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Birsa Agricultural University, Ranchi

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2002 - 2009192010 - 201854
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Subject: Biotechnology × End date: 2018 × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    Protocol Development of In- Vitro Cultivation of Bamboo ( Dendrocolamus As per )
    (Birsa Agricultural University, Ranchi, 2002) Peddy Srikanth; Z.A. Haither
    Bamboos are perennial, woody and evergreen monocotyledonous arborescent grasses belonging to the family Poaceae (Graminae) and Sub-family Bambusoideae. There are about 75 genera and 1250 species of bamboos. Dendrocalamus asper is one of the economically important and widely cultivated bamboo species. The tender shoots of this plant can be consumed as food and is a good source of foreign exchange to various countries. Mature culms of this plant are used for pulp and paper manufacture. Bamboos are propagated artificially by different methods, including through seed. But utilization of seeds as propagating material is difficult and unreliable due to long and unpredictable flowering habit, short dormancy period of seed, poor viability, inborn microbial infestation, poor seed set during off-season flowering, seed sterility and large scale. Consumption of seeds by rodents and wild animals. The vegetative methods, on the other hand, are costly, lobour intensive, cumbersome and time. Consuming. These vegetative propagates are bulky, difficult to transport to distant places and their survival rates are also not very high. This limits large scale cultivation of bamboos in general. Under the situation, propagation through tissue culture seem to be a viable method for large scale propagation of the bamboo species. Therefore the present project was undertaken to establish a protocol for in-vitro propagation of Dendrocalamus asper. In the present study nodal segments (3-4cm) with axillary buds from young juvenile mother plant was used as explants. Surface sterilization using 0.1% (w/v) mercuric chloride (Hgcl₂) for 10 minutes followed by 3-4 times subsequent washing with sterile distilled water proved the best as it resulted the highest percentage (92.68%) of bud break after two weeks. The sterilized nodal segments were cultured aseptically on MS medium supplemented with 0-15 mgl-1 BAP and maximum shoot proliferation. (14-15 shoots per propgule) was achieved on medium supplemented with 12mgl¹ BAP. These proliferated axillary shoots were excised and subcultured on MS liquid medium +3 mgl BAP for the first two subcultures to increase the number of shoots. The shoot multiplication was achieved on both MS solid as well as liquid medium supplemented with 1-5 mgl¹! BAP Highest rate of shoot multiplication (fold) i.e., 15.77 was obtained on MS liquid medium supplemented with 3 mg l-¹ BAP in four weeks. MS solid medium supplemented with 3 mgl¹ BAP resulted only 8.55 fold. Incorporation of NAA (0.2-1.0 mgl ¹) to the medium along with BAP did not increase the rate of shoot multiplication and shoot length but it resulted in better quality erect shoots. MS medium in its full strength (1x) was found to be the most effective basal nutrient medium for shoot multiplication. The studies on sucrose concentration in the medium showed that 3% sucrose was essential for rapid multiplication of shoots. The effect of pH reflected that shoot multiplication occured even on acidic medium and highest rate of shoot multiplication (15.88) was obtained at pH 5.8. A regular subculture cycle at an interval of 4 weeks resulted in healthy cultures devoid of brown leaves and high rate of shoot multiplication. For in-vitro root regeneration on MS medium supplemented with 10mgl-¹ IBA yielded 90% rooting, 19.66 roots per propagule in four weeks, while 3 mg NAA supplemented MS medium resulted 91.66% rooting with 10 roots per propagule NAA resulted short roots while IBA resulted long roots. Addition of BAP (0.1-0.5 mgl ¹) to the rooting medium, neither enhanced root regeneration percentage nor improved the number of the in-vitro roots The cultured plantlets were successfully hardened under high humidity on sterilized soil sand FYM(1:1:1) mixtured with 1/2 strength MS nutrient medium irrigations (without organics).
