Agrobacterium and other methods of plant transformation including gene gun, inplanta
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Genetic transformation is a powerful tool and a significant strategy for studying plant functional genomics, i.e. gene exploration, new insights into gene regulation, and the analysis of genetically regulated characteristics. Furthermore, the work of isolated genes utilizing map-based cloning of mutant alleles has been verified through functional complementation via genetic transformation. In addition, genetic engineering allows the insertion of alien genes into crop plants and the accelerated creation of new genetically modified organisms.
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Determination of chromosome shape:
Chromosome shape can also be defined in terms of the centromeric index or the arm ratio.
The centromeric index is the length of the shorter arm divided by the total chromosome length, and thus varies from 0.5 for a truly metacentric chromosome to zero for a telocentric one.
The arm ratio is the length of the long arm divided by the length of the short arm, and thus ranges from unity for a truly metacentric chromosome to infinity for a truly telocentric chromosome.
Quantification of degree of asymmetry:
Four ratios between the sizes of smallest and longest chromosomes (1,2,3,4) and three different proportions of metacentric chromosomes (A,B,C,D) in the karyotype.
So that 12 classes of karyotypes are possible in the increasing order of asymmetry
Increased karyotype asymmetry was associated with specialised zygomorphic flowers
Ideogram: it is a schematic diagrammatic representation of a karyotype that shows all the homologous pairs of chromosomes in the nucleus.
Advantages of karyotyping:
• Reveals the structural features of each chromosome
• Helps in studying chromosome banding pattern
• Helps in identifying chromosomal aberrations
• Detection of prenatal genetic disorders
• Aids in studying evolutionary changes
Chromosome banding
Chromosome banding: When subjected to different treatments before staining, the chromosomes develop different dark and light regions in form of bands – chromosome banding.
• This is a technique for the identification of chromosomes and its structural abnormalities in the chromosome complement.
• Chromosome identification depends on their morphological characteristics such as relative length, arm ratio, presence and absence of secondary constrictions on the chromosome arms.
• It is an additional and useful tool for the identification of individual chromosome within the chromosome complement.
• It could be used for identification of chromosome segments that predominantly consist of either GC or AT rich regions or constitutive heterochromatin.
• On banded chromosome, darkly stained or brightly fluorescent transverse bands (positive bands) alternate with the lightly stained or less fluorescent (negative bands).
D2 analysis & it's Interpretation
- The document discusses D2 analysis, a technique used to assess genetic diversity among plant genotypes.
- D2 analysis involves calculating distances between genotypes based on multiple quantitative traits and clustering genotypes based on these distances.
- The document provides details on the steps involved in D2 analysis, including data collection, calculation of variances/covariances, computation of D2 values, clustering genotypes, and interpretation.
- An example application of D2 analysis to assess genetic diversity among litchi hybrids is described. Five clusters were identified among 18 hybrids based on quantitative traits.
Hardy weinberg equilibrium
This document provides an introduction to population genetics concepts including:
- What is a population and how it relates to evolution
- Key terms like gene pool, allele frequency, and genotype frequency
- Mendelian populations and how they interact
- Hardy-Weinberg equilibrium and how evolutionary forces can affect equilibrium
- Factors like mutation, migration, non-random mating, natural selection, and genetic drift that influence population genetics
Salinity stress Resistant Mechanisms
The document discusses mechanisms of salinity tolerance in plants. It describes how plants can tolerate high salt concentrations through avoidance, tissue tolerance, and salt dilution. Avoidance mechanisms keep salt ions away from sensitive plant tissues through exclusion, excretion, or compartmentalization of ions. Tissue tolerance allows plants to tolerate accumulated ions through compartmentalization at the cellular and intracellular levels. Salt dilution increases plant storage volume to dilute ion concentrations. The document provides examples of plant species that exhibit different tolerance mechanisms and strategies to mitigate the effects of salinity on crops.
Somatic hybridization and Protoplast Isolation
The document discusses protoplast isolation and somatic hybridization. It defines somatic hybridization as the development of hybrid plants through the fusion of somatic protoplasts from different plant species or varieties. The key steps in somatic hybridization are isolating protoplasts, fusing the protoplasts from desired species, identifying and selecting hybrid cells, culturing the hybrid cells, and regenerating hybrid plants. Protoplasts can be isolated using enzymatic or mechanical methods, and their viability and ability to form cell walls can be tested. Various factors like enzyme concentration, temperature, and pH affect protoplast isolation and viability. Protoplast fusion can occur spontaneously or be induced using methods like chemical treatment, electro
Root excudates and their role in nutrition
Root exudates play an important role in plant nutrition by interacting with soil microbes and influencing the rhizosphere. Root exudates include primary metabolites like amino acids, carbohydrates, organic acids, and secondary metabolites like flavonoids, lignins, coumarins, and fatty acids. These exudates are released from the root tip through passive diffusion and active transport processes. They function to attract beneficial microbes, chelate nutrients, and defend against pathogens in the rhizosphere. The composition and concentration of root exudates varies depending on plant species and environmental conditions like nutrient availability.
stability for grain yield in Finger millet
The document discusses stability for grain yield in finger millet. It begins by introducing the challenges plant breeders face in developing stable cultivars across environments due to genotype by environment (GxE) interactions. It then defines stability and the agronomic and biological concepts of stability. Different statistical models for analyzing stability are described, including the Additive Main effects and Multiplicative Interaction (AMMI) model. The AMMI model uses analysis of variance and principal component analysis to evaluate GxE interactions and identify stable genotypes. Biplots are used to graphically represent GxE interactions identified through AMMI analysis. Specifically, biplots allow visualization of genotypes and environments as well as patterns of GxE interactions.
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1
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- 3. Non-biological based transformation Particle BOMBARDMENT/BIOLISTICS • Gene constructs for biolistics can be in the form of circular or linear plasmids or a linear expression cassette. • Embryogenic cell cultures are likely the best explants because they can be spread out as uniform targets of cells and have high recovery capacity. • Rice transformation has also been successfully achieved via the bombardment of embryogenic calli, in which transformation efficiency has been raised to 50% (Li et al., 1993). • The transformation efficiency might be lower than with Agrobacterium mediated transformation and it is more costly, as well. • Intracellular targets are random and DNA is not protected from damage. As a result, many researchers have avoided particle bombardment method because of the high frequency of complex integration patterns and multiple copy insertions that could cause gene silencing and variation of transgene expression (Dai et al., 2001; Darbani et al., 2008).
- 13. laser beam is focused on cell membrane to ablate the cell membrane, form a transient hole, and the exogenous nucleic acid was delivered into cell. High intensity laser irradiate the cell and produce plasma, which create high pressure and act on cell membrane to enhance the permeability of cell membrane (Noack & Vogel, 1995). Also, plasma can generate bubbles that ultimately collapse and produce secondary shock waves to perforate the cell membrane
- 23. A) The cell membrane is composed of a lipid bilayer, with a hydrophobic interior and hydrophilic exterior. (B) Liposomes are also composed of a lipid bilayer arranged as a spherical shell. (C) A scientist performs a brief reaction that allows liposomes to form around DNA. (D) Cells in culture can endocytose the liposome, digesting it within vesicles to release DNA. (E) Alternatively, liposomes can directly fuse with the plasma membrane, directly releasing DNA into cells.
- 28. Biological gene transfer (Indirect method) Agrobacterium mediated transformation