Vol. 5(3) August 2010
Role of Single Fungal Isolates and Consortia as Plant
Growth Promoters under Saline Conditions
Pandya Urja and Saraf Meenu
Seven fungal isolates were screened from soil samples
of different regions of Khambat and Bhavnagar. They were Aspergillus sp. (SR2, S11,
N3, N4 and S17), Penicillium sp. (S12B), Fusarium sp. (S217). At 2% NaCl concentration,
all fungal isolates had maximum growth after 72 h of incubation. Seven fungal isolates
showed zone of phosphate solubilization on agar ranging from 15 to 30 mm within
72 h of incubation. Siderophore production was studied after 15 d of incubation
on succinic acid medium. Maximum zone of siderophore production showed by isolate
SR2 was 63 mm followed by isolate S17. Pot study was carried out to observe the
effect of fungal inoculation on chickpea plant. Inoculation with all the fungal
isolates resulted in increase in plant height, number of leaves, number of lateral
roots, fresh weight, dry weight and total chlorophyll as compared to control. Penicillium
sp. (S12B) and Aspergillus sp. (S11) showed increase in plant growth compared to
other fungal isolates in 2% saline soils. S11 and S17 were biocompatible and showed
maximum increase in all vegetative parameters compared to single culture inoculation
and control suggesting that the combination could be more effective than individual
strains at 2% level of salinity.
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Effect of Tillage Systems on Glomalin-Related Soil
Protein in an Aquic Brown Soil
Liang W.J.
Conservation agriculture such as no tillage may increase
soil carbon sequestration. The main objective of this study was to examine the profile
distribution of soil organic carbon (SOC) and glomalin-related soil protein (GRSP)
under no-tillage (NT), conventional tillage (CT, mouldboard ploughing) and fallow
(FF, unmanaged field lasting for 6 years grown with weeds) treatments to a 1 m depth
based on a 6-year tillage experiment in Northeast China. The results showed that
NT enhanced SOC concentration only at the surface depths of 0–5 and 5–15 cm, but
not at deeper depths. SOC concentration at the top 0–5 cm depth was significantly
higher in FF than in NT treatments, but significantly lower at the 5–15 and 15–30
cm depths. Compared to CT, NT significanlly enhanced the concentrations of TG and
EEG throughout the soil profile. In addition, SOC was positively correlated with
TG and EEG in present study. Our study indicates that GRSP may be a useful indicator
for distiguishing tillage systems on soil carbon sequestration across soil horizons.
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Impact of Cultivation Conditions on Type of Polyhydroxyalkanoate
Biopolyester Production
Kanchanasuta Suwimon, Boonyawanich Sirione and Pisutpaisal Nipon
The production of polyhydroxyalkanoate biopolyester
from three bacterial strains, Alcaligenes euthophus, Alcaligenes faecalis and Pseudomonas
oleovorans was investigated. The effect of cultivation and operational conditions;
pH, temperature, nitrogen content, aeration condition and type of co-substrate
on the yield and type of polyester produced were examined. A. euthophus yields the
maximal extent of the biopolyester under the aerobic cultivation with pH 5 and 30o
C when glucose was used as a main and sole carbon source. Structural analysis using
1H NMR indicated that the accumulated biopolyester was poly (3-hydroxybutyrate-co-3-hydroxyvalerate)
when propionic acid was used as co-substrate whereas poly (3-hydroxybutyrate-co-4-hydroxyutyrate)
was synthesized when acetate or butyrate were used as a co-substrate.
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Effect of C/N Ratio and Physicochemical Conditions
on the production of Rhamnolipids by Pseudomonas Aeruginosa LBI
Lovaglio R. B., Costa S. G. V. A. O., Lima C.J.B., Cortezi M. and Contiero Jonas
Waste utilization for biosurfactant production is an
important approach contributing to the reduction of waste treatment costs, while
increasing the economic value of residues and reducing biosurfactant production
costs. In an attempt to optimize rhamnolipid production by Pseudomonas aeruginosa
LBI, the effects of the oil refinery waste as carbon source, C/N ratio, agitation
speed, aeration rate, pH control and fed-batch culture conditions on the production
of this biosurfactant were assessed in a bioreactor. Maximum production (16.9 g
L-1) was attained at a C/N ratio of 23, agitation at 800 rpm and aeration at 2.0
vvm. The rhamnolipid solution (0.1%, w/v) exhibited surface and interface tensions
of 25 mN/m and 0.63 mN/m respectively. The critical micelle concentration (CMC)
was 73.9 mg L-1. The optimal parameters used in this study promoted the formation
of a useful biosurfactant with potential for many industrial applications.
