biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 56:43-49, 2012 | DOI: 10.1007/s10535-012-0014-5

Molecular cloning and characterization of a novel stress responsive gene in alfalfa

R. Long2,3, Q. Yang2, J. Kang2, Y. Chao2, P. Wang3, M. Wu2, Z. Qin3, Y. Sun1,*
1 College of Animal Science and Technology, China Agriculture University, Beijing, P.R. China
2 Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, P.R. China
3 Bioengineering College of Chongqing University, Chongqing, P.R. China

A suppression subtraction hybridization (SSH) cDNA library of alfalfa (Medicago sativa L.) cv. Zhongmu NO.1 had been constructed to identify differentially expressed genes under stress. Based on the sequence of a 460 bp expressed sequence tags (ESTs), a cDNA of 1652 bp was cloned by rapid amplification of cDNA ends (RACE) method. This gene (MsPBL) was predicted to encode a 434-amino-acid protein, which contained a Phox and Bem1 (PB1) domain. PB1 domain is a functional domain comprising about 80 amino acid residues, which exists in many signal transduction proteins and mediates dimerization in the proteins. PB1 domain is mostly involved in two cell signal transduction pathways: MAPK and NF-KB. When fused to the green fluorescent protein, we found MsPBL localization in the nucleus of onion (Allium cepa L.) epidermal cells. The transcripts of MsPBL rose significantly when alfalfa was treated with 300 mM NaCl, 0.1 mM ABA, and 20 % polyethylene glycol (PEG-6000). These results indicated that MsPBL may be functional within the nucleus as a signal transduction protein to allow alfalfa to rapidly respond to the environmental stress signals.

Keywords: Allium cepa; gene expression; Medicago sativa; PB1 domain; phylogenetic tree; RACE; subcellular localization
Subjects: gene expression; PB1 domain; phylogenetic tree; RACE; subcellular localization; expressed sequence tag; PCR; nucleotide sequence; amino acid sequence; NaCl; abscisic acid; polyethylene glycol

