Immunological characterization of various Rhizobium isolates from Pigeon pea (Cajanus Cajan (L.) Millsp.) isolated from acidic soils of the State of Jharkhand
Loading...
Date
2023
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Birsa Agricultural University, Ranchi
Abstract
When there is an accumulation of acid in the soil, soil acidity results. Almost
90% of the land in Jharkhand is covered with acidic soil. For optimal growth,
most leguminous plants need neutral or slightly acidic soil, particularly when
relying on symbiotic nitrogen fixation.
Pigeon peas are legumes that improve soil fertility by biologically fixing
nitrogen. According to reports, soil acidity is one of the major issues that affects
the establishment of Rhizobia in the soil of Jharkhand and contributes around
40 kg N ha-1. Gram-negative, soil-dwelling bacteria are called rhizobium.
Symbiosis is the relationship between the rhizobium, which delivers organic
nitrogenous compound to the plant, and the plant, which produces compound
through photosynthesis.
An effective and popular technique for analyzing complex protein
mixtures isolated from cells, tissues, or other biological materials is twodimensional
electrophoresis (2-D electrophoresis). In two distinct processes,
this method isolates proteins based on two independent properties: their
isoelectric points (pI) in the first-dimension step of Isoelectric focusing (1EF),
and their molecular weights in the second-dimension step of SDS
polyacrylamide gel electrophoresis (SDS-PAGE). The resultant twodimensional
array's spots each represent a distinct protein species from the
sample. A list of experimental peptide masses, often known as "mass
fingerprints," is produced by MALDI after it weighs peptides synthesized from
trypsinized parent proteins. An ion's m/z ratio is calculated during MALDI-TOF
analysis by timing how long it takes for it to go down the flight tube.
A method for the specific histochemical demonstration of antibody in
cells and parts of cells is described. It consists of carrying out a two stage
immunological reaction on frozen sections of tissues: (a) allowing reaction
between antibody in the tissue and dilute antigen applied in vitro, and (b) the
detection of those areas where this antigen has been specifically absorbed by
means of a precipitin reaction carried out with fluorescein-labelled antibody.
The western blotting technique is a rapid and sensitive assay for detection and
characterization of proteins. In this method labeled antibody against a particular
protein is used to identify the desired protein, so it is a specific test. ELISA
(Enzyme Immuno Sorbent Assay) is a technique used for detection of antigen or
antibody. It is based on the formation of antigen-antibody complex, which is
detected by chromogenic detection using enzyme. ELISA was performed at
405nm for pre immune sera, 1" and 3rd Bleed. HRP properties of the secondary
antibodies were used to develop colour reactions. The ELISA results showed
that 1st and 3rd bleed has a great titer for the antibodies produced. The ELISA
also confirmed that the color reaction was obtained, thus showing the crossreactivity
to the protein specific to the acidic-soil regime in soybean crop
The aim of the work is immunological characterization of various
Rhizobium isolates from Pigeon pea (Cajanus cajan (L.) Millsp.) isolated from
acidic soils of the State of Jharkhand. The present study was undertaken with
the following broad objectives in mind:
1. ELISA/Western Blot analysis was performed for proteins of selected
Rhizobium isolates from acidic-soils of state of Jharkhand,
2. Generation of antibody profiles.