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ThesisItem Open Access TRANSCRIPTOME AND HORMONE PROFILING OF SESAME (Sesamum indicum L.) DURING INTERACTION WITH Macrophomina phaseolina 3035(JAU, JUNAGADH, 2019-07) RADADIYA NIDHI GHANSHYAMBHA; B. A. GolakiyaSesame (Sesamum indicum L.; Pedaliaceae) is a diploid (2n = 26) dicotyledonous and one of the oldest oil seed crop grown widely in tropical and subtropical areas for its edible oil and food products. Beside of large land covered for cultivation of sesame there is a wide demand–supply gap as its production is constrained by various biotic and abiotic stresses which leads to less productivity. Biotic stresses such as diseases, insects, and pests affect sesame crops adversely causing substantial yield losses. Among the major diseases, charcoal rot caused by Macrophomina phaseolina affects severely at the all stages of the crop growth. In India, this disease incidence was recorded up to 50%. Keeping this constrain in mind present study was undertaken to understand the molecular responses of the host cell against a fungal infection and the putative role of regulatory players. In present study, two different genotypes GT-10 [resistant(R)] and RT-373 [susceptible(S)] were selected to perform transcriptome and four genotypes GT-10, Rama (resistant) and RT-373, AT-306 (susceptible) for biochemical characterization. Total 8 samples were collected at post flowering stage at four different post inoculation stages. Root samples of resistant and susceptible genotypes were collected at 0 hpi (hours post inoculation), 24 hpi, 48 hpi, 72 hpi. The mRNA was isolated from all the collected samples and was sequenced in IonS5 next generation sequencer. After completions of sequencing run the total raw sequence generated were assessed through FastQC quality control tool in which all 8 (eight) samples having good quality sequence were selected for further analysis. Trimming of raw reads yielded a total of 8443423, 8381018, 10143124, 11552102 reads in R1, R2, R3 and R4 respectively and 8713438, 8149237, 11265254, 9281491reads yielded in S1, S2, S3 and S4 respectively. This HQ data was mapped on M. phaseolina genome to remove possible contamination of fungus genome. Unmapped data was extracted and mapped on sesame genome which was downloaded from NCBI. Clear reads were used to carry out differential gene expression analysis. Differential expression analysis of both the genotypes yielded top 10 up and down regulated transcripts expressed at each PIS which were selected for the Gene Ontology enrichment analysis. GO terms like response to auxin (GO:0009733), integral component of membrane (GO:0016021), lipid metabolic process (GO:0006629) were found to be significantly over-represented in sequences having higher expression in GT-10. Moreover, glutathione transferase activity (GO:0004364), oxidoreductase activity (GO:0016491), alpha,alpha-trehalose-phosphate synthase (UDP-forming) activity (GO:0003825), oxidoreductase activity (GO:0016491), integral component of membrane (GO:0016021), beta-fructofuranosidase activity ii (GO:0004564), malate transport(GO:0015743) were found in up regulated sequence in GT-10 over the RT-373. In pathway enrichment analysis, common KEGG pathways like starch and sucrose metabolism and phenyl propanoid biosynthesis were enriched in up-regulated transcripts, however their fold expression was different in both the genotypes. Fatty acid degradation, Arachidonic acid metabolism, Glycerophospholipid metabolism, Tryptophan metabolism, Glyoxylate and dicarboxylate metabolism pathways were enriched in down regulated transcripts in susceptible genotype. Inositol phosphate metabolism, Ascorbate and aldarate metabolism and zeatin biosynthesis were enriched in up regulated transcripts of susceptible genotypes. The DEGs obtained from Deseq software were validated by qRT PCR. The expression patterns of the randomly selected DEGs in to RT-qPCR were in agreement with those obtained by the RNA-Seq, suggesting that the RNA-seq data reflected the real expression patterns of the sesame genes in the compatible interaction. The leaves and roots were analyzed at the interval of 24 hours up to 72 hours after pathogen inoculation. The β-1,3 glucanase and chitinase activities were increased up to 48 hpi thereafter decreased at 72 hpi. The activation of chitinase was more rapid and higher in plants of resistant genotypes than in susceptible genotypes. The results of phenol determination using LC-MS/MS reveals that in both the resistant genotypes, Rama and GT-10, vanillic acid, chlorogenic acid and salicylic acid were found to be higher in diseased condition which indicates these phenolics might play important role in defense mechanism. While in susceptible genotype (RT-373) syringic acid, catechin, coumaric acid, salicylic acid, ferulic acid and cinnamic acid were high in post inoculation stages. In AT-306 vanllic acid, catechin, gallic acid and cinnamic acid were increased during interaction with pathogen. In phytohormone profiling, the levels of IAA, Zeatin, ABA, ACC, JA and SA in the plant extracts were successfully quantified in MRM mode, but this was not possible for GA because of the low level of this phytohormone in plant tissues. At 48 hpi the level of IAA increased in resistant genotypes. The concentration of JA was found higher at 24 hpi in leaves and roots of both susceptible genotypes. No significant change in JA concentration was found in both resistant genotypes among all PIS. The level of SA increased at 48 hpi in all genotypes however its concentration varied among the all. The level of ACC increased in all genotypes after fungus inoculation in both the tissues. Zeatin slightly increased in leaves and root of GT-10 at 48 hpi. It drastically increased in root of Rama at 72 hpi.ThesisItem Open Access GENETIC ARCHITECTURE OF SEED YIELD AND ITS COMPONENT CHARACTERS IN F1 AND F2 GENERATIONS OF CASTOR (Ricinus communis L.) 2951(JAU, JUNAGADH, 2019-09) BARAD HITESH R.; K. L. DOBARIYAThe experimental material, consisting of 101 entries including 10 parents, 45 crosses, 45 F2s and one check hybrid (GCH 7) was raised in a Randomized Block Design with three replications at the Main Oilseeds Research Station, JAU, Junagadh during kharif 2016-17. The objectives of this investigation were to study the nature and magnitude of heterosis and inbreeding depression, general combining ability effect of parents, specific combining ability effect of hybrids, nature and magnitude of gene action involved and genetic components of variation for seed yield per plant and its attributing characters. The results revealed that the mean squares due to genotypes were highly significant for all the characters. The mean squares due to parents, hybrids and F2s were highly significant for all the characters except for number of nodes up to primary raceme in parents. The mean squares due to parents vs hybrids differed significantly for all the characters, while the mean squares due to parents vs F2s were found significant for days to 50% flowering of primary raceme, days to