Development of 11 polymorphic microsatellite markers in a macrophyte of conservation concern, Vallisneria americana Michaux (Hydrocharitaceae)

Abstract

Vallisneria americana Michaux (wild celery) is currently a target of submersed aquatic vegetation restoration efforts in the Chesapeake Bay watershed. To aid these efforts, we have developed 11 polymorphic microsatellite markers to assess the distribution and degree of genetic diversity in both restored and naturally occurring populations in the Chesapeake Bay. In 59 individuals from two populations, we detected two to 10 total alleles per locus. Observed heterozygosity ranged from 0.125 to 0.929, and two loci exhibited significant deviations from Hardy-Weinberg equilibrium in at least one of the populations assayed.

The structure of population genetic diversity in Vallisneria americana in the Chesapeake Bay: implications for restoration

“…We genotyped 11 microsatellite loci representing tri-nucleotide repeats from each sample using robust primers with specific amplification that were developed for the species (Burnett et al 2009). Polymerase chain reactions (PCR) were performed Burnett et al (2009) with the exception of the locus Vaam_AAG004, for which we added dimethyl sulfoxide and Q-Solution (QIAGEN) to each reaction for optimal specificity. PCR products were separated and measured on an ABI 3730xl DNA Analyzer with GeneScan TM -500 ROX TM or 500 LIZ TM Size Standard (Applied Biosystems) after tagging the PCR product with fluorescent labeled forward primers (Applied Biosystems).…”

“…Genomic DNA was isolated and purified using methods described in (Burnett et al 2009). We genotyped 11 microsatellite loci representing tri-nucleotide repeats from each sample using robust primers with specific amplification that were developed for the species (Burnett et al 2009).…”

Does genetic diversity of restored sites differ from natural sites? A comparison of Vallisneria americana (Hydrocharitaceae) populations within the Chesapeake Bay

“…We genotyped 10 microsatellite loci from each shoot using robust primers with specific amplification that we developed for the species (Burnett et al 2009). Polymerase chain reactions (PCR) were performed as in Burnett et al (2009), with the exception of the locus Vaam_AAG004, for which we added dimethyl sulfoxide and Q-Solution (QIAGEN) to reactions to optimize specificity.…”

“…We genotyped 10 microsatellite loci from each shoot using robust primers with specific amplification that we developed for the species (Burnett et al 2009). Polymerase chain reactions (PCR) were performed as in Burnett et al (2009), with the exception of the locus Vaam_AAG004, for which we added dimethyl sulfoxide and Q-Solution (QIAGEN) to reactions to optimize specificity. PCR products were separated, measured, and peaks analyzed using identical methods and quality control procedures, which included repeated analyses to ensure high reproducibility of PCR reactions, as detailed in Lloyd et al (2011).…”

“…Genomic DNA was isolated and purified using methods outlined in Burnett et al (2009). We genotyped 10 microsatellite loci from each shoot using robust primers with specific amplification that we developed for the species (Burnett et al 2009).…”