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ThesisItem Open Access DIFFERENTIAL GENE EXPRESSION IN RICE (Oryza sativa L.) IN RESPONSE TO SALINITY STRESS THROUGH cDNA-AFLP(Plant Molecular Biology and Biotechnology Dept., NMCA, NAU, Navasari, 2016-07) Patel, Bhumi J.; Singh, DiwakarRice is a staple food crop and over half of the world’s population depends on it and in Asian diet 30-80% daily calories consumed from rice. Various abiotic stresses limit rice production in rainfed environments, which comprise about 45% of the global rice area. Soil salinity limits the rice plant’s growth and development, resulting in yield losses of more than 50 percent. Rice yields are reduced by 12 percent for every unit of salinity (dSm-1). Different morpho-physio parameters were studied under different level of salt treatment. Under saline condition the germination percentage, shoot length, root length, shoot & root fresh and dry weight were studied. The magnitude of reduction in germination percent increased with increasing salinity levels. Shoot length showed decreasing trend with the increasing salinity levels. Root length showed increasing trend with the increasing salinity levels. The roots of IR 29 genotype (salt sensitive) were affected maximum with salt stress as a result root fresh weight was found minimum. Different biochemical constituents were analyzed at different time intervals and at different salinity levels. Rice seedling showed decline in chlorophyll content with salinity. For carotenoid content also decline was recorded ABSTRACT with increasing salinity. MDA content was also studied from rice shoot as well as root. MDA content increased with increasing salinity level. The MDA content in rice seedlings represents the peroxidation level of membrane lipids, which is used as an effective index of cellular damage. Antioxidant activity (%) was studied from root and shoot and maximum antioxidant activity was recorded in Pokkali (salt tolerant genotype) and minimum in IR 29 (salt sensitive genotype). The antioxidant activity shows an increasing trend with increasing salt concentration. After analysis for Na+ and K+ the maximum Na+/K+ ratio in shoot and root was measured in Pokkali. Dandi showed maximum Na+/K+ ratio in root at highest salinity level (200mM). High NaCl salinity leads to growth suppression in most of the glycophytes due to the decrease in water and nutrient uptake and the toxicity of Na+ and/or Cl-. Enzyme assay was done from shoot and root at different time interval and at different salt concentrations. Enzyme acitivity of Apox, Gpox, Catalase, GR and SOD was studied. In all cases higher enzyme activity was recorded for salt tolerant genotype (Pokkali and Dandi), whereas, lower for salt sensitive genotype (IR 29). Polyamine content was also studied in shoot only using HPLC and it was found maximum in salt tolerant genotype (Dandi) and minimum in salt sensitive genotype IR 29. Differential gene expression analysis was carried out using the technique cDNA AFLP and several fragments were uniquely identified as up-regulated in shoot as well as root. Most of the up-regulated fragments were reported from salt tolerant varieties Dandi and Pokkali, whereas only few up-regulated fragments were reported from salt sensitive genotype IR 29. After elution and sequencing of nine specific fragments it was blast using the Rice Genome Database Project or NCBI Data Base. After bioinformatic analysis plasma membrane-associated cation-binding protein, proline-rich extensin-like protein, Retrotransposon protein, RNA recognition motif containing protein, transposon protein, BZIP protein, NADH dehydrogenase 1 alpha subcomplex subunit 13 and OsFBD9 - F-box and FBD domain containing protein were uniquely expressed. Thus, it can be concluded that cDNA AFLP could identify several differentially expressed genes by studying contrasting genotypes for salinity stress. Cell wall-related genes, especially plasma membrane-associated cation binding gene, proline-rich extension-like gene were up-regulated in salt tolerant genotypes, suggesting cell wall restructuring is a general adaptive mechanism during salinity stress. Further the differentially expressed genes need to be validated using real-time PCR analysis.ThesisItem Open Access Comparative Protein Expression Analysis of Bacterial Wilt Susceptible and Resistant Brinjal(Plant Molecular Biology and Biotechnology Dept., NMCA, NAU, Navasari, 2016-06) Gandhi, Anjali