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Cloning and expression analysis of 2-on-2 hemoglobin from soybean

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Abstract

Hemoglobins (Hbs) are heme proteins found in all five kingdoms of living organisms. In plants, three different classes of Hbs have been identified-nonsymbiotic Hbs from diverse species, symbiotic Hbs from nitrogen-fixing plants, and so-called 2-on-2 Hbs. Here, we report the cloning and expression analysis of the 2-on-2 Hb gene,GmGLB3, from soybean. TheGmGLB3 cDNA clone encodes a protein for 172 amino acid residues. Its deduced amino acid sequence shows the highest identity (74%) with 2-on-2 Hb fromMedicago truncatula. Multiple sequence alignment confirms the conserved and signature amino acid residues previously reported with plant 2-on-2 Hbs. Genomic Southern hybridization demonstrates thatGmGLB3 has two copies in the soybean genome. Based on our northern hybridization, theGmGLB3 gene is specifically expressed in root nodules, with levels increasing in the late stage during nodule development. Its transcript level is also increased under flooding and kinetin treatments in the roots, or under flooding and 2-iP treatments in the stems. However, no transcript is detected in the leaves regardless of treatment. Therefore, we propose that theGmGLB3 gene is specifically expressed in root nodules and that its expression in other plant organs is regulated by cytokinin and/or flooding.

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Literature cited

  • Altschul SF, Gish W, Miller W, Meyers EW, Lipman D (1990) Basic local alignment search tool. J Mol Biol 215: 403–410

    PubMed  CAS  Google Scholar 

  • Andersson CR, Jensen EO, Llewellyn DJ, Dennis ES, Peacock WJ (1996) A new hemoglobin gene from soybean: A role for hemoglobin in all plants. Proc Natl Acad Sci USA 93: 5682–5687

    Article  CAS  Google Scholar 

  • Appleby CA (1992) The origin and functions of hemoglobin in plants. Sci Prog 76: 365–398

    CAS  Google Scholar 

  • Dehio C, de Bruijn FJ (1992) The early nodulin gene SrEnod2 fromSesbania rostrata is inducible by cytokinin. Plant J 2: 117–128

    PubMed  CAS  Google Scholar 

  • Doyle JJ, Doyle Jl (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13–15

    Google Scholar 

  • Ehrhardt DW, Atkinson EM, Long SR (1992) Depolarization of alfalfa root hair membrane potential byRhizobium meliloti Nod factors. Science 256: 998–1000

    Article  PubMed  CAS  Google Scholar 

  • Ellfolk N, Sievers G (1973) The primary structure of soybean leghemoglobin. IV Fractionation and sequence of thermolytic peptides of the apoprotein of the slow component (Lba). Acta Chem Scand 27: 3986–3992

    Article  PubMed  CAS  Google Scholar 

  • Fang Y, Hirsch AM (1998) Studying early nodulin gene ENOD40 expression and induction by nodulation factor and cytokinin in transgenic alfalfa. Plant Physiol 116: 53–68

    Article  PubMed  CAS  Google Scholar 

  • Fedorova M, van de Mortel J, Matsumoto PA, Cho J, Town CD, VandenBosch KA, Gantt JS, Vance CP (2002) Genome-wide identification of nodule-specific transcripts in the model legumeMedicago truncatula. Plant Physiol 130: 519–537

    Article  PubMed  CAS  Google Scholar 

  • He CY, Zhang JS, Chen SY (2002) A soybean gene encoding a proline-rich protein is regulated by salicylic acid, an endogenous circadian rhythm and by various stresses. Theor Appl Genet 104: 1125–1131

    Article  PubMed  CAS  Google Scholar 

  • Hill RD (1998) What are hemoglobins doing in plants? Can J Bot 76: 707–712

    Article  CAS  Google Scholar 

  • Hurrell JG, Leach SJ (1977) The amino acid sequence of soybean leghemoglobin c2. FEBS Lett 80: 23–26

    Article  PubMed  CAS  Google Scholar 

  • Hyldig-Nielsen JJ, Jensen EO, Paludan K, Wiborg O, Garrett R, lorgensen P, Marcker KA (1982) The primary structures of two leghemoglobin genes from soybean. Nucleic Acids Res 10: 689–701

    Article  PubMed  CAS  Google Scholar 

  • Jimenez-Zurdo Jl, Frugier F, Crespi MD, Kondorosi A (2000) Expression profiles of 22 novel molecular markers for organogenetic pathways acting in alfalfa nodule development. Mol Plant Microbe Interact 13: 96–106

