I would like to thank my husband (Monodip Das) and my new family for their support and motivation. I am grateful to all my laboratory members and friends for their continued support and love.
Summary and Future prospects
RFO Pamilia ti Rafinosa Oligosakarido ACC 1-aminosiklopropano 1-karboksiliko nga asido NGPP46 Amianan a Guwahati Pongamia pinnata IITG Indiano nga Instituto ti Teknolohia Guwahati IGEPAL Octylphenoxypoliethoxyethanol. PPTY Pongamia pinnata Dagiti Ty1-kopia a klono JCTY Jatropha curcas Ty1-kopia a klono RCTY Ricinus communis Ty1-kopia a klono MFTY Mesua ferrea Ty1-kopia a klono.
UNITS
LIST OF TABLES
4B.7 Phylogenetic relationships between the nucleotide sequences of the RT-RH domain of Ty1-copia clones of J. 4B.8 Phylogenetic relationships between the nucleotide sequences of the RT-RH domain of Ty1-copia clones of R.
ABSTRACT
INTRODUCTION
INTRODUCTION
In previous reports, with genome size estimation, intra- and interspecies variations have been reported (Zedek et al. 2010). In true diploid species, variation in genome size exists either due to repetitive elements or due to environmental factors.
Specific Objectives
Based on the background information, the current study is therefore focused on the following objectives.
REVIEW LITERATURE
REVIEW OF LITERATURE
- INTRODUCTION
- Source of biofuel production
- Environmental aspects
- Industrial applications
- Plant development aspects
- Pharmacological aspect
- Nuclear genome content
- Chromosome study
- Retrotransposons in correlation with genome size
- Abiotic stress evaluation in non-edible oil crops
- Study of seed markers during development and germination
Genome size estimates are also crucial for the construction and screening of genomic and cDNA libraries (Bennett et al., 2000). Transposable elements (TEs) are found to be present in all eukaryotic genomes (Wicker et al., 2007).
GENOME SIZE ESTIMATION AND CYTOLOGICAL ANALYSIS IN NON-EDIBLE OIL CROPS
GENOME SIZE ESTIMATION AND CYTOLOGICAL ANALYSIS IN NON-EDIBLE OIL CROPS
INTRODUCTION
Estimation of nuclear DNA content and chromosome number could be useful for understanding genome organization and evolution. Thus, in this study, an attempt has been made to use flow cytometric analysis to determine the nuclear DNA content and chromosome number in plants collected from different locations in Assam, India.
MATERIALS AND METHODS .1 Plant material
- Flow cytometric analyses
- Statistical analysis
- Cytological analysis
- Collection and storage of root tips
- Pre-treatment and fixation
- Hydrolysis and staining
- Slide preparation, squash, and observation
Mature tender leaves were used for the preparation of the suspension of intact kernels. The analysis was repeated to check if the coefficient of variation of the sample was >5.0.
RESULTS AND DISCUSSION
- Buffer optimization
- Reference standards for flow cytometry analysis
- Estimation of Nuclear DNA content
- Cytological analysis .1 Collection of root tips
- Hydrolysis and staining
- Slide preparation and observation
In this study, the plants used as reference standards to estimate the genome size were P. A reference standard should have a genome size larger than the sample genome size, but not more than 4 times (Suda & Leitch, 2010). The reliability of the estimated nuclear DNA content was compromised by the presence of inhibitors.
A reported study confirms that greatly exaggerated changes in fluorochrome fluorescence may not be due to differences in nuclear DNA content (Price et al., 2000). Taking into account the minimum deviation of the MFI standards, the estimated nuclear DNA content for the test samples according to external, internal and pseudo-internal standardization was calculated. Further experiments in the four plants under study are warranted for proper justification of the above assessment regarding intra-species variations in genome size and its relation to temperature.
CONCLUSION
GENOME SIZE CORRELATION TO CELL PHENOTYPIC TRAITS AND MOBILE ELEMENT
GENOME SIZE CORRELATION WITH CELL PHENOTYPIC TRAITS AND MOBILE ELEMENT
Sectioning of the paraffin blocks (10 µm thick) containing the samples was performed using a vibratome series (Leica RM 2245, Germany). Copy number = (haploid genome size x average proportion of nuclear genomic DNA hybridizing to probe) / probe element size. 88 Figure 4B.7: Phylogenetic relationships among RT-RH domain nucleotide sequences of Ty1-copia clones from J .
