Development of male-specific SCAR marker in date palm (Phoenix dactylifera L.)

Genetics of control mechanisms that underlies sex differentiation in date palm is not known. Sex of the plants becomes known only at the time of first flowering, which takes around 5 years. In comparison, molecular diagnosis (if available/feasible) promises quick and reliable identification of sex t...

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Bibliographic Details
Published in:Tree genetics & genomes 2013-10, Vol.9 (5), p.1143-1150
Main Authors: Dhawan, Charu, Kharb, Pushpa, Sharma, Richa, Uppal, Sanjogta, Aggarwal, Ramesh K
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Biotechnology
Deoxyribonucleic acid
DNA
flowering
Forestry
Genetic testing
Genetics
Genomes
Genomics
genotype
Genotypes
Horticulture
Identification
Life Sciences
Molecular biology
Original Paper
Phoenix dactylifera
Plant Breeding/Biotechnology
Plant Genetics and Genomics
Plant reproduction
plantlets
random amplified polymorphic DNA technique
seedbeds
sexual development
Tree Biology
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Summary:Genetics of control mechanisms that underlies sex differentiation in date palm is not known. Sex of the plants becomes known only at the time of first flowering, which takes around 5 years. In comparison, molecular diagnosis (if available/feasible) promises quick and reliable identification of sex types very early when plantlets are growing in seedbeds. To develop such an assay, genomic DNA from 45 individual plants (25 female and 20 male) belonging to different varieties of date palm was subjected to PCR amplification using 100 random amplified polymorphic DNA (RAPD) and 104 intersimple sequence repeat (ISSR) primers. Initially, two bulk genomic DNA samples (each made by pooling DNA from ten male and female plants, separately) were used. A primer showing sex-specific band in bulked samples was further used for amplification of the genomic DNA of the individual samples of that bulk. Only one RAPD primer, OPA-02, amplified a fragment of ~1.0 kb in all the individual samples of male genotypes, whereas this fragment was absent in all the female genotypes. This male-specific fragment was cloned and sequenced (GenBank accession no. JN123357), and a sequence-characterized amplified region (SCAR) primer pair was designed that amplified a 406-bp fragment in both female and male genotypes and a unique fragment of 354 bp in only male genotypes. The SCAR marker was further validated using 25 female and ten male date palm plants belonging to different varieties collected from different locations.
ISSN:1614-2942
1614-2950
DOI:10.1007/s11295-013-0617-9