B.; Kapadia, ChintanBrinjal (Solanum melongena L.) is an important vegetable crop and growing in various tropical, sub tropical and warm temperate regions of the world. Bacterial wilt caused by Ralstonia solanacerum has become a major problem causing an important damage in production of Brinjal. Bacterial wilt disease is very difficult to control because the pathogen is soil-borne and has a wide host range. Infected stems and roots of brinjal samples were collected from different areas of NAU, Navsari. Isolation were performed on Triphenyl Tetrazolium Chloride (TZC) medium. Bacteria with similar characteristics were isolated which produced fluidal colonies that were entirely white or white with pink/red centre colonies after incubation period. They found positive test for KOH, Catalase, Kovacs oxidation, Production of Fluorescent pigment, Oxidation/fermentation of glucose and negative test for Levan production, Lipase activity, Arginine dihydrolase, Gelatin hydrolysis, Salt tolerance and Lecithinase activity test. Wilting and necrosis were observed for their pathgenicity test. Thus, isolates were identified as Ralstonia solanacerum. Moreover, molecular analysis of the organism using 759/760 primer gave 282 bp band indicated positive results. An efficient protein extraction method is a prerequisite for successful implementation of proteomics. Roots of brinjal were infected with Ralstonia solanacerum bacterial strain and proteins were extracted after 24 hrs, 48 hrs, 72 hrs and 96 hrs. Subsequently, different protocols of protein extraction were applied to roots of brinjal for comparing extraction efficiency by conducting two-dimensional electrophoresis (2-DE). The results indicated that precipitation of Ammonium acetate in Methanol after Phenol/Methanol extraction method provided the best effect. Less impurity was contained in extracted proteins and the 2-DE gel images showed less horizontal and vertical stripes, the most proteinaceous spots were extracted. Trichloroacetic acid (TCA) with Acetone method and Acetone method exhibited relatively less quality of gel images compared to other method. Results showed that a staining protocol using sensitized solution containg Glacial acetic acid, Methanol and Coomassie brilliant blue G-250 demonstrated the superior staining effect. Higher resolution was achieved when 11 cm of IPG gel strip with pH range of 3-10 was used and optimal loading quantity was determined as 250 µg for 11 cm strip. This study is the first to monitor protein expression patterns of the Brinjal responding to challenge by Ralstonia solanacerum and to detect response differences between resistant and susceptible plant.ThesisItem Open Access Analysis of defence metabolites production in brinjal infected by the bacterial pathogen Ralstonia solancearum L(Plant Molecular Biology and Biotechnology Dept., ACHF, NAU, Navasari, 2016-05) Vasava, Divyesh K.; Kapadia, ChintanPhytopathogen infection leads to changes in primary metabolism as well as secondary metabolism based on the modification in the metabolites levels leads to change in the growth and development of the plant. Therefore, pathogen attack causes crop yield losses even in interactions, which do not end up with disease or death of the plant. While the regulation of defense responses has been intensively studied for decades, less is known about the effects of pathogen infection on primary metabolism. Qualitative changes in the amino acid, Phenolic acids, phytohormones, polyamines and organic acid contents of the Eggplant (Solanum melongena L.) during pathogenesis caused by Ralstonia Solanacearum were investigated chromatographically. There was remarkable alteration in the level of various metabolites during interaction. Total in all nine different amino acids namely, Aspartic acid, Glutamic acid, Histidine, Glycine, Cysteine, Arginine, Serine, Leucine, and Phenylalanine, were detected in the Arka Nidhi and Pusa Purple Long during different time intervals after infection. Lysine, Isoleucine, Tyrosine, Threonine, Alanine, Methionine, and Valine were not found in the susceptible, as well as resistant plant, on the other hand, Proline was found only in the susceptible plant but not in resistant plant. Similarly, Arginine and leucine were not in any time point found in resistant plant. The organic acids, which detected in both the plant, were Fumaric acid, Malonic acid, Malic acid, Succinic acid and Citric acid, during the given incubation period, in all the given incubation periods. There were no remarkable changes in the concentration and composition of detected organic acids in diseased and control plant during various incubation periods. The concentration of Malic acid was the only factor that could be considered here in Eggplant (Solanum melongena L.)