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    ThesisItemOpen Access
    Protocol Development of invitro Clonal Propagation of Orchid ( Vanda Spa)
    (Birsa Agricultural University, Ranchi, 2002) Ranjeet Kumar Sinha; Z.A Haither
    Orchids, one of the most beautiful group of flowering plants belong to the family Orchidaceae (Monocotyledons). The exquisite beauty of Orchid flowers due to brilliance in colour, remarkable range of sizes, manifold shapes, and variation in the form and wide range of distribution has aroused highest admiration throughout the world. The Orchid comprises about 800 genera with around 35,000 species. In India, about 1300 species of Orchids are found in Himalayas and others scattered in eastern and western Ghats. A vast majority of Indian Orchids are confined to mountain where they are distributed from base of hill to the elevation of 4300 m above mean sea level in climates ranging from tropical to temperate. Orchids are terrestrial, epiphytic, lithophytic or saprophytic but no Orchid is aquatic. The cut flower trade of Orchid involves 3% internationally. Major suppliers, like, Thailand, Netherlands and Singapore export flowers worth of US $ 80.0, 77.4 and 20.0 millions, in order per year. Due to their alkaloid, flavanoid, glycosides and other phytochemical constituents Orchids have high therapeutic value. The flower juice of Vanda coerulea is used to cure eye diseases. Cymbidium elegans, Cymbidium pubescens, Epicactic latifolia are used as local medicines. for treatment of nervous disorders. Orchids are also used in many countries as food or for making refreshing drinks. Unfortunately the natural population of Orchid is fast declining due to excessive collection and over harvesting by traders and botanical explorers. So there is need to cultivate and conserve the endangered Orchids. The conventional method of propagation is tedious and time taking. The alternative means of propagation is in vitro clonal propagation. Keeping this in mind the present experiment. on developing a viable protocol for in-vitro clonal propagation of Orchid (Vanda Miss Joaquim) was undertaken. The explant, like, shoot apex and shoot node were washed with detergent and teepol and then sterilized with 0.2% mercurio chloride for 10 minutes. The explants were cut in small pieces under laminar flow hood and subsequently inoculated in Murashige and Skoog (1962) medium modified with different plant growth regulators. The inoculated materials were cultured under aseptic condition at 25+2°C with 16 hours photoperiod of 3000 lux. The medium containing 2% sucrose, 2 mgl¹ BA+ 0.2 mgl¹ NAA was best for shoot node culture and developed 10 shoots/node and 4 leaves per shoot. Protocorm like bodies were developed in cytokinins combination. The combination 1 mgl BA + 0.3 mgl kinetin proved better for getting higher number of buds. However, 7.67 buds/node were found with 1mgl kinetin + 0.1 mgl¹ 2,4-D in around 46 days. It is worthy to note here that 2 mg1¹ kinetin in absence of 2,4-D yielded 7.56 buds/node which is statistically at par with the combination treatment 1mgl kinetin + 0.1 mgl 2,4-D. The shoot apex culture gave significant results on MS. medium supplemented with 2 mgl¹ BA + 0.5 mgl'¹ NAA, 1 mgl¹ BA + 0.2mgl kinetin and 1mgl kinetin + 0.1 mgl 2,4-D. Sub-culturing of plantlet on 2 mgl¹ BA and 0.5 mgl¹ NAA gave about 70-100 shoots. Best result on rooting was achieved on MS medium supplemented with 1 mgr¹ IBA+ 0.5 mgl¹ NAA, 1 mgr¹ NAA + 0.1mgl¹ BA. The maximum root length (49.5 mm and 60 mm) was obtained on medium supplemented with 1 mgr¹ NAA+ 0.1mgr¹ BA and 2 mgl¹¹ NAA + 1 mg/¹¹ IBA respectively. The cultured shoots were hardened successfully in pots containing bark, brick pieces and charcoal in 1:1:1 ratio.
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    ThesisItemOpen 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, Himanshu
    The 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.
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    ThesisItemOpen 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, Himanshu
    Agriculture 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.
