Pushpa Kharb
CCS Haryana Agricultural University, India
Title: Development of SCAR markers in date palm (Phoenix dactylifera L.) and jojoba (Simmondsia chinensis Schneider) for early sex identification
Biography
Biography: Pushpa Kharb
Abstract
The genetic control and mechanisms leading to sex differentiation in date palm (Phoenix dactylifera L.) and jojoba (Simmondsia chinensis Link Schneider) are not known. Genomic DNA from a total of 45 genotypes (25 female and 20 male) of date palm was subjected to PCR amplification using 100 RAPD and 104 ISSR primers. Only one RAPD primer OPA-02 amplified a fragment of 1.0 kb in all the male genotypes. This male specific fragment was sequenced and the sequence was deposited in GenBank (accession no. JN123357). SCAR dpF (forward) and SCAR dpR (reverse) primers were designed based on this male-specific sequence. This primer pair amplified a 406 bp fragment in both female and male genotypes and a unique allele of 354 bp in only male genotypes (patent filed; application no. 1513/DEL/2010 dated 29/6/10). The SCAR marker was further validated using 25 female and 10 male date palm plants belonging to different varieties. Similarly in jojoba, an ISSR marker (already identified by our group) amplifying a unique allele of 1100 bp in only male genotypes was first confirmed using 10 male and 10 female genotypes. This male-specific fragment was cloned and sequenced. The sequence was deposited in the GenBank (accession no. HQ166029). Primers SCAR scF (forward) and SCAR scR (reverse) was designed based on this sequence amplified a unique allele of 1000 bp in male genotypes only (patent Application no. 1563/DEL/2010 dated 02/7/10). The SCAR marker was further validated using five male and five female jojoba genotypes which was not used earlier.