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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (03): 440-448.doi: 10.3724/SP.J.1006.2013.00440

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Isolation and Characterization of a Rape MAP Kinase BnMPK9 from Group D

YU Shun-Wu1,ZHANG Li-Da2,LIU Guo-Lan1,LUO Li-Jun1,*   

  1. 1 Shanghai Agrobiological Gene Center, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; 2 Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China
  • Received:2012-07-26 Revised:2012-11-16 Online:2013-03-12 Published:2012-12-11
  • Contact: 罗利军, E-mail: lijun@sagc.org.cn, Tel: 021-52230526

Abstract:

Under environmental stresses, plant has developed complex signaling networks to respond environmental signals and adapt to unfavorable conditions. Mitogen-activated protein kinase (MAPK) signaling pathways are one of the most important and conserved system to control cellular response and growth. A gene encoding Mitogen-activated protein kinase was isolated from oilseed rape (Brassica napus) treated with drought based on RT-PCR, designated as BnMPK9 (GenBank accession number AY737714). The cloned gene contains a Pkinase domain and a conserved CD domain. BnMPK9 is highly homologous to AtMPK9 gene from Arabidopsis thaliana and belongs to the D-Subgroup of mitogen-activated protein kinase with phosphorylation motif (TDY). Southern bolt analysis revealed that there were at least two copies in rape genome. Northern bolt analyses showed that BnMPK9 could express in different tissues. The expression level of BnMPK9 transcript was up-regulated by mannitol, UV, and H2O2,but down-regulated by low temperature and salicylic acid (SA). Through Real-time RT-PCR, mannitol and H2O2 treatments could induce its high expression under different time. In root, mannitol treatment could enhance its expression. BnMPK9 was ligated into pYES2.1 and transformed into yeasts, and BnMPK9s expression enhanced yeasts’ resistance to mannitol and peroxide in 600 m mol L–1 mannitol and 0.2 mmol L–1 tBuOOH drug assays. The present studies show that BnMPK9 is a member of MAPK family involved in eukaryotic responses and resistance to osmotic and ROS stresses.

Key words: BnMPK9, Brassica napus, Abiotic stress, Yeast expression

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