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ThesisItem Open Access SoybPeroatne [iGn lcyhcainrea cmtearxi z(aLt.i)o Mn efrrorimll] R frhoizmo baicuimdic is soolailste osf oSft ate of Jharkhand by utilizing immunological approach(Birsa Agricultural University, Ranchi, 2023) Srishti Kandulna; Himanshu Dubey(Al3+) cation (Positively charged ions) in soils, which also serves as an indicator of soil acidity. Agricultural practices and climate changes increase the amount of land affected by acidity and thus limit legume crop productivity. Worldwide, more than 1.5 Gha of acid soil limit agriculture production and as much as 25% of the earth’s crop lands are impacted by problems associated leguminous plants require a neutral or slightly acidic soil for growth, especially when depending on symbiotic nitrogen fixation. Out of several gases present in the atmosphere nitrogen share the major portion (about 71%) and is found in the di-nitrogen (an inert) form. It is a component of many bio-molecules required for the growth and development of all organisms. Most of the eukaryotes are capable of utilizing nitrogen directly from the environment; only a certain group of prokaryotes are genetically feasible to fix the atmospheric nitrogen into the biologically useful from like ammonia which is further utilized by eukaryotes. Rhizobia, a gram negative bacteria associates symbiotically with legume crop and are genetically feasible in reducing (fixing) atmospheric nitrogen for leguminous crop. The plant in turn provides the bacteria with organic compounds made by photosynthesis. The symbiotic factor like soil pH, temperature etc., which affects the survival of Rhizobia as well as the nodulation process and thus the nitrogen fixation. Soybean is scientifically known as Glycine max, are a species of legume that have become one of the most widely consumed foods in the world. Soybeans are rich in various bioactive plant compounds like Isoflavones, phytic acid, and saponins. Soybeans are high in protein, and are also a decent source of both carbs and fat. They are a rich source of various vitamins, minerals and beneficial plant compounds, such as Isoflavones. Proteomics is a large scale study of proteins. Two-dimensional electrophoresis (2-D electrophoresis) is a powerful and widely used method for the analysis of complex proteins mixtures extracted from cells, tissues, or other biological samples. This technique sort’s protein according to two independent properties in two discrete steps: the first-dimension step, isoelectric focusing (IEF), separates proteins according to their Isoelectric Point (pI); the second-dimension step, SDS-polyacrylamide gel electrophoresis (SDS-PAGE), separates proteins according to their molecular weight. Each spot on the resulting two-dimensional array corresponds to a single protein species in the sample. Thousands of different proteins can thus be separated, and information such as the protein pI, the apparent molecular weight, and the amount of each protein is obtained. 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, 1st 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 cross-reactivity to the protein specific to the acidic-soil regime in soybean crop. The purified antibodies can be used as bio-marker for the acidic soil regimes. The aim of present work is Protein characterization from Rhizobium isolates of Soybean [Glycine max (L.) Merrill] from acidic soils of State of Jharkhand by utilizing immunological approach. The present study was undertaken with the following broad objectives in mind: 1. ELISA analysis was performed for proteins of selected Rhizobium isolates from acidic-soils of State of Jharkhand, and 2. Generation of antibody profiles.ThesisItem Open Access Diversity analysis of endophytes of finger millet (Eleusinecoracana L. Gaertn.) using RAPD markers(Birsa Agricultural University, Ranchi, 2023) SWATI KUMARI; Anita PandeThe present study could show the closeness between the endophytes derived from one host versus those from a different host. The relatedness could help devise consortia of different endohytes for colonisation of other hosts once the role of the individual endophytes has been elucidated.ThesisItem Open Access Immunological characterization of various Rhizobium isolates from Pigeon pea (Cajanus Cajan (L.) Millsp.) isolated from acidic soils of the State of Jharkhand(Birsa Agricultural University, Ranchi, 2023) Aryan Raj; Himanshu DubeyWhen 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.ThesisItem Open Access Immunological Characterization of Rhizobium isoIlamtmesu fnroolmog cichaicl kcpheaar a(Ccitceerirz aartiioenti nouf mRh Liz.o) bisioulmat ed from acidic soils of the State of Jharkhand(Birsa Agricultural University, Ranchi, 2023) Poonam Priyanka TirkeySoil is a living, dynamic matrix that is vital to the terrestrial ecology. It is an essential resource for agricultural production and food security, as well as the preservation of most biological processes. Decomposition and nutrient cycling rely heavily on the operations of soil biota. As a result, the majority of microbial activity is restricted to the rhizosphere's "hot-spot," i.e., aggregates with accumulated organic matter. When there is a build-up of acid in the soil, it is known as soil acidity. Acid formation in soils is a natural process, and many soils in high-rainfall areas are acidic by nature. Acidification is a slow process, but agriculture hastens it. Plants that aren't tolerant of acidic conditions don't thrive in acidic soils, and productivity suffers as a result. Nitrogen is the most common deficient nutrient in many soils which are absorbed in the form of salts of nitrogen. Rhizobium are gram negative soil bacteria, motile, non-sporulating rods that fix atmospheric nitrogen i.e., convert atmospheric N2 into ammonia for their legume host plant once the symbiosis is established. Rhizobia are considered to be the most important nitrogen fixation agent in agriculture. N2 fixing systems are the symbiotic systems which play a significant role in improving the fertility and productivity of low- N soils. Chickpea (Cicer arietinum L.) is grain legumes grown mainly in areas with temperate and semiarid climate and about 400 millimetres of annual rainfall. It is characterized by a high content of protein, fat, vitamins, fibre, and a lower content of carbohydrates than flour of wheat. These are of two types “desi” and “kabuli” which are currently grown in India. Proteomics is a large scale study of proteins. Proteome techniques bring up new options to examine the complex functions of model plants and crop species at different levels. Proteomics include two-dimensional electrophoresis (2-D electrophoresis), separates proteins according to isoelectric points (pI) and SDS- Polyacrylamide gel electrophoresis (SDS-PAGE), and separates proteins according to molecular weight. These methods are used to analyse complex proteins mixture extracted from cells, tissues or other biological samples. Besides 2-D electrophoresis and SDS-PAGE, Western blot and ELISA are also used. ELISA is used to detect and measure antibodies, antigens, proteins, glycoproteins, and hormones, among other things. Antibodies and antigens are combined to create a measurable outcome in detection of these products. Western blot can use an antibody to specifically recognize its antigen. It separates different proteins in a sample using SDS-polyacrylamide gel electrophoresis (SDS-PAGE). In this study, soils from different locations were taken and landmark protein was identified. The two rabbits were employed through which antibodies were produced. ELISA analysis was done for antibodies detection. These antibodies can be used as biomarkers and the findings of this study provide a basis for further research on the use of Rhizobium inoculants in sustainable agriculture. The aim of present work is “Immunological characterization of Rhizobium isolates from chickpea (Cicer arietinum L.) isolated from acidic soils of the State Of Jharkhand”. The present study was undertaken with the following broad objectives in mind:ThesisItem Open Access Identification of drought specific cDNA in finger millet (Eleusine coracana L. Gaertn.) variety BM 3(Birsa Agricultural University, Ranchi, 2023) ANISHA KERKETTA; Anita PandeInformation generated in the present study will lead to greater understanding of the regulation of gene expression of finger millet under drought stress. It also shows that there may be some level of conservation or similarity between these different organisms at the genetic level when it comes to stress response. However, more research is needed to determine the significance of these findings and their potential impact on the biology of finger millet. It would be worth investigating further to determine the specific mechanisms by which these genetic similarities might be playing a role in the stress response of finger millet plants. This could potentially lead to the development of new techniques or technologies for improving crop resilience and yield under stress conditions.ThesisItem Open Access Diversity analysis of brinjal (Solanum melongena L.) genotypes using RAPD markers(Birsa Agricultural University, Ranchi, 2023) SUPRIYA BHARATI; Anita Pande; SUPRIYA BHARATIRAPD pattern can help to depict similarities and dissimilarities between different brinjal genotypes and support of phenotypic characterization. The present study shows that brinjal genotypes studied have a narrow genetic base. The use of RAPD markers for diversity analysis has been successfully demonstrated.ThesisItem Open Access Fingerprinting of Brinjal (Solanum melongena L.) genotypes using microsatellite markers(Birsa Agricultural University, Ranchi, 2023) BHARTI KUMARI; Anita PandeThe present study showed a very low degree of polymorphic present in the genotypes. This could be due to the selection of genotypes from a narrow base. Also, mostly compound microsatellites were used in present study the occurance of which may be infrequent. The use of simple repeats SSR and other markers may help in uncovering a larger number of polymorphic sites which these could then be used for generation of specific fringerprints.ThesisItem Open Access Effect of meta-Topolin on enhancement of in-vitro multiplication rate in