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    Melania Lilik Swaningtias
    1) Chemical analysis found higher levels of protein, amino acids, and ash in the stems of red kidney bean plants treated with 2,4-D compared to untreated plants. 2) The treated plant stems had lower levels of carbohydrates like sugars and starch, indicating carbohydrates were used for protein synthesis. 3) The composition of proteins in treated plant stems changed, with some amino acids increasing and others decreasing compared to controls.

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    1) Chemical analysis found higher levels of protein, amino acids, and ash in the stems of red kidney bean plants treated with 2,4-D compared to untreated plants. 2) The treated plant stems had lower levels of carbohydrates like sugars and starch, indicating carbohydrates were used for protein synthesis. 3) The composition of proteins in treated plant stems changed, with some amino acids increasing and others decreasing compared to controls.

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    1) Chemical analysis found higher levels of protein, amino acids, and ash in the stems of red kidney bean plants treated with 2,4-D compared to untreated plants. 2) The treated plant stems had lower levels of carbohydrates like sugars and starch, indicating carbohydrates were used for protein synthesis. 3) The composition of proteins in treated plant stems changed, with some amino acids increasing and others decreasing compared to controls.

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    35 views5 pages

    IN OF: (7) - Tissue

    Uploaded by

    Melania Lilik Swaningtias
    1) Chemical analysis found higher levels of protein, amino acids, and ash in the stems of red kidney bean plants treated with 2,4-D compared to untreated plants. 2) The treated plant stems had lower levels of carbohydrates like sugars and starch, indicating carbohydrates were used for protein synthesis. 3) The composition of proteins in treated plant stems changed, with some amino acids increasing and others decreasing compared to controls.

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    CHANGES IN CHEMICAL COMPOSITION OF THE STEMS OF RED

    KIDNEY BEAN PLANTS TREATED WITH 2,


    4-DICHLOROPHENOXYACETIC ACID'
    HAROLD M. SELL, RICHARD W. LIUECKE, BETTY M.
    TAYLO R and CHARLES L. HAMNER
    Received January 3, 1949

    Introduction
    It has been shown (2, 5, 8, 11) that treatment of plants with 2, 4-D re-
    sults in a reduction in carbohydrate content and an accumulation of nitrogen.
    Investigators (4, 6,) have reported that some plants sprayed with solutions
    of 2, 4-D develop enlarged and proliferated tissues, noted primarily in the
    stems. It has also been observed that some plants which were sprayed with
    2, 4-D and which developed proliferated tissue of this type were eaten more
    readily by animals (7). Since little is known about the chemical composi-
    tion of this abnormal tissue, the work reported in this paper was projected
    to include the analysis for amino acids, ether extract, carbohydrates, ash and
    crude fiber. Results of chemical analyses are presented for the stems of un-
    treated red kidney bean plants and for those treated with 2, 4-D.
    Materials and methods
    SAMPLES OF STEM TISSUE
    Seeds of the red kidney bean were selected for uniformity of size and
    planted in 4-inch pots in the greenhouse. Each pot contained two plants
    that were treated when the first trifoliate leaf was expanding. Two repli-
    cations of 200 plants each were used from which to obtain material of
    treated and non-treated plants (controls). Application of 2, 4-D was made
    by applying one drop (.05 ml.) of a 1000-p.p.m. solution to the base of the
    blade of one of the primary leaves (6). The plants were harvested six days
    after treatment at the time the stem tissue had proliferated considerably but
    yet showed no signs of necrosis. The material was dried according to the
    procedure given by LINK (3) and then segregated into the various parts.
    The hypocotyl, first internode, and leaf petioles were grouped together as
    stem tissue, and analyses were made of this material.
    ANALYTICAL METHODS
    Amino acids were determined microbiologically with the organisms Lac-
    tobacillus arabinosus, Streptococcus faecalis and Leuconostic ntesenteroides.
    The media used in the various determinations was essentially the same as
    that described by SAUBERLICH and BAUMANN (9). Samples were prepared
    for assay according to the method of STOKES et at. (12), with the exception
    1 Journal Article No. 1008 (n.s.), Michigan Agricultural Experiment Station, East
    Lansing, Michigan.
    295
    296 PLANT PHYSIOLOGY

    of the tryptophan sample which was prepared according to the method of


    WOOLEY and SEBRELL (13). The analyses for carbohydrates were made by
    the procedure outlined by SELL, JOHNSON and LAGASSE (10) and the nitro-
    gen, crude fiber, ether extract and ash were determined by A. 0. A. C. pro-
    cedures (1).
    Results and discussion
    The results of the chemical analyses are summarized in tables I and II.
    Each entry is the average of two determinations on each of two separate sam-
    ples.
    The data in table I, based both on the percentage of amino acid or pro-
    tein in the sample and on the milligrams per stem, show the following
    trends: The stems of the plants treated with 2, 4-D contain approximately
    twice as much protein as the stems of the non-treated plants. A similar ob-
    AMINO ACID CONTENT OF THE STEMS OF RED KIDNEY BEAN PLANTS TREATED
    WITH 2, 4-DICHLOROPHENOXYACETIC ACID
    (EXPRESSED ON A LIPIDE-FREE* DRY WEIGHT BASIS)

