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Use of genome sequence data in the design and testing of SSR markers for Phytophthora species

Identifieur interne : 000241 ( Pmc/Checkpoint ); précédent : 000240; suivant : 000242

Use of genome sequence data in the design and testing of SSR markers for Phytophthora species

Auteurs : Leonardo Schena [Italie] ; Linda Cardle [Royaume-Uni] ; David El Cooke [Royaume-Uni]

Source :

RBID : PMC:2647557

Abstract

Background

Microsatellites or single sequence repeats (SSRs) are a powerful choice of marker in the study of Phytophthora population biology, epidemiology, ecology, genetics and evolution. A strategy was tested in which the publicly available unigene datasets extracted from genome sequences of P. infestans, P. sojae and P. ramorum were mined for candidate SSR markers that could be applied to a wide range of Phytophthora species.

Results

A first approach, aimed at the identification of polymorphic SSR loci common to many Phytophthora species, yielded 171 reliable sequences containing 211 SSRs. Microsatellites were identified from 16 target species representing the breadth of diversity across the genus. Repeat number ranged from 3 to 16 with most having seven repeats or less and four being the most commonly found. Trinucleotide repeats such as (AAG)n, (AGG)n and (AGC)n were the most common followed by pentanucleotide, tetranucleotide and dinucleotide repeats. A second approach was aimed at the identification of useful loci common to a restricted number of species more closely related to P. sojae (P. alni, P. cambivora, P. europaea and P. fragariae). This analysis yielded 10 trinucleotide and 2 tetranucleotide SSRs which were repeated 4, 5 or 6 times.

Conclusion

Key studies on inter- and intra-specific variation of selected microsatellites remain. Despite the screening of conserved gene coding regions, the sequence diversity between species was high and the identification of useful SSR loci applicable to anything other than the most closely related pairs of Phytophthora species was challenging. That said, many novel SSR loci for species other than the three 'source species' (P. infestans, P. sojae and P. ramorum) are reported, offering great potential for the investigation of Phytophthora populations. In addition to the presence of microsatellites, many of the amplified regions may represent useful molecular marker regions for other studies as they are highly variable and easily amplifiable from different Phytophthora species.


Url:
DOI: 10.1186/1471-2164-9-620
PubMed: 19099584
PubMed Central: 2647557


Affiliations:


