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Birsa Agricultural University, Ranchi
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ThesisItem Open Access Optimization of protocol for in vitro regeneration in Hylocereus polyrhyzus (Weber) Britton and rose (Dragon fruit)(Birsa Agricultural University, Ranchi, 2019) RANI, NEHA; Banerjee, MadhuparnaDragon fruit is also known as pitaya or the strawberry pear, and is a beautiful tropical fruit that is sweet and crunchy in taste. Dragon fruit is actually a type of cactus which includes about 20 different species. Hylocereus polyrhyzus is one of the species in which the fruit is red fleshed. This plant is mainly popular in Southeast Asia and Latin America, however, dragon fruit is now grown and enjoyed all over the world. Dragon fruit was named after its appearance which is somewhat similar to an artichoke. The pointy scales around the oval shaped fruit is reminiscent of a dragon. The fruit comes in four varieties three are pink-skinned, one with white flesh, one with red flesh, and the third with purple flesh. The fourth variety has yellow skin with white flesh. All have tiny black seeds that are edible, just like kiwifruit. It is also rich with potassium, protein, fiber, Sodium and Calcium which is good for health than other fruit. Dragon fruit is usually propagated by seeds or cuttings. The in vitro culture is one of the best techniques for mass propagation and crop improvement to increase productivity leading to full supply of the demand. In the present work, an efficient in vitro method for plantlet regeneration explants of H. polyrhyzus (Variety- Red Singhal) was developed. The best survival percentage during surface sterilization of H. polyrhyzus (Variety- Red Singhal) explants were achieved by treating explant with 0.05% of HgCl2 for 20 minutes. Shoot multiplication was induced on MS media supplemented with 3.0 mg/l of BAP, 100 mg/l AdSO4 and 100 mg/l ascorbic acid. Highest number of shoots per explant (7 shoots) was observed after 45 days of inoculation in the same media.ThesisItem Open Access Growth curves of finger millet endophytes and maximum antimicrobial substance production(Birsa Agricultural University, Ranchi, 2019) ., ANURADHA; Pande, AnitaThe growth curves generated for the fourteen endophytes will serve as a reference point for any future studies on these endophytes. The endophytes should be preserved at harvest and not subjected to frequent subculture to prevent loss of production of important substances. Alternatively, the genes involved in the production of economically important substances can be cloned and expressed with a constitutive promoter.ThesisItem Open Access Allele mining for drought tolerance associated DREB genes in finger millet (Eleusine coracana (L.) Gaertn.) genotypes(Birsa Agricultural University, Ranchi, 2019) SINGH, AAKANSHA; Pande, AnitaThe information generated in the study will help discover novel genes involved in drought stress tolerance. It will also help in identification of endophytes which help the host plant combat various stresses. Though finger millet is an inherently drought tolerant crop, in depth studies are required to understand how it combats this important abiotic stress. The present study is a step in this direction.ThesisItem Open Access Characterization of Rhizobium isolates of Pigeon pea collected from the acidic soils of the state of Jharkhand.(Birsa Agricultural University, Ranchi, 2019) KUMARI, RANJEETA; Dubey, HimanshuAgriculture sector is the backbone of Indian economy however; agricultural productivity is very low in some countries. The low productivity is due to decreasing soil fertility many factors responsible for decreasing soil fertility such as soil acidity, soil erosion, and continuous cropping. Soil acidity o.ccurs when there is a buildup of acid forming element. Acidic soil in Jharkhand occupies approximately 90 percent area of the land. Most leguminous plant requires neutral or slightly acidic soil for growth, especially when depending on symbiotic nitrogen fixitation. Legumes, improves soil fertility through biological nitrogen fixation. It is reported to contribute approximately 40 kg N ha-1 soil acidity is one of the serious problem which affect growth of Rhizobia in the soil of Jharkhand Rhizobium is a gram negative bacteria, soil living bacteria. Rhizobium provides organic nitrogenous compound to the plant and plant provide organic compound made by photosynthesis, this relation is known as symbiosis. The present study focuses on the proteomic analysis of Rhizobium isolates of pigeon pea. This objective was achieved by performing the 2 DE using fourteen (14) different isolate of Rhizobium collected from different regimes of state of Jharkhand. We analysed unique spots with the help of Two Dimensional Gel Electrophoresis. ‘Unique' protein spots identified the genes implicated in the acid-soil tolerance. Two Dimensional Polyacrylamide Gel Electrophoresis of proteins is a robust and reproducible technique. It is the most widely used separation tool in proteomics, by utilizing MALDI-TOF-TOF approach subsequently genes which might be implicated to play crucial roles in imparting acid soil-tolerance to Rhizobium isolates of pigeon pea were identified from the isolates collected acid soils of the state of Jharkhand various spots of Rhizobium isolates were analyzed Protein play a major role to cope with abiotic stress. In this study we perform 2-DE electrophoresis to establish the reproducibility of the protein and MALDI TOF/TOF analysis to identify gene identity of the protein spot. Synthetic peptide was prepared against the protein spot which is expressed in the crop in response to the acid-soil tolerance. Subsequently, antibody is produced by against this peptide as a follow-up. This antibody is employed to perform the Western Blot Analysis