Abstract
A fast and reliable method for the extraction of cellular polyamines and major inorganic ions (Ca, Mg, Mn, K, and P) from several plant tissues is described. The method involves repeated freezing and thawing of samples instead of homogenization. The efficiency of extraction of both the polyamines and inorganic ions by these two methods was compared for 10 different tissues. In each case, the freeze-thaw procedure resulted in a precise and quantitatively equal, or greater, yield than homogenization. Freeze-thawing not only eliminates the need for various tissue homogenizers (such as polytrons, tissumizers, and mortars and pestles), but it is so simple that a large number of samples can be processed simultaneously. We routinely processed 50–80 samples for quantitation of polyamines and inorganic ions. Freeze-thawing was equally useful for the extraction of polyamines from liver, spleen, and kidney tissues of mice.
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References
Anderson DL, Henderson LJ (1986) Sealed chamber digestion for plant nutrient analysis. Agron J 78:937–939
Birecka H, Birecki M, Cohen EJ, Bitonti AJ, McCann PP (1988) Ornithine decarboxylase, polyamines, and pyrrolizidine alkaloids in Senecio and Crotalaria. Plant Physiol 86:224–230
De Scenzo RA, Minocha SC (1993) Modulation of cellular polyamines in tobacco by transfer and expression of mouse ornithine decarboxylase cDNA. Plant Mol Biol 22:113–127
Faure O, Mengoli M, Nougarede A, Bagni N (1991) Polyamine pattern and biosynthesis in zygotic and somatic embryo stages of Vitis vinifera. J Plant Physiol 138:545–549
Flores HE, Arteca RN, Shannon JC (1990) Polyamines and ethylene: Biochemistry, physiology and interactions. American Society of Plant Physiologists, pp 1–425
Isaac RA, Johnson WC (1976) Determination of total nitrogen in plant tissue using a block digestor. J AOAC 59:98–100
Kingston HM, Jassie LB (1986) Microwave energy for acid decomposition at elevated temperatures and pressures using biological and botanical samples. Anal Chem 58:2534–2541
Kuennen RW, Wolnik KA, Fricke FL (1982) Pressure dissolution and real sample matrix calibration for multielement analysis of raw agricultural crops by inductively coupled plasma atomic emission spectrometry. Anal Chem 54:2146–2150
Kushad MM, Yelenosky G (1987) Evaluation of polyamine and proline levels during low temperature acclimation of citrus. Plant Physiol 84:692–695
Maki H, Ando S, Kodama J, Komamine A (1991) Polyamines and the cell cycle of Catharanthus roseus cells in culture. Plant Physiol 96:801–806
Meijer EGM, Simmonds J (1988) Polyamine levels in relation to growth and somatic embryogenesis in tissue cultures of Medicago saliva L. (alfalfa). J Exp Bot 39:787–794
Minocha R, Kvaalen H, Minocha SC, Long SL (1993) Polyamines in embryogenic cultures of Norway spruce (Picea abies) and red spruce (Picea rubens). Tree Physiol 13:365–377
Minocha R, Minocha SC, Komamine A, Shortle WC (1991) Regulation of DNA synthesis and cell division by polyamines in Catharanthus roseus suspension cultures. Plant Cell Rep 10:126–130
Minocha R, Minocha SC, Long SL, Shortle WC (1992) Effects of aluminum on DNA synthesis, cellular polyamines, polyamine biosynthetic enzymes, and inorganic ions in cell suspension cultures of a woody plant, Catharanthus roseus. Physiol Plant 85:417–424
Minocha SC, Minocha R, Robie CA (1990) High-performance liquid chromatographic method for the determination of dansyl-polyamines. J Chromatogr 511:177–183
Minocha R, Shortle WC (1993) Fast, safe, and reliable methods for extraction of major inorganic cations from small quantities of woody plant tissues. Can J For Res 23:1645–1654
Nielsen KA (1990) Polyamine content in relation to embryo growth and dedifferentiation in barley (Hordeum vulgare L.). J Exp Bot 41:849–854
Noh EW, Minocha SC (1990) Pigment and isozyme variation in aspen shoots regenerated from callus culture. Plant Cell Tiss Org Cult 23:39–44
Rastogi R, Davies PJ (1989) Polyamine metabolism in ripening tomato fruit. Plant Physiol 94:1449–1455
Robie CA, Minocha SC (1989) Polyamines and somatic embryogenesis. I. The effects of difluoromethylornithine and difluoromethylarginine. Plant Sci 65:45–54
Slocum RD, Flores HE (1991) Biochemistry and physiology of polyamines in plants. CRC Press, Boca Raton
Smith TA (1985) Polyamines. Ann Rev Plant Physiol 36:117–143
Smith MA (1991) Chromatographic methods for the identification and quantitation of polyamines. In: Slocum RD, HE Flores (eds) Biochemistry and physiology of polyamines in plants. CRC Press, Boca Raton, pp 229–242
Snedecor GW, Cochran WG (1956) Statistical methods. The Iowa State University Press, Ames
Sun DY, Minocha R, Minocha SC (1994) Genetic transformation of Populus to modulate polyamine metabolism. In: Ahuja MR (ed) Proceedings of the IUFRO workshop on biotechnology of trees, Valsain, Spain, 18–22 October 1993. (in press)
Torrigiani P, Serafini-Fracassini D, Bagni N (1987) Polyamine biosynthesis and effect of dicyclohexylamine during the cell cycle of Helianthus tuberosus tuber. Plant Physiol 84:148–152
Wikoff LR, Moraghan JT (1986) Determination of plant iron, manganese and zinc by wet ashing procedures. J Sci Food Agric 37:839–844
Wolf B (1982) A comprehensive system of leaf analyses and its use for diagnostic crop nutrient status. Commun Soil Sci Plant Anal 13:1035–1059
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New Hampshire Agricultural Experiment Station, scientific contribution no. 1845.
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Minocha, R., Shortle, W.C., Long, S.L. et al. A rapid and reliable procedure for extraction of cellular polyamines and inorganic ions from plant tissues. J Plant Growth Regul 13, 187–193 (1994). https://doi.org/10.1007/BF00226036
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DOI: https://doi.org/10.1007/BF00226036