Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (5): 889-897.doi: 10.3864/j.issn.0578-1752.2013.05.003

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

The Expression Analysis and Screening of Interaction Protein of Mitogen-Activated Protein Kinase (CaMAPKK2) in Salt-Stress Signal Pathways of Chenopodium album

 CHEN  Sha-Sha, HE  Zhuan-Zhuan, JIANG  Sheng-Xiu, LI  Xiao-Rong, XING  Jia-Jia, 吕Xiu-Yun , LAN  Hai-Yan   

  1. College of Life Science and Technology, Xinjiang University/Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046
  • Received:2012-11-14 Online:2013-03-01 Published:2013-01-16

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Abstract: 【Objective】The aim of the present study is to understand the mechanism of stress signal transduction to induce the response of adverse tolerance with Chenopodium album, an adverse-tolerant plant species, through analysis of the expression pattern of mitogen-activated protein kinase gene (MAPKK) under different abiotic stresses and screening of interaction proteins of MAPKK in MAPK signal transduction pathways. 【Method】 Using the total RNA from the leaf of C. album as the template, the expression patterns of MAPKK under NaCl, H2O2 and ABA were analyzed by quantitative PCR technique. The full-length cDNA sequence of MAPKK was obtained by combined reverse transcription-PCR (RT-PCR) and RACE techniques. The yeast two-hybrid technique was employed to analyze the interaction components of MAPKK in salt-stress signal pathway. 【Result】 A full length cDNA fragment of MAPKK gene from C. album included a 1 089 bp ORF, which encodes with 362 putative amino acids, named as CaMAPKK2. qPCR analysis showed that CaMAPKK2 gene was significantly induced by salt and H2O2 but not by ABA stress, at least in the present study. DPI or Na2WO4 (inhibitor of H2O2 or ABA synthesis) significantly inhibited the expression of CaMAPKK2 in C. album when exposed to 300 mmol.L-1 NaCl stress. By using the CaMAPKK2 as bait protein, five potential clones which may interact with CaMAPKK2 were obtained by the first screening of yeast two hybridization. Sequencing result indicated that one 794 bp fragment out of 5 cDNA showed correct reading frame, and shared identities of 79% and 78% in nucleotide sequence to AgTHI1 and AtTHI1 (thiazole biosynthetic enzyme) from Alnus glutinosa and Arabidopsis thaliana, respectively, while other 4 cDNA sequences had no correct reading frame. 【Conclusion】CaMAPKK2 gene was significantly induced by salt and H2O2 stress, suggesting that salt stress might induce the accumulation of H2O2 and ABA, which in turn leads to expression increasing of MAPKK. More positive clones should be acquired and the relevant experiments (e.g. Pull down, CoIP) should be performed for screening and verifying the interaction component of CaMAPKK2.

Key words: Chenopodium album , salt stress , signal transduction pathway , MAPKK , yeast two-hybrid technique , quantitative PCR

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