Identification and expression analysis of pineapple AcCAX1
[Objective]This study aimed to clone and identify the Ca2+/H+antiport protein AcCAX1 in pineapple,analyze its full-length sequence and expression profile,and establish a foundation for functional studies of this gene.[Methods]The Ac-CAX1 sequence was obtained through PCR amplification techniques,and its protein structure,sequence alignment,and phylo-genetic relationships were analyzed using bioinformatics tools.Real-time fluorescence quantitative PCR(qPCR)was employed to assess the response of AcCAX1 to different ion,temperature stress,and plant hormones.[Results]AcCAX1 has an open reading frame of 1428 base pairs and encodes a stable hydrophobic protein consisting of 514 amino acids with a relative molecu-lar mass of 50 801.11 Da and a theoretical isoelectric point of 6.24.Sequence analysis revealed that the amino acid sequence of AcCAX1 contained 11 typical trans-membrane domains,including two Ca2+/H+antiporter domains.Secondary structure predi-cations revealed that the α-helix constituted the largest proportion of the CAX1 protein structure at 58.74%,followed by irregu-lar coils(21.68%),elongation chains(14.95%),and β-sheets(4.63%).Real-time qPCR analysis showed that the tran-script level of AcCAX1 was upregulated by exogenous CaCl2 and NaCl treatments,while MnCl2 treatment led to a decrease in transcript levels.Spatial expression analysis revealed that AcCAX1 exhibited higher expression levels in older leaves and stems but lower levels in fruits and flower buds.Subcellular localization in tobacco showed that AcCAX1 was expressed in the cell membrane,cytoplasm,and nucleus.In addition,the promoter region of AcCAX1 contained a large number of abiotic stress re-sponse elements.The transcript level of AcCAX1 was significantly down-regulated under both cold and heat stress conditions.[Conclusion]The Ca2+/H+antiport protein AcCAX1 in pineapple likely plays a significant role in the plant's response to abiot-ic stresses.
万方数据
维普