89 Figure 4B.8: Phylogenetic relationships among the nucleotide sequences of the RT-RH domain of Ty1-copia clones from R. 90 Figure 4B.9 Phylogenetic relationships among the nucleotide sequences of the RT-RH domain of Ty1-copia clones from M The high number of copies of retrotransposons, which have been found to be the cause in most of the previous reports (Khaliq et al., 2012).
This may be a sign of the importance of Ty1-copia retrotransposons in the evolution of the plant genome and its size. Among all retrotransposons, Ty1-copia is considered one of the causes of genome obesity.
STRESS EVALUATION IN NON-EDIBLE OIL CROPS - ANATOMICAL, PHYSIOLOGICAL AND PHYTOCHEMICAL
STUDIES
STRESS EVALUATION IN NON-EDIBLE OIL CROPS - ANATOMICAL, PHYSIOLOGICAL AND PHYTOCHEMICAL
INTRODUCTION
The seeds contain oil that can be processed to produce a high-quality biodiesel usable in a standard diesel engine (Mardhiah et al., 2017; Openshaw, 2000). As the four crops have the potential to produce high oil-yielding seeds, they are considered pioneers of second-generation sustainable biofuel crops. Number of reports are available on the oil aspect of these four biodiesel crops (Achten et al., 2007; Aydin et al., 2010; Bora et al., 2013; Kesari et al., 2008) but no information is available on the anatomical , physiological and biochemical changes in the plant during abiotic stress.
The aim of the present work was therefore to investigate the effect of salinity and drought on various anatomical, physiological and phytochemical parameters required for the commercial production of plant products.
MATERIALS AND METHODS
- Growth of the plants and stress application
- Stomatal parameters and epidermal cell area measurement
- Plant physiological study
- Chlorophyll content
- Determination of root: shoot ratio
- Determination of relative growth rate
- Phytochemical study of leaves
- Quantitive analysis of phytochemicals
- Statistical analysis
Methanolic leaf extracts (0.1 ml) were taken in test tubes and the volume was made up to 5 ml with distilled water, followed by the addition of 0.3 ml of 5% NaNO2. About 20 g of plant leaf powder from the four plants under study were taken in 4 different 500 ml conical containers and 200 ml of 20% ethanol was added to them. The mixtures were filtered and the obtained residue was then re-extracted with another 200 ml of 20% ethanol.
The residue was washed with double distilled water and then dissolved in 10 ml of 67% (v/v) H2SO4, followed by incubation for 1 hour. To estimate the neutral detergent fiber (NDF), the powder leaves (1 g) of the four plants were placed separately in a reflux flask and 100 mL of cold neutral detergent solution was added. To estimate acidic detergent fiber (ADF), 1 g of powdered leaves of the four plants was transferred to a reflux flask and 100 mL of cold acidic detergent solution was added.
RESULTS AND DISCUSSION .1 Plant anatomical study
- Plant physiological study
- Abiotic stress induced phytochemical study in leaves .1 Qualitative analysis of phytochemicals in leaves
- Quantitative analysis of phytoconstituents
Stress exposure resulted in a decrease in total chlorophyll content in drought- and salinity-affected leaves compared to control leaves. The difference in the physiological values of the test plant compared to the control plant is considered statistically significant (P < 0.05) for salinity (250 mM NaCl) and 100% high drought stress. The diverse uses of plants in the treatment of different diseases are due to the presence of the phytoconstituents (Lekhak & Yadav, 2012).
The results show that plants under stress had significantly reduced values of cellulose, hemicellulose, lignin and reducing sugar compared to the control. This indicates that the effect of salinity was greater in the production of these metabolites relative to these plants. There was a gradual decrease in most anatomical and physiological parameters with increasing stress.
