Abstract
The broad bean late nodulins, Nvf-28/32, are composed of two types of repetitively occurring sequence modules flanked by unique N- and C-terminal modules. Six isoforms of these nodulins were characterized by a specific modular structure resulting from a different individual order of repetitive sequence modules. A detailed analysis of genomic PCR fragments revealed that the repetitive modules and the N-terminal unique module exactly corresponded to exons, whereas the C-terminal module was specified by two exons. Since those exons encoding the repetitive modules missing in specific Nvf-28/32 isoforms were consistently present within genomic sequences, a post-transcriptional generation of VfNOD28/32 transcripts specifying six Nvf-28/32 nodulins was concluded. Using tissue-print hybridizations, these transcripts were localized in the interzone II–III and the nitrogen-fixing zone III of root nodules. From this and from cDNA-cDNA hybridizations demonstrating a comparable timing of expression of VfNOD28/32 and of leghemoglobin transcripts in root nodules, a function of the modular nodulins Nvf-28/32 in late developmental stages of broad bean nodules was inferred.
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References
Arnold W, Pühler A (1988) A family of high-copy-number plasmid vectors with single end-label sites for rapid nucleotide sequencing. Gene 70:171–179
Benson D, Lipman DJ, Ostell J (1993) GenBank. Nucleic Acids Res 21:2963–2965
Brewin NJ (1991) Development of the legume root nodule. Annu Rev Cell Biol 7:191–226
Bullock WC, Fernandez JM, Short JM (1987) XL1-Blue: a high efficiency plasmid transformingrecAEscherichia coli strain with beta-galactosidase selection. Biotechniques 5:376–379
Caetano-Annollés G, Gresshoff PM (1991) Plant genetic control of nodulation. Annu Rev Microbiol 45:345–382
Delauney AJ, Verma DPS (1988) Cloned nodulin genes for symbiontic nitrogen fixation. Plant Mol Biol Rep 6:279–285
Downie AJ (1994) Signalling strategies for nodulation of legumes by rhizobia. Trends Microbiol 2:318–324
Franssen HJ, Yiyn I, Yang WC, Bisseling T (1992) Developmental aspects of theRhizobium-legume symbiosis. Plant Mol Biol 19:89–107
Gumpel NJ, Smith AG (1992) A novel calcium-binding protein fromEuglena gracilis. Characterization of a cDNA encoding a 74-kDa acidic-repeat protein targeted across the endoplasmic reticulum. Eur J Biochem 210:721–727
Harrison J, Molloy PL, Clark SJ (1994) Direct cloning of polymerase chain reaction products in anXcmI T-vector. Anal Biochem 216:235–236
Heijne G von (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14:4683–4690
Kammen A van (1984) Suggested nomenclature for plant genes involved in nodulation and symbiosis. Plant Mol Biol Rep 2:43–45
Kannenberg EL, Brewin NJ (1994) Host-plant invasion byRhizobium. The role of cell-surface components. Trends Microbiol 2:277–283
Kessler C (1992) Nonradioactive labeling and detection of biomolecules. Springer, Berlin Heidelberg New York
Kijne JW (1992) TheRhizobium infection process. In: Stacey G, Burris RH, Hardd J (eds) Biological nitrogen fixation. Chapman and Hall, New York, pp 349–398
Kiss GB, Vincze É, Végh Z, Tóth G, Soós J (1990) Identification and cDNA cloning of a new nodule-specific gene, Nms-25 (nodulin 25) ofMedicago sativa. Plant Mol Biol 14:467–475
Küster H, Frühling M, Perlick AM, Pühler A (1993) The sucrose synthase gene is predominantly expressed in the root nodule tissue ofVicia faba. Mol Plant-Microbe Interact 6:507–514
Küster H, Perlick AM, Pühler A (1994) Members of a broad bean nodulin family with partial homologies to the alfalfa nodulin 25 are composed of two types of amino acid repeats flanked by unique sequence termini. Plant Mol Biol 24:143–157
Küster H, Schröder G, Frühling M, Pich U, Rieping M, Schubert I, Perlick AM, Pühler A (1995) The nodule-specificVfENOD-GRP3 gene encoding a glycine-rich early nodulin is located on chromosome I ofVicia faba L. and is predominantly expressed in the interzone II–III of root nodules. Plant Mol Biol 28:405–421
Luehrsen KR, Taha S, Walbot V (1994) Nuclear pre-mRNA processing in higher plants. Prog Nucleic Acid Res Mol Biol 47:149–193