- Ralstonia Solanacearum interaction. Polyamines are implicated in the regulation of many processes in the plant cell, including functioning of ion channels, DNA replication, gene transcription, mRNA translation, cell proliferation and programmed cell death. Plant polyamines occur either in free form, covalently bound to proteins, or conjugated to hydroxycinnamic acids forming phenol amides. Our data revealed a striking increase in brinjal polyamine putrescine and spermidine in susceptible Pusa Purple Long than resistant Arka Nidhi. In this addendum, we describe that changes in polyamine metabolism take place even in earlier stages of brinjal plant infection with Ralstonia Solanacearum. However, after 24 hpi the levels of both hormones go in opposite direction in susceptible plant. Phenolic compounds were measured in roots of brinjal in control and inoculated plants. Phenolic were analyzed by high performance liquid chromatography. 4-dihydroxybenzoic acid, vanillic acid and Ferulic acid were not found in the resistant; while vanillic acid was absent in the susceptible plant at all the time point of study. Gallic acid, 3, 4-dihydroxybenzoic acid, Catechol, ferulic and p-coumaric were very much higher in susceptible control plant than resistant plants. However, there was gradual decline in the all the phenolic acid after pathogen inoculation in susceptible plant. The plant hormone abscisic acid (ABA) is involved in a wide variety of plant processes, including the initiation of stress adaptive responses to various environmental cues. In our study, ABA, IAA and GA3 emerged as a central factor in the regulation and integration of plant immune responses, although little is known about the underlying mechanisms. Aiming to advance our understanding of cross talk of these hormones during disease resistance/susceptibility, we have analyzed the quantity of SA, JA, ABA, IAA and GA3 during infection process of Ralstonia Solanacearum and defence process by plants. Whereas chemical and genetic disruption of ABA biosynthesis and signaling, respectively, led to enhanced Xoo resistance. In addition, as in our case, we have detected increase concentration of these phytohormones in susceptible plant than resistant plant during pathogenesis. Our results clearly indicate that, there was suppression of SA mediated defence pathway. There was reduction of JA in susceptible inoculated plant than resistant inoculated plant, indicated their role in imparting susceptibility towards disease at higher concentration.ThesisItem Open Access Biochemical and Molecular Characterization of Cellulase Producing Microorganisms from Termite(Plant Molecular Biology and Biotechnology Dept., ACHF, NAU, Navasari, 2016-07) Gadhiya, Komal J.; Mahatma, LalitThree bacterial isolates viz., Pseudomonas viridivida NAULK-1, Pseudomonas fluorescens NAULK-2 & Serratia plymuthica NAULK-3 were isolated from the termite (Heterotermes tenuis) gut. These were Gram negative, short rod shaped occurring singly and/or cluster. P. fluorescens NAULK-2 was short rod forming capsule. No capsule was seen in other two isolates. All three isolates were motile. All three bacteria can grow at 5-8 pH, however, showed distinct cultural characteristics on Luria Burtani (LB) agar plate. Three cellulose degrading enzyme viz., endoglucanase, exoglucanase and β-glucosidase were produced by all the three microorganism. Thereby they were capable to hydrolyze long chains of the cellulose in a progressive process because of exoglucanases, cleaves intramolecular β-1,4-glucosidic linkages and decreases the specific viscosity because of endoglucanases enzyme. Wood principally contains Cellulose, Hemicellulose and Lignin. Lignins are particularly important in the formation of cell walls, especially in wood and bark. Lignin fills the spaces in the cell wall between cellulose, hemicellulose, and pectin components, especially in xylem tracheids, vessel elements and sclereid cells. It is covalently linked to hemicellulose and therefore crosslinks different plant polysaccharides, conferring mechanical strength to the cell wall and by extension the plant ABSTRACT Abstract…. as a whole. Therefore, the bacteria having