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    ThesisItemOpen Access
    Analysis and Characterization of “Landmark Protein” from Rhizobium isolates of Soybean collected from the acid soils of the state of Jharkhand
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Kumar, Manish; Dubey, Himanshu
    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 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. The aim of my work is to check the reproducibility of two unique spots from Rhizobium isolates of Soybean crop and performing 2-dimensional gel electrophoresis. 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 twodimensional gel analysis reveals a host of proteins which are found to be up-regulated or downregulated 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.
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    ThesisItemOpen Access
    Cadmium as a remedy for Fusarium wilt in Tomato (Solanum lycopersicum L.)
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Kumari, Namita; Banerjee, Madhuparna
    Tomato is the highly commercially used edible crop affected by a disease called Fusarium wilt caused by Fusarium oxysporum f.sp. lycopersici. Cadmium induces various functional-based alterations in plants. In the present study, efficacy of cadmium against F. oxysporum in the tomato plant was checked under in vitro conditions by optimizing cadmium concentrations. The result revealed that 100 ppm of cadmium was effective and used to perform the different biochemical analysis. Effective cadmium treatment (supplemented in liquid Murashige and Skoog (MS) media) given to tomato plant grown in- vitro in cocopeat for 5days, after 5 days recovery also given..The effect of cadmium was studied with biochemical analysis of anthocyanin, proline, antioxydative enzyme assay, lipid peroxidation and molecular analysis of total protein content in contrast to control, treated plants and recovered plants. The data so obtained shows an increase in anthocyanin, proline, antioxydative enzyme assay, lipid peroxidation and protein content of treated as well as recovered plant as compared to the control. The maximum increase of biochemical are observed in the sample which are treated with cadmium as well as infected with Fusarium.This increase was related as a defence mechanism to cope up the cadmium induced stress condition by different mechanism like osmoregulation, full utilisation of source and sink, antioxidant action, etc. These aspects can be utilised further in production of transgenic resistant variety of tomato.
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    ThesisItemOpen Access
    Antagonistic effect of Trichoderma for resistance of Fusarium wilt on in vitro grown Grand Nain Banana plant (Musa acuminata)
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Singh, Dikchha; Banerjee, Madhuparna
    The antagonistic activity of Trichoderma against Fusarium oxysporum were examined on potato dextrose agar plates by dual culture method. The results revealed that, all the antagonists showed inhibitory effect on growth of the test fungus and were effective in controlling the growth of pathogen. There were two species of Trichoderma, Trichoderma harzianum (2 different strains viz. 1 and 6) and Trichoderma viride, as there was no specific strains of Trichoderma viride present. Among the strains of Trichoderma sp. ,TH6 was found to be most effective in inhibiting the growth of Fusarium oxysporum of about 50 and 72 per cent of test fungus on 5th and 9th day respectively. The result revealed that both species are effective in antagonizing Fusarium sp. by mycoparasitic activity and also by competing for both rhizosphere colonization and nutrient. Plants treated with Trichoderma have altered metabolic activities as they undergo a stress condition. Thus any changes in the metabolites production was analysed by performing some biochemical assays. Among them the most effected stress related parameters were anthocyanin content, proline content, Antioxidative (CAT & SOD) and ROS by Lipid peroxidation activity. Anthocyanin content was found to be increased significantly upto 10g/F.W. in the T8 treated plants as compared to control (3.525g/F.W.). Similarly results also reflect increased accumulation of proline in banana T8 treated plants upto 1.367μM in vitro as compared to control plants (0.454μM). Also in case of antioxidative enzyme assay, we found that significant 10 fold increase in CAT and 0.166 Unit mg-1 increase in SOD activity was observed as the enzyme concentration decrease and increase in ROS was observed in infected plants upto 0.164mM mg-1 as compared to control (0.0358mM mg-1) due to activation of defense mechanism of plant, which was then maintained on recovery of pretreated infected plants approximately 0.084mM mg-1.On the basis of these results, T8 treated plant was found to be the most suitable for resisting Fusarium. Molecular estimation of total protein content was also performed. The quantified protein so obtained shows an increase in concentration of protein upto 3.327μg/μl in T4 treated plants as compared to control (1.991μg/μl).
