IMMUNOCHEMICAL CHARACTERIZATION OF ALPHA-AMYLASES IN WHEAT SEEDS AT DIFFERENT ONTOGENICAL STEPS

  • 1 January 1978
    • journal article
    • research article
    • Vol. C129  (2-3) , 215-+
Abstract
Protein extracts of wheat seeds taken at an early stage of development, at maturation and after 7 days of germination were investigated by using immunochemical techniques with immune sera prepared against .alpha.-amylases purified from developing seeds and germinated seeds. After immunochemical analyses carried out in agarose gel, .alpha.-amylase characterization was performed by using .beta.-limit dextrin followed by iodine staining. Detection of 3 antigenic .alpha.-amylases separated by agarose immunoelectrophoretic analysis at pH 8.6 and called I, II, III from the anode to the cathode, as well as an antigenic relationship between the anodic enzymes I and II were confirmed. Three constituents in I and 4 in II were further distinguished by using long duration electrophoresis in agarose gel. The immune sera reacted with all of these constituents. Thus with these immune sera, a quantitative determination of all anodic .alpha.-amylase proteins can be attempted as quantitation of 2 antigens. During this identification, constituents with activity on .beta.-limit dextrin, but delivering incomplete unstained substrate, were detected. These constituents found in developing seeds have electrophoretic mobility close to that of constituent I, but differ antigenically from .alpha.-amylases I and II. Combination of rocket-line-immunoelectrophoresis and .alpha.-amylase characterization reaction on precipitin bands was developed for comparing amounts of each of 3 .alpha.-amylase antigens in different seed extracts. The use of this technique for quantitating, at the same time, 2 antigen groups having a certain cross reactivity was discussed. Preliminary results of a quantitative study on each of these antigens in developing, mature and germinating seeds were reported.

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