Nuclear Microsatellite Primers for the Endangered Relict Fir, Abies pinsapo (Pinaceae) and Cross-Amplification in Related Mediterranean Species

Sánchez‐Robles, Jose M.; Balao, Francisco; García-Castaño, Juan Luis; Terrab, Anass; Navarro‐Sampedro, Laura; Lozano, Salvador Talavera

doi:10.3390/ijms131114243

Cited by 18 publications

(15 citation statements)

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“…), and five were recently developed for A. pinsapo (Sánchez‐Robles et al . ). Polymerase chain reaction (PCR) was conducted on a Veriti Thermal Cycler (Applied Biosystems, Foster City, CA, USA).…”

Section: Methodsmentioning

confidence: 97%

“…Amplification conditions and PCR cycle profiles of A. pinsapo ‐specific nSSRs can be found in Sánchez‐Robles et al . (). For the other nSSRs, PCRs were performed in a final volume of 10 μL using 9 μL of a 1.1× Reddy Mix PCR Master Mix 2.5 m m MgCl 2 (Thermo Scientific Inc., Bremen, Germany), 0.2 μL of each primer (10 m m ) and 0.6 μL of template DNA (~50 ng); for PCR cycle profiles, see Table .…”

Section: Methodsmentioning

confidence: 97%

“… Significant inbreeding coefficient (adjusted P ‐value for 5% nominal level = 0.00455); 1 Abies pinsapo (Sánchez‐Robles et al . ); 2 Abies alba (Cremer et al . ); 3 Abies nordmanniana (Hansen et al .…”

Section: Methodsmentioning

confidence: 99%

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Effects of tree architecture on pollen dispersal and mating patterns inAbies pinsapoBoiss. (Pinaceae)

et al. 2014

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Plant architecture is crucial to pollination and mating in wind-pollinated species. We investigated the effect of crown architecture on pollen dispersal, mating system and offspring quality, combining phenotypic and genotypic analyses in a low-density population of the endangered species Abies pinsapo. A total of 598 embryos from three relative crown height levels (bottom, middle and top) in five mother plants were genotyped using eleven nuclear microsatellite markers (nSSRs). Paternity analysis and mating system models were used to infer mating and pollen dispersal parameters. In addition, seeds were weighed (N = 16 110) and germinated (N = 736), and seedling vigour was measured to assess inbreeding depression. Overall, A. pinsapo shows a fat-tailed dispersal kernel, with an average pollen dispersal distance of 113-227 m, an immigration rate of 0.84-26.92%, and a number of effective pollen donors (Nep ) ranging between 3.5 and 11.9. We found an effect of tree height and relative crown height levels on mating parameters. A higher proportion of seeds with embryo (about 50%) and a higher rate of self-fertilization (about 60%) were found at the bottom level in comparison with the top level. Seed weight and seedling vigour are positively related. Nevertheless, no differences were found in seed weight or in seedling-related variables such as weight and length of aerial and subterranean parts among the different relative crown height levels, suggesting that seeds from the more strongly inbred bottom level are not affected by inbreeding depression. Our results point to vertical isotropy for outcross-pollen and they suggest that self-pollen may ensure fertilization when outcross-pollen is not available in low-density population.

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Section: Methodsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 97%

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Effects of tree architecture on pollen dispersal and mating patterns inAbies pinsapoBoiss. (Pinaceae)

et al. 2014

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“…canariensis ( Appendix 1 ). Size-selected microsatellite enrichment was performed following a Dynabeads-based protocol ( Glenn and Schable, 2005 ) that has been successfully applied in other plants ( Sánchez-Robles et al, 2012 ; Jiménez-López et al, 2015 ). Genomic DNA was digested with Rsa I and Bst UI (New England Biolabs, Ipswich, Massachusetts, USA) and enriched for (AC) 12 , (AG) 12 , (AAC) 6 , (AAG) 8 , (AAT) 12 , (ACT) 12 , (ATC) 8 , (AAAG) 6 , (ACCT) 6 , (ACTC) 6 , (AATC) 6 , (ACAG) 6 , (ACTG) 6 , (AAAC) 6 , (AATG) 6 , (AGAT) 8 , (AACT) 8 , (AAGT) 8 , (AAAT) 8 , and (ACAT) 8 .…”

