Microsatellite primer development for the invasive perennial herb Gypsophila paniculata (Caryophyllaceae)

Abstract

Premise of the Study Gypsophila paniculata (baby's breath; Caryophyllaceae) is a herbaceous perennial that has invaded much of northern and western United States and Canada, outcompeting and crowding out native and endemic species. Microsatellite primers were developed to analyze the genetic structure of invasive populations.Methods and ResultsWe identified 16 polymorphic nuclear microsatellite loci for G. paniculata out of 73 loci that successfully amplified from a primer library created using Illumina sequencing technology. Microsatellite primers were developed to amplify di‐, tri‐, and tetranucleotide repeats and tested in three invasive populations in Michigan.ConclusionsThese markers will be useful in characterizing the genetic structure of invasive populations throughout North America to aid targeted management efforts, and in native Eurasian populations to better understand invasion history. Five of these developed primers also amplified in G. elegans.

Old Meets New: Combining Herbarium Databases with Genetic Methods to Evaluate the Invasion Status of Baby’s Breath (Gypsophila paniculata) in North America

“…Details for 14 microsatellite (nSSR) loci specific to G. paniculata developed byLeimbach-Maus et al (2018b) and used in this study.…”

Genetic Structure of Invasive Baby’s Breath (Gypsophila paniculata L.) Populations in a Michigan Dune System

“…We amplified samples at 14 polymorphic nuclear microsatellite loci (hereafter nSSRs) that were developed specifically for G. paniculata ( Table S2 ) [ 52 ]. We conducted polymerase chain reactions (PCR) using a forward primer with a 5′-fluorescent labeled dye (6-FAM, VIC, NED, or PET) and an unlabeled reverse primer.…”

“…We conducted polymerase chain reactions (PCR) using a forward primer with a 5′-fluorescent labeled dye (6-FAM, VIC, NED, or PET) and an unlabeled reverse primer. PCRs consisted of 1× Taq buffer with KCl (Thermo Scientific, Waltham, MA, USA), 2.0–2.5 mM MgCl 2 depending on the locus ( Table S2 ); 300 µM dNTP; 0.08 mg/mL bovine serum albumin (BSA); 0.4 µM forward primer fluorescently labeled with either FAM, VIC, NED, or PET; 0.4 µM reverse primer; 0.25 units of Taq polymerase (Thermo Scientific, Waltham, MA, USA); and a minimum of 50 ng DNA template, all in a 10.0 µL reaction volume [ 52 ]. The thermal cycle profile consisted of denaturation at 94 °C for 5 min followed by 35 cycles of 94 °C for 1 min, annealing at 62 °C for 1 min, extension at 72 °C for 1 min, and a final elongation step of 72 °C for 10 min.…”

“…We checked nSSR genotypes for null alleles and potential scoring errors due to stuttering and large allele dropout using the software Micro-Checker v2.2.3 [ 57 , 58 ]. Prior to marker selection, the loci used in this study were previously checked for linkage disequilibrium [ 52 ]. We checked for heterozygote deficiencies in the package STRATAG in the R statistical program.…”