STUDY OF BIOMARKER FOR SEED GERMINATION IN NON-EDIBLE OIL CROPS
STUDY OF BIOMARKER FOR SEED GERMINATION IN NON- EDIBLE OIL CROPS
INTRODUCTION
Several biomarkers such as antioxidants, membrane integrity, ethylene as an indicator, raffinose family oligosaccharides and cell cycle markers have been reported as remarkable markers for seed quality and germination (Abts et al., 2013; Souza et al., 2016; Rewers & Sliwinska, 2012; Ventura et al. , 2012). Synthesis of reactive oxygen species (ROS) occurs during the metabolic process of seeds; thus, seed quality maintenance is an important factor (Kumar et al., 2016). This damages the integrity of cell membranes, affecting permeability and fluidity through lipid peroxidation (Avila et al., 2007).
Thus, it has been hypothesized that the decrease in ROS concentration acts as a messenger in seed germination (Jeevan Kumar et al., 2015). Given the fact that the active cells complete their cell cycle within a short span, the 4C/2C ratio may be a good marker for seed germination in addition to its quality assessment (Chamberlin et al., 1993a). This chapter highlights the study of ROS intensity and cell cycle activity, which can be used as a good marker for seed germination.
MATERIALS AND METHODS .1 Sample collection
- Determination of ROS accumulation intensity
- Cell cycle activity
- Sample preparation and isolation of nuclei
- Flow cytometric analyses
- Statistical analysis
Collected seeds were thoroughly washed with distilled water (3-4 times) and were used fresh for the experiments. Seeds of different stages of the candidate plants were isolated from their pods and crushed in 10 ml of 10 mM Potassium phosphate buffer in an ice-cold mortar. Fluorescence of the samples and the positive control was measured using a spectrofluorometer (Fluoromax-4, Horiba scientific) at
All four stages of seeds belonging to the candidate plants were used for the preparation of the suspension of intact nuclei. For the preliminary study, propidium iodide/hypotonic citrate buffer (Krishan, 1975) was used for the extraction and staining of the cells. The embryo axis was minced using surgical scissors dipped in 1 ml propidium iodide (PI) hypotonic citrate buffer according to the protocol described by Ramesh et al., (2014).
RESULTS AND DISCUSSION
- Reactive oxygen species concentration
- Cell cycle activity
According to the measured intensity, phase 4 (late ripening) has the lowest ROS intensity than the rest of the phases. According to the measured intensity, stage 3 (precocious) has the lowest ROS intensity than the rest of the stages. This result complements the previous research on sunflower seeds, which found that ROS content was higher in the initial stages of seed development (Kibinza et al., 2006).
Thus, the 4C/2C ratio was determined for all seed stages of the four plants and the values were tabulated in Table 6.1. This indicates that the rate of cell division is faster in the germination stage of the seed. In this study, the 4C/2C ratio was found to be highest in the late ripening seeds of Ricinus and Pongamia, whereas in the case of Jatropha, the 4C/2C ratio was more in early ripening seeds.
SUMMARY AND FUTURE PROSPECTS
SUMMARY
Genome size estimates have been made for these four biofuel crops, which may be useful in the search for a powerful trait in this field. The repetitive DNA content covers a large portion of the nuclear genome, leading to the variation in genome size among different species. Thus, calculating the copy number of Ty1-copia and understanding its likely role in genetic diversity will serve as an important background for further understanding of retrotransposons in the genomic study.
Thus, a study was conducted to investigate the effects of the abiotic stresses on several anatomical, physiological and biochemical parameters. Furthermore, no significant variation in Ty1-copia copy number was observed, so it is concluded that the variation in genome size is mainly due to environmental variation. Although the plants experience abiotic stress, minimal difference in the value of phytoconstituents was observed.
FUTURE PROSPECTS
High frequency plant regeneration from leaf disc cultures of Jatropha curcas L.: an important biodiesel plant. Origin, evolution, and proposed stabilization of the terms "genome size" and "C-value" to describe nuclear DNA content. Lineage-specific differential amplification of transposable elements is responsible for genome size variation in Gossypium.
Leaves and seeds as materials for flow cytometric estimation of genome size of 11 Rosaceae woody species containing DNA staining inhibitors. Antiulcer activity of methanolic extract of Jatropha curcas (Linn.) on aspirin-induced gastric lesions in Wistar rats. Genome size doubling without polyploidization: dynamics of retrotransposition-driven genomic expansions in Oryza australiensis, a wild relative of rice.