    NON-TREATED TREATED
    CONSTITUENT IN THE PER IN CRUDE IN THE PER IN CRUDE
    SAMPLE STEM PROTEIN SAMPLE STEM PROTEIN
    per cent. milligrams per cent. per cent. milligrams per cent.
    Protein (NX6.25) 16.89 17.40 30.54 32.98
    Leucine .72 .74 4.23 1.44 1.56 4.70
    Isoleucine .77 .79 4.53 1.56 1.68 5.11
    Valine .73 .75 4.28 1.75 1.89 5.21
    Phenylalanine .47 .48 2.80 1.04 1.12 3.39
    Histidine .43 .44 2.51 .76 .82 2.48
    Arginine .84 .87 4.94 1.49 1.61 4.82
    Lysine .51 .53 3.04 1.48 1.60 4.82
    Tryptophan .12 .12 .69 .17 .18 .55
    Methionine .07 .07 .38 .20 .22 .65
    Threonine .67 .69 3.96 1.14 1.23 3.73
    Aspartic 1.48 1.52 11.31 1.88 2.03 6.37
    soluble in diethyl ether.
    * Substances
    servation was made by SMITH, HAMNER and CARLSON (11). The same
    trend as noted in the protein content was observed for leucine, isoleucine,
    valine, phenylalanine, histidine, arginine and threonine. The lysine and
    methionine content of the stems of the treated plants was approximately
    three times that of the controls. The tryptophan and asparic acid content
    were only slightly greater in treated than in non-treated plants.
    In calculating the data in table I as per cent. of amino acid in the protein,
    the following results were obtained: The amount of histidine is almost the
    same in the stems of the treated plants as in the controls. The content of
    tryptophan, threonine, and arginine is slightly less in the stems of the
    treated plants than in the stems of those not treated. Some of the greatest
    differences in the quantity of amino acids in the stem tissue were found in
    the content of aspartic acid, lysine, valine, methionine and phenylalanine.
    It is noted that the stems of the non-treated plants contain 43.7 per cent.
    SELL ET AL.: BEANS TREATED 297
    more aspartic acid than the stems of the treated plants. However, the stems
    of the treated plants contain 17.9, 36.9, 41.5 and 17.4 per cent. more valine,
    lysine, methionine, and phenylalanine, respectively, than the controls.
    The data presented on the basis of percentage of amino acid or protein in
    the sample and of milligrams per stem show that there is an accumulation
    of protein and amino acids in the stems of plants treated with 2, 4-D. When
    the data are expressed as per cent. amino acid in the crude protein, the re-
    sults indicate that the character of the protein in the stems of the treated
    plants has changed as compared to that of the controls.
    In table II, the stems of the plants treated with 2, 4-D show a depletion of
    reducing and non-reducing sugars. A similar observation has been noted
    by RASMUSSEN (8) in dandelion roots. SMITH et al. (11) and MITCHELL
    CARBOHYDRATE, CRUDE FIBER, ETHER EXTRACT AND ASH CONTENT OF THE STEMS OF RED
    KIDNEY BEAN PLANTS TREATED WITH 2, 4-DICHLOROPHENOXYACETIC ACID
    (EXPRESSED ON A LIPIDE-FREEt DRY WEIGHT BASIS)

