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 well-known ubiquitous intracellular secondary messenger in plants. Calcium-dependent protein kinases (CPKs) are Ser/Thr protein kinases found only in plants and some protozoans. CPKs are Ca2+ sensors that have both Ca2+ sensing function and kinase activity within a single protein and play crucial roles in plant development and responses to various environmental stresses.ResultsIn this study, we mined the available expressed sequence tags (ESTs) of B. napus and identified a total of 25 CPK genes, among which cDNA sequences of 23 genes were successfully cloned from a double haploid cultivar of canola. Phylogenetic analysis demonstrated that they could be clustered into four subgroups. The subcellular localization of five selected BnaCPKs was determined using green fluorescence protein (GFP) as the reporter. Furthermore, the expression levels of 21 BnaCPK genes in response to salt, drought, cold, heat, abscisic acid (ABA), low potassium (LK) and oxidative stress were studied by quantitative RT-PCR and were found to respond to multiple stimuli, suggesting that canola CPKs may be convergence points of different signaling pathways. We also identified and cloned five and eight Clade A basic leucine zipper (bZIP) and protein phosphatase type 2C (PP2C) genes from canola and, using yeast two-hybrid and bimolecular fluorescence complementation (BiFC), determined the interaction between individual BnaCPKs and BnabZIPs or BnaPP2Cs (Clade A). We identified novel, interesting interaction partners for some of the BnaCPK proteins.ConclusionWe present the sequences and characterization of CPK gene family members in canola for the first time. This work provides a foundation for further crop improvement and improved understanding of signal transduction in plants.
Highlights
Canola (Brassica napus L.) is one of the most important oil-producing crops in China and worldwide
As the first step to understand the roles of Calcium-dependent protein kinases (CPKs) genes in canola growth and development, as well as in response to abiotic and biotic stress conditions, we initiated a project to identify and clone the cDNA sequences of CPK genes from canola
No systematic analysis of the CPK family genes has been reported in the important oil crop canola
Summary
Canola (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. Plants have developed complex signal transduction pathways to cope with fluctuating environmental changes throughout their life cycles. Environmental stresses, such as high salinity, drought, cold and heat affect crop yield and quality. CPKs contain four characteristic functional domains including an N-terminal domain, a protein kinase domain, an autoinhibitory domain and a calmodulin-like domain [5,6,7]. Previous studies identified a total of 34 CPK genes in the model plant Arabidopsis [5,7]. Phylogenetic analysis of CPKs from both Arabidopsis and rice demonstrated that they can be clustered into four distinct subgroups [15]
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