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    ThesisItemOpen Access
    Antagonistic effect of Trichoderma for resistance against Fusarium wilt in Chickpea (Cicer arietinum L.)
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Nag, Ritu; Haider, Z. A.
    Chickpea provide high quality protein to large population sectors in South and West Asia, and the Mediterranean Basin. This crop has a significant role in farming systems as substitute for fallow in cereal rotations. Fusarium wilt, caused by the soil borne fungus Fusarium oxysporum f. sp. ciceries, has become a major factor limiting chickpea production worldwide. The pathogen long survival in soil and high pathogenic variability, with eight races 0, 1A, 1B/C, 2, 3, 4, 5, and 6 having been identified so far, are key elements in the development and management of the disease. The antagonistic activity of Trichoderma against Fusarium oxysporum were examined on potato dextrose agar plates by dual culture method. The results revealed that, all the antagonists showed inhibitory effect on growth of the test fungus and were effective in controlling the growth of pathogen .There were two species of Trichoderma, Trichoderma harzianum and Trichoderma viride, and these two strain was selected for the analysis .Among the two strains of Trichoderma viride was found to be most effective in inhibiting the growth of Fusarium oxysporum of about 35 and 57.124 per cent growth of test fungus on 3th and 7th day respectively, whereas Trichoderma harzanium6 inhibited 12.5 and 20 per cent on 3th and 7th day respectively. The results revealed that Trichoderma viride species were found effective against Fusarium oxysporum and thus Trichoderma viride were selected for infecting the chickpea seedlings. The antagonistic effect of Trichoderma was studied with biochemical analysis of anthocyanin, proline, Antioxidant enzyme assay, Lipid peroxidation and molecular analysis of total protein in contrast to control. The data so obtained shows an increase in anthocyanin, proline, Antioxidant enzyme activity and protein content of treated as well as recovered plant however insignicant change in ROS activity was observed.. Molecular estimation of total protein content was also performed. The quantified protein so obtained shows a decrease in concentration of protein in treated plants as compared to control. As per our analysis the treated as well as recovered plant were transfer from in vitro to in vivo condition are found to be still healthy which shows the antagonistic effect of Trichoderma viride against Fusarium f. Sp ciceries..Our study revealed that the biological control agent such as Trichoderma can be useful component of integrated Chickpea Fusarium wilt management.
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    ThesisItemOpen Access
    Antagonistic effect of Trichoderma for resistance against Fusarium wilt of Tomato (Solanum lycopersicum L.)
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) Shahab, Samrah; Banerjee, Madhuparna
    Tomato (Solanum lycopersicum L.) is a widely grown vegetable crop acquiring the status of world’s most popular vegetable due to its importance for nutritive value, processing and export potential. Tomato is affected by many diseases among which wilt caused by Fusarium oxysporum f.sp. lycopersici is one of the important disease, resulting in 25-55% yield loss around the world. Fusarium oxysporum f.sp. lycopersici is a serious soil-borne pathogen and persists for longer periods in the soil. The antagonistic activity of Trichoderma against Fusarium oxysporum were examined on potato dextrose agar plates by dual culture method. The results revealed that, all the antagonists showed inhibitory effect on growth of the test fungus and were effective in controlling the growth of pathogen. There were two species of Trichoderma, Trichoderma harzianum (TH) and Trichoderma viridae (TV). Among the strains of Trichoderma harzianum and Trichoderma viridae Trichoderma viridae was found to be most effective in inhibiting the growth of Fusarium oxysporum f.sp. lycopersici of about 40-60% growth of test fungus on 3rd and 7th day respectively, whereas Trichoderma harzianum inhibited 14.5-20% on 3rd and 7th day respectively.