizing  10  different  carbon
sources. DNA was isolated from potent producer of EPA and IAA and amplified by using
16S r-DNA primer and then sequenced. By using bioinformatics tool BLAST the nucleotide
sequence of isolate was aliened and isolate was found to be Bradyrhizobium yuanmingense.
The nucleotide sequence was submitted to NCBI gene bank data and the accession number
provided by NCBI for this sequence is JN 575481.

A total of 21 free living diazotrophs were isolated from different rhizospheric Soils of North
Gujarat. Out of them one individual isolate was selected for the production of indole acetic
acid (IAA) in a medium with 1, 2 and 5 mg/ml of tryptophan. The isolate showed high level
(31.4  mg/l)  production  of  IAA  at  5  mg/ml  of  tryptophan  while  at  1  and  2  mg/ml  the
production was 8.76 and 18.92  mg/l respectively. Pot experiment study  indicates  increased
root length and shoot height indicates efficient IAA production by the isolate. The isolate was
also analyzed  for  its  nitrogen  fixing ability. The  DNA of  bacterial  isolate was  isolated and
amplified using 16S r-DNA primers. Amplified PCR product was purified and sequenced for
its  identification.  The  sequence  of  bacterial  isolate  was  used  to  identify  the  species  of
organism  using  BLAST.  The  organism  was  identified  as  a  Azotobacter  troerpicalis. The
sequence  of  isolate  was  submitted  to  NCBI  gene  bank  data  and  the  accession  number
provided by NCBI is JN591767.

A total of 21 isolate of nitrogen-fixing bacteria were isolated from rhizospheric soils of different agricultural
plant species, including rye, maize and wheat, from North Gujarat, using nitrogen-deprived selective isolation
conditions. Ability to fix nitrogen was verified by the acetylene reduction assay. Out of them one individual
isolate was found to produce biosurfactant. Biosurfactant production was confirmed by conventional screening
methods, including haemolytic, drop collapsing and lipase production activity. These studies revealed that
the isolate produced surface active agents which degrade oil and hence, find its importance in oil contaminated
soils to  maintain soil fertility  by performing dual  role of nitrogen  fixation and oil  degradation. Molecular
identification results from 16S r-DNA analysis  were also corroborated by morphological  and  biochemical  data.

The North Gujarat region is geographically having varied terrestrial ecosystem which is comprised of semiarid to arid and highly arid saline soil types. It also has tropical dry deciduous Forest patches. Thus, considering these ecoclimatic conditions, it is thought to have different diazotrophic populations which might be explored for potential biofertilizer to improve crop productivity in the region. In this context, the long-lasting challenges in the development of effective methods to know the types of microorganisms present in soils, and to determine functions which the microbes perform in situ. Total 68 free living nitrogen fixing bacteria having ability to grow on N2 free media were categorized in different group and identified at species level by ARDRA analysis and 16S r-RNA sequences. One isolate is closely related to uncultured bacteria based on complete sequence analysis. Our result indicated that higher diversity of free-living nitrogen-fixers was mainly affected by the p...

Phosphorus and nitrogen are very much crucial for growth of Plant. Nitrogen fixing bacteria and phosphate solubilizing bacteria (PSB) promote plant growth and enhance various cellular processes like root elongation, proliferation and changes of root architecture, seed development and normal crop maturity. In present study the inoculation effect of phosphate solubilizing and Nitrogen fixing bacteria, isolated from rhizosphere and non-rhizosphere soil of North Gujarat, India are investigated. The isolated bacteria were tested for indole acetic acid (IAA), ammonia production and phosphate solubilization in vitro. Various parameters like root and shoot height, number of root hair, length, width and weight of leaves and chlorophyll content under greenhouse condition was measured. The study shows highest IAA production 0.1822 µg mL-1 by isolate N8 and highest phosphate production 0.029 µg mL-1 by isolate P6. The DNA of effective bacterial isolates was amplified using 16S r-DNA primers. Amplified PCR product were purified and sequenced to identify the isolates up to species level using BLAST. The isolates were identified as Azotobacter vinelandii and Bacillus niabensis, respectively (Accession number: JX564632 and KF535156).

The source of soil nitrogen is the atmosphere where Nitrogen gas occupies about 79 per cent of the total atmospheric gases. Living organisms that are present in the soil have profound effect on N 2 transformation, which provide food and fibre for an expanding world population. Although nitrogen is very abundant in nature; it often limits plant productivity because atmospheric nitrogen is only available to a very narrow range of organisms belonging to diazotrophs, symbiotically and non-symbiotically. Biosurfactants are microbial surface active agents produced by certain microorganisms during their growth phase. They may be extracellular or intracellular in nature (Chen et al., 2007). Substrates for biosurfactant production are sugars, oils, alkanes and waste materials (Lin, 1996). Biosurfactants are amphiphilic, non-toxic and biodegradable molecules with high Corresponding author : P.H. PATEL, Mehsana Urban Institute of Biosciences, Ganpat University, KHERVA (GUJARAT) INDIA Email: Pr...