maturity of primary raceme, plant height up to primary raceme, number of effective branches per plant, 100-seed weight and seed yield per plant. The results thus, indicated the presence of considerable amount of genetic variability in the material studied and its suitability for the study of manifestation of heterosis and genetic parameters involved in the inheritance of different traits in castor. The results revealed that the best three hybrids identified on the basis of per se performance and standard heterosis for seed yield per plant viz., JI 368 x RG 2787, DPC 17 x SKI 343 and DPC 17 x JI 353 also depicted significant positive standard heterosis over GCH 7 for important yield contributing traits i.e. JI 368 x RG 2787 manifested significant standard heterosis for days to 50 % flowering of primary raceme, days to maturity of primary raceme, oil content and seed yield per plant; DPC 17 x SKI 343 for days to 50 % flowering of primary raceme, days to maturity of primary raceme, number of nodes up to primary raceme and seed yield per plant and DPC 17 x JI 353 for days to 50% flowering of primary raceme, days to maturity of primary raceme, plant height, number of nodes up to primary raceme, number of capsules on primary raceme and seed yield per plant. This emphasized that high degree of heterosis for seed yield might be attributed to the heterosis observed for the component Abstract ii characters. In the present study, five crosses viz., DPC 17 x JI 368, DPC 17 x JI 372, JI 411 x JI 372, DCS 85 x JI 372 and DCS 85 x PCS 124 displayed significant negative heterobeltiosis and standard heterosis simultaneously for days to 50% flowering of primary raceme, days to maturity of primary raceme, plant height up to primary raceme and number of nodes up to primary raceme. These high heterotic crosses may be considered as potential and be exploited in breeding for the development of early maturity and dwarf genotypes in castor. In the present study, positive significant (desirable) inbreeding depression was observed only for number of nodes up to primary raceme. While, negative significant (desirable) inbreeding depression was observed for some the characters viz., length of primary raceme, effective length of primary raceme, number of effective branches per plant, number of capsules per plant and 100-seed weight. None of the crosses showed significant inbreeding depression for seed yield per plant which is highly desirable. The analysis of variance for combining ability revealed that GCA and SCA variances were significant for all the eleven characters in both the generations reflecting the importance of both additive and non-additive genetic variances for controlling these traits. The ratio of GCA/SCA variances was found more than unity for effective length of primary raceme in F1 indicating the predominance of additive genetic components of variation. Days to 50 % flowering of primary raceme, days to maturity of primary raceme, plant height up to primary raceme, number of nodes upto primary raceme, length of primary raceme, number of capsules on primary raceme, number of effective branches per plant, 100-seed weight, oil content and seed yield per plant, the GCA/SCA ratio was less than unity in both the generations, suggesting the predominance of non-additive gene action for the inheritance of these traits and emphasized the utility of hybrid breeding approach to exploit existing heterosis in the castor. None of the parents was found good general combiner simultaneously for all the traits in both the generations. Among the parents, PCS 124, JI 353 and JI 368 in F1 generation and JI 353 and DPC 17 in F2 generation exhibited good general combining ability effect for seed yield per plant. The best three hybrids on the basis of significant positive sca effect for seed yield per plant were JI 368 x RG 2787 (good x average combiners), DPC 17 x SKI 343 (average x average combiners) and JI 411 x SKI 215 (poor x poor combiners) in F1 generation. While in F2 generation, the top high yielding hybrids DPC 17 × SKI 215 (good x average combiners), DPC 17 × SKI 343 (good x average combiners) and DPC 17 x RG 2787 (good x average combiners), with significant heterobeltiosis as well as standard heterosis, also depicted significant sca effect for seed yield per plant. In facts, in most of the crosses, the best specific combinations for different characters were either good x good, good x poor, average x average, average x poor or poor x poor general combiners. It indicated additive x additive, additive x dominance and dominance x dominance type of gene interactions, which could produce desirable transgressive segregants in subsequent generations. The overall results of genetic components of variances for seed yield and other characters revealed that the estimates of dominance component of genetic Abstract iii variance (D) were significant for days to 50% flowering of primary raceme, days to maturity of primary raceme, number of nodes up to primary raceme, length of primary raceme, effective length of primary raceme, number of capsules per plant, 100-seed weight, oil content and seed yield per plant in both F1 and F2 generations except for number of nodes up to primary raceme in F2 and oil content and seed yield per plant in F1 generation. This suggested the involvement of dominance gene action in the inheritance of these traits. The components of dominance effects (H1 and H2) were significant for all the characters in both the generations except for H2 in case of effective length of primary raceme in F2 and number of effective branches per plant in F1 generation suggesting the prevalence of both these dominance components in the inheritance of all these characters. Further, the magnitude of H1 and H2 components was higher as compared to D component for all the characters in both the generations except H2 for 100-seed weight in F1, which was suggested that dominance components played a pivotal role in the genetic control of all these traits. The results obtained for genetic components of variance are also in confirmation of predominant role of non-additive gene action observed under the combining ability analysis in the present study. Average degree of dominance (H1/D)1/2 was in the range of over dominance for all the characters. The ratio of H2/4H1 indicated the asymmetrical distribution of positive and negative alleles in the parents for all the characters studied in F1 and F2 generations. However, proportion of recessive genes was higher than that of dominant genes for all the traits except days to 50% flowering of primary raceme for which dominant genes was higher than recessive genes. The ratio of total number of dominant to recessive genes in the parents (KD/KR) indicated unequal frequency of recessive and dominant with more number of dominant genes in all the traits except for oil content in both the generations in which proportion of recessive genes was more. Narrow sense heritability estimate was high for effective length of primary raceme and 100-seed weight in F1 generation suggesting that selection