banana (Musa spp.) var. Behula(Birsa Agricultural University, Ranchi, 2023) Dharitri Patar; Madhuparna BanerjeeBanana is one of the healthiest fruits on the planet, and as such, it is popular all over the world. India is one of the world's largest banana producer. Most preferred technology for banana propagation is through plant tissue culture, which overcomes the limitations of conventional propagation. Behula is a vegetative type of banana that produces a high yield and is a superior selection from the popular cooking banana of West Bengal. In the present study, MS media supplemented with various concentrations of meta-Topolin (1.0-4.0 mg/1) and 6.0 mg/l BAP + 100 mg/l AdSO4 + 0.2 mg/1 IAA (established media) were used for comparing the efficacy of these media on multiplication of banana (Musa spp.) var. Behula. After 21 days, the highest bud breaking 88.8 ± 7.87 percent was observed on MS media supplemented with 2.0 mg/1 meta- Topolin. It was 75.1 ±5.82 percent after 21 days, when inoculated in established media. The rate of multiplication was found to vary for different hormonal regime combinations. Highest rate of shoot multiplication was observed 14.5±0.04 shootlets/explant in media supplemented with 2.0 mg/1 meta-Topolin after 45 days of inoculation, whereas 8.30.20 shootlets/explant was observed in established media after 45 days of inoculation. Therefore, MS media containing 2.0 mg/1 meta-Topolin was found to be more efficient for both bud breaking as well as inducing shoot multiplication. Further, MS media with meta- Topolin and established media grown shootlets were excised and transferred into two different auxin rich rooting media. The highest number of roots was produced in MS media containing 0.5 mg 1 IAA, which was 4,3 ± 0.18 roots explant, after 15 days of inoculation. MS media containing 0.5 mg/1 IBA produced 3.4-0.40 roots after 15 days of inoculation. Longer root length of an average of 8.3-0.43cm was produced in media supplemented with 0.5 mg/1 IAA whereas 2.5 0.59 cm in media supplemented with 0.5 mg/1 IBA after 15 days of inoculation. Therefore, in the present study, for micropropagation of banana (Musa spp.) var, Behula, MS media supplemented with 2.0 ing 1 meta-Topolin was found best for bud breaking and shoot multiplication followed by rooting in MS media supplemented with 2.0 mg 1 BAP-50 mg 1 AdSO-05 mg 1 IAA-281 Activated charcoal.ThesisItem Open Access In-vitro regeneration in Gladiolus hybrida L.(Birsa Agricultural University, Ranchi, 2023) Mahasweta Mandal; Manigopa ChakrabortyGladiolus hybrida L., a perennial bulbous flowering plant belongs to monocot family Iridaceae. It is one of the few plants which produce pleasant cut flowers with long spikes. The application of tissue culture techniques in the propagation of ornamental plants has become widely accepted as a standard practice in the agricultural industry. meta-Topolin [6-(3- hydroxybenzylamino)purine] is an aromatic cytokinin. It was first isolated from poplar leaves. Its name is derived from “topol”, the Czech word for poplar. The metabolism of meta-Topolin is similar to that of other cytokinins. Just as zeatin and BAP, meta-Topolin may undergo ribosylation at position 9 without a significant effect on the activity. This study was conduct on G. hybrida in two variety var. Darshan and var. Arka pratham in which among different combinations as well as concentrations of phytohormones, highest bud breaking in Darshan was 84.3±3.72 % and in var. Arka Pratham was 86.3±6.26 %, on MS media supplemented with 2 mg/l meta-Topolin after 21 days of inoculation. It was 45.2±4.15 % in Darshan and 64.4±6.92 % in Arka pratham after 21 days when inoculated in MS media supplemented with 2.0 mg/l BAP and 25 mg/l AdSO4 (established media). Bud breaking declined when cultured for long period in the same media due to accumulation of phenolic compound leading to darkening of culture media. Shoot multiplication rate was slower in established media in comparison to MS media supplemented with meta-Topolin. Rate of shoot multiplication was found highest when explants were grown in MS media + 2 mg/l mT (6.2±1.05 shootlets per explant in Darshan and 7.06±0.80 shootlets per explant in Arka pratham) after 45th days of inoculation. Whereas inoculation in established media resulted in 3.46±0.73 shootlets per explant in Darshan and 3.5±0.20 shootlets per explant in Arka pratham. For root initiation, of the two combinations, MS media + 2mg/l NAA (1-Naphthalene acetic acid) +1mg/l BAP and MS media + 2mg/l IAA + 2g/l BAP used, maximum numbers of roots per explant (4.6±0.20 in Darshan and 5.3±0.76 in Arka pratham) and mean root length per explant (4.08±0.21 in Darshan and 6.56±0.75 in Arka pratham) in NAA containing media, when derived from shootlets regenerated in media containing 2mg/l meta-Topolin after 30 days of inoculation. Therefore, in the present study for micro propagation of Gladiolus hybrida, MS media supplemented with 2mg/l mT was found effective for shoot multiplication followed by rooting in MS media supplemented with 2mg/l NAA (1-Naphthalene acetic acid) +1mg/l BAP for both the varieties (Darshan and Arka pratham).
