Cloning and Expression Characteristic Analysis of SsCBL4 gene from Sugarcane (Saccharum spp.)
LING Qiu-Ping, WU Jia-Yun, YANG Zhan-Duan, ZHOU Wen-Ling, HUANG Ying, LI Qi-Wei, ZENG Qiao-Ying*
Guangzhou Bioengineering Institute (Guangzhou Sugarcane Industry Research Institute) / Guangdong Key Lab of Sugarcane Improvement and Biorefinery/Guangdong Sugarcane Improvement Engineering Center), Guangzhou 510316, China
Abstract：The calcineurin B-like protein (CBL), which is a unique group of calcium sensors in plant, play a key role in stress response. Sugarcane (Saccharum spp.) is an important crop for sugar and bio-energy in China. According to the datasets which are transcription sequencing of sugarcane genes under abiotic stress, a calcineurin B-like gene named SsCBL4 (GenBank No. KY674 987.1), was cloned from root in sugarcane under low potassium stress by RT-PCR (reverse transcription-PCR). SsCBL4 contained a 636 bp complete open reading frame that encodes 211 amino acids. Its molecular weight was 23. 97 903 kD. And the isoelectric point of SsCBL4 was 4.88. The deduced polypeptide of SsCBL2 had 2 transmembrane domains which could bind calcium ions. The SsCBL4 protein contain 3 EF-hand domains (elongation factor hand) that are the typical domain of CBL family, a conserved N-myristoylation motif and FPSF motif which interacted with CBL-interacting protein kinase (CIPK) kinases. The homology analysis indicated that SsCBL4 has high homology to the CBL4 from Sorghum bicolor, Zea mays, Oryza sativa and Triticum aestivum. The changes in expression of SsCBL4 under low nitrogen, low phosphorus, low potassium, drought or salt stresses were detected by qPCR analysis. The expressions of SsCBL4 were commonly up-regulated under low potassium, drought or salt stresses. Under low nitrogen or low phosphorus stress, the fluctuations in SsCBL4 gene expression levels were observed, with the highest level at 96 h, followed by a significant decrease. Under low potassium stress from 8 to 48 h, the expression of SsCBL4 gradually decreased. SsCBL4 was induced to express at relatively high levels at 8, 24 and 96 h after drought treatment, and reached the highest level at 96 h, which was 20.5-fold as high as that of control. Under salt stress from 24 h to 96 h, the expression of SsCBL4 gradually increased, and reached the highest inducible expression level at 96 h, which was 9.1-fold as high as control. These results suggest that SsCBL4 gene may play an important role in different stresses. This study provides a reference for further study of SsCBL4 stress resistance and related molecular mechanisms.
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