Genome-wide identification and expression analysis of the molecular chaperone binding protein BiP genes in Citrus
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Here, we report for the first time the genome-wide identification and expression analysis of the molecular chaperone BiP genes in Citrus. Six genes encoding the conserved protein domain family GPR78/BiP/KAR2 were identified in the genome of Citrus sinensis and C. clementina. Two of them, named here as CsBiP1 and CsBiP2, were classified as true BiPs based on their deduced amino acid sequences. Alignment of the deduced amino acid sequences of CsBiP1 and CsBiP2 with BiP homologs from soybean and Arabidopsis showed that they contain all the conserved functional motifs of BiPs. Analysis of the promoter region of CsBiPs revealed the existence of cis-acting regulatory sequences involved in abiotic, heat-shock, and endoplasmic reticulum (ER) stress responses. Publicly available RNA-seq data indicated that CsBiP1 is abundantly expressed in leaf, flower, fruit, and callus, whereas CsBiP2 expression is rarely detected in any tissues under normal conditions. Comparative quantitative real-time PCR (qPCR) analysis of expression of these genes between C. sinensis grafted on the drought-tolerant “Rangpur” lime (C. limonia) and -sensitive “Flying Dragon” trifoliate orange (Poncirus trifoliata) rootstocks showed that CsBiP1 was upregulated by drought stress on the former but downregulated on the latter, whereas the CsBiP2 mRNA levels were downregulated on drought-stressed “Flying Dragon,” but remained constant on “Rangpur.” CsBiP2 upregulation was only observed in C. sinensis seedlings subjected to osmotic and cold treatments. Taken together, these results indicate the existence of two highly conserved BiP genes in Citrus that are differentially regulated in the different tissues and in response to abiotic stresses.
KeywordsAbiotic stress Drought tolerance Endoplasmic reticulum HSP70 Immunoglobulin-heavy-chain-binding protein Mandarin Sweet orange UPR pathway
R.F.G., L.P.G., J.C.C.M., and G.S.S. conducted the experiments. R.F.G., L.R.C., A.O.S., and A.M.M.B. analyzed the data and drafted the manuscript. F.C.A., M.G.C.C., and C.P.P. supported the project and designed the experiments. M.G.C.C. polished the manuscript. All authors read and approved the manuscript.
This work was supported by research grants from CNPq (Process # 306667/2014-2), Instituto Nacional de Ciência e Tecnologia (INCT) de Genômica para Melhoramento de Citros (CNPq Process # 465440/2014-2, and FAPESP Process # 2008/2014/50880-0) and Embrapa (Macroprograma 2). L.R.C. is recipient of a CAPES/PNPD postdoctoral fellowship. M.G.C.C. and C.P.P. are CNPq Research Fellows. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Conflict of interest
The authors declare that they have no competing interests.
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