only the three cellulose degrading enzyme viz., endoglucanase, exoglucanase and β-glucosidase cannot degrade wood independently. S. plymuthica NAULK-3 had only these three enzymes. Isolate P fluorescens NAULK-2 produce all the three cellulose degrading and xylan degrading enzymes viz., endoglucanase, exoglucanase, β-glucosidase and xylanase. It was more efficient then the S. plymuthica NAULK-3 in degradation of wood, however, were not capable to degrade fully. P. viridivida NAULK-1 produced four different enzymes produced by the P. fluorescens NAULK-2 and additionally it produced laccase to digest lignin. Accordingly, it would be most capable to digest the wood as it have almost all the enzymes to degrade all the principal components of the plant wood. All the three isolates required neutral pH, however, P. viridivida NAULK-1 required 45oC temperature, P. fluorescens NAULK2 required 350C temperature and S. plymuthica NAULK-3 required 30oC temperature for their optimum expression. Expression of all these were maximum at 28 days post inoculation on the CMC and cellulose. Microorganism began the utilization of the natural agricultural/ agroindustrial wastes viz., wheat bran/straw, saw dust and rice straw within 7 days and the optimum time to degrade different substrate varied according to the substrate utilized and microorganism used. Among the different substrates, what straw/bran and rice straw was easiest to degrade by all the isolates, almost equal time was taken by all the isolated to degrade both the natural products, however, among the different isolates P. viridivida NAULK-1 degraded it earliest (21 days for the 50 per cent digestion and 28 days for the complete digestion), followed by P. fluorescens NAULK-2 (28 days for the 50 per cent digestion and 35 days for the complete digestion). Isolate S. plymuthica NAULK-3 took 35 days for the 50 per cent digestion and 42 days for the complete digestion. In the case of saw dust as substrate P. viridivida NAULK-1 took 28 days for Abstract…. the 50 per cent digestion and 35 days for the complete digestion. Isolate P. fluorescens NAULK-2 took 35 days for the 50 per cent digestion and 49 days for the complete digestion and isolate S. plymuthica NAULK-3 took 42 days for the 50 per cent digestion and 56 days for the complete digestion. The data were in accordance to the synthetic media, however, the long time to degrade the same over the synthetic media might be because of the complex structure of these natural molecules. For bacterial genomic DNA analysis 27F & 1525R primer were used to amplify ~1.5 kB DNA band size at 500C annealing temperature. In PCR all the isolates gave ~1.5 kB DNA band.ThesisItem Open Access Molecular and Biochemical Characterization of Capsicum annuum L. Varieties(PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY, ASPEE COLLEGE OF HORTICULTURE AND FORESTRY, ACHF, NAVASARI, 2015-09) Gor, Dimple V.; Singh, DiwakarChilli (Capsicum annuum L.) is an important vegetable and spice crop, consumed in a small amount to enrich the diet. It is a source of minerals, vitamins, antioxidants and other food components. The species Capsicum annuum is considered to have a wide diversity but not potentially characterized and evaluated for identification as well as crop improvement programme. Present study was planned to characterize 20 chilli genotypes on the basis of morphological, biochemical and molecular characters. Twenty chilli genotypes were characterized using total 6 quantitative characters (plant height, fruit length, fruit width, fruit weight, number of seeds per fruit and test weight of seeds) and 3 qualitative characters (plant branching type, fruit colour and seed colour). The plant height was found in the range of 41.93 to 97.33 cm, maximum in genotype 2011/CHIVAR-2 and minimum in KTPL-19. The fruit length was observed in the range of 4.19 to 9.44 cm, highest in genotype 2011/CHIVAR-2 and least in 2011/CHIVAR-5. The range of fruit width was obtained from 0.61 to 2.09 cm, maximum in genotype KTPL-19 and minimum in Utkal yellow. The fresh fruit weight was found in the range of 1.13 to 5.18 g, maximum in genotype 2011/CHIVAR-7 and minimum in KTPL-19. The number of seeds from fruit was in the range of 16.67 to 146.67, maximum in genotype SKAU-P-7 and minimum in Pant chilli-3. The test weight of seed was obtained in the range of 0.29 to 0.71 g, maximum in genotype ACS-2000-2 ABSTRACT and minimum in 2011/CHIVAR-9. The plant branching type, i.e. intermediate