[Cloning and expression of a class II chitin synthase gene PstChs II from the rust fungus Puccinia striiformis].
OBJECTIVE
To clone the chitin synthase gene PstChs II from Puccinia striiformis and to analyze its expression pattern.


METHODS
We isolated the cDNA and genomic DNA of PstChs II via RT-PCR and PCR,analyzed the sequences with different bioinformatic tools, characterized the gene expression pattern via real-time PCR.


RESULTS
The coding region of PstChs II (Genbank accession no: GQ329851), interrupted by 15 introns, corresponded to a 2727 bp open reading frame encoding 908 amino acids. PstChs II showed a highest 94% similarity to PgtChs II from Puccinia graminis. PstChs II had 7 transmembrance regions and several conserved chitin synthase domains and motifs such as "QXRRW", "GXGPL" and "DXD". PstChs II belonged to the class II sub-family and had a closer phylogenetic relationship to its homologs from basidiomycetes than those from ascomycetes. PstChs II expression level increased by 10-fold at the urediospore germination stage.


CONCLUSIONS
PstChs II might be involved in the cell wall synthesis during the germ tube elongation process. The cloning and expression analysis of PstChs II served as a good foundation for further analyzing the role of this gene in the pathogenesis process.
Liang
Xiaofei
X
College of Plant Protection, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling 712100, China. xiaofei123@nwsuaf.edu.cn
Liu
Bo
B
Zhu
Lin
L
Zhang
Xiaoyu
X
Wang
Xiaojie
X
Huang
Lili
L
Kang
Zhensheng
Z
D000595:Amino Acid Sequence; D001487:Basidiomycota / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D002687:Chitin Synthase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment
D005656:Fungal Proteins; D002687:Chitin Synthase