McKeown M (1992) Alternative mRNA splicing. Annu Rev Cell Biol 8:133–155
Mead DA, Szczesna-Skorupa E, Kemper B (1986) Single stranded DNA “blue” T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng 1:67–74
Mellor RB (1989) Bacteroids in theRhizobium-legume symbiosis inhabit a plant lytic compartment: implications for other microbial endosymbioses. J Exp Bot 40:831–839
Mellor RB, Werner D (1987) Peribacteroid membrane biogenesis in mature legume root nodules. Symbiosis 3:75–100
Miao GH, Verma DPS (1993) Soybean nodulin-26 gene encoding a channel protein is expressed only in the infected cells of nodules and is regulated differently in roots of homologous and heterologous plants. Plant Cell 5:781–794
Mylona P, Pawlowski K, Bisseling T (1995) Symbiotic nitrogen fixation. Plant Cell 7:869–885
Nakai K, Kanehisa M (1992) A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics 14:897–911
Nap JP, Bisseling T (1990a) The roots of nodulins. Physiol Plantarum 79:404–414
Nap JP, Bisseling T (1990b) Developmental biology of a plant-prokaryote symbiosis: the legume root nodule. Science 250:948–954
Perlick AM, Pühler A (1993) A survey of transcripts expressed specifically in root nodules of broad bean (Vicia faba L.). Plant Mol Biol 22:957–970
Perlick AM, Frühling M, Küster H, Schröder G, Pühler A (1994) Analysis of nodulin genes expressed in root nodules ofVicia faba L. In: Endre G, Kiss GB (eds) Proceedings of the first european nitrogen fixation conference. Officina Press, Szeged, pp 225–228
Perlick AM, Frühling M, Schröder G, Frosch SC, Pühler A (1996) The broad bean gene VfNOD32 encodes a nodulin with sequence similarities to chitinases that is homologous to (α/β)8-barrel-type seed proteins. Plant Physiol 110:147–154
Pich U, Schubert I (1993) Midiprep method for isolation of DNA from plants with a high content of polyphenolics. Nucleic Acids Res 21:3328
Pridmore RD (1987) New and versatile cloning vectors with kanamycin-resistance marker. Gene 56:309–312
Priefer U (1989) Genes involved in lipopolysaccharide production and symbiosis are clustered on the chromosome ofRhizobium leguminosarum biovarviciae VF39. J Bacteriol 171:6161–6168
Quandt H-J, Pühler A, Broer I (1993) Transgenic root nodules ofVicia hirsuta: a fast and efficient system for the study of gene expression in indeterminate-type nodules. Mol Plant-Microbe Interact 6:699–703
Rhijn P van, Vanderleyden J (1995) TheRhizobium-plant symbiosis. Microbiol Rev 59:124–142
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Sanchez F, Padilla JE, Perez H, Lara M (1991) Control of nodulin genes in root-nodule development and metabolism. Annu Rev Plant Physiol Plant Mol Biol 42:507–528
Scheres B, Engelen F van, Knaap E van der, Wiel van de C, Kammen A van, Bisseling T (1990) Sequential induction of nodulin gene expression in the developing pea nodule. Plant Cell 2:687–700
Shapiro MB, Senapathy P (1987) RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res 15:7155–7174
Staden R (1986) The current status and portability of our sequence handling software. Nucleic Acids Res 14:217–231
Vasse J, Billy F de, Camut S, Truchet G (1990) Correlation between ultrastructural differentiation of bacteroids and nitrogen fixation in alfalfa nodules. J Bacteriol 172:4295–4306
Végh Z, Vincze É, Kadirov R, Tóth G, Kiss GB (1990) The nucleotide sequence of a nodule-specific gene, Nms-25 ofMedicago sativa: its primary evolution via exon-shuffling and retro-transposon-mediated DNA rearrangements. Plant Mol Biol 15:295–306
Verma DPS (1992) Signals in root nodule organogenesis and endocytosis ofRhizobium. Plant Cell 4:373–382
Zimmermann J, Voss H, Schwager C, Stegemann J, Erfle H, Stucky K, Kristensen T, Ansorge W (1990) A simplified protocol for fast plasmid DNA sequencing. Nucleic Acids Res 18:1067
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Küster, H., Frühling, M., Pühler, A. et al. The modular nodulins Nvf-28/32 of broad bean (Vicia faba L.): alternative exon combinations account for different modular structures. Molec. Gen. Genet. 252, 648–657 (1996). https://doi.org/10.1007/BF02173970
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DOI: https://doi.org/10.1007/BF02173970