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Antibacterial Activity of Aleurites Moluccana (Euphorbiaceae)
against Some Clinical Isolates
Samah Othman Abd and Razar Rasyidah Mohamad
Aleurites moluccana extracts were found to inhibit the
growth of Staphylococcus aureus and Pseudomonas aeruginosa in a previous study.11
The antibacterial activities of acetonitrile, methanol and water extracts of husk,
leaf and stem bark of A. moluccana were further investigated against 8 clinical
isolates. The disc diffusion assays showed that crude extract of A. moluccana inhibited
the growth of Staphylococcus aureus (S 277), Streptococcus pyogenes (S 526), Pseudomonas
aeruginosa (P 84) and Klebsiella pneumoniae (K 6). The lowest MIC for P. aeruginosa
(P 84) was exhibited by methanol extract of stem bark at concentration 98 µg/ml
and S. pyogenes (S 526) at 781 µg/ml whilst both S. aureus (S 277) and K. pneumoniae
(K 6) exhibited in acetonitrile of husk at 195 µg/ml and 6.25 µg/ml, respectively.
The antibacterial action was bactericidal for all bacteria except for S. pyogenes
(S 526) where the action was bacteriostatic. The highest polyphenols content was
observed in methanol extract of stem bark. The overall results indicate that the
extracts from the stem bark are potential source of phytomedicine agents for both
Gram-positive and Gram-negative bacteria.
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Characterization of Commercial Microbial Products
by Polymorphic DNA Markers and Enzymatic Activity Diversity: Occurrence and Potential
Effects on Freshwater Mussels exposed to Municipal Effluents
Douville M., Gagné F., Zhu B., Fortier M. and Fournier M.
The increasing commercial applications of microbial products
and consortia have raised concerns about their release to the environment and the
potential for toxic effects on aquatic biota. This study characterized 25 bacteria,
one yeast and one fungus using a combination of random amplified polymorphic DNA
and 19 hydrolytic enzyme activities, with the purpose of determining their occurrence
and potential effects on the immune systems of mussels exposed to a primary-treated
municipal effluent for seven weeks. The amplification product (552-bp) of cpn60
gene by using universal primers was sequenced for each micro-organism and available
in GenBank. The microbial community structures in the digestive gland of mussels
were also characterized using 16s ribosomal gene analysis by denaturing gradient
gel electrophoresis (DGGE). The results revealed that each micro-organism was easily
cultivated with usual growth media and the enzyme activity profiles permitted to
propose an identification method using hierarchical tree and backpropagation neural
networks classifications. The bacterial community structure based on DGGE of 16S
rDNA in mussels changed with the concentration of the municipal effluent. This was
also revealed by a characteristic change in enzyme activity profiles with increased
frequency in acid and amide phosphatase, leucine aminopeptidase and β-galactosidase
in the isolated heterotrophic colonies in mussels. The analysis of heterotrophic
bacteria colonies in unexposed mussels revealed that most colonies (6/10) were of
unkown origin and only three bacterial colonies shared close similarities with B.
subtilisis 11685, E. aerogenes, P. putida 12633. In mussels exposed to the municipal
effluents, about half of the colonies were classified as unknowns while the other
half was composed of bacteria showing close similarites with E. aerogenes, P. putida
12633 and 31800. These bacteria differed from those found in forming biofilms which
was mainly associated to B. subtilis 11685 or P. fluorescens 13525 functional properties.
The immune system of the mussels was also compromised with decreased hemocyte density,
increased phagocytosis activity and cytotoxic activity. While no trend between phagocytosis
activity and the enzyme activity profiles was found, the increased frequency in
leucine aminopeptidase and acid phosphatase were negatively correlated with the
hemocyte density and cytotoxic activity. In conclusion, a methodology to track for
the occurrence, bioavailability and potential immunotoxic effects of commercial
microorganisms is proposed based on a combination of genetic and phenotypic (enzyme
activity) markers and immunocompetence assessment. Preliminary results indicate
mussels contained only three bacteria out of 25 bacteria that bear high functional
similarity and the characteristic pertubations with the enzyme activity profiles
were associated with immunocompetence changes.
