Abstract
Calmodulin is a ubiquitous transducer of calcium signals in eukaryotes. In diploid plant species, several isoforms of calmodulin have been described. Here, we report on the isolation and characterization of calmodulin cDNAs corresponding to 10 genes from hexaploid (bread) wheat (Triticum aestivum). These genes encode three distinct calmodulin isoforms; one isoform is novel in that it lacks a conserved calcium binding site. Based on their nucleotide sequences, the 10 cDNAs were classified into four subfamilies. Using subfamily-specific DNA probes, calmodulin genes were identified and the chromosomal location of each subfamily was determined by Southern analysis of selected aneuploid lines. The data suggest that hexaploid wheat possesses at least 13 calmodulin-related genes. Subfamilies 1 and 2 were both localized to the short arms of homoeologous-group 3 chromosomes; subfamily 2 is located on all three homoeologous short arms (3AS, 3BS and 3DS), whereas subfamily 1 is located only on 3AS and 3BS but not on 3DS. Further analysis revealed thatAegilops tauschii, the presumed diploid donor of the D-genome of hexaploid wheat, lacks a subfamily-1 calmodulin gene homologue, whereas diploid species related to the progenitors of the A and B genomes do contain such genes. Subfamily 3 was localized to the short arm of homoeologous chromosomes 2A, 2B and 2D, and subfamily 4 was mapped to the proximal regions of 4AS, 4BL and 4DL. These findings suggest that the calmodulin genes within each subfamily in hexaploid wheat represent homoeoallelic loci. Furthermore, they also suggest that calmodulin genes diversified into subfamilies before speciation ofTriticum andAegilops diploid species.
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References
Ahn S, Anderson JA, Sorrells ME, Tanksley SD (1993) Homoeologous relationships of rice, wheat and maize chromosomes. Mol Gen Genet 241:483–490
Antosiewicz DM, Polisensky DH, Braam J (1995) Cellular localization of the Ca2+ binding TCH3 protein ofArabidopsis. Plant J 8:623–636
Baum G, Lev-Yadun S, Fridmann Y, Arazi T, Katsnelson H, Zik M, Fromm H (1996) Calmodulin binding to glutamate decarboxylase is required for regulation of glutamate and GABA metabolism and normal development in plants. EMBO J 15:2988–2996
Braam J (1992) Regulated expression of the calmodulin-relatedTCH genes in culturedArabidopsis cells: induced by calcium and heat shock. Proc Natl Acad Sci USA 89:3213–3216
Braam J, Davis RW (1990) Rain-, wind-, and touch-induced expression of the calmodulin-related genes inArabidopsis. Cell 60:357–364
Bush DS (1995) Calcium regulation in plant cells and its role in signaling. Annu Rev Plant Physiol Plant Mol Biol 46:95–122
Chao S, Raines CA, Longstaff M, Sharp PJ, Gale MD, Dyer TA (1989) Chromosomal location and copy number in wheat and some of its close relatives of genes for enzymes involved in photosynthesis. Mol Gen Genet 218:423–430
Delaney DE, Nasuda S, Endo TR, Gill BS (1995) Cytologically based physical maps of the group 3 chromosomes of wheat. Theor Appl Genet 91:780–782
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Devos KM, Dubcovsky J, Dvorak J, Shinoy CN (1995) Structural evolution of wheat chromosomes 4A, 5A, and 7B and its impact on recombination. Theor Appl Genet 91:282–288
Doyle KE, Kovalik GE, Lee E, Becknigham K (1990)Drosophila melanogaster contains a single calmodulin gene: further structure and expression studies. J Mol Biol 213:599–605
Dvorak J (1988) Cytogenetical and molecular inferences about the evolution of wheat. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genet Symp. Cambridge, UK, pp 187–192
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Feldman M, Lupton FGH, Miller TE (1995) Wheats,Triticum spp. (Gramineae-Triticinae). In: Smartt J, Simmonds NW (eds) Evolution of Crop Plants. Longman, London, pp 184–192
Fischer R, Koller M, Flura M, Mathews S, Strehler-Page MA, Krebs J, Penniston JT, Carafoli E and Strehler EE (1988) Multiple divergent mRNAs code for a single human calmodulin. J Biol Chem 263:17055–17062
Galili G, Feldman M (1984) Mapping of glutenin and gliadin genes located on chromosome 1B of common wheat. Mol Gen Genet 193:293–298
Galili S, Galili G, Feldman M (1991) Chromosomal location of genes for Rubisco small subunit and rubisco-binding protein in common wheat. Theor Appl Genet 81:98–104.