in order to detect the proteins induced specifically in response to the acid soil tolerance regimes.ThesisItem Open Access Molecular analysis of antagonistic effect of Trichoderma to Fusarium in Grand Nain Banana plant(Birsa Agricultural University, Ranchi, 2019) Kumar Mahato, BharatProduction of banana is highly affected due to Fusarium Wilt also called Panama disease, caused by Fusarium oxysporum all around the world, leading to higher economic loss. The disease now is widespread and destructive in almost all the banana-growing states in India, Trichoderma harzianum was found to be most effective in inhibiting the growth of Fusarium oxysporum . plants treated with Trchoderma harzianum with Fusarium oxysporum at different interval 3 and 5 days ( Treated, Recovery + Fusarium). The result revealed that Trichoderma harzianum is effective in antagonizing Fusarium spp. by mycoparasitic activity and also by competing for rhizosphere colonization and nutrient. Plants treated with Trichoderma have some altered metabolic activities. Thus, any changes was analyzed by visual observation and marker analysis. The OPA series of primer was found to amplify the DNA sample of Trichoderma harzianum, Fusarium oxysporum and Banana. In standardization process the best temperature (annealing and extension) was selected with the better primers which amplified all the DNA samples. Distinct bands were observed. OPA 02 at 72⁰C extension 55⁰C annealing temperature showed some bands but the distinct bands were not observed. However, further refinement of the protocol is needed.ThesisItem Open Access Characterization of Rhizobium isolates of Chickpea from acidic soil of Jharkhand(Birsa Agricultural University, Ranchi, 2019) GOPE, SAPNA; Dubey, HimanshuOut 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 the component of many bio-molecules required for the growth and development of all organisms. Most of the eukaryotes are incapable 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 form like ammonia which is further utilized by eukaryotes. Rhizobium a gram negative bacteria associates symbiotically with legume crop and are genetically feasible in reducing (fixing) atmospheric nitrogen for leguminous crop. Legumes in turn provide shelter and energy to them. The specificity of Rhizobia to inoculate legume falls either in broad range host specificity to narrow range host specificity. Several abiotic stresses adversely influence the activity of Rhizobium. Soil pH is one of the stresses which hamper the symbiotic association between the two. As per the reports soil pH in the range of 6.5-7.0 are considered best in the case of leguminous crop for the optimal activity of the bacteria. Soil pH below or above this range minimizes the Biological Nitrogen Fixation (BNF) through Rhizobia. Two-dimensional electrophoresis (2-D electrophoresis) is a powerful and widely used method for the analysis of complex protein 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 points (pI); the second-dimension step, SDS-polyacrylamide gel electrophoresis (SDS-PAGE), separates proteins according to their molecular weights. 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 aim of my work is to perform 2-D Electrophoretic profiling of Rhizobium isolates of Chickpea collected from various soil regimes. Chickpea plant has been taken as a model system for the collection of nodules. Chickpea , as a legume, improves soil fertility through (BNF) biological nitrogen fixation. Chickpea is a crop that provides cash income from its grain. It requires no N fertilizers owing to its ability to fix atmospheric N, and in rotation can improve the N nutrition and yield of subsequent cereals, one of the most important factors that affect the efficiency of symbiosis between Rhizobia and plants is the Ph of the soil in which they interact. The host plant to any symbiotic Rhizobium appears to be the limiting factor for growth in extreme pH, as most legumes require a neutral or slightly acidic soil for growth especially when they depend on symbiotic nitrogen fixation. The present work is associated with the comparison of the soluble protein fraction of the Rhizobium from the nodules of the legume chickpea grown in the normal environmental condition to the soluble protein fraction of the Rhizobium from the nodules of the legume chickpea grown in the acidic soil by employing Two- Dimensional (2-D) Gel Electrophoresis. Gel analysis shows differences in the expression of protein between the two types of Rhizobia.ThesisItem Open Access Characterization of Rhizobium isolates of Soybean collected from the acidic soils of the state of Jharkhand(Birsa Agricultural University, Ranchi, 2019) KUMAR, MIRTYUNJAY; Dubey, HimanshuOut 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 the component of many bio-molecules required for the growth and development of all organisms. Most of the eukaryotes are incapable 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 form like ammonia which is further utilized by eukaryotes. Rhizobium, a gram negative bacteria associates symbiotically with legume crop and are genetically feasible in reducing (fixing) atmospheric nitrogen for leguminous crop. Legumes in turn provide shelter and energy to them. The specificity of Rhizobia to inoculate legume falls either in broad range host specificity to narrow range host specificity. Several abiotic stresses adversely influence the activity of Rhizobium. Soil pH is one of the stresses which hamper the symbiotic association between the two. As per the reports soil pH in the range of 6.5-7.0 are considered best in the case of leguminous crop for the optimal activity of the bacteria. Soil pH below or above this range minimizes the Biological Nitrogen Fixation (BNF) through Rhizobia. My work is to characterize the Rhizobium isolates of soybean collected from acidic soils