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<title>Background</title>
<p>Microsatellites or single sequence repeats (SSRs) are a powerful choice of marker in the study of
<italic>Phytophthora </italic>
population biology, epidemiology, ecology, genetics and evolution. A strategy was tested in which the publicly available unigene datasets extracted from genome sequences of
<italic>P. infestans</italic>
,
<italic>P. sojae </italic>
and
<italic>P. ramorum </italic>
were mined for candidate SSR markers that could be applied to a wide range of
<italic>Phytophthora </italic>
species.</p>
</sec>
<sec>
<title>Results</title>
<p>A first approach, aimed at the identification of polymorphic SSR loci common to many
<italic>Phytophthora </italic>
species, yielded 171 reliable sequences containing 211 SSRs. Microsatellites were identified from 16 target species representing the breadth of diversity across the genus. Repeat number ranged from 3 to 16 with most having seven repeats or less and four being the most commonly found. Trinucleotide repeats such as (AAG)n, (AGG)n and (AGC)n were the most common followed by pentanucleotide, tetranucleotide and dinucleotide repeats. A second approach was aimed at the identification of useful loci common to a restricted number of species more closely related to
<italic>P. sojae </italic>
(
<italic>P. alni, P. cambivora, P. europaea </italic>
and
<italic>P. fragariae</italic>
). This analysis yielded 10 trinucleotide and 2 tetranucleotide SSRs which were repeated 4, 5 or 6 times.</p>
</sec>
<sec>
<title>Conclusion</title>
<p>Key studies on inter- and intra-specific variation of selected microsatellites remain. Despite the screening of conserved gene coding regions, the sequence diversity between species was high and the identification of useful SSR loci applicable to anything other than the most closely related pairs of
<italic>Phytophthora </italic>
species was challenging. That said, many novel SSR loci for species other than the three 'source species' (
<italic>P. infestans</italic>
,
<italic>P. sojae </italic>
and
<italic>P. ramorum</italic>
) are reported, offering great potential for the investigation of
<italic>Phytophthora </italic>
populations. In addition to the presence of microsatellites, many of the amplified regions may represent useful molecular marker regions for other studies as they are highly variable and easily amplifiable from different
<italic>Phytophthora </italic>
species.</p>
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<article-title>Use of genome sequence data in the design and testing of SSR markers for
<italic>Phytophthora </italic>
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<given-names>Leonardo</given-names>
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<email>lschena@unirc.it</email>
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<surname>Cardle</surname>
<given-names>Linda</given-names>
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<xref ref-type="aff" rid="I2">2</xref>
<email>Linda.Milne@scri.ac.uk</email>
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<name>
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<email>David.Cooke@scri.ac.uk</email>
</contrib>
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Department of "Gestione dei Sistemi Agrari e Forestali", Mediterranean University of Reggio Calabria, Località Feo di Vito, 89124 Reggio Calabria, Italy</aff>
<aff id="I2">
<label>2</label>
Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, Scotland, UK</aff>
<pub-date pub-type="collection">
<year>2008</year>
</pub-date>
<pub-date pub-type="epub">
<day>19</day>
<month>12</month>
<year>2008</year>
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<volume>9</volume>
<fpage>620</fpage>
<lpage>620</lpage>
<ext-link ext-link-type="uri" xlink:href="http://www.biomedcentral.com/1471-2164/9/620"></ext-link>
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<date date-type="received">
<day>6</day>
<month>6</month>
<year>2008</year>
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<date date-type="accepted">
<day>19</day>
<month>12</month>
<year>2008</year>
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<permissions>
<copyright-statement>Copyright © 2008 Schena et al; licensee BioMed Central Ltd.</copyright-statement>
<copyright-year>2008</copyright-year>
<copyright-holder>Schena et al; licensee BioMed Central Ltd.</copyright-holder>
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<pmc-comment> Schena Leonardo lschena@unirc.it Use of genome sequence data in the design and testing of SSR markers for Phytophthora species 2008BMC Genomics 9(1): 620-. (2008)1471-2164(2008)9:1<620>urn:ISSN:1471-2164</pmc-comment>
</license>
</permissions>
<abstract>
<sec>
<title>Background</title>
<p>Microsatellites or single sequence repeats (SSRs) are a powerful choice of marker in the study of
<italic>Phytophthora </italic>
population biology, epidemiology, ecology, genetics and evolution. A strategy was tested in which the publicly available unigene datasets extracted from genome sequences of
<italic>P. infestans</italic>
,
<italic>P. sojae </italic>
and
<italic>P. ramorum </italic>
were mined for candidate SSR markers that could be applied to a wide range of
<italic>Phytophthora </italic>
species.</p>
</sec>
<sec>
<title>Results</title>
<p>A first approach, aimed at the identification of polymorphic SSR loci common to many
<italic>Phytophthora </italic>
species, yielded 171 reliable sequences containing 211 SSRs. Microsatellites were identified from 16 target species representing the breadth of diversity across the genus. Repeat number ranged from 3 to 16 with most having seven repeats or less and four being the most commonly found. Trinucleotide repeats such as (AAG)n, (AGG)n and (AGC)n were the most common followed by pentanucleotide, tetranucleotide and dinucleotide repeats. A second approach was aimed at the identification of useful loci common to a restricted number of species more closely related to
<italic>P. sojae </italic>
(
<italic>P. alni, P. cambivora, P. europaea </italic>
and
<italic>P. fragariae</italic>
). This analysis yielded 10 trinucleotide and 2 tetranucleotide SSRs which were repeated 4, 5 or 6 times.</p>
</sec>
<sec>
<title>Conclusion</title>
<p>Key studies on inter- and intra-specific variation of selected microsatellites remain. Despite the screening of conserved gene coding regions, the sequence diversity between species was high and the identification of useful SSR loci applicable to anything other than the most closely related pairs of
<italic>Phytophthora </italic>
species was challenging. That said, many novel SSR loci for species other than the three 'source species' (
<italic>P. infestans</italic>
,
<italic>P. sojae </italic>
and
<italic>P. ramorum</italic>
) are reported, offering great potential for the investigation of
<italic>Phytophthora </italic>
populations. In addition to the presence of microsatellites, many of the amplified regions may represent useful molecular marker regions for other studies as they are highly variable and easily amplifiable from different
<italic>Phytophthora </italic>
species.</p>
</sec>
</abstract>
</article-meta>
</front>
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