was obtained with highest frequency (40%) and dense type with lowest (25%). The green colour of fruit was obtained with maximum frequency (55%) and dark green with minimum (10%). The straw colour of seed was found with maximum frequency (60%) and brown colour with minimum (40%). Based on morphological characters the genetic diversity was assessed. The highest genetic dissimilarity (24%) was found between genotypes Utkal yellow and KTPL-19. Based on squared Euclidean distance 20 genotypes could be divided into two clusters, Cluster I included only two genotypes KTPL-19 and O7 whereas all other 18 genotypes fall under Cluster II. All 20 chilli genotypes were also subjected to biochemical characterization. The ascorbic acid was obtained in the range of 22.57 to 388.89 mg 100g-1, maximum in genotype 2011/CHIVAR-8 and minimum in 2011/CHIVAR-1. The total protein content was found in the range of 16.83 to 48.71 µg ml-1, maximum in genotype 2011/CHIVAR-9 and minimum in X-235. The range of capsaicin content was from 0.09 to 0.73 mg g-1, highest in genotype KTPL-19 and lowest in 2011/CHIVAR-9. The chlorophyll content was found in the range of 0.12-1.41 mg g-1, maximum in genotype ACS-92-4 and minimum in 2011/CHIVAR-9. The range for carotenoid content was found from 0.02 to 0.21 µg ml-1, maximum in the genotype KTPL-19 and minimum in 2011/CHIVAR-9. Genetic diversity and clustering analysis was also done based on biochemical characters. The highest genetic dissimilarity (19%) was obtained between genotypes KTPL-19 and 2011/CHIVAR-3 & genotype KTPL19 and 2011/CHIVAR-9. The cluster analysis generated two clusters in which Cluster I included only one genotype ACS-2000-2, whereas all other 19 genotypes fall under Cluster II. Based on combined analysis of morphological and biochemical data, highest genetic dissimilarity (36%) was observed between genotypes 2011/CHIVAR-7 and KTPL-19, whereas combined characters produced clustering pattern with three clusters. Only three genotypes, KTPL-19, O7 and ACS-2000-2 were included in Cluster I, six genotypes were included in Cluster III and Cluster II included maximum (11) genotypes. The molecular characterization of 20 chilli genotypes was done using 13 RAPD and 10 EST-SSR markers. Based on RAPD analysis total 143 loci were produced out of which 104 loci were polymorphic and showed 72.22 per cent polymorphism. The RAPD primer OPI 11 revealed highest polymorphism i.e. 92.3 per cent. The average PIC (Polymorphism Information Content) value for RAPD primer was 0.88. The highest PIC value was 0.91 for RAPD primer OPG 12 and OPH 12, thus considered as highly useful for Capsicum annuum L. The highest genetic dissimilarity (53%) was found between genotypes 2011/CHIVAR-1 and Pant chilli-3. From the cluster analysis three clusters were formed from which Cluster I included single genotype, 2011/CHIVAR-2, Cluster II included 4 genotypes and maximum number of genotypes were included in Cluster III. The molecular characterization was also assessed using 10 newly developed EST-SSR primers which showed alleles in the range of 2 to 8 with mean of 3.8 alleles per primer. The average PIC value was 0.56 and the highest PIC was 0.82 for primer DiwCA 27 and DiwCA 30, thus these two primers are considered to be highly useful for Capsicum annuum L. The highest genetic dissimilarity (25%) was obtained between genotypes 2011/CHIVAR-2 and 2011/CHIVAR-9. Based on EST-SSR, two clusters were generated. Cluster I included genotypes 2011/CHIVAR-2 and 2011/CHIVAR-7 whereas Cluster II included the rest 18 genotypes. The combination of both molecular maker data, RAPD and EST-SSR, gave the highest dissimilarity (72%) compared to that from all other characters and was found between genotypes 2011/CHIVAR-3 and 2011/CHIVAR-7 and, thus can be used for chilli breeding programme. The cluster analysis with combined molecular data generated two main clusters in which Cluster I included only two genotypes 2011/CHIVAR-2 and 2011/CHIVAR-7 and the rest 18 genotypes were classified under Cluster II. Thus, it can be concluded that several type of markers like morphological, biochemical and molecular markers could be used to characterize different chilli genotypes. The combined study of morphological and biochemical characters could produce more comprehensive clusters, but these characters are mostly affected by environmental factors. Molecular characterization based on RAPD and EST-SSR could find out precise genetic diversity