based on these attributes would be confer rapid improvement. Moderate heritability was noticed for number of nodes up to primary raceme, length of primary raceme, number of effective branches per plant, number of capsules on primary raceme and oil content in F1 generation, suggesting that breeder can get moderate response while considering above said traits in selection programme. Remaining characters possessed low estimate of narrow sense heritability. It can be concluded that sufficient variation was present in the material for seed yield and its components. Considerable amount of heterobeltiosis and standard heterosis was observed for most of the characters studied. The crosses JI 368 x RG 2787, DPC 17 x SKI 343 and DPC 17 x JI 353 displayed high magnitude of standard heterosis, high sca effect along with high per se performance for seed yield per plant and some of its components. These crosses could be exploited further for obtaining desirable types in castor. Both additive and non-additive genetic variances were found important in the expression of all the traits. The non-additive gene action was more important for all the characters. This suggested that heterosis breeding or biparental mating would be more suitable for the improvement of these traits in castor.ThesisItem Open Access DEVELOPMENT OF TRACTOR OPERATED AUTOMATED INTER AND INTRA ROW WEEDER 2938(JAU, JUNAGADH, 2019-07) JAKASANIA RONAK GORDHANBHAI; R. YadavWeed management is a tedious task especially in organic crop production since synthetic herbicides are prohibited, and manual weeding is relatively costly and time consuming. However, mechanical weed control is one of the options that can be implemented by growers to control weeds but the main challenge to both practical farmers and researchers is the selective control of the intra-row weeds. Based on this challenge the objective of the study was to develop a tractor operated inter and intra row weeder. In this research work static structural analysis of inter row weeder was carried out using Creo and ANSYS software. If a inter row weeder is unable to handle with high magnitude reaction forces from the soil during weeding operation, they become useless due to plastic deformation or parts breaking. Therefore, proper design of inter row weeder is necessary in order to increase their working life time and reduce the farming costs. 3D models of inter row weeder was made using Creo software and static structural analysis was carried out using ANSYS software. Main components of the automated weeder was ultrasonic sensor, D.C. motor, microcontroller, battery, frame, tine, inter and intra row blade. Ultrasonic sensor was used to differentiate crop plant from weed. When the ultrasonic sensor detect a crop plant then intra row blades rotate in anti-clockwise direction and when there are no plants then intra row blades rotate in clockwise direction and come into the working position to remove the particular weed. The performance of automated weeder was evaluated in the field for determining weeding efficiency, percentage of plant damage and field efficiency. These parameters are mainly affected by the forward speed of the tractor and the rake angle of the blade. Different forward speeds of the tractor were 1.5, 2.0 and 2.5 km/h where, three different rake angles 18º, 20º and 22º were selected for evaluating it in laboratory as well as in field. Results of simulation of inter row weeder showed that maximum deformation was observed as 0.65 mm at the given boundary conditions while maximum equivalent (von-mises) stresses of 42.39 MPa was observed. Maximum equivalent stresses was found to be very low as compared to yield strength of material so inter row weeder satisfy the safety conditions. Weeding efficiency, plant damage percentage and field efficiency were found 93.65%, 11.48% and 94.04%, respectively. The optimum forward speed and best rake angle were found as 1.5 km/h and 22º respectively, looking to minimum plant damage and maximum field efficiency. From the experimental results, the depth and with of cut of automated inter and intra row weeder was found as 30 mm and 600 mm, respectively. The draft requirement was 49.13 kgf at 1.50 km/h speed of operation. The power required for weeding operation was 0.93 hp with fuel consumption of 1.21 l/h and 9.57% wheel slip at 12.07% moisture content (d.b.). The theoretical field capacity of weeder was found as 0.09 ha/h while effective field capacity was 0.084 ha/h with the field efficiency of 94.03%. By adapting this automated weeder, problems of the labor crises can be reduced in terms of time reduction 88.44% and cost reduction 35.73% as compared to manually weeding. The payback period and B-C ratio of developed weeder was found as 4.93 years and 2.02, respectively. The results indicate that the tractor operated automated inter and intra row weeder is viable proposition and usful in reducing time as well as cost of operation.ThesisItem Open Access MAPPING AND VALIDATION OF SSR MARKERS ASSOCIATED WITH FOLIAR-FUNGAL DISEASE RESISTANCE QTLs AND DEVELOPMENT OF RILs IN GROUNDNUT (Arachis hypogaea L.) 2928(JAU, JUNAGADH, 2019-08) SUHAIL AHMAD A. HAKEEM; H. P. GajeraGroundnut (Arachis hypogaea L.) is an economically important oilseed crop which is widely cultivated in semi-arid tropic regions of Asia, Africa and America. Two devastating foliar-fungal diseases, viz., late leaf spot (LLS, Phaseoisariopsis personata) and rust (Puccinia arachidis), which often occurs together causes heavy losses along with adverse effects on the quality of kernel and fodder. The objective of the present study was to identify and map the biotic stress relevant candidate EST-SSR markers in the major QTL regions controlling resistance genes for LLS and rust diseases and validate the identified linked markers in diverse groundnut genotypes. The F2 mapping population consisting of 328 lines was developed from diverse genotypes i.e. GJG 17 and GPBD 4. In the parental polymorphism survey, 84 out of 1311 SSR markers were found polymorphic (which includes 900 newly developed EST-SSR at ICAR-DGR and SSRs from the previous reports). A total of 70 out of 84 markers were mapped on 14 linkage groups (LGs) covering a total map distance of 797.55 cM with an average inter-marker distance of 11.39 cM. In the bulk segregant analysis (BSA) four new EST-SSR markers namely DGR329, DGR508, DGR800 and DGR2409 were identified as putatively linked to LLS and rust resistance genomic region. A total of seven new EST-SSR markers, including newly identified 4 markers with BSA, were successfully mapped in the LLS and rust fine QTL region. Among them, DGR259 was found flanked to major LLS QTL. One common QTL for LLS and rust was identified in the map interval of 1.41 cM (between SSR_GO340445 and FRS72) on linkage group (LG-A03) which contributed 47.45% and 70.52% PVE variation, respectively. These two markers, SSR_GO340445 and FRS72, were tightly linked to LLS and rust resistance. Another major QTL for LLS was found on same linkage group (LG_A03), explained 29.06% PVE flanked by the marker