    Article  PubMed  CAS  Google Scholar 

  • Kim HJ, An CS (2003) Molecular characterization and expression analysis of hemoglobin cDNA from small radish (Raphanus sativusL. var. Sativus). J Plant Biol 46: 161–166

    Article  Google Scholar 

  • Larsen K (2003) Molecular cloning and characterization of cDNAs encoding hemoglobin from wheat (Thticum aestivum) and potato (Solarium tuberosum). Biochim Biophys Acta 1621: 299–305

    PubMed  CAS  Google Scholar 

  • Lira-Ruan V, Sarath G, Klucas RV, Arredondo-Peter R (2001) Synthesis of hemoglobins in rice (Oryza sativa var. Jackson) plants growing in normal and stress conditions. Plant Sci 161: 279–287

    Article  PubMed  CAS  Google Scholar 

  • Mathesius U, Charon C, Rolfe BG, Kondorosi A, Crespi M (2000) Temporal and spatial order of events during the induction of cortical cell divisions in white clover byRhizobium leguminosarum bv. Trifolii inoculation or localized cytokinin addition. Mol Plant Microbe Interact 13: 617–628

    Article  PubMed  CAS  Google Scholar 

  • Pesce A, Couture M, Dewilde S, Guertin M, Yamauchi K, Ascenzi P, Moens L, Bolognesi M (2000) A novel two-over-two alpha-helical sandwich fold is characteristic of the truncated hemoglobin family. EMBO J 19: 2424–2434

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Seregelyes C, Mustardy L, Ayaydin F, Sass L, Kovacs L, Endre G, Lukacs N, Kovacs I, Vass I, Kiss GB, Horvath GV, Dudits D (2000) Nuclear localization of a hypoxia-inducible novel non-symbiotic hemoglobin in cultured alfalfa cells. FEBS Lett 29: 125–130

    Article  Google Scholar 

  • Sowa AW, Duff SMG, Guy PA, Hill RD (1998) Altering hemoglobin levels changes energy status in maize cells under hypoxia. Proc Natl Acad Sci USA 18: 10317-KB21

    Article  Google Scholar 

  • Taylor ER, Nie XZ, MacGregor AW, Hill RD (1994) A cereal hemoglobin gene is expressed in seed and root tissues under anaerobic conditions. Plant Mol Biol 24: 853–862

    Article  PubMed  CAS  Google Scholar 

  • Trevaskis B, Watts RA, Andersson CR, Llewellyn DJ, Hargrove MS, Olson JS, Dennis ES, Peacock WJ (1997) Two hemoglobin genes inArabidopsis thaliana: The evolutionary origins of leghemoglobins. Proc Natl Acad Sci USA 28: 12230–12234

    Article  Google Scholar 

  • Uchiumi T, Shimoda Y, Tsuruta T, Mukoyoshi Y, Suzuki A, Senoo K, Sato S, Kato T, Tabata S, Higashi S, Abe M (2002) Expression of symbiotic and nonsymbiotic globin genes responding to microsymbionts onLotus japonicus. Plant Cell Physiol 43: 1351–1358

    Article  PubMed  CAS  Google Scholar 

  • Uhde-Stone C, Zinn KE, Ramirez-Yanez M, Li A, Vance CP, Allan DL (2003) Nylon filter arrays reveal differential gene expression in proteoid roots of white lupin in response to phosphorus deficiency. Plant Physiol 131: 1064–1079

    Article  PubMed  CAS  Google Scholar 

  • Vincent JM (1970) A Manual for the Practical Study of Root-Nodule Bacteria, I.B.R Handbook, No. 15. Black-well, Oxford

    Google Scholar 

  • Watts RA, Hunt PW, Hvitved AN, Hargrove MS, Peacock WJ, Dennis ES (2001) A hemoglobin from plants homologous to truncated hemoglobins of microorganisms. Proc Natl Acad Sci USA 28: 10119–10124

    Article  Google Scholar 

  • Weber RE, Vinogradov SN (2001) Nonvertebrate hemoglobins: Functions and molecular adaptations. Physiol Rev 81: 569–628

    PubMed  CAS  Google Scholar 

  • Wiborg O, Hyldig-Nielsen JJ, Jensen EO, Paludan K, Marcker KA (1982) The nucleotide sequences of two leghemoglobin genes from soybean. Nucleic Acids Res 10: 3487–3494

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Chung Sun An.

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Lee, H., Kim, H. & An, C.S. Cloning and expression analysis of 2-on-2 hemoglobin from soybean. J. Plant Biol. 47, 92–98 (2004). https://doi.org/10.1007/BF03030637

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