Screening and Testing Primers for Use in Sex Identification of Date Palm (Phoenix dactylifera L.) by PCR Technique

Abstract

Date palm (Phoenix dactylifera L.) is a dioecious plant with a very slow growth rate and late reproductive phase. The sex of plants became known only at the time of first flowering that takes around 3-7 years after seeding. This research aims to study the DNA markers for sex determination in the seedling stage by PCR technique. Thirty-nine primer pairs from reporting and new designing were used to screen and examine of date palm sex. Results, the primers, DpDOAmale5F and DpDOAmale4R could identify sex of plants. The PCR product was generated with the length approximately 450 bp as specific to male plant. DNA extraction steps were developed to doing faster through comparison the methods between metal beads with Homogenizer and CTAB with liquid nitrogen. Both methods presented similar of DNA purity, while metal beads with Homogenizer methods was reduced time and cost. To validate male-specific primer, genomic DNA 120 individual known sex plants belonging to KL1 and Bahee varieties of date palm were amplified. The primers displayed specific to male plant in 100 percentages of individuals. Furthermore, the male-specific marker used to investigate the sex ratio in seedling stage with 4-12 months of age. The sex detection of 483 seedling samples was showed 42 percentages of male plant. However, this DNA marker could be used for sex determination of date palm in seedling stage and useful to decrease time and cost for crop managements.