Cloning And Expression Analysis Of CBL1 From Ammopiptanthus Mongolicus

Ca2+ is the critical component in the signal transduction pathways that lead to stress gene expression in plants, the cellular calcium changes are perceived by calcium sensors. The CBL (calcineurin B-like) protein family represents a unique group of calcium sensors in plants. A member of this family, CBL1, can be induced by multiple stresses, and can positively enhance the tolerance of Arabidopsis thaliana to salt and drought. CBL1 was only cloned in Arabidopsis thaliana. Ammopiptanthus mongolicus, as the unique evergreen shrub in Chinese hungriness area, develops high endurance to drought, salt, and low temperature. The aim of our study is to isolate the CBL1 gene from A. mongolicus, so as to provide gene foundation for the later gene transformation to improve their stress resistance.In our research, A.momgolicus seedlings growing in MS medium for 2 weeks were firstly treated with modulate drought and salt stress. And then, the full-length open reading frame of CBLl cDNA was isolated by RT-PCR with RACE strategy from A.mongolicus treated materials. Analysis with the cloned AmCBL1, which has a complete 642bp ORF, and encodes 213 amino acid, including 4 EF-hand motif structures. Semi-quantity RT-PCR Southern blot was made for the total RNA of 0,2,4,12,24h drought and 300mM NaCl gradient treated respectively, using AmCBLl cDNA as the probe and Actin2 as an equal loading control. The result that AmCBLl can response to drought and salt stress treatment, and respectively reaches the maximum expression in 4h point and 2h point, indicates AmCBLl may mediate in responsing to drought and salt in Ammopiptanthus mongolicus and play a significant role.