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Published on: May 10, 2016
[Cloning and transcriptional profiling of PsNCS1 from Puccinia striiformis f. sp. tritici]
1College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling 712100, China. guojunwgq@nwsuaf.edu.cn
Objective:
We cloned PsNCS1 encoding neuronal calcium sensor from Pst and analyzed its transcriptional profile.
Methods:
A full-length cDNA of PsNCS1 was cloned by using RT-PCR in combination with cDNA library screening, the sequence was analyzed with different bioinformatic tools and the gene expression pattern was characterized via real-time RT-PCR.
Results:
PsNCS1 (Genbank accession no. GU134621) encoded 190 amino acids, with a molecular weight of 22.17 KDa and a pI of 4.96. PsNCS1 contained four conserved EF-hand domains and was N-terminally myristoylated. Phylogenetic analysis indicated that PsNCS1 was highly similar to the NCS from Basidiomycetes and the highest similarity was with that from Puccinia graminis (96%). Real-time RT-PCR analysis indicated the amount of PsNCS1 transcripts of urediospores and of germinated urediospores were doubled or more comparing with those of fungal bodies at other different developmental stages.
Conclusion:
PsNCS1 might be involved in the process of urediospore formation and germ tube elongation. The present results may provide basic data for further analysis of the role of PsCNS1 in pathogenesis process and calcium signaling.