of the state of Jharkhand. Through studying the proteome of Rhizobium in acidic soil condition, the response of the isolates towards acidity of soil is being analyzed. At the molecular level, we find that the two dimensional gel analysis reveals a host of proteins which are found to be up-regulated or down regulated in response to different pH conditions. We hypothesize that the protein changes observed on two-dimensional electrophoresis in response to different pH of acidic soil reflected the molecular adaptation mechanism taking place in progress in soybean to combat and recover in response to abiotic stress such as acidic soil.ThesisItem Open Access Identification and characterization of “Landmark Protein” from Rhizobium isolates of Cicer arietinum from acidic soil of Jharkhand(Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Kumari, Neha; Dubey, HimanshuThe soil pH is a measure of the acidity or alkalinity in soils. Soil pH is considered as an important factor of soils as it controls many chemical processes that take place in plants and microorganisms that live in soil. Nitrogen is the most common deficient nutrient in many soils around the world and supply of nitrogen through fertilizers has severe environmental concerns. Nitrogen cannot be absorbed by plants directly from atmosphere, but only in the form of salts of nitrogen. Rhizobia are soil bacteria and are gram negative, motile, non sporulating rods that fix nitrogen i.e. convert atmospheric nitrogen into ammonia (ammonia acts as a natural fertilizer for the plant) after becoming established inside root nodules of legumes (Fabaceae). The plant in turn provides the bacteria with organic compounds made by photosynthesis. The symbiotic relationship implies a signal exchange between both partners. There are various environmental factors like soil pH, temperature etc. which affects the survival of Rhizobia as well as the nodulation process and thus the N2 fixation. Chickpea (Cicer arietinum L.) is one of the important pulses cultivated in India during winter season. It is a good source of carbohydrates and proteins which together constitute about 80% of the total dry seed mass and the protein quality is considered to be better than other pulses. It has significant amounts of all the essential amino acids except sulfur containing types. It is cholesterol free and is a good source of dietary fiber, vitamins and minerals. 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 protein mixtures extracted from cells, tissues, or other biological samples. This technique sorts proteins according to two independent properties in two discrete steps: the first-dimension step, isoelectric focusing (IEF), separates proteins according to their Isoelectric Points (pI); the second-dimension step, SDSpolyacrylamide gel electrophoresis (SDS-PAGE), separates proteins according to their molecular weights. 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 aim of my work is to identify and characterize of “Landmark Protein” from Rhizobium isolates of Chickpea (Cicer arietinum L.) collected from various soil regimes of Jharkhand.ThesisItem Open Access Characterization of “Land mark Protein” from Rhizobium isolate of Pigeon pea collected from the acidic soil regimes of the State Of Jharkhand(Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Mahto, Priyanka; Dubey, HimanshuAgriculture sector is the backbone of Indian economy however; agricultural productivity is very low in some countries. The low productivity is due to decreasing soil fertility many factors responsible for decreasing soil fertility such as soil acidity, soil erosion, and continuous cropping. Soil acidity occurs when there is a buildup of acid forming element. Acidic soil in Jharkhand occupies approximately 90 percent area of the land. Most leguminous plant requires neutral or slightly acidic soil for growth, especially when depending on symbiotic nitrogen fixation. Pigeon pea is a legume, improves soil fertility through biological nitrogen fixation. It is reported to contribute approximately 40 kg N ha-1 soil acidity is one of the serious problem which affect growth of rhizobia in the soil of Jharkhand Rhizobium is a gram negative bacteria, soil living bacteria. Rhizobium provides organic nitrogenous compound to the plant and plant provide organic compound made by photosynthesis, this relation is known as symbiosis. Proteomics is a large scale study of proteins. Two-dimensional gel electrophoresis is commonly used to analyse proteins. In 2-D gel electrophoresis proteins are separated in two dimensions on the basis of two independent properties i.e. Isoelectric point [pI] (1st Dimension (Isoelectric focusing (IEF)) and Molecular weight [MW] (2nd Dimension (SDS-PAGE)) In this study we perform 2-DE electrophoresis to establish the spots reproducibility of the protein In our present study we find that protein changes between isolates from normal pH and acidic pH intervals. Lot of proteins change in form of prominent spots which were altered across the various isolates collected from various pH regimes. Such changes reflect the adaptative changes which are under play at protein level for helping the bacteria survive under harsh acidic pH regimes. On the other hand, it is also a possibility that such changes reflect many other protein changes which arise out of processes, such as post-translational modification and the presence of isoforms. Reproducible land mark protein spots were observed in the gels by 2D electrophoresis by utilizing 2-DE approach subsequently protein which might be implicated to play crucial roles in imparting acid soil-tolerance to Rhizobium isolates of pigeon pea were identified from the isolates collected acid soils of the state of Jharkhand, various spots of Rhizobium isolates were analyzed Protein play a major role to cope with abiotic stress.