DGR259 and FRS59 close to LLS and rust common QTL. The common QTL region identified for LLS and rust resistance carries one Resistance ABSTRACTgene (R gene) and five resistance-related genes which shown to have role in inducing a programmed cell death or hypersensitive response (HR) to combat pathogen attack. A total of 7 new biotic stress relevant EST-SSR markers (DGR259, DGR312, DGR329, DGR361, DGR508, DGR800 and DGR2409) were integrated in the major QTL hotspot region, where one of it flanking the major LLS QTL region and six markers showed tight association with major QTL region. Most EST-SSR markers were from the genes involved in the disease resistance. Initial validation carried out in selected genotypes showed that these markers were able to distinguish resistant genotypes more clearly and efficiently. Further validation of all the 24 SSR markers saturated in LLS and rust QTL region were validated in 177 diverse Indian genotypes. Of these, only 12 genotypes resistant to both LLS and rust carries resistant alleles of markers flanking the LLS and rust QTLs and eight more markers present in the surrounding region of these QTLs. Moreover, the new identified EST-SSR markers were validated in the introgression lines. All the markers were able to distinguish resistant introgression line from susceptible recurrent parent and susceptible genotypes. These markers could be used to introgress this fine QTL regions to improve LLS and rust resistance in the groundnut breeding program more efficiently by minimizing the risk of linkage drag. Furthermore, these markers will be of great help to the groundnut breeders for the selection of resistant genotype from the segregating population, advanced breeding lines and varieties even in the absence of a disease nursery or epiphytotics and will be useful in MAS under crop improvement programThesisItem Open Access DEVELOPMENT AND CHARACTERIZATION OF FISH CRACKERS MADE FROM THE BULL’S EYE (Priacanthus hamrur, Forsskal, 1775) FISH MEAT 2914(JAU, JUNAGADH, 2019-07) CHUDASAMA BHARATKUMAR GOVINDJI; S. M. ZOFAIRThe bull’s eye fish (Priacanthus hamrur), which is present in a large biomass in the South-East coast of India, is generally considered an underutilized fish species. In an attempt to add value, it was used to develop fish crackers using this fish. It is the most popular snack food in Southeast Asian countries. Different starches flour (Rice, Sago and Tapioca), as a functional ingredient, was used to prepare fish cracker at 50:50 ratio in Experiment – 1 as prelim trial for fish cracker. The products was subjected analysed for proximate as well as comparative physicochemical, microbiological and sensory evaluation so as to determine the nutritive value and its quality attributes for general acceptance. Dried cracker had moisture content of 10 to 12 %, protein content of 22 to 24 % whereas, lipid content found low but it was significantly increased after frying due to oil absorption. Maximum linear expansion was observed in tapioca based fish crackers (83.03 %). Water absorption index (WAI) was decreased while water solubility index (WSI) increase for tapioca based fish cracker. Maximum lightness and lower redness as well as yellowness was noticed with sago based fish cracker after frying. However, TMA, TVBN, PV and TPC were found well within the critical limit for rice, sago and tapioca based crackers respectively. Fish crackers incorporated with tapioca starch had maximum score for sensory aspect of crispiness (4.6), texture (4.4), odour (3.4) taste (3.8) and overall acceptability (4.4). In general tapioca starch was suitable for development of fish cracker. In Experiment – 2, fish crackers were prepared by blending lean fish (Priacanthus hamrur) along with tapioca starch flour in five different proportions as treatments T1 – (C) (50:50), T2 (40:60), T3 (30:70), T4 (20:80) and T5 (10:90) respectively. The samples were examined for their physicochemical, microbiological and sensory properties upon dried and fried condition. Protein, lipid, and ash contents increased whereas, moisture and carbohydrate decreased with an increase in the proportion of fish in formulation of fish cracker. The linear expansion of the fried crackers increased with the increased proportion of fish. The sensory evaluation tests showed that the most acceptable formulations for the crackers were obtained using 40:60 fish to tapioca starch (T2) combination. ii The physicochemical properties like linear expansion, oil absorption and hardness decreased whereas bulk density increased with the increase in the fish concentration. A sharp decrease in the Water Absorption Index (WAI) and a sharp increase in the Water Solubility Index (WSI) were noticed. The sensory evaluation conducted by the expert panels suggested that 40% fish incorporated crackers with a protein content of 21.94 % is having a better overall quality and is considered to be the best formulation. The objective of following Experiment – 3 study was to develop a standard recipe of fish cracker production by optimization of tapioca flour-based fish cracker using Response Surface Methodology (RSM). The fish:starch ratio in the selected formula (40:60) was optimised so that a product of maximum linear expansion, lesser oil absorption and best organoleptic qualities was obtained. The gelatinization conditions of the fish crackers were optimized in order to improve the linear expansion, bulk density, hardness and oil absorption using RSM. The process variables were steaming time (40, 50, 60 min); gel setting time (12, 18, 24 hrs.) and drying temperature (40, 50, 60 ⁰C) and the responses taken were linear expansion, bulk density, hardness and oil absorption. Quadratic models expressed the relationship between the dependent and independent variables. The average value found was 88.58 % for linear expansion, 0.0827 g/cm³ for bulk density, 1216.90 N/cm2 for hardness and 25.75 % for oil absorption of the products. The optimum condition predicted by RSM to have high linear expansion of products was at a steaming time of 50 min, a gel setting time of 18 hrs. and drying temperature of 55 ⁰C. Higher expansion of samples resulted in decreased hardness. This study enabled to optimize few important process conditions that are not examined earlier to improve the linear expansion, lower the hardness and reducing the oil uptake of the fish crackers during formulation. Fish cracker developed under optimized conditions contained 10.00 % of moisture, 22.70 % of protein, 1.98 % of lipid, 5.62 % of ash, 59.71 % of carbohydrate and 347 kcal/kg of energy value noticed for the dried crackers. Whereas, linear expansion was improved to 102.62 % after optimization. However, hardness and oil absorption was slightly increased after optimization using RSM. The storage studies conducted on both dried and fried samples suggested that the samples are safe and shelf stable for a period of 180 days. The objective of study was to found the effect of selected hydrocolloids i.e. xanthan gum and guar gum at 1 % of incorporation, on nutritional value, linear