Received: March 19, 2010; Accepted: November 8, 2010; Published: March 1, 2012  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Long, R., Yang, Q., Kang, J., Chao, Y., Wang, P., Wu, M., Qin, Z., & Sun, Y. (2012). Molecular cloning and characterization of a novel stress responsive gene in alfalfa. Biologia plantarum, 56(1), 43-49. doi: 10.1007/s10535-012-0014-5
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Agarwal, P.K., Jha, B.: Transcription factors in plants and ABA dependent and independent abiotic stress signalling. - Biol Plant. 54: 201-212, 2010. Go to original source...
    2. Boyer, J.S.: Plant productivity and environment. - Science 218: 443-448, 1982. Go to original source...
    3. English, J.M., Vanderbilt, C.A., Xu, S., Marcus, S., Cobb, M.H.: Isolation of MEK5 and differential expression of alternatively spliced forms. - J. biol. Chem. 270: 28897-28902, 1995. Go to original source...
    4. Finkelstein, R., Gampala, S.S., Lynch, T.J., Thomas, T.L., Rock, C.D.: Redundant and distinct functions of the ABA response loci ABA-INSENSITIVE (ABI) 5 and ABREBINDING FACTOR (ABF) 3. - Plant mol. Biol. 59: 253-267, 2005. Go to original source...
    5. Gonzalez-Guzman, M., Abia, D., Salinas, J., Serrano, R., Rodriguez, P.L.: Two new alleles of the abscisic aldehyde oxidase 3 gene reveal its role in abscisic acid biosynthesis in seeds. - Plant Physiol. 135: 325-333, 2004. Go to original source...
    6. Heidenreich, T., Wollers, S., Mendel, R.R., Bittner, F.: Characterization of the NifS-like domain of ABA3 from Arabidopsis thaliana provides insight into the mechanism of molybdenum cofactor sulfuration. - J. biol. Chem. 280: 4213-4218, 2005. Go to original source...
    7. Hirano, Y., Yoshinaga, S., Ogura, K., Yokochi, M., Noda, Y., Sumimoto, H., Inagaki, F.: Solution structure of atypical protein kinase C PB1 domain and its mode of interaction with ZIP/p62 and MEK5. - J. biol. Chem. 279: 31883-31890, 2004. Go to original source...
    8. Hunter, T.: Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. - Cell 80: 225-236, 1995. Go to original source...
    9. Ito, T., Matsui, Y., Ago, T., Ota, K., Sumimoto, H.: Novel modular domain PB1 recognizes PC motif to mediate functional protein-protein interactions. - EMBO J. 20: 3938-3946, 2001. Go to original source...
    10. Jin, H., Sun, Y., Yang, Q., Chao, Y., Kang, J., Li, Y.: Screening of genes induced by salt stress from alfalfa. - Mol. biol. Rep. 37: 745-753, 2010. Go to original source...
    11. Leon-Kloosterziel, K.M., Gil, M.A., Ruijs, G.J., Jacobsen, S.E., Olszewski, N.E., Schwartz, S.H., Zeevaart, J.A., Koornneef, M.: Isolation and characterization of abscisic acid-deficient Arabidopsis mutants at two new loci. - Plant J. 10: 655-661, 1996. Go to original source...
    12. Long, S.P., Ainsworth, E.A., Leakey, A.D., Nosberger, J., Ort, D.R.: Food for thought: lower-than-expected crop yield stimulation with rising CO2 concentrations. - Science 312: 1918-1921, 2006. Go to original source...
    13. Moscat, J., Diaz-Meco, M.T., Albert, A., Campuzano, S.: Cell signaling and function organized by PB1 domain interactions. - Mol. Cell 23: 631-640, 2006. Go to original source...
    14. Munnik, T., Ligterink, W., Meskiene, I.I., Calderini, O., Beyerly, J., Musgrave, A., Hirt, H.: Distinct osmo-sensing protein kinase pathways are involved in signalling moderate and severe hyper-osmotic stress. - Plant J. 20: 381-388, 1999. Go to original source...
    15. Munns, R., Tester, M.: Mechanisms of salinity tolerance. - Annu. Rev. Plant Biol. 59: 651-681, 2008. Go to original source...
    16. Pfaffl, M.W.: A new mathematical model for relative quantification in real-time RT-PCR. - Nucl. Acids Res. 29(Suppl): e45, 2001. Go to original source...
    17. Ponting, C.P., Ito, T., Moscat, J., Diaz-Meco, M.T., Inagaki, F., Sumimoto, H.: OPR, PC and AID: all in the PB1 family. - Trends Biochem. Sci. 27: 10, 2002. Go to original source...
    18. Robinson, M.J., Cobb, M.H.: Mitogen-activated protein kinase pathways. - Curr. Opin. Cell Biol. 9: 180-186, 1997. Go to original source...
    19. Ruggiero, B., Koiwa, H., Manabe, Y., Quist, T.M., Inan, G., Saccardo, F., Joly, R.J., Hasegawa, P.M., Bressan, R.A., Maggio, A.: Uncoupling the effects of abscisic acid on plant growth and water relations. Analysis of sto1/nced3, an abscisic acid-deficient but salt stress-tolerant mutant in Arabidopsis. - Plant Physiol. 136: 3134-3147, 2004. Go to original source...
    20. Saio, T., Yokochi, M., Inagaki, F.: The NMR structure of the p62 PB1 domain, a key protein in autophagy and NFkappaB signaling pathway. - J. Biomol. NMR 45: 335-341, 2009. Go to original source...
    21. Terasawa, H., Noda, Y., Ito, T., Hatanaka, H., Ichikawa, S., Ogura, K., Sumimoto, H., Inagaki, F.: Structure and ligand recognition of the PB1 domain: a novel protein module binding to the PC motif. - EMBO J. 20: 3947-3956, 2001. Go to original source...
    22. Waskiewicz, A.J., Cooper, J.A.: Mitogen and stress response pathways: MAP kinase cascades and phosphatase regulation in mammals and yeast. - Curr. Opin. Cell Biol. 7: 798-805, 1995. Go to original source...
    23. Zhu, J.K.: Genetic analysis of plant salt tolerance using Arabidopsis. - Plant Physiol. 124: 941-948, 2000. Go to original source...

    Return to the content