Galili S, Galili G, Avivi Y, Feldman M (1992) Identification and chromosomal location of four subfamilies of the Rubisco small subunit genes in common wheat. Theor Appl Genet 83:385–391
Gawienowski MC, Szymanski D, Perera IY, Zielinski RE (1993) Calmodulin isoforms inArabidopsis encoded by multiple divergent mRNAs. Plant Mol Biol 22:215–225
Harcourt RL, Gale MD (1991) A chromosome-specific DNA sequence which reveals a high level of RFLP in wheat. Theor Appl Genet 81:397–400
Ito T, Hirano M, Akama K, Shimura Y, Okada K (1995) Touch-inducible genes for calmodulin and a calmodulin-related protein are located in tandem on a chromosome ofArabidopsis thaliana. Plant Cell Physiol 36:1369–1373
James P, Vorherr T, Carafoli E (1995) Calmodulin-binding domains: just two-faced or multifaceted? Trends Biochem Sci 20:38–42
Jena PK, Reddy ASN, Poovaih BW (1989) Molecular cloning and sequencing of a cDNA for plant calmodulin: signal-induced changes in the expression of calmodulin. Proc Natl Acad Sci USA 86:3644–3648
Kidwell KK, Osborn TC (1992) Simple plant DNA isolation procedures. In: Berkmann JS, Osborn TC (eds) Plant genomes: methods for genetics and physical mapping. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 1–13
Kurata N, Moore G, Nagamura Y, Foote T, Yano M, Minobe Y, Gale MD (1994) Conservation of genome structure between rice and wheat. Bio/Technology 12:276–278
Lee SH, Kim JC, Lee MS, Heo WD, Seo HS, Yoon HW, Hong JC, Lee SY, Bahk JD, Hwang I, Cho MJ (1995) Identification of a novel divergent calmodulin isoform from soybean which has differential ability to activate calmodulin-dependent enzymes. J Biol Chem 270:21806–21812
Ling V, Zielinski RE (1989) Cloning of cDNA sequences encoding the calcium-binding protein, calmodulin, from barley (Hordeum vulgare L.). Plant Physiol 90:714–719
Ling V, Perera I, Zielinski RE (1991) Primary structure ofArabidopsis calmodulin isoforms deduced from the sequences of cDNA clones. Plant Physiol 96:1196–1202
McFadden ES, Sears ER (1946) The origin ofTriticum spelta and its free-threshing hexaploid relatives. J Hered 37:81–89, 107–116
McIntosh RA, Hart GE, Gale MD (1993) Catalogue of gene symbols for wheat. In: Li ZS, Xin ZY (eds) Proc 8th Int Wheat Genet Symp. China Agricultural Scientech Press, Beijing, China, pp 1333–1500
Mickelson-Young L, Endo TR, Gill BS (1995) A cytogenetic ladder-map of the wheat homoeologous group-4 chromosomes. Theor Appl Genet 90:1007–1011
Nishikawa K (1983) Species relationship of wheat and its putative ancestors as viewed from isozyme variation. In: Sakamoto S (ed) Proc 6th Int Wheat Genet Symp. Kyoto, Japan, pp 59–63
Ogihara Y, Shimiza H, Hasegawa K, Tsujimoto H, Sasakuma T (1994) Chromosome assignment of four photosynthesis-related genes and their variability in wheat species. Theor Appl Genet 88:383–394
Perera I, Zielinski RE (1992) Structure and expression of theArabidopsis CaM-3 calmodulin gene. Plant Mol Biol 19:649–664
Roberts DM, Harmon AC (1992) Calcium-modulated proteins: targets of intracellular calcium signals in higher plants. Annu Rev Plant Physiol Plant Mol Biol 43:375–414
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning a laboratory manual (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sears ER (1954) The aneuploids of common wheat. Mo Agric Exp Sta Res Bull 572:1–58
Sears ER (1966) Nullisomic-tetrasomic combinations in hexaploid wheat. In: Riley R, Lews KR (eds) Chromosome manipulation and plant genetics. Oliver and Boyd, Edinburgh, UK, pp 29–45
Sears ER (1982) A wheat mutation conditioning an intermediate level of homoeologous chromosome pairing. Can J Genet Cytol 24:715–719
Sears ER, Sears LMS (1978) the telocentric chromosomes of common wheats. In: Ramanujam S (ed) Proc 5th Int Wheat Genet Symp. Indian Soc Genet Plant Breed, New Delhi, pp 389–407
Sistrunk M, Antosiewicz DM, Purugganan MM, Braam J (1994)Arabidopsis TCH3 encodes a novel Ca2+ binding protein and shows environmentally induced and tissue-specific regulation. Plant Cell 6:1553–1565
Snedden WA, Koutsia N, Baum G, Fromm H (1996) Activation of a recombinant petunia glutamate decarboxylase by calcium/calmodulin or by a monoclonal antibody which recognizes the calmodulin-binding domain. J Biol Chem 271:4148–4153
Takezawa D, Liu ZH, An G, Poovaiah BW (1995) Calmodulin gene family in potato — developmental and touch-induced expression of the messenger-RNA encoding a novel isoform. Plant Mol Biol 27:693–703
Toda H, Yazawa M, Sakiyama F (1985) Amino acid sequence of calmodulin from wheat germ. J Biochem 98:833–842
Van Deynze AE, Nelson JC, Yglesias ES, Harrington SE, Braga DP, McCouch SR, Sorrells ME (1995) Comparative mapping in grasses. Wheat relationships. Mol Gen Genet 248:744–754
Werner JE, Endo TR, Gill BS (1992) Toward a cytogenetically based physical map of the wheat genome. Proc Natl Acad Sci USA 89:11307–11311
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Yang, T., Segal, G., Abbo, S. et al. Characterization of the calmodulin gene family in wheat: structure, chromosomal location, and evolutionary aspects. Molec. Gen. Genet. 252, 684–694 (1996). https://doi.org/10.1007/BF02173974
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DOI: https://doi.org/10.1007/BF02173974