Identification and characterization of CBL and CIPK gene families in canola (Brassica napus L.)

Abstract

BackgroundCanola (Brassica napus L.) is one of the most important oil-producing crops in China and worldwide. The yield and quality of canola is frequently threatened by environmental stresses including drought, cold and high salinity. Calcium is a ubiquitous intracellular secondary messenger in plants. Calcineurin B-like proteins (CBLs) are Ca2+ sensors and regulate a group of Ser/Thr protein kinases called CBL-interacting protein kinases (CIPKs). Although the CBL-CIPK network has been demonstrated to play crucial roles in plant development and responses to various environmental stresses in Arabidopsis, little is known about their function in canola.ResultsIn the present study, we identified seven CBL and 23 CIPK genes from canola by database mining and cloning of cDNA sequences of six CBLs and 17 CIPKs. Phylogenetic analysis of CBL and CIPK gene families across a variety of species suggested genome duplication and diversification. The subcellular localization of three BnaCBLs and two BnaCIPKs were determined using green fluorescence protein (GFP) as the reporter. We also demonstrated interactions between six BnaCBLs and 17 BnaCIPKs using yeast two-hybrid assay, and a subset of interactions were further confirmed by bimolecular fluorescence complementation (BiFC). Furthermore, the expression levels of six selected BnaCBL and 12 BnaCIPK genes in response to salt, drought, cold, heat, ABA, methyl viologen (MV) and low potassium were examined by quantitative RT-PCR and these CBL or CIPK genes were found to respond to multiple stimuli, suggesting that the canola CBL-CIPK network may be a point of convergence for several different signaling pathways. We also performed a comparison of interaction patterns and expression profiles of CBL and CIPK in Arabidospsis, canola and rice, to examine the differences between orthologs, highlighting the importance of studying CBL-CIPK in canola as a prerequisite for improvement of this crop.ConclusionsOur findings indicate that CBL and CIPK family members may form a dynamic complex to respond to different abiotic or hormone signaling. Our comparative analyses of the CBL-CIPK network between canola, Arabidopsis and rice highlight functional differences and the necessity to study CBL-CIPK gene functions in canola. Our data constitute a valuable resource for CBL and CPK genomics.

Genome-wide analysis of drought induced gene expression changes in flax (Linum usitatissimum)

“…Conventional approaches along with more recent genomic technology based tools are driving the advances in this field. While priority crops like corn, 37 soybean, 38 rice, 39 wheat, 40 and canola 41,42 along with Arabidopsis model system [43][44][45][46] have witnessed significant recent advances in drought research including the application of molecular breeding tools in new varietal development; so far very limited progress has been made in the flax crop. To address this gap, in the present study, we performed genome-wide gene expression analysis to identify genetic programs associated with drought in flax.…”

Evolution of an intron-poor cluster of the CIPK gene family and expression in response to drought stress in soybean

“…The CIPK gene family has been analyzed in Arabidopsis 2122, poplar ( Populus trichocarpa )22, maize ( Zea mays )23, rice ( Oryza sativa )24, and canola ( Brassica napus )25. These studies reported that the CIPK gene family could be divided into an intron-rich clade and an intron-poor clade, in which a subgroup in Arabidopsis , maize, rice and canola CIPK genes were induced by drought23242526.…”

“…These studies reported that the CIPK gene family could be divided into an intron-rich clade and an intron-poor clade, in which a subgroup in Arabidopsis , maize, rice and canola CIPK genes were induced by drought23242526. Although soybean ( Glycine max var.…”

A CBL-Interacting Protein Kinase TaCIPK2 Confers Drought Tolerance in Transgenic Tobacco Plants through Regulating the Stomatal Movement

“…To date, CIPK family genes have been identified in Arabidopsis, rice, canola, maize and wheat, and some CIPKs have been functionally characterized [5–9]. The AtCBL4-CIPK24 (SOS3-SOS2) complex specifically regulates Na + efflux by the Na + /H + antiporter and enhances salt tolerance [10, 11].…”