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    Auxin Bioassay

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    Shreeya mohapatra
    This document describes four common bioassays used to test for the presence and concentration of the plant hormone auxin: 1. Avena sative Coleoptile Curvature Test measures the curvature of oat seedling coleoptiles exposed to auxin to determine concentration. 2. Coleoptile Section Straight Growth Test measures the growth of cut coleoptile sections in varying auxin concentrations. 3. Mesocotyl Straight Growth Test similarly measures the growth of oat mesocotyl sections. 4. Split Pea Stem Curvature Test induces curvature in pea stem segments based on auxin concentration measured. These bioassays allow quantification of auxin levels through

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    Auxin Bioassay

    Uploaded by

    Shreeya mohapatra
    484 views22 pages
    This document describes four common bioassays used to test for the presence and concentration of the plant hormone auxin: 1. Avena sative Coleoptile Curvature Test measures the curvature of oat seedling coleoptiles exposed to auxin to determine concentration. 2. Coleoptile Section Straight Growth Test measures the growth of cut coleoptile sections in varying auxin concentrations. 3. Mesocotyl Straight Growth Test similarly measures the growth of oat mesocotyl sections. 4. Split Pea Stem Curvature Test induces curvature in pea stem segments based on auxin concentration measured. These bioassays allow quantification of auxin levels through

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    This document describes four common bioassays used to test for the presence and concentration of the plant hormone auxin: 1. Avena sative Coleoptile Curvature Test measures the curvature of oat seedling coleoptiles exposed to auxin to determine concentration. 2. Coleoptile Section Straight Growth Test measures the growth of cut coleoptile sections in varying auxin concentrations. 3. Mesocotyl Straight Growth Test similarly measures the growth of oat mesocotyl sections. 4. Split Pea Stem Curvature Test induces curvature in pea stem segments based on auxin concentration measured. These bioassays allow quantification of auxin levels through

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    484 views22 pages

    Auxin Bioassay

    Uploaded by

    Shreeya mohapatra
    This document describes four common bioassays used to test for the presence and concentration of the plant hormone auxin: 1. Avena sative Coleoptile Curvature Test measures the curvature of oat seedling coleoptiles exposed to auxin to determine concentration. 2. Coleoptile Section Straight Growth Test measures the growth of cut coleoptile sections in varying auxin concentrations. 3. Mesocotyl Straight Growth Test similarly measures the growth of oat mesocotyl sections. 4. Split Pea Stem Curvature Test induces curvature in pea stem segments based on auxin concentration measured. These bioassays allow quantification of auxin levels through

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 22

    Auxin Bioassay

    Created by:- Guided by:-


    Shreeya Mohapatra Khusbu Agarwal
    What is bioassay?
    • Analytical method
    • Determines concentration of a substance.
    • How? By measuring its effect on a living organism in standard
    conditions.
    • ‘Bio’ = Living
    • It involves :-
    • A stimulus applied to a subject
    • A response is triggered and measured.
    Bioassay for auxin
    • Several bioassays have been devised for Auxin.
    • Avena sative Coleoptile Curvature Test
    • Coleoptile Section Straight Growth Test
    • Mesocotyl Straight Growth Test
    • Split Pea Stem Curvature Test
    Avena sative Coleoptile Curvature Test:
    • In 1928, Frits Went worked on identifying the factor that
    caused the plants to bend towards light.
    • By building on the work of Darwin and Boysen-Jensen, he was
    able to isolate the factor and show how it worked.
    Procedure
    • Avena grains are germinated and grown in total darkness.
    Procedure
    • The seedlings are exposed to short periods(2-4 hrs) of red light
    two days after germination.

    After 2 days
    Procedure
    • When roots = 2mm long, the seedlings are planted in special
    glass holders (using water culture method).

    2 mm
    Procedure
    • The straight coleoptiles are selected.
    Procedure
    • The tips of the coleoptile (about 1mm) are removed without
    injuring the primary leaf and placed on agar- agar.
    • The tips contain auxin.

    Cut tip
    Primary leaf

    B
    Straight coleoptile

    A
    C
    Procedure
    • 3 hours later, a second decapitation of the coleoptile is made.
    • The first leaf is pulled so that its connection from base is
    removed.
    Procedure
    • An agar block containing auxin is now placed on one side of
    the cut coleoptile.
    • The projecting primary leaf gives support to the auxin block.
    Procedure
    • After 90 minutes, the shadowgraphs of seedlings are taken.
    • The angle of curvature is measured by vertical line and a line
    parallel to the curved portion of the coleoptile.
    Conclusion
    • Within limits the curvature of the coleoptile is directly
    proportional to the concentration of auxin in agar block.
    • In IAA the maximum response is at about 0.2 mg/litre.
    • This concludes
    • Auxin responsible for growth
    • It induces growth on opposite side.
    Split pea stem curvature test

    • Pea seedlings are grown upto 8 days


    • 6cm of stem from 3rd internode taken
    1

    • An incison of 3cm is made in one end


    • Kept (24)in petridishes with varying
    2 auxin concentrations
    Conclusion

    • Auxin enters the entire


    length.
    • Auxin induces higher
    growth in epidermal cells
    • A curvature is obtained at
    the middle of the stem
    segment.
    Coleoptile section Straight
    Growth Test
    Avena seeds are germinated

    Seedlings are grown in week red light

    2mm below the tip of coleoptile is cut


    (6mm)
    Incubated in petri dishes with increasing amounts of
    auxin
    24 hrs later, length of coleoptiles are
    measured

    Graph is plotted
    Conclusion
    • Optimum concentration of auxin for max length of the section
    can be measured.
    • The auxin content of an unknown sample can be known.
    Mesocotyl Straight Growth Test
    • Mesocotyl is the tubular, white, stem-like tissue connecting
    the seed and the base of coleoptile.
    • In order to grow mesocotyl in oats seedlings, the plant is
    grown in complete darkness without any red light.
    • 4mm sections of mesocotyl is cut below the nodal zone
    • Incubated in petri dishes containing varying
    concentrations of auxin.
    • Growth of mesocotyl is measured after 24 hours.

    • Conclusion :
    • Growth of mesocotyl was seen even in low
    concentration of auxin.
    Cress Root Inhibition Test:
    Sterilized seeds of cress are allowed to grow on
    moist filter paper.

    Roots reach 2 cm growth.

    50 % are taken in test solution containing auxin

    remaining kept in
    moist paper

    48hrs later, length of


    roots measured.
    Conclusion
    • Roots in test solution do not show any growth
    • Those in controlled conditions show normal growth.

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