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Background  Plant calcium (Ca 2+ ) signals are involved in a wide array of intracellular signalling pathways following pathogen invasion. Ca 2+ -binding sensory proteins such as Ca 2+ -dependent protein kinases (CPKs) have been predicted to mediate signalling following Ca 2+  influx after pathogen infection. However, to date this prediction has remained elusive.    Results  We conducted a genome-wide identification of the  Malus x domestica CPK  ( MdCPK ) gene family and identified 30  CPK  genes. Comparative phylogenetic analysis of  Malus  CPKs with CPKs of  Arabidopsis thaliana  ( AtCPKs ),  Oryza sativa  ( OsCPKs ),  Populous trichocarpa  ( PtCPKs ) and  Zea mays  ( ZmCPKs ) revealed four different groups. From the phylogenetic tree, we found that  MdCPKs  are closely related to  AtCPKs  and  PtCPKs  rather than  OsCPKs  and  ZmCPKs , indicating their dicot-specific origin. Furthermore, comparative quantitative real time PCR and intracellular cytosolic calcium ([Ca 2+ ] cyt ) analysis were carried out on fire blight resistant and susceptible  M . x  domestica  apple cultivars following infection with a pathogen ( Erwinia amylovora ) and/or mechanical damage. Calcium analysis showed an increased [Ca 2+ ] cyt  over time in resistant cultivars as compared to susceptible cultivars. Gene expression studies showed that 11 out of the 30  MdCPKs  were differentially expressed following pathogen infection.    Conclusions  We studied the genome-wide analysis of  MdCPK  gene family in  Malus  x  domestica  and analyzed their differential gene expression along with cytosolic calcium variation upon pathogen infection. There was a striking difference in  MdCPKs  gene expressions and [Ca 2+ ] cyt  variations between resistant and susceptible  M .  x domestica  cultivars in response to  E .  amylovora  and mechanical wounding. Our genomic and bioinformatic analysis provided an important insight about the role of  MdCPKs  in modulating defence responses in susceptible and resistant apple cultivars. It also provided further information on early signalling and downstream signalling cascades in response to pathogenic and mechanical stress.

bmc genomics

Differential expression of CPKs and cytosolic Ca2+ variation in resistant and susceptible apple cultivars (Malus x domestica) in response to the pathogen Erwinia amylovora and mechanical wounding