ABSTRACT: We cloned a cell division cycle PsCdc2 from Puccinia striiformis f. sp. tritici (Pst) and analyze its expression profile.
Using PCR and RT-PCR methods, we isolated the cDNA and genomic DNA sequences of PsCdc2. We analyzed the amino acid sequence of PsCdc2 using bioinformatic softwares. In addition, Real time RT-PCR was used to analyze the gene expression pattern of PsCdc2 at different time points after wheat inoculated.
A 2279 bp DNA sequence of PsCdc2 was cloned and comprised of 11 extrons and 10 introns. The cDNA sequence of PsCdc2 included a complete 885 bp open reading frame and encoded a putative protein composed of 294 amino acids, with a molecular weight of 33.14 KDa and a pI of 6.26. PsCdc2 contained two conserved kinase domains and a transmembrane domain. Phylogenetic analysis indicated that PsCdc2 showed high similarity with Cdc2 from Puccinia graminis (73.1%), Cryptococcus neoformans (72.4%) and Ustilago maydis (70.4%), respectively. Real time RT-PCR analysis showed that in compatible interaction between Pst and wheat, PsCdc2 was up-regulated at early stage of infection. The maximum induction occurred at 12 hpi, at which transcripts were 1.62 fold over that in urediniospore. From 24 to 268 hpi, the accumulation of transcripts decreased steadily. The minimum accumulation occurred at 96 h, at which transcripts were only 0.07 fold of that in uredinisopore. During incompatible interaction between Pst and wheat, PsCdc2 was down-regulated and its accumulation was lower than that in urediniospore. The maximum induction occurred at 12 h, at which transcripts were 0.34 fold of that in urediniospore. The minimum accumulation occurred at 96 h, whose transcript was only 0.02 fold of that in urediniospore.
PsCdc2 might be involved in primary hyphal growth and haustorium formation during early infection by regulating cell cycle of Pst. The present study would be helpful for understanding the essence of cell cycle control and provided basis for new chemical control of Pst.
ACTA MICROBIOLOGICA SINICA 02/2010; 50(2):174-81.