Identification Of Interacting Proteins With AtGluRS By Using Yeast Two-hybrid System And Physiological Analysis Of Transgenic Arabidopsis Treated With Different Plant Hormones

The yeast two-hybrid system has become an important method to analyze the protein-protein interactions. The interactions between unknown proteins have revealed with it. ABA plays a major role in regulating plant growth and development including seed maturation, germination, dormancy, as well as mediating adaptations to environmental stresses such as cold, drought and salinity.AtGluRS belongs to aminoacyl-tRNA synthetase family I, the structure with much subunit, the fuctions involve combining tRNA to amino acid,accelerating tRNA maturation and emendation. In addition, the protein is concerned with ABA signal transduction. AtGluRS is confirmed as a factor of ABA signal pathway. Its major effect: the first is the reciprocity with ABI1 and ABI2. ABI1 and ABI2 are as negative regulators in stomata closure induced by ABA. AtGluRS can combine with ABI1 to restrain its activity and affect the signal transduction; at the level of transcription, the transcription of AtGluRS depends on ABI1 and ABI2, AtGluRS seems to be involved in the processing of ABI1 and ABI2 by regulating splicing of the pre-mRNA to mature mRNA. Another effect is that it can separately impact biosynehesis of ABA by controlling some key enzyme, for example NCED3.ABA, ethylene, GA are important plant hormones,the cross-talk perhaps exists in these signaling pathways. As one of important factors in ABA signaling, AtGluRS may play a role in ethylene and GA signaling.Objective is to screen interacting proteins with AtGluRS in Arabidosis by using yeast-two hybrid technique. The physiological parameter is detected in order to reveal whether AtGluRS participates in other plant hormone signal pathway. Methods include extracting Arabidosis cDNA, and retesting the positive copies, then sequencing them. Conclusion indicate that the bait plasmid pGBKT7-AtGluRS(fc) and pGBKT7-AtGluRS( A C) are successfully constructed in the yeast two-hybrid system, and we detect the length of culm and root treated with ethephon. We also caculate the germination rate treated with GA3. As a result, AtGluRS sense lines and AtGluRS anti-sense lines are not sensitive to ethylene and GA3. AtGluRS sense lines germinate prior to 35S-GUS transformant lines and AtGluRS anti-sense lines...