    NON-TREATED TREATED
    CONSTITUENT MILLIGRAMS MILLIGRAMS
    PER CENT. PER STEM PER CENT. PER STEM

    Reducing Sugar 1.66 1.71 .00 .00


    Non-reducing Sugar 4.78 4.92 .00 .00
    Starch 7.22 7.44 2.08 2.25
    Polysaccharide* 12.16 12.52 10.09 10.09
    Crude Fiber 30.34 31.25 19.99 21.59
    Ether Extract (lipides, etc.) 1.64 1.69 2.26 2.44
    Unsaponifiable Residue
    of the Ether Extract .98 1.01 1.25 1.35
    Fatty Acids (By difference)
    of Ether Extract .61 .68 1.01 1.09
    Ash 11.41 11.75 15.98 17.26
    *
    Acid hydrolyzable residue from starch determinations.
    t Substances soluble in diethyl ether.
    and BROWN (5) observed simliar trends in bindweed and the morning glory.
    Considerable reduction in starch and a decrease in acid hydrolyzable poly-
    saccharides were noted in stems of treated plants. Similar results have been
    obtained by other workers (5, 8, 11).
    The depletion of reducing and non-reducing sugar and the utilization of
    the starch reserves and the large increase in protein in the stems of plants
    treated with 2, 4-D indicate that most of the carbohydrates are used in pro-
    tein synthesis.
    The quantity of crude fiber in the stems of the treated plants was consid-
    erably less than that of the controls. The decrease in acid hydrolyzable poly-
    saccharide and crude fiber suggests that the simple sugars are not utilized to
    a great extent in the formation of structural constituents in the stems of
    plants treated with 2, 4-D. The amount of ash in the stems of the treated
    plants increased as compared to the non-treated plants. The accumulation
    of organic acids or an increase of free carboxyl groups in the protein mole-
    cule of the treated tissue may explain the increase in ash content.
    298 PLANT PHYSIOLOGY
    The ether soluble substances (lipides, etc.) were slightly greater in the
    stems of the treated plants than in the controls. This same trend was noted
    in the contents of unsaponifiable material and fatty acid content in the ether
    extract. An increase in the amount of ether extract may be explained on
    the assumption that some of the carbohydrates are transformed into lipide
    material.
    Summary
    1. Both protein and amino acids accumulated in greater quantities in
    the stems of plants treated with 2, 4-D than in the stems of the non-treated
    plants. The amino acids, calculated as per cent. of crude protein, showed
    the greatest differences in the quantity of amino acids of the stem tissue to
    occur in aspartic acid, lysine, valine, methionine and phenylalanine. The
    variations in the content of amino acid indicate that the character of the
    protein is different in the treated stems than in the controls.
    2. The reducing and non-reducing sugars were depleted in the 2, 4-D
    treated plants. The treated plants also showed a considerable reduction in
    carbohydrate reserves and a decrease in acid hydrolyzable polysaccharides.
    This decrease in sugar content and the tremendous increase in total protein
    suggests that a large portion of the carbohydrates is utilized in protein syn-
    thesis.
    3. The amount of crude fiber decreased in the stems of 2, 4-D treated
    plants as compared to the controls. The decrease in crude fiber and acid
    hydrolyzable polysaccharides indicates that the simple sugars are not uti-
    lized in the formation of structural constituents in 2, 4-D treated plants.
    The ash content was greater in the stems of the treated plants than in the
    stems of the non-treated plants. This increase of ash may be due to an ac-
    cumulation of organic acids or an increase in free carboxyl groups in the
    protein of the stems of the treated tissue.
    4. The amounts of ether extract, unsaponifiable material and fatty acids
    of the ether extract were slightly greater in the stems of the treated plants
    than in those of the non-treated plants. These differences may be due to the
    conversion of some of the carbohydrates to lipide material.
    LITERATURE CITED
    1. AssOCIATION OF OFFICIAL AGRICULTURAL CHEMISTS. Official and tenta-
    tive methods of Analysis. Sixth edition: 26-27, 408-409, 504.
    1945.
    2. ERICKSON, L. C., SEELY, C. I., and KLAGES, K. H. Effect of 2,4-D upon
    the protein content of wheats. Jour. Am. Soc. Agron. 40: 659-660.
    1948.
    3. LINK, K. P. Effects of the method of desiccation of the carbohydrates
    of plant tissue. Jour. Am. Chem. Soc. 47: 470-476. 1925.
    4. MARTH, P. C., and MITCHELL, J. W. 2,4-Dichlorophenoxyacetic Acid
    as a differential herbicide. Bot. Gaz. 106: 224-232. 1944.
    SELL ET AL.: BEANS TREATED 299
    5. MITCHELL, J. W., and BROWN, J. W. Effect of 2,4-dichlorophenoxy-
    acetic acid on the readily available carbohydrate constituents in
    annual morning-glory. Bot. Gaz. 107: 120-129. 1945.
    6. MITcHsEui, J. W., and HAMNER, C. L. Polyethylene glycols as carriers
    for growth regulating substances. Bot. Gaz. 104: 474-483. 1944.
    7. QUISENBERRY, S. K. Private communication to C. L. Hamner.
    8. RASMUSSEN, L. W. The physiological action of 2,4-dichlorophenoxy-
    acetic acid on dandelion, Taraxacum officinale. Plant Physiol. 22:
    377-392. 1947.
    9. SAUBERLICH, H. E., and BAUMANN, C. A. The effect of dietary protein
    upon amino acid excretion by rats and mice. Jour. Biol. Chem.
    166: 417. 1946.
    10. SELL, H. M., JOHNSON, F. A., LAGASSE, F. S. Changes in the chemical
    composition of the tung fruit and its component parts. Jour. Agr.
    Res. 73: 319-334. 1946.
    11. SMITH, F. G., HAMNER, C. L., and CARLSON, R. F. Changes in food
    reserves and respiratory capacity of bindweed tissues accompany-
    ing herbicidal action of 2,4-dichlorophenoxyacetic acid. Plant
    Physiol. 22: 58-65. 1947.
    12. STOKES, J. L., GUNNESS, M., DwYn, I. M., and CASWELL, M. C. Micro-
    biological methods for the determination of amino acids. Jour.
    Biol. 160: 35. 1945.
    13. WOOLEY, J. G., and SEBRELL, W. H. Two microbiological methods for
    the determination of 1 (-) tryptophan in proteins and other com-
    plex substances. Jour. Biol. Chem. 157: 141. 1945.

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