expansion, oil absorption, overall acceptability and shelf life of fish cracker were studied. As a result, xanthan gum significantly (p < 0.05) decreased the hardness and oil absorption as well as retained nutritional value and improve linear expansion of the fried crackers during storage. Linear expansion showed decreasing trends with increasing storage period. However, higher linear expansion (108.38 %) was observed with treatment T7 (1% xanthan added cracker) which changed to 73.47 % at the end of storage. Fried fish crackers had hardness of 1617.29, 913.68 and 1156.45 (N/cm2 ) initially for treatments T6 (C), T7 and T8 respectively which was increased as period of storage increase. Sharp raising in hardness value was noticed after 120 days of storage period. Oil absorption found within the ranged of 23.35 to 27 .12, 19.49 to 22.50 and 22.06 to 25.76 % for the treatments T6 (C), T7 and T8 respectively during storage. Among the two hydrocolloids evaluated in the present study, xanthan gum (T7) was iii the most effective in decreasing the oil absorption property while increasing the water binding ability and viscoelastic properties, and it decreased the sizes and numbers of the cavities in the gel that influenced the textural characteristics of the final product. However, guar gum interfered with the gelation process and significantly decreased the viscoelastic properties of the product and also retaining more oil after frying of the cracker during subsequent storage. With storage period TMA, TVBN, peroxide value, moisture content, colour a* values and overall change in colour (ΔE) increases but colour L* and b* values decreased. However, TPC decreased in subsequent storage. Overall acceptability of fish crackers formulated with 1% of xanthan gum was rated highest (4.2 to 2.8) compared to control (3.8 to 2.4) and guar gum (3.4 to 2.0) treated sample throughout the storage period. Based on physicochemical and microbiological quality, hydrocolloids added fish cracker was found to have a shelf life of 180 days while based on sensory characteristics the shelf life was 150 days under LDPE pouch packaging at ambient temperature.The bull’s eye fish (Priacanthus hamrur), which is present in a large biomass in the South-East coast of India, is generally considered an underutilized fish species. In an attempt to add value, it was used to develop fish crackers using this fish. It is the most popular snack food in Southeast Asian countries. Different starches flour (Rice, Sago and Tapioca), as a functional ingredient, was used to prepare fish cracker at 50:50 ratio in Experiment – 1 as prelim trial for fish cracker. The products was subjected analysed for proximate as well as comparative physicochemical, microbiological and sensory evaluation so as to determine the nutritive value and its quality attributes for general acceptance. Dried cracker had moisture content of 10 to 12 %, protein content of 22 to 24 % whereas, lipid content found low but it was significantly increased after frying due to oil absorption. Maximum linear expansion was observed in tapioca based fish crackers (83.03 %). Water absorption index (WAI) was decreased while water solubility index (WSI) increase for tapioca based fish cracker. Maximum lightness and lower redness as well as yellowness was noticed with sago based fish cracker after frying. However, TMA, TVBN, PV and TPC were found well within the critical limit for rice, sago and tapioca based crackers respectively. Fish crackers incorporated with tapioca starch had maximum score for sensory aspect of crispiness (4.6), texture (4.4), odour (3.4) taste (3.8) and overall acceptability (4.4). In general tapioca starch was suitable for development of fish cracker. In Experiment – 2, fish crackers were prepared by blending lean fish (Priacanthus hamrur) along with tapioca starch flour in five different proportions as treatments T1 – (C) (50:50), T2 (40:60), T3 (30:70), T4 (20:80) and T5 (10:90) respectively. The samples were examined for their physicochemical, microbiological and sensory properties upon dried and fried condition. Protein, lipid, and ash contents increased whereas, moisture and carbohydrate decreased with an increase in the proportion of fish in formulation of fish cracker. The linear expansion of the fried crackers increased with the increased proportion of fish. The sensory evaluation tests showed that the most acceptable formulations for the crackers were obtained using 40:60 fish to tapioca starch (T2) combination. ii The physicochemical properties like linear expansion, oil absorption and hardness decreased whereas bulk density increased with the increase in the fish concentration. A sharp decrease in the Water Absorption Index (WAI) and a sharp increase in the Water Solubility Index (WSI) were noticed. The sensory evaluation conducted by the expert panels suggested that 40% fish incorporated crackers with a protein content of 21.94 % is having a better overall quality and is considered to be the best formulation. The objective of following Experiment – 3 study was to develop a standard recipe of fish cracker production by optimization of tapioca flour-based fish cracker using Response Surface Methodology (RSM). The fish:starch ratio in the selected formula (40:60) was optimised so that a product of maximum linear expansion, lesser oil absorption and best organoleptic qualities was obtained. The gelatinization conditions of the fish crackers were optimized in order to improve the linear expansion, bulk density, hardness and oil absorption using RSM. The process variables were steaming time (40, 50, 60 min); gel setting time (12, 18, 24 hrs.) and drying temperature (40, 50, 60 ⁰C) and the responses taken were linear expansion, bulk density, hardness and oil absorption. Quadratic models expressed the relationship between the dependent and independent variables. The average value found was 88.58 % for linear expansion, 0.0827 g/cm³ for bulk density, 1216.90 N/cm2 for hardness and 25.75 % for oil absorption of the products. The optimum condition predicted by RSM to have high linear expansion of products was at a steaming time of 50 min, a gel setting time of 18 hrs. and drying temperature of 55 ⁰C. Higher expansion of samples resulted in decreased hardness. This study enabled to optimize few important process conditions that are not examined earlier to improve the linear expansion, lower the hardness and reducing the oil uptake of the fish crackers during formulation. Fish cracker developed under optimized conditions contained 10.00 % of moisture, 22.70 % of protein, 1.98 % of lipid, 5.62 % of ash, 59.71 % of carbohydrate and 347 kcal/kg of energy value noticed for the dried crackers. Whereas, linear expansion was improved to 102.62 % after optimization. However, hardness and oil absorption was slightly increased after optimization using RSM. The storage studies conducted on both dried and fried samples suggested that the samples are safe and shelf stable for a period of 180 days. The objective of study was to found the effect of selected hydrocolloids