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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (10): 1756-1766.

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

Characterization and Functional Analysis of a Small GTP-binding Protein AtRAB Interacting with H+-Pyrophosphatase AVP1 in Arabidopsis thaliana

LIU Rong-Bang1,2,CHEN Ming2,*,GUO Meng-Meng2,SI Qing-Lin2,GAO Shi-Qing3,XU Zhao-Shi2,LI Lian-Cheng2,MA You-Zhi2,YIN Jun1,*   

  1. 1 Henan Agricultural University / National Key Laboratory of Wheat and Maize Crop Science / Collaborative Innovation Center of Henan Grain Crops, Zhengzhou 450002, China; 2 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture, Beijing 100081, China; 3 Beijing Hybrid Wheat Engineering and Technology Research Center, Beijing 100097, China
  • Received:2014-03-10 Revised:2014-06-16 Online:2014-10-12 Published:2014-07-23
  • Contact: 尹钧, E-mail: xmzxyj@126.com, Tel: 0371-63558203; 陈明, E-mail: chenming02@caas.cn, Tel: 010-82108789

Abstract:

H+-Pyrophosphatase (H+-PPase) is an important proton transporter in plants. It cooperates with H+-ATPase and transport protons in vacuole or extracellular area to maintain a constant H+ gradient, which enables the transport of ions and other components (e.g. amino acids, carbohydrates). In the current research, AVP1 was applied to membrane proteins-based yeast two-hybrid system, and a small GTP-binding protein AtRAB was identified by screening the Arabidopsis cDNA library. The interaction between AVP1 and AtRAB was confirmed by interaction analysis in yeast. Bimolecular fluorescent complementation (BiFC) analysis suggested that AtRAB and AVP1 interaction took place in the plasma membrane and the nucleus. Phenotypes of wild type (WT) and Arabidopsis mutants of avp1 (AVP1) and rab (AtRAB) were compared under high-salt, low-phosphorus and low-potassium conditions. Under the high salt stress, avp1 and rab presented similar phenotypes along with the increase of NaCl concentration, which were characterized by shortened root length and reduced lateral root number in comparison with the WT. The decreasing phosphorus concentration also led to the phenotypes of shortened root length, reduced lateral root number and reduced total root area in the two mutants. Particularly, stronger response was observed in rab than in avp1 under the same phosphorus concentration. The potassium treatments resulted in similar phenotypes to those in phosphorus treatment. The results indicated that AVP1 can interact with AtRAB on the plasma membrane and the nucleus and further influence the absorption of ions in plants. The two mutants of avp1 and rab showed similar phenotypes, suggesting that both AVP1 and AtRAB positively regulate plant response to high salt, low phosphorus and low potassium stresses in the same signaling pathway.

Key words: H+-Pyrophosphatase, Small GTP-binding protein AtRAB, Yeast two-hybrid system, Protein interaction, stresses response

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