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Role of Chaperon Co-Expression System in the Production
of Recombinant Unnatural Proteins in Escherichia Coli
Ayyadurai Niraikulam , Yoon Hyungdon and Lee Sun-Gu
Here we evaluated the chaperon co-expression system for
the production of unnatural recombinant proteins through selective pressure incorporation.
In this study green fluorescent protein (GFP) was utilized as model proteins and
L-homopropargylglycine (Hpg) was used as a model unnatural amino acid which is a
methionine surrogate and can be incorporated into the methionine residues of recombinant
protein. Initial experiments showed yields of L-Hpg containing GFP proteins were
almost negligible compared to those of canonical amino acid incorporated natural
proteins. These results indicated that incorporation of unnatural amino acid into
protein affects the normal protein folding which immediately affects the protein
functionality and productivity. Co-expression of the chaperons has been well reported
to enhance the protein folding and preventing the aggregation from harsh environment
condition. Hence, we expected that chaperon co-expression system will facilitate
the production of unnatural recombinant protein production. Coexpression of DnaK-DnaJ-GrpE
chaperons systems shows that the unnatural amino acid protein production was slightly
higher than the wild type. However, chaperon systems GroEL and GroES completely
failed to rescue the unnatural amino containing protein so the production yield
of the unnatural protein was not improved. These results demonstrated that incorporation
of unnatural amino acid tremendously influences the production yield of unnatural
containing proteins in Escherichia coli. As well as chaperon teams are not critically
involved in protein folding of unnatural amino acid containing protein and they
failed to assist stability and prevent the disaggregation of proteins.
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Growth Responses of Calotropis Procera R.Br. - A Medicinal
Plant exposed to coal-smoke pollution
Kumar Sanjeev *1 and Narula Alka 2
The present work was carried out to assess the impact
of coal-smoke pollution (fly ash) on growth response of Calotropis procera, a shrub
of family Asclepiadaceae growing in the vicinity of coal based Badarpur Thermal
Power Plant, which releases a mixture of pollutants like SO2, NO2, CO2, HF, fly
ash and particulate matter. Plants growing in close vicinity with highly polluted
environment showed changes in physio-chemical characteristics like increase in
shoot length, leaf area, biomass but considerable decline in chlorophyll and protein
content, nitrate reductase activity, rate of photosynthesis etc. Visible symptoms
on the foliage were chlorosis and necrosis coupled with reduced growth and yield
followed by premature death of plants in extreme cases.
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A Study by Absorption Spectrophotometry of the Combined
Effect of Temperature and Copper Sulphate on Bilirubin Concentrations
Pillay Avin E., Salih Fadhil M. and Maleek Muthana I.
Dilute solutions of unconjugated bilirubin in aqueous
NaOH were spiked with trace levels of CuSO4. The dependence of the specimen on the
level of the spike and on concomitant increases in temperature was closely examined
by high resolution absorption spectrophotometry. We found that the characteristic
absorbance (A) of bilirubin, in the region of 453 nm dropped systematically in the
presence of 1-25 mM CuSO4. This depletion was accompanied by a corresponding growth
of biliverdin around 365 nm. It was discovered that the influence of temperature
on the spiked solutions intensified the phenomenon. The absorbances of the species
were investigated as a function of CuSO4 concentration and temperature in the range
25 – 65 oC. Appropriate statistical treatment of the data provided a means of indirectly
estimating the variation in bioanalytical concentration. Consideration of ΔA vs
ΔT and ΔA vs Δ [CuSO4] revealed that the thermal effect competes with the “copper
sulphate effect.”
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Technology Development for Ethanol Production from
the Wild Fruits of Mimusops Hexandra
Daripkar Nilambari and Jadhav B. L.*
Present investigation was carried out to develop technology
for the ethanol production from the wild fruits of Mimusops hexandra using two different
yeast strains. The wild fruits used in the study were collected from different parts
of Maharashtra. The chemical analysis of fresh fruit showed protein, lipid, total
carbohydrates and moisture content about 3.53%, 2.6%, 22% and 71.87% respectively.