among chilli genotypes and are not affected by environmental factors. The results produced in this study may be further used by a chilli breeder for genetic improvement programme of chilli genotypes.ThesisItem Open Access MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF CHICKPEA GENOTYPES FOR Fusarium oxysporum f. sp. ciceri RESISTANCE(PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY, ASPEE COLLEGE OF HORTICULTURE AND FORESTRY, ACHF, NAVASARI, 2014-07) Maisuria, Hemanshukumar J.; Patel, R.M.Chickpea (Cicer arietinum L.,) is the third most important pulse crop in area under cultivation worldwide after dry bean and pea, and also ranks third in terms of production. The major constraints in chickpea production are biotic and abiotic stresses. Among the biotic stresses a prime factor is wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. ciceri which may cause up to 90 per cent yield loss. The present study was carried out at Department of Plant Molecular Biology and Biotechnology, ASPEE College of Horticulture and Forestry, Navsari Agricultural University, Navsari. Marker assisted characterization of fifteen chickpea genotypes differing in their reaction to Fusarium wilt was carried out using different molecular markers linked to wilt resistance viz., random amplified polymorphic DNA, sequence characterized amplified region, DNA amplification fingerprinting, inter simple sequence repeat, sequence-tagged microsatellite sites, reported by earlier workers linked to disease resistance or susceptibility and ten random RAPD primers. Seed storage protein profiling was also done for biochemical characterization of chickpea genotypes using sodium dodecyl sulfate polyacrylamide gel electrophoresis as well as to identify possible markers for resistance or susceptibility. Seed storage protein profiling data was valid for geographical distribution analysis. In the present study, four different markers (viz., CS-27, CS-27A, UBC-825 and OP-U17-1) linked to susceptibility and four different markers (UBC-170, TA-59, TA-96 and TR-19) linked to resistance allele were validated. It was observed that one RAPD primer, CS- 27 gave amplification of 700 bp in susceptible genotypes and other RAPD primer UBC-170 gave amplification in resistance genotypes. One SCAR marker (CS-27A) developed from RAPD (CS-27) also amplified 700 bp in susceptible genotypes. The ISSR marker UBC-825 produced amplification of 1200 bp in susceptible genotypes. One DAF primer OP-U17-1 gave amplification of 1014 bp in susceptible genotypes only. Three STMS primers (TA-59, TA-96 and TR-19) gave allele specific amplification in wilt resistant genotypes. The STMS primer TA-59 was amplified 258 bp allele in resistance genotypes. The alleles of 265 bp amplified by primer TA-96 was present only in resistance genotypes. Also, TR-19 was amplified 227 bp allele in resistance genotypes. In present study, the reported markers linked to susceptibility and resistance proved their effectiveness and further can be exploited for maker assisted selection (MAS) of wilt resistance breeding in chickpea. Ten random primers were used for RAPD analysis in chickpea genotypes. The average polymorphism recorded by the RAPD loci was 24.40 per cent. The size range of amplified fragment was from 100 to 1000 bp for ten RAPD primers. The highest polymorphism 42.85 per cent was exhibited by primer OPH-03. The polymorphic information content value was developed in OPH-19 primer and the highest marker index value was found in OPH-20. A combined dendrogram developed by NTYSYS-pc system for all 10 primers which formed three major clusters. Seed storage protein profiling analysis of fifteen chickpea genotypes using SDS-PAGE showed minor difference in protein banding pattern. Total twenty four polypeptide bands were observed in SDS-PAGE gel. Total number of bands present in genotypes ranged from 15 to 20. The relative mobility values ranged from 0.04 to 0.97 among these genotypes. Dendrogram was prepared NTYSYS-pc system binary data of banding pattern. Dendrogram showed three major clusters. But, the polypeptide banding pattern did not show any correlation between resistance and susceptible genotypes for Fusarium wilt. So, seed storage protein profiling could not used for distinction of susceptible and resistant genotypes to wilt. The RAPD marker found to be more informative for chickpea diversity analysis as compared to SDS-PAGE of seed storage protein, further the difference in clustering