i.e. xanthan gum and guar gum at 1 % of incorporation, on nutritional value, linear expansion, oil absorption, overall acceptability and shelf life of fish cracker were studied. As a result, xanthan gum significantly (p < 0.05) decreased the hardness and oil absorption as well as retained nutritional value and improve linear expansion of the fried crackers during storage. Linear expansion showed decreasing trends with increasing storage period. However, higher linear expansion (108.38 %) was observed with treatment T7 (1% xanthan added cracker) which changed to 73.47 % at the end of storage. Fried fish crackers had hardness of 1617.29, 913.68 and 1156.45 (N/cm2 ) initially for treatments T6 (C), T7 and T8 respectively which was increased as period of storage increase. Sharp raising in hardness value was noticed after 120 days of storage period. Oil absorption found within the ranged of 23.35 to 27 .12, 19.49 to 22.50 and 22.06 to 25.76 % for the treatments T6 (C), T7 and T8 respectively during storage. Among the two hydrocolloids evaluated in the present study, xanthan gum (T7) was iii the most effective in decreasing the oil absorption property while increasing the water binding ability and viscoelastic properties, and it decreased the sizes and numbers of the cavities in the gel that influenced the textural characteristics of the final product. However, guar gum interfered with the gelation process and significantly decreased the viscoelastic properties of the product and also retaining more oil after frying of the cracker during subsequent storage. With storage period TMA, TVBN, peroxide value, moisture content, colour a* values and overall change in colour (ΔE) increases but colour L* and b* values decreased. However, TPC decreased in subsequent storage. Overall acceptability of fish crackers formulated with 1% of xanthan gum was rated highest (4.2 to 2.8) compared to control (3.8 to 2.4) and guar gum (3.4 to 2.0) treated sample throughout the storage period. Based on physicochemical and microbiological quality, hydrocolloids added fish cracker was found to have a shelf life of 180 days while based on sensory characteristics the shelf life was 150 days under LDPE pouch packaging at ambient temperature.ThesisItem Open Access SEASONAL INFLUENCE ON SEXUAL BEHAVIOUR AND PROTEIN PROFILING OF MAJOR HSP (HSP 70, 90) IN RELATION TO SEMEN QUALITY AND FERTILITY OF GIR BULL 2893(JAU, JUNAGADH, 2019-07) SOLANKI GAJENDRASINGH BHAWARSINGH; F. S. KAVANIFor successful AI in cattle and buffalo, bull fertility is essential to assure optimal reproductive efficiency. Routinely used traditional semen quality tests and even CASA provide limited information about the potential fertility of bulls. Several studies have been conducted in the past decades in the search of exact and potential fertility markers in bulls. So determining the seminal proteomic pattern of bulls with high semen freezability and low semen freezability, we can use this technique as a prediction tool to choose the semen donor bulls. Studies on sexual behaviour, semen characteristics, fertility and potential biomarkers of fertility, viz., heat shock proteins (HSP 70, 90) in semen as such, and the documented reports on the influence of seasons on these aspects are meager in Gir bulls, the world famous milch purpose cattle breed of India. These indices could be used in selection and culling of Gir bulls in the breeding program. Hence these investigations were undertaken at Cattle Breeding Farm, JAU, Junagadh during two seasons (summer and winter) from Nov 2017 to Oct 2018. The study was carried out to know the seasonal variation and correlations amongst and between the sexual behaviour, semen quality traits, conception rates and major heat shock proteins expression in six mature Gir bulls. The effect of season on individual bulls for these parameters was also studied. A total of 96 observations were availed at fortnightly interval for sexual behaviour study, while 384 semen samples at twice-weekly interval during each season from six bulls were utilized. The first AI conception rate was found by inseminating a total of 1200 Gir cows, equally dividing for each bull and season. The protein profiling of major HSP proteins (70, 90) in sperm from neat as well as post-thawed semen was carried out. The mean temperament (0-5 scale), libido (0-9 scale), penile erection (0-4 scale), protrusion (0-4 scale), intensity of thrust (0-4 scale), reaction time (sec.) and Flehmen reaction (0, 1) during summer were 1.77±0.08, 5.23±0.22, 2.73±0.07, 2.68±0.07, 3.19±0.11, 153.54±4.82 and 0.46±0.07, respectively, while the respective values in winter season were 2.08±0.06, 5.96±0.22, 3.02±0.06, 2.96±0.43, 3.40±0.39, 128.09±4.24 and 0.56±0.07. There were significant differences among bulls for these traits in both the seasons. However, in general, summer significantly exerted an adverse effect on the sexual behaviour of Gir bulls, while winter was the favoured season.Abstract ii The neat seminal characteristics of Gir bulls, viz., volume (ml), colour (score), concentration (million/ml), mass activity (score), initial motility (%), live sperm (%), abnormal sperm (%), HOST reactive sperm (%) and acrosomal integrity (%) observed in summer season were 5.02±0.27, 3.06±0.11, 1.070.46±49.56, 3.83±0.05, 79.58±0.05, 87.23±0.75, 6.73±0.47, 76.02±0.57, 90.35±0.35, respectively, whereas the respective values in winter were 5.81±0.26, 3.35±0.11, 1282.98±58.08, 3.94±0.25, 82.40±0.56, 90.17±0.71, 5.13±0.51, 78.23±0.49, 92.42±0.31. The percentages of post-thaw motility, post-thaw live sperm, post-thaw abnormal sperm, post-thaw HOST reactive sperm, post-thaw acrosomal integrity and first AI conception rate for semen cryopreserved during summer were 53.19±0.32, 65.90±0.92, 9.94±0.57, 55.27±0.82, 74.38±0.38 and 49.33±0.32, respectively, whereas the corresponding values for semen cryopreserved during winter were 55.29±0.39, 69.06±1.09, 8.19±0.54, 58.58±0.86, 75.17±0.35 and 55.04±0.35. There were significant differences among bulls for all of these traits in both the seasons. Despite the fact that the sperm motility, concentration, viability, abnormality, membrane integrity and acrosomal integrity were within permissible limit for semen processing throughout the year, the quality was significantly better during the winter than the summer season under Junagadh Agro-climatic conditions. Temperament, libido, penile erection, penile protrusion, and intensity of thrust were significantly and positively correlated among each other, and negatively with reaction time and Flehmen reaction. Sperm motility traits like mass activity, initial motility and post-thaw motility showed significant positive correlations among themselves (r=0.562 to 0.689) and with first AI conception rates (r=0.133 to 0.357). Sperm viability was positively correlated with post-thaw motility (r=0.164) and conception rate (r=0.266). Membrane integrity (r=0.188) and acrosomal integrity (0.373) were positively correlated with the conception rate. Reaction time was significantly negatively correlated with mass activity (r=-0.203) and initial motility (r=-0.181). Conception rate was significantly positively correlated with temperament (r=0.261), penile erection (r=0.253), penile protrusion (r= 0.277) and thrust (r=0.324), while negatively with reaction time (r=-0.293). The reaction time was also negatively correlated with initial live sperm, acrosomal integrity and post-thaw motility. The mean protein concentrations (ng/mg of protein) of major heat shock protein 70 (HSP70) were significantly higher during summer as compared to winter season both at a pre-freeze ((56.68 ±43.19 vs. 40.25±3.07) and post-thaw stage (118.24±6.44 vs. 80.81±5.11). While the concentration (ng/mg of protein) of HSP90P at the pre freeze stage was non-significantly higher during summer than winter (1.27±0.05 vs. 1.19±0.07) season, while the respective values at the post-freeze stage were 2.36±0.08 and 2.37±0.13. This was also confirmed qualitatively by western blotting. The HSP70P in neat semen had a significant positive correlation with mass activity (r=0.326), initial motility (r=0.463), sperm viability (0.565), acrosomal integrity (r=0.330), first AI conception rate (r=0.431) and HSP70P in post-thaw semen (r=0.474). The HSP90P had a significant positive correlation with sperm motility (r=0.322), viability (r=0.301) and conception rate (r=0.409). It could be concluded from the present study that the Gir bulls had good sexual behaviour, better semen quality and quantity throughout the year. However, semen quality was better during winter as compared to the summer season. The protein expression of major HSPs (70, 90) may be utilized as markers for heat tolerance, sexual behaviour, semen quality and conception rate for the Gir bull.ThesisItem Open Access EFFECTS OF PROBIOTIC Lactobcillus spp. GRWOTH, SURVIVAL STRESS TOLERANCE AND RESISTANCE AGAINST Vibrio Spp. ON JUVENILES OF PACIFIC WHITE SHRIMP Litopenause vannamei (BOONE, 1931) 2829(JAU, JUNAGADH, 2019-05) PATEL MUKESHKUMAR PURUSHOTTAMBHAI; K. H. VadherThesisItem Open Access GENETICS OF SEED YIELD AND ITS COMPONENT TRAITS IN CASTOR (Ricinus communis L.), 2802(JAU, JUNAGADH, 2019-06) MORI KIRAN KALA; J. B. PatelThe present investigation was undertaken with a view to investigate genetic basis of seed yield and its component traits, to study the extent of heterosis and inbreeding depression and underlying genetic causes for different traits in castor. The experimental materials consisted of twelve generations, namely P1, P2, F1, F2, B1, B2, B11, B12, B21, B22, B1s and B2s of four crosses of castor viz., JP 104 x JI 433 (cross 1), SKP 84 x JI 433 (cross 2), SKP 84 x JI 437 (cross 3) and SKP 84 x JI 441 (cross 4). Experiment was laid-out in Compact Family Block Design with three replications during Kharif 2017 at Sagdividi Farm, Department of Seed Science and Technology, College of Agriculture, Junagadh Agricultural University, Junagadh. The observations were recorded on twelve characters viz., days to flowering of primary raceme, days to maturity of primary raceme, plant height up to primary raceme (cm), number of nodes up to primary raceme, length of primary raceme (cm), effective length primary raceme (cm), number of effective branches per plant, number of capsules on primary raceme, shelling out turn (%), 100 seed weight (g), seed yield per plant (g) and oil content (%) on five randomly selected plants in each replication for P1, P2 and F1; fifteen plants for B1 and B2; and twenty five plants for each of F2, B11, B12, B21, B22, B1s and B2s. The analysis of variance between families (crosses) revealed that the mean squares due to crosses were significant for all the characters studied except oil content. The analysis of variance among progenies within each family indicated significant differences among twelve generation means for all the characters studied in all the four crosses. Mean performance of F1 hybrids exceeded the value of their better parent in desired direction for length of primary raceme and effective length of primary raceme in JP 104 x JI 433 and SKP 84 x JI 441; for number of effective branches per plant and 100 seed weight in SKP 84 x JI 441; for oil content in JP 104 x JI 433 and SKP 84 x JI 433; for number of nodes up to primary raceme in SKP 84 x JI 433, SKP 84 x JI 437 and SKP 84 x JI 441; and for number of capsules on primary raceme and shelling out turn in SKP 84 x JI 437. None of the F1 exceeded the seed yield per plant over their better parent in positive direction. The per se performance of F1 was lower than its better parent for plant height up to primary raceme in crosses SKP 84 x JI 437 and SKP 84 x JI 441 and for number of nodes up to primary raceme in cross JP 104 x JI 433, while for days to flowering of primary raceme and days to maturity of primary raceme, none of the F1 per se was lower than its better parent. Mean performance of backcross progenies was not consistent in different crosses for different traits, however, several back crosses exceeded the mean performance over their better parent in desirable direction for seed yield per plant and its component traits. With respect to seed yield per plant, back cross progenies B11, B21 and B1s in cross JP 104 x JI 433; B1, B12, B21, B22 and B1s in cross SKP 84 x JI 433; all the backcross progenies in crosses SKP 84 x JI 437; and all the backcross progenies except B1 in cross SKP 84 x JI 441 exceeded the mean performance over their better parents. Significance of simple scaling tests and Cavali’s joint scaling test indicated the presence of gene interactions for all the traits in all four crosses. Based on six parameter model of Hill (1966), a valid conclusion can be drawn by the 𝜒 2 (2) value at six degrees of freedom about the presence of higher order epistasis. The significant 𝜒 2 (2) pointed out presence of trigenic or higher order epistasis in all the crosses for all the characters. The trigenic ten-parameter model was found to be adequate in cross JP 104 x JI 433 for effective length primary raceme; in crosses JP 104 x JI 433 and SKP 84 x JI 437 for number of capsules on primary raceme; and in cross SKP 84 x JI 441 for number of effective branches per plant. On the other hand, 𝜒 2 (3) with two degrees of freedom was found significant for remaining traits in all the four crosses showing the presence of higher order epistasis and/or linkage. All the ten-parameters were significant for shelling out turn percentage in two crosses SKP 84 x JI 437 and SKP 84 x JI 441 and for days to maturity of primary raceme in one cross SKP 84 x JI 433. The heterosis over better parent was significant and positive in all four crosses for days to flowering of primary raceme and days to maturity of primary raceme, indicating delay in flowering and maturity in hybrid combinations. The heterosis over better parent was significant and negative for dwarf stature in cross SKP 84 x JI 437. The heterosis over better parent was significant and positive for longer length of primary raceme in crosses JP 104 x JI 433 and SKP 84 x JI 441, for effective length of primary raceme in cross JP 104 x JI 433, for shelling out turn