The fermentation of the pulp of fresh fruit was carried out by commercial MTCC –
173 strain and wild strain of S. cerevisiae. The ethanol content of the ferment
was determined by Gas Chromatography. The MTCC – 173 strain produced 21% ethanol
while it was 7.05% by wild strain.
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Detection of Low Molecular Weight Protein Biomarker
by Serum Protein Profiling of Acute Myeloid Leukemic Patients
Sharma Chandra Kant and Sharma Vinay
Mostly cancer cells contain genetic damage for event
leading to tumor genesis. The genetic damage present in a parental tumorigenic cell
is maintained (i.e. not correctable) such that it is a heritable trait of all cells
of subsequent generations. One of the causes of genetic damage found in cancer
cells is due to Proto-oncogenes, a protein capable of cellular transformation.
The process of activation of Proto-oncogenes to oncogenes can include retroviral
transduction or retroviral integration, point mutations, insertion mutations, gene
amplification, chromosomal translocation and/or protein-protein interactions.
These proteins are generally absent in healthy persons. These anomalous proteins
may be growth factor, growth factor receptor, intra-cellular signaling molecule
or enzyme of diverse signaling pathways and can be taken as biomarker for the identification
of cancer in the vicinity of the begining stages so that it can be treated appropriately
and cured utterly. In our study the molecular weight of the extra protein in AML
patients is approximately 16.21 kDa. This abnormal serum protein can be utilized
as biomarker for the early detection of AML
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Direct Shoot Organogenesis of Indian Cotton G. Arboreum
CV PA 402 from Embryonic Axis Explants
Talegaokar A. P. and Dangat S.S.
Cotton is multipurpose crop serving as an engine of economic
growth in both developing and developed countries across six continents. India is
the only country to grow all four cultivated (both diploid as well as tetraploid)
species on commercial scale. The diploid cotton species can serve as valuable gene
pool for the agronomically desirable tetraploid cultivars and offer better opportunities
to study gene structure and function through gene knockouts. Development of transgenic
lines is one of the ways for value addition in terms of transfer of different agronomic
traits as well as study of different gene functions using different tetraploid and
diploid germplasm backgrounds. Regeneration protocol through somatic embryogenesis
is primary requisite to exploit valuable traits available through plant transformation.
In present investigation successful direct shoot organogenesis was attempted in
PA 402 variety of diploid cotton using MS media supplemented with Myo-inositol
100 mg/L Thiamine 10 mg /L Glucose 30 gm/L and agar 7 gm/L .The hormone kinetin
was used in the range of 0.1 –2.5 mg/L in five different levels. Growth of shoots
on MS medium supplemented with Myo-inositol 100 mg/L thiamine 10 mg /L Glucose 30
g/L and NAA 0.1 mg/L resulted in regeneration of plantlets. Among different levels
of kinetin 0.1 mg/L was found the most suitable concentration for direct shoot organogenesis
from embryonic Axis explants.
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Review Paper: The Utilisation of Microbial Poly-Hydroxy
Alkanoates (PHA) in Food Industry
Arzu Akpinar-Bayizit, Tulay Ozcan, Oya Irmak Sahin and Lutfiye Yilmaz-Ersan
Biodegradability is defined as the capacity of to be
broken down by the action of living things, such as bacteria and fungi being the
main participants in the process of biodegradation in the natural world. The breakdown
of materials provides them with precursors for cell components and energy for energy-requiring
processes. Biodegradation is thus defined as catabolism. Some synthetic polymers,
having chemical compositions resisting enzymatic attacks, can be microbially degraded
with slower processes. Bioplastics is a term used for packaging materials derived
from renewable resources and are considered safe to be used in food applications.
In comparison to conventional plastics, the monomer composition of bio-based polymers
can be differed by varying the producing strains, substrates and cosubstrates, and
biopolymers such as polyhydroxyalkanoates (PHA), polylactic acid (PLA), polysaccahrides.
PHAs are carbon-energy storage material accumulated
intracellularly in a variety of microorganisms under controlled concentrations.
Their biodegradability and possibility to be synthesised from renewable agricultural
or even industrial wastes, make PHAs of considerable interest. Their main applications
in food industry are serving as waterproof layer for the production of disposable
food containers and utensils, cast films and cast sheets for thermoforming and utilisation
as good odour and oxygen barriers.
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