Section: Methods and Resultsmentioning

confidence: 99%

Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing

et al. 2015

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Premise of the study:Nuclear microsatellite markers were developed in Rumex bucephalophorus subsp. canariensis (Polygonaceae) to investigate its genetic diversity and structure.Methods and Results:Sixteen polymorphic microsatellite markers were obtained using 454 next-generation sequencing with di-, tri-, and tetranucleotide repeats. The average number of alleles was 5.688 and 3.813 for R. bucephalophorus subsp. canariensis var. canariensis and var. fruticescens, respectively. Slightly higher levels of mean genetic diversity were found in var. canariensis (expected heterozygosity = 0.600) than in var. fruticescens (expected heterozygosity = 0.514). Cross-amplifications in related taxa within R. bucephalophorus showed good amplification and polymorphic patterns.Conclusions:These 16 novel nuclear microsatellite markers are the first in the genus Rumex and may serve as valuable tools to carry out studies on genetic diversity and structure as well as progeny studies.

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“…In the present study, the high level of polymorphism and heterozygosity of 22 microsatellite loci would be a valuable to investigate further population studies with more extensive samplings from A. koreana and / or A. nephrolepis. The values of H E in A. koreana and A. nephrolepis were moderate in the Abies genus (Saito et al, 2005;Cremer et al, 2006;Josserand et al, 2006;Lian et al, 2007;Rasmussen et al, 2008;Sánchez-Robles et al, 2012;Zhan et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of novel microsatellites for Abies koreana and A. nephrolepis (Pinaceae)

Jin¹,

J²,

By³

et al. 2016

Genet. Mol. Res.

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ABSTRACT.Abies koreana is an endemic and rare species from Korea and is classified as endangered by the International Union for Conservation of Nature. Although the genetic diversity assessment for current population of A. koreana needs to be performed urgently, no microsatellite markers have been developed for this species. In the present study, we developed 22 novel polymorphic microsatellite loci and the characteristics of these loci were determined in A. koreana as well as in Abies nephrolepis, the most closely related species, and these loci were compared with previously reported microsatellite markers developed for the Abies genus. Genomic sequence (161 Mbp; 325,776 reads) was obtained from one individual of A. koreana using Roche 454 GS-FLX Titanium sequencing and 19,258 repeat motifs were identified from it. A total of 288 primer pairs with high copy numbers of direpeat motifs were evaluated for amplification in A. koreana and A. nephrolepis. A total of 71 primer pairs successfully amplified fragments, of which 22 showed polymorphisms in A. koreana and A. nephrolepis. The average expected diversity was 0.767 and 0.717 in A. koreana and A. nephrolepis, respectively; these heterozygosity levels were moderate compared to the previously reported microsatellite loci from Abies species. This is the first set of microsatellite markers developed for A. koreana as well as A. nephrolepis and further population genetic studies of both species and genetic delimitation can be carried out for the species conservation and management.

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Nuclear Microsatellite Primers for the Endangered Relict Fir, Abies pinsapo (Pinaceae) and Cross-Amplification in Related Mediterranean Species

Cited by 18 publications

References 18 publications

Effects of tree architecture on pollen dispersal and mating patterns inAbies pinsapoBoiss. (Pinaceae)

Effects of tree architecture on pollen dispersal and mating patterns inAbies pinsapoBoiss. (Pinaceae)

Characterization of nuclear microsatellite markers for Rumex bucephalophorus (Polygonaceae) using 454 sequencing

Isolation and characterization of novel microsatellites for Abies koreana and A. nephrolepis (Pinaceae)

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