in cross SKP 84 x JI 437, for test weight in SKP 84 x JI 441, and for oil content in JP 104 x JI 433 and SKP 84 x JI 433. The heterobeltiosis was significant and negative in crosses JP 104 x JI 433, SKP 84 x JI 433 and SKP 84 x JI 441 for seed yield per plant. Moderate inbreeding depression was observed in the present study as a whole. The observed and the expected estimates for heterosis over mid parent, over better parent and inbreeding depression were in close agreement with one another for days to flowering of primary raceme, days to maturity of primary raceme, plant height up to primary raceme, number of nodes up to primary raceme, 100 seed weight and oil content in all four crosses. Overall from the present study, it could be concluded that seed yield per plant and its component traits recorded in four castor crosses were governed by additive, dominance and digenic and/or trigenic epistasis gene effects. Reciprocal recurrent selection could be followed which would facilitate exploitation of both additive and non-additive gene effects simultaneously for genetic improvement in castor. Importance of duplicate type of gene action was observed for all of the characters studied in all four crosses showing digenic/trigenic interaction. Under a situation of this type, breeding procedures involving either multiple crosses or biparental crosses may be restored to get transgressive segregants.ThesisItem Open Access MORPHOMETRIC AND HYDROLOGIC ANALYSIS OF BHADAR BASIN OF GUJARAT USING REMOTE SENSING & GIS TECHNIQUES, 2801(JAU, JUNAGADH, 2019-01) KELAIYA JAINISH HIMANSHUBHAI; H. D. RankThe natural resources planning and management can be successful if it is based on the drainage basin scale. The feasible and viable techno-economic planning for the soil and water conservation requires the hydrologic response to climate and various land uses. Morphometric analysis of a drainage basin expresses fully the state of dynamic balance that has been attained due to dealings between matter and energy. Morphometric analysis based either in GIS environment or otherwise is a competent tool for geo-hydrological studies and are very useful for identifying and planning the groundwater potential zones and watershed management, including the whole gamut of natural resources connected with the watershed. It becomes necessary to understand the topography, erosion status and drainage patterns of the region. The SWAT model is the tool to assess the water balance components of the hydrological cycle like rainfall, runoff, evapotranspiration and groundwater recharge under given land use, management and climate. The runoff and groundwater recharge assessment are very useful for the water resources development and planning. This study was undertaken for the Bhadar River basin divided into 16 watersheds and total drainage area of 7330.40 km2 . Morphometric analysis, such as linear, aerial and relief aspects of the Bhadar River basin has been done with the help of remote sensing and GIS techniques. The climatic, discharge and sediment yield data were divided into calibration period (1981-2000) and validation period (2001- 2010). The climatic and discharge data, sediment data, SRTM-DEM imagery, soil maps and land use/cover classification from LISS IV imagery are used as primary inputs for SWAT model. The SWAT model was used to assess the hydrologic response of the basin after sensitivity analysis, calibration and validation. In this study SWAT model was developed for sediment yield simulation at the outlet of the basin and this model applied to the every watersheds of basin for the estimation of sediment yield. The watershed prioritization was made using the morphometric parameters and sediment yield from all watersheds. The groundwater recharge was estimated using water table fluctuation and SWAT Model and compared. Hydrologic analysis is an analysis of interrelated components, including the processes of precipitation, evaporation, transpiration, infiltration, groundwater flow, streamflow, sediment yield etc., in addition to those structures and devices that are used to manage the hydrologic system. The capabilities of remote sensing and GIS techniques are explored in this study to simulate sediment yield in the basin. The ground water recharge was also estimated using empirical equations. Precipitation measured at 37 stations over 1981−2010 is used as an input dataset to establish hydrologic analysis. 30 years of climatic and discharge data along with different remote sensing imagery and soil map were also used. The empirical model was developed and proposed to assess the groundwater recharge using the seasonal rainfall data. The performance indices like NSE and goodness of fit were used in the modeling to assess the accuracy in the predictions. The river basin is designated as 7th order basin. The 1 st order streams are mostly dominating the basin. The moderate drainage density indicates that weak subsurface material and belongs to medium texture. The study reveals that the basin has less elongated to elongated shape and associated with steep ground slopes. The ranges of Rb are high as mean Rb of the basin is 5.38, and it has been characterized by the influences of underlying structures and lithology. Relief having a value of 0.303 km indicates low gravity of water flow as well as infiltration and high runoff conditions. The watershed 5G1B15 should be treated first while 5G1B4 at last. Highest priority indicates the greater degree of erosion in the particular watershed and it becomes potential candidate for applying soil conservation measures first. The developed soil theme map shows that clay and fine soil are dominating in the Bhadar river basin. The land use/cover analysis shows that agricultural land is dominating in the basin. The sediment yield obtained from different watershed using SWAT model also gives the same highest priority to watershed 5G1B15 having highest sediment yield among all watersheds. The performance of SWAT model is satisfactory in terms of different performance indices such as NSE and goodness of fit for calibration(NSE=64.6%, R2 =0.762) and validation(NSE=68.5%, R2 =0.798). SWAT model provides a better description of water balance of the watershed. The average annual rainfall, runoff, evapotranspiration, percolation to shallow aquifer and recharge to deep aquifer in the basin are 569.3 mm, 197.19 mm, 244.6 mm, 111.64 mm and 5.58 mm respectively. The annual average runoff is 36.37% of the average annual rainfall. The average annual sediment yield during calibration and validation periods are 14.13 t/ha and 19.40 t/ha. Among the developed models, the non-linear empirical ground water recharge model as R = 1.642(P – 230.054)0.583 (with NSE=86.59%, where R and P are the seasonal groundwater recharge, mm for the seasonal rainfall amount, mm of P) was found best. Performance of SWAT, a physically based model and empirical models were compared to simulate the ground water recharge of the Bhadar river basin. The best fit empirical model performs better than and SWAT model (NSE = 74.94%, R2 =0.898).
