BIOL114 - Lecture 7

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  • Created by: Katherine
  • Created on: 16-03-16 14:39
What is plant biotechnology?
It is the application of labatory based techniques for plant propagation or genetic improvement
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Most plant biotechnology is based on the technique of...
Tissue culture (the culture of plant organs, tissues, cells or protoplasts on nutrient media under sterile conditions).
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What is a cell suspension culture?
It is he growth of plant cells under sterile conditions in a liquid medium, with shaking.
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What are cell suspension cultures used for?
Research (convenient method of obtaining homogeneous mass of cells in the lab), commercial production of high value secondary metabolites and other compounds e.g. antimicrobials, vitamins and food flavours.
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What is a protoplast?
A plant cell without a cell wall.
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What is somatic hybridization?
The production of novel hybrids between sexually incompatible plant species through protoplast fusion.
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How does somatic hybridization occur?
Protoplasts from the two species are mixed and induced to fuse to produce hybrid protoplasts. Plants are regenerated from the hybrid protoplasts by tissue culture.
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Give an example of somatic hybridization:
Shanrong No.3 wheat (salt and drought resistant wheat)
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What is micropropagation?
It is commercially important for propagting individual plant genotypes where other methods of propagation are difficult.
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How is micropropagation conducted?
You start with an explant which is cultured under sterile conditions designed to promote regeneration of whole plants.
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The ability to regenerate whole plants from small pieces of plant material depends on what property of the plant cell?
The totipotency
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What is totipotency?
The ability of an individual cell to divide and form all parts of the mature organism
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What is dedifferentitation
It is
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What is redifferenetiation?
It is
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How is regeneration by organogenesis conducted?
Isolation of a small piece of plant material under sterile conditions. Callus production on nutrient medium containing plant hormones, Organogenesis stage I: the generateion of new shoots from the undifferentiated callus. Organogeneis II.
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What happens in organogenesis stage ii:
The generation of roots from the shoots created in stage i.
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What is somatic embryogenesis?
It is development of embryos and whole plants directly from the comatic cells.
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Is soatic embryogenesis easy or difficult?
Difficult in most species
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What is somaclonal variation?
The phenomenon of phenotypic variability between individual plants derived from plant tissue culture.
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What does it mean if something is polyploidy?
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What does it mean if something is aneuploidy?
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What are the advantages of somaclonal variation?
Creation of additional genetic variability for plant improvement
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What are the disadvantages of somaclonal variation?
Lack of uniformity is a problem in micropropagation for horticulure and forestry industries.
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How many steps are there in creating genetically engineered or transgenic plants?
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What are the steps in creating genetically engineered or transgenic plants
Production of a DNA construct that harbours the gene of interest, transformation of plant cells with the construct, selection of transformed cells using a selectable marker, regeneration of whole plants from the transformed cells (using organogenesis
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What kind of promoter is required in this?
Constitutive (on all the time throughout the plant), Tissue specific (only expressed in root hairs), Developmentally regulated, Inducible
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What are two cell specific promoters?
Pollen specific and pericycle specific.
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The promoters direct the expression of a ...
reporter gene 'Green Fluorescent Protein'
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What is a selectable marker?
Usually genes of bacterial origin that confer resistance to compounds that are toxic to plants
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Selectable marker genes usually have:
Antibiotic resistance e.g. E.coli nptll gene for neomycin phosphotransferase which inactivates a range of aminoglycoside antibiotics such as kanamycin and neomycin
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How do you get foreign DNA into the plant?
Naked DNA delivery system e.g. particle bombardment, electroporation., or Natural delivery systems. These are mainly used for transformation of dicots but also used successfully with some rice genotypes.
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What happens in naked DNA delivery by particle bombardment?
Fire DNA coated particles into plant cells at high velocity using particle gun. Particles penetrate cell walls and membrane and the DNA is released and incorporated into the plant chromosomes.
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What is electroporation?
It is use of a short high voltage electrical discharge to make the plasma membrane permeable to DNA (or other polar molecules). The high voltage disrupts areas of the membrane temporarily, allowing DNA to pass through. The membrane reseals.
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What is agrobacterium tumefaciens?
A plant pathogen the infects wound sites and produces tumours in many plant species. It acts as a natural genetic engineer - transferring a small segment of its DNA (the T-DNA) into the plant genome.
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T DNA carries genes that:
Induce uncontrolled cell division (tumorigenic), direct the synthesis of opines - amino acids the bacterium can use as nutrients, but not the plant.
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Which plasmid is the T-DNA carried on?
The Ti plasmid.
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What does Ti mean?
Tumour inducing
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The T-DNA is bounded by two regions called...
Left (Tl) ad right (Tr) borders
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For integration into the plant DNA, what is required?
The borders and repeats are the only sequences necessary for integration
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What is the floral dipping method of plant transformation?
A simple transformation method in which developing flowers are immersed in a suspension of Agrobacterium, requires no tissue culture.
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Describe the process of Floral dipping:
Developing flowers are dipped in Agrobcterium suspension, Transformation of germ line cells occurs at a frequency of
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Card 2


Most plant biotechnology is based on the technique of...


Tissue culture (the culture of plant organs, tissues, cells or protoplasts on nutrient media under sterile conditions).

Card 3


What is a cell suspension culture?


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Card 4


What are cell suspension cultures used for?


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Card 5


What is a protoplast?


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