Abstract
Using primers synthesized from two conserved regions and employing PCR, a DNA segment coding for part of the apoprotein of assimilatory nitrate reductase could be amplified from the fungi Aspergillus nidulans, Pythium intermedium, Phytophthora infestans, Phytophthora megasperma and Glomus D13. Sequencing of the amplificates as well as DNA hybridization revealed strong homologies with the nitrate reductase gene in all cases. The digoxigenin-labeled amplificate from Glomus hybridized with DNA isolated from Glomus spores. The data from these gene probing experiments are generally in accord with the published results from enzyme measurements. Thus assimilatory nitrate reductase occurs in saprophytic, parasitic as well as arbuscular mycorrhizal fungi. No amplificates with these primers were obtained with DNA isolated from Mucor mucedo and Saprolegnia ferax. Such results agree with the failure to detect nitrate assimilation physiologically in these two organisms.
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Azcon R, Gomez M, Tobar R (1992) Effects of nitrogen source on growth, nutrition, photosynthetic rate and nitrogen metabolism of mycorrhizal and phosphorus-fertilized plants of Lactuca sativa L. New Phytol 121:227–234
Calza R, Huttner E, Vincentz M, Rouzé P, Galagan F, Vauchert H, Cherel I, Meyer C, Kronenberger J, Caboche M (1987) Cloning of DNA fragments complementary to tobacco nitrate reductase mRNA and encoding epitopes common to the nitrate reductase from higher plants. Mol Gen Genet 209:552–562
Choi HK Kleinhofs A, An G (1989) Nucleotide sequence of rice nitrate reductase genes. Plant Mol Biol 13:731–733
Cummings B, Wood T (1989) A simple and efficient method for isolating genomic DNA from endomycorrhizal spores. Gene Anal Technol 6:89–92
Danneberg G, Latus C, Zimmer W, Hundeshagen B, Schneider-Poetsch HJ, Bothe H (1992) Influence of vesicular-arbuscular mycorrhiza on phytohormone balances in maize (Zea mays L.). J Plant Physiol 141:33–39
Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissues. Focus 12:13–15
Esch H, Hundeshagen B, Schneider-Poetsch HJ, Bothe H (1994) Demonstration of abscisic acid in spores and hyphae of the arbuscular mycorrhizal fungus Glomus and in the N2-fixing cyanobacterium Anabaena variabilis. Plant Sci 99:9–16
Gowri G, Campbell WH (1989) cDNA clones for corn leaf NADH: nitrate reductase and chloroplast NAD(P) +: glyceraldehyde-3-phosphate dehydrogenase. Plant Physiol 90:792–798
Harley JL, Smith SE (1983) Mycorrhizal symbiosis. Academic Press, London
Hayman DS (1983) The physiology of vesicular-arbuscular mycorrhizal fungi. Can J Bot 61:944–963
Ho I, Trappe JM (1975) Nitrate reducing capacity of two vesicular-arbuscular mycorrhizal fungi. Mycologia 67:886–888
Johansen A, Jakobsen I, Jensen ES (1992) Hyphal transport of 15N-labelled nitrogen by a vesicular-arbuscular mycorrhizal fungus and its effect on depletion of inorganic soil. N. New Phytol 122:281–288
Johnstone IL, McCabe PC, Greaves P, Gurr SJ, Cole GE, Brow MAD, Unkles SE, Clutterbuck AJ, Kinghorn JR, Innis MA (1990) Isolation and characterization of the crnA-niiA-niaD gene cluster for nitrate assimilation in Aspergillus nidulans. Gene 90:181–192
Lilly VG, Barnett HL (1951) Physiology of the fungi. McGraw-Hill, London
Okamoto PM, Fu YH, Marzluf GA (1991) Nit-3, the structural gene of nitrate reductase in Neurospora crassa: nucleotide sequence and regulation of mRNA synthesis and turnover. Mol Gen Genet 227:213–223
Prosser JM, Lazarus CM (1990) Nucleotide sequence of spinach nitrate reductase cDNA. Plant Mol Biol 15:187–190
Raeder U, Broda P (1985) Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol 1:17–20
Sambrock J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Schmitz O, Danneberg G, Hundeshagen B, Klingner A, Bothe H (1991) Quantification of vesicular-arbuscular mycorrhiza by biochemical parameters. J Plant Physiol 139:106–114
Smith SE, Gianinazzi-Pearson V (1988) Physiological interactions between symbionts in vesicular-arbuscular mycorrhizal plants. Annu Rev Plant Physiol Plant Mol Biol 39:221–244
Stewart V (1988) Nitrate respiration in relation to facultative metabolism in enterobacteria. Microb Rev 52:190–232
Sundaresan P, Rata NV, Gunaseka P, Laksman N (1988) Studies on nitrate reduction by VAM fungal spores. Curr Sci 57:84–85
Unkles SE, Campbell EI, Punt PJ, Hawker KL, Contreras R, Hawkins AR, Van den Hondel CAM, Kinghorn JR (1992) The Aspergillus niger niaD gene encoding nitrate reductase: upstream nucleotide and amino acid sequence comparison. Gene 11:149–155
Webster J (1983) Pilze. Springer, Berlin Heidelberg New York
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Kaldorf, M., Zimmer, W. & Bothe, H. Genetic evidence for the occurrence of assimilatory nitrate reductase in arbuscular mycorrhizal and other fungi. Mycorrhiza 5, 23–28 (1994). https://doi.org/10.1007/BF00204016
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DOI: https://doi.org/10.1007/BF00204016