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Master Degree Theses

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2010 - 20124
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Subject: Plant Biochemistry ×

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    ThesisItemOpen Access
    Molecular and biochemical Characterization of Pigeonpea (Cajanus cajan L.) Genotypes for Fusarium wilt Resistance
    (Biotechnology Department, N. M. college of Agriculture Navsari Agricultural University, 2012-02) Mohanlal, Swami Praksh; Mahatma, Mahesh Kumar
    Pigeonpea belongs to genus cajanus and species cajun under sub-family papilionoidae of family Leguminosae, tribe-phaseo/eae, sub-tribe cajaninue. It has a diploid genome comprising 11 pairs of chromosomes (2n = 22) with a physical size estimated at about 0.853 pg (Greilhuber and Obermayer, 1988). Pigeonpea is short day plant. Its grains are highly nutritious and rich in protein content (21-23%), carbohydrates, fiber and minerals. It contributes to the C, Nand P economy of the soil enhancing its performance even under marginal input. Globally pulses are the second most important group of crops after cereals. Developing countries are the major source of pulses contributing 74 per cent of its production . • India is the largest producer and consumer of pulses in the world accounting 33 per cent of the area and 25 per cent of the global production. In India. pigeonpea accounts for 20 % of the total pulse production. India ranks first with about 90 % production in the world. It is the second most important pulse crop after chickpea in area as well as productivity in India. It occupies about 15.14 % of total area under pulses and contributes 14.91 % of the total pulse production in India. In India. it occupies 3.38 million hectares with the production of 2.27 million tonnes with the productivity of 671 kg/ha. In Gujarat, during kharif 2009 area under pigeon pea was 0.27 million hectares and production was 0.26 million tonnes with the productivity of 989 kg/ha (Anonymous. 2009). The major pigeonpea growing states are Unar Pradesh. Madhya Pradesh, Karnataka. Maharashtra, Gujarat. Rajasthan. Haryana. Punjab. Tamilnadu. Orissa and Bihar. Fusarium wilt is an important biotic constraint in pigeonpea production in the Indian subcontinent, which results in 16-47% crop losses. The fungus enters the host vascular system at the root tips through wounds or invasion made by nematodes. leading to progressive chlorosis of leaves, branches. wilting and collapse of the root system. In India alone. the loss due to this disease is estimated to be US $71 million and the percentage of disease incidence • varies from 5.3 to 22.6% (Raju el a/" 2010).
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    ThesisItemOpen Access
    Molecular variation Analysis in Date palm (Phonix dactylifera L)
    (Biotechnology N. M. College of Agriculture Navsari, 2010) Chandra, Shrivastav Vishal Subhash; Mahatma, Mahesh
    In other case study of 5 Date palm genotypes (Ghanshyam, Early maturing, Late maturing, Male 2 small and Male! tall), five type of markers with one best primer for each marker (RAPD, ISSR, SSR, ITS and 18s rRNA) were used. Amplification of genomic DNA of 5 Date palm genotypes yielded 39 scorable fragments out of which 22 were polymorphic, with an average of 4.4 polymorphic fragments per primer. Number of amplified fragments ranged from 3 (BNL-448) to 10 (OPG-2 and ITS-3) and varied from -200bp to -4kb. Percentage of polymorphism ranged from 20% (ITS-3) to a maximum of 100% (BNL-448).Cluster analysis using UPGMA contains two clusters A and B with Jaccard's similarity matrix with low value of 0.550 in that Early maturing genotype was distinct. On the other hand, cluster A contains Male 2 small and Male 1 tall with highest value of 0.910 and Late maturing genotype having similarity value of 0.784. In cluster Al Ghanshyam genotype with similarity value of 0.770 was found diverse in cluster A. In Principal Coordinate Analysis Male 2 small and Male I tall were grouped while other genotypes were randomly distributed. In 14 seedling samples raised from Ghanshyam, banding pattern showed all bands similar; except seedling sample no. 4 in which one band of MW -750bp was absent.
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    ThesisItemOpen Access
    Molecular and Biochemical Characterization of Karanja (Derris indica L.)
