Novel polymorphic nuclear microsatellites in Cupressus sempervirens L.

Abstract

We report the characterization and optimization of nine polymorphic nuclear microsatellites from Cupressus sempervirens L. using an enriched library method. A total of 24 individuals from four different populations were used to estimate genetic diversity parameters. High level of allelic diversity was found with a number of alleles ranging from two to 13. These markers will prove very useful in screening diversity at different geographical scales and in monitoring gene flow in cypress orchards.

Molecular evidence for the natural production of homozygous Cupressus sempervirens L. lines by Cupressus dupreziana seed trees

“…Genetic diversity in French cypress populations is similar to that in Italian populations. The null allele frequencies estimated in this study for French populations are of the same magnitude as those found by Sebastiani et al (2005) (from 0.06 to 0.19 for loci Cyp52, Cyp139, Cyp258 and Cyp293).…”

“…Inheritance of the SSR markers The analysis of allele segregations among the 77 C. sempervirens progeny produced by controlled crosses of C. sempervirens parents confirmed the putative Mendelian inheritance of the SSR markers developed by Sebastiani et al (2005).…”

“…Seedlings were genotyped using seven nuclear microsatellites (SSRs) developed for C. sempervirens: Cyp52, Cyp84, Cyp101, Cyp174, Cyp257, Cyp258 and Cyp293 (Sebastiani et al, 2005). These markers were highly polymorphic in C. sempervirens populations (Valgimigli, 2005), providing high resolution for paternity analysis and heterozygote detection, but their putative nuclear inheritance had never been verified.…”

“…The forward primers were labeled with FAM, HEX or TAMRA. The PCR thermal profile (except for Cyp258, see Sebastiani et al (2005)) was as follows: denaturation at 94 1C for 5 min followed by 30 cycles of 94 1C for 30 s, 50 1C for 30 s, 72 1C for 30 s and a final extension of 72 1C for 8 min. PCRs were carried out using a Perkin-Elmer GeneAmp 9700 thermal cycler (Applied Biosystems, Foster City, CA, USA).…”

“…It is represented by only one natural population in the Tassili N'Ajjer desert, Algeria, and the number of surviving individuals is limited to 233 (Abdoun and Beddiaf, 2002). This species can easily be distinguished from C. sempervirens L. (common cypress or Mediterranean cypress) on the basis of a multi-locus profile obtained using AFLP (Pichot et al, 2008) and nuclear microsatellites (transferred from Sebastiani et al, 2005). The precarious state of the species results from a combination of extreme climatic conditions, long-term human pressure and extremely scarce regeneration.…”