A mutation screening and genotyping by high-resolution melting (HRM) to detect delta-6-desaturase (D-6-D) variants in Argania spinosa,


Louati M., UÇARLI C., ARIKAN B., Hannach, A. S. , Tetlow I., TURGUT KARA N.

2018 Eastern Regional Meeting, London, Canada, 24 November 2018, pp.32

  • Publication Type: Conference Paper / Full Text
  • City: London
  • Country: Canada
  • Page Numbers: pp.32
  • Istanbul University Affiliated: Yes

Abstract

Theargantreeisamultipurposetreeofgreatsocio-economicinterest.Itsoilisusedinfood,medicine,
and in cosmetics, the tree is considered a good buffer against desertification and erosion and provides a
goodqualitywoodforvarious purposes.It’sraretofindthisspeciesoutsideofMorocco buthundreds
of treesexist in Tunisia indifferent locations. Fewargangenes aresequenced, and suchsequenced
geneisthedelta-6-desaturase(D-6-D)akeyenzymetooilbiosyntheticpathway.Thepurposeofthis
study is to report the use of hight resolution melting (HRM) analysis for scanning and genotyping
Argania spinosa individuals which has never been reported before, to detect delta-6-desaturase (D-6-D)
variants. The present study describes an HRM analysis of natural sequence variation within a small
regionof D-6-Dgenefrom60argantreesbelongingtofourdifferentsitesinTunisia.MethylatedDNA
and unmethylated DNA acquire different sequences after bisulphite treatment resulting in PCR
productswithmarkedlydifferentmeltingprofiles,theefficiencyoftheHRMprocedurewasaffectedby
various factors, including specificity and efficiency of PCR, amplicon length and position, and DNA
templatequality.Inadditiontothesefactors,theuseofPCRproductratherthangenomicDNAinHRM
increasedthequalityofmeltingcurves,thusaffectingtheaccuracyandsensitivityoftheassay.Using
theshapeofthemeltingcurves,wecouldassignthedifferencesbetweencurvesofthedifferenttrees
under investigation infactHRMclassified 10 groupsof variants carrying deletion mutants and single and multiple SNPs.