    (Biotechnology Department, N. M. college of Agriculture Navsari Agricultural University, 2011-07) Arjun, Hake Anil; Jha, Sanjay
    Genetic diversity of 30 Karanja accessions were evaluated with Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (lSSR) markers. Total 160 and 73 scorable bands were generated by 20 RAPD and 10 ISSR primers respectively. RAPD primer OPE-9 showed maximum polymorphism (100%) while OPE-20 and OPH-18 showed minimum polymorphism (33.33%). A total of 107 and 28 (total 135) polymorphic bands were generated by RAPD and ISSR primers respectively. The Jaccard similarity coefficients with ISSR marker varied from 0.82 to 1.00 with mean value of 0.91, for RAPD primers the similarity coefficient range from 0.72 to 0.93 with • mean of 0.82. In present study low level of genetic diversity was observed among the Karanja genotypes. Average RPI, PIC and MI value for RAPD primers were 2.09, 0.25 and 18.06 respectively; while for ISSR marker it was 0.66, 0.08 and 4.48 respectively. Genetic diversity analysis by Isozyme marker viz., POX (peroxidase). PPO (polyphenol oxidase), EST (esterase) and SOD (superoxide di smutase) scores total 19 isozyme loci in 30 accessions of Karanja. Highest loci was scored by PPO (6 loci) followed by POX (5 loci) ',. . While 4 loci scored by SOD and EST. Highest polymorphism and highest number of polymorphic loci was shown by PPO isozyme (83.33%) while lowest polymorphism and lowest number of polymorphic loci was shown by SOD isozyme (SO %). The Jaccard similarity coefficients for isozyme varied from 0.44 to 1.0 with the mean of 0.72. The average Isozyme Polymorphic Index, PIC and MI for Isozyme marker is 2.0. 0.42 and 28.81 ,. I respectively. Jaccard similarity coefficients between the accessions for composite RAPD, ISSR and isozyme displayed a mean of 0.893. The results indicated that the genetic diversity harbored among var. NAUK-16, NAUK-29 and NAUK-21 for RAPD and for ISSR, NAUK-21 genotype shows diverse. In combination of ISSR, RAPD and isozyme NAUK-21 showed distinctness from other genotypes which could be further exploited for • • • karanJa genetic Improvement. In biochemical characterization the analysis of four different isozymes. estimation of six enzymes and primary metabolites was done to distinguish 30 accessions. Highest GR (248.Q2 l1M/minlg protein) and PPO (19.06 ~M/minlg protein) activity was found in NAUK-II while maximum catalase (14.02 l1M/minlg protein) and APX activty (255.17 ~M/minlg protein) exhibited by NAUK-24. Two genotypes NAUK-2 and NAUK- 13, expressed highest GPOD (165.16 ~M/minlg protein), SOD (84.01 U/g protein). In metabolites highest total soluble sugar (5.96 mg/g FW leal) and total phenol content (10.23 mg/g FW leal) was observed in NAUK-24 while highest amount free amino acid (5.03 mg/g FW leal) was found in NAUK-30. The maximum amount of total chlorophyll (3.13mg/gm FW leaf) and carotenoid content (1.95 mg/g FW leaf) was observed in NAUK-2 and NAUK-22 respectively . In morphological characterization, in temns of height, tallest tree and average longest branch was recorded by NAUK-15 and NAUK-16 respectively. Average highest number of branches and largest leaflet width was recorded by NAUK-9 while longest leaflet length recorded by NAUK-I . Largest average collar diameter and branch diameter recorded in NAUK-6 and NAUK-7 respectively. Smallest size of leaflet length and leaflet width recorded by NAUK-22 while smallest average branch height and branch diameter recorded byNAUK-4.
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    ThesisItemOpen Access
    In Planta Transformation in Cotton variety G Cot. 10
    (Biotechnology N. M. College of Agriculture Navsari, 2010-07) Harkishansinh, Solanki Vanrajsinh; Khandelwal, Vikas
    In this study, genotype independent in planta transformation of cotton (Gossypium hirsutum) variety G.Cot.l0, using injection method and cut method has been reported. Agrobacterium tumefaciens strain containing construct pBIN 1 F carrying the nptII and cry 1 F gene was used for plant transformation. The integration of the gene was confirmed by using PCR. The transformation experiment was performed by optimizing two important parameters: age of the seedlings and cocultivation duration with Agrobacterium. When the effect of the age of the seedlings and co-cultivation duration evaluated three days old seedlings and cocultivated for three days yielded higher survival percentage as well the frequency of transformation in both methods (injection method, • 1. e. 6% and cut method, • 1. e. 4%). Overall higher frequency of transformation was yielded in injection method • 1. e. 1.16% while • In cut method overall frequency of transformation obtained was 0.83%. To confirm the effectiveness of the Cry 1 F protein insect feeding assay was performed using second instar larvae of Spodoptera litura. Leaf feeding bioassay of the putative transform ants showed 53.00 to 100% mortality and 42 to 90% reduction in the weight of surviving larvae. Present results of insect bioassay indicated that plant produced significant amount of protein which was sufficient to cause the mortality and lessened the damage to the plants caused by S. litura.