of genotypes was observed among Dendrogram generated by RAPD and seed storage protein profiling.ThesisItem Open Access MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF CHICKPEA GENOTYPES FOR Fusarium oxysporum f. sp. ciceri RESISTANCE(PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY, ASPEE COLLEGE OF HORTICULTURE AND FORESTRY, ACHF, NAVASARI, 2014-07) Maisuria, Hemanshukumar J.; Patel, R.M.Chickpea (Cicer arietinum L.,) is the third most important pulse crop in area under cultivation worldwide after dry bean and pea, and also ranks third in terms of production. The major constraints in chickpea production are biotic and abiotic stresses. Among the biotic stresses a prime factor is wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. ciceri which may cause up to 90 per cent yield loss. The present study was carried out at Department of Plant Molecular Biology and Biotechnology, ASPEE College of Horticulture and Forestry, Navsari Agricultural University, Navsari. Marker assisted characterization of fifteen chickpea genotypes differing in their reaction to Fusarium wilt was carried out using different molecular markers linked to wilt resistance viz., random amplified polymorphic DNA, sequence characterized amplified region, DNA amplification fingerprinting, inter simple sequence repeat, sequence-tagged microsatellite sites, reported by earlier workers linked to disease resistance or susceptibility and ten random RAPD primers. Seed storage protein profiling was also done for biochemical characterization of chickpea genotypes using sodium dodecyl sulfate polyacrylamide gel electrophoresis as well as to identify possible markers for resistance or susceptibility. Seed storage protein profiling data was valid for geographical distribution analysis. In the present study, four different markers (viz., CS-27, CS-27A, UBC-825 and OP-U17-1) linked to susceptibility and four different markers (UBC-170, TA-59, TA-96 and TR-19) linked to resistance allele were validated. It was observed that one RAPD primer, CS- 27 gave amplification of 700 bp in susceptible genotypes and other RAPD primer UBC-170 gave amplification in resistance genotypes. One SCAR marker (CS-27A) developed from RAPD (CS-27) also amplified 700 bp in susceptible genotypes. The ISSR marker UBC-825 produced amplification of 1200 bp in susceptible genotypes. One DAF primer OP-U17-1 gave amplification of 1014 bp in susceptible genotypes only. Three STMS primers (TA-59, TA-96 and TR-19) gave allele specific amplification in wilt resistant genotypes. The STMS primer TA-59 was amplified 258 bp allele in resistance genotypes. The alleles of 265 bp amplified by primer TA-96 was present only in resistance genotypes. Also, TR-19 was amplified 227 bp allele in resistance genotypes. In present study, the reported markers linked to susceptibility and resistance proved their effectiveness and further can be exploited for maker assisted selection (MAS) of wilt resistance breeding in chickpea. Ten random primers were used for RAPD analysis in chickpea genotypes. The average polymorphism recorded by the RAPD loci was 24.40 per cent. The size range of amplified fragment was from 100 to 1000 bp for ten RAPD primers. The highest polymorphism 42.85 per cent was exhibited by primer OPH-03. The polymorphic information content value was developed in OPH-19 primer and the highest marker index value was found in OPH-20. A combined dendrogram developed by NTYSYS-pc system for all 10 primers which formed three major clusters. Seed storage protein profiling analysis of fifteen chickpea genotypes using SDS-PAGE showed minor difference in protein banding pattern. Total twenty four polypeptide bands were observed in SDS-PAGE gel. Total number of bands present in genotypes ranged from 15 to 20. The relative mobility values ranged from 0.04 to 0.97 among these genotypes. Dendrogram was prepared NTYSYS-pc system binary data of banding pattern. Dendrogram showed three major clusters. But, the polypeptide banding pattern did not show any correlation between resistance and susceptible genotypes for Fusarium wilt. So, seed storage protein profiling could not used for distinction of susceptible and resistant genotypes to wilt. The RAPD marker found to be more informative for chickpea diversity analysis as compared to SDS-PAGE of seed storage protein, further the difference in clustering of genotypes was observed among Dendrogram generated by RAPD and seed storage protein profiling.