Limits...
Constructing Physical and Genomic Maps for Puccinia striiformis f. sp. tritici, the Wheat Stripe Rust Pathogen, by Comparing Its EST Sequences to the Genomic Sequence of P. graminis f. sp. tritici, the Wheat Stem Rust Pathogen.

Ma J, Chen X, Wang M, Kang Z - Comp. Funct. Genomics (2010)

Bottom Line: The percentages of homologous genes varied greatly among different Pst libraries with 54.51%, 51.21%, and 13.61% for the urediniospore, germinated urediniospore, and haustorial libraries, respectively, with an average of 33.92%.The 1,432 Pst genes with significant homology with Pgt sequences were grouped into physical groups corresponding to 237 Pgt supercontigs.The physical relationship was demonstrated by 12 pairs (57%), out of 21 selected Pst gene pairs, through PCR screening of a Pst BAC library.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, China.

ABSTRACT
The wheat stripe rust fungus, Puccinia striiformis f. sp. tritici (Pst), does not have a known alternate host for sexual reproduction, which makes it impossible to study gene linkages through classic genetic and molecular mapping approaches. In this study, we compared 4,219 Pst expression sequence tags (ESTs) to the genomic sequence of P. graminis f. sp. tritici (Pgt), the wheat stem rust fungus, using BLAST searches. The percentages of homologous genes varied greatly among different Pst libraries with 54.51%, 51.21%, and 13.61% for the urediniospore, germinated urediniospore, and haustorial libraries, respectively, with an average of 33.92%. The 1,432 Pst genes with significant homology with Pgt sequences were grouped into physical groups corresponding to 237 Pgt supercontigs. The physical relationship was demonstrated by 12 pairs (57%), out of 21 selected Pst gene pairs, through PCR screening of a Pst BAC library. The results indicate that the Pgt genome sequence is useful in constructing Pst physical maps.

No MeSH data available.


Related in: MedlinePlus

Agarose gels showing positive amplification of the Pst BAC clones using multiplex PCR with primer pairs PSTCY32GT1071F/R (upper bands) and PST78SP3H2F/R (lower bands).  (a) Amplification of 58 BAC plate pools to identify positive pools.  (b) Amplification of row and column pools of a positive plate to identify individual positive clones.  The arrow in (a) indicates the bands amplified with Pst genomic DNA and in (b) indicates molecular size marker.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2821759&req=5

fig3: Agarose gels showing positive amplification of the Pst BAC clones using multiplex PCR with primer pairs PSTCY32GT1071F/R (upper bands) and PST78SP3H2F/R (lower bands). (a) Amplification of 58 BAC plate pools to identify positive pools. (b) Amplification of row and column pools of a positive plate to identify individual positive clones. The arrow in (a) indicates the bands amplified with Pst genomic DNA and in (b) indicates molecular size marker.

Mentions: To validate the physical relationships of Pst genes, a total of 84 forward and reverse primers were designed for 42 genes to form 21 pairs (Table 1). The genes in each pair were selected based on their proximity within 50 Kb in the physic map. Clones that were positively amplified with the first pair of primers resulted from the three-dimensional pooling screening were amplified with the second pair of primers, as illustrated in Figure 3. Of the 21 pairs of genes tested, 12 pairs (57%) were successfully identified in same BAC clones. The results clearly showed that these genes in pairs were truly colocated in the Pst genome.


Constructing Physical and Genomic Maps for Puccinia striiformis f. sp. tritici, the Wheat Stripe Rust Pathogen, by Comparing Its EST Sequences to the Genomic Sequence of P. graminis f. sp. tritici, the Wheat Stem Rust Pathogen.

Ma J, Chen X, Wang M, Kang Z - Comp. Funct. Genomics (2010)

Agarose gels showing positive amplification of the Pst BAC clones using multiplex PCR with primer pairs PSTCY32GT1071F/R (upper bands) and PST78SP3H2F/R (lower bands).  (a) Amplification of 58 BAC plate pools to identify positive pools.  (b) Amplification of row and column pools of a positive plate to identify individual positive clones.  The arrow in (a) indicates the bands amplified with Pst genomic DNA and in (b) indicates molecular size marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2821759&req=5

fig3: Agarose gels showing positive amplification of the Pst BAC clones using multiplex PCR with primer pairs PSTCY32GT1071F/R (upper bands) and PST78SP3H2F/R (lower bands). (a) Amplification of 58 BAC plate pools to identify positive pools. (b) Amplification of row and column pools of a positive plate to identify individual positive clones. The arrow in (a) indicates the bands amplified with Pst genomic DNA and in (b) indicates molecular size marker.
Mentions: To validate the physical relationships of Pst genes, a total of 84 forward and reverse primers were designed for 42 genes to form 21 pairs (Table 1). The genes in each pair were selected based on their proximity within 50 Kb in the physic map. Clones that were positively amplified with the first pair of primers resulted from the three-dimensional pooling screening were amplified with the second pair of primers, as illustrated in Figure 3. Of the 21 pairs of genes tested, 12 pairs (57%) were successfully identified in same BAC clones. The results clearly showed that these genes in pairs were truly colocated in the Pst genome.

Bottom Line: The percentages of homologous genes varied greatly among different Pst libraries with 54.51%, 51.21%, and 13.61% for the urediniospore, germinated urediniospore, and haustorial libraries, respectively, with an average of 33.92%.The 1,432 Pst genes with significant homology with Pgt sequences were grouped into physical groups corresponding to 237 Pgt supercontigs.The physical relationship was demonstrated by 12 pairs (57%), out of 21 selected Pst gene pairs, through PCR screening of a Pst BAC library.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, China.

ABSTRACT
The wheat stripe rust fungus, Puccinia striiformis f. sp. tritici (Pst), does not have a known alternate host for sexual reproduction, which makes it impossible to study gene linkages through classic genetic and molecular mapping approaches. In this study, we compared 4,219 Pst expression sequence tags (ESTs) to the genomic sequence of P. graminis f. sp. tritici (Pgt), the wheat stem rust fungus, using BLAST searches. The percentages of homologous genes varied greatly among different Pst libraries with 54.51%, 51.21%, and 13.61% for the urediniospore, germinated urediniospore, and haustorial libraries, respectively, with an average of 33.92%. The 1,432 Pst genes with significant homology with Pgt sequences were grouped into physical groups corresponding to 237 Pgt supercontigs. The physical relationship was demonstrated by 12 pairs (57%), out of 21 selected Pst gene pairs, through PCR screening of a Pst BAC library. The results indicate that the Pgt genome sequence is useful in constructing Pst physical maps.

No MeSH data available.


Related in: MedlinePlus