PJB-2024-88
Development of GPPS gene derived Microsatellites markers related to quinone and phenolics biosynthesis in diverse Nigella sativa L. germplasm
Arusa Aftab
Abstract
Nigella sativa L. (Ranunculaceae) is a widely cultivated plant for its therapeutic purposes. Its seeds are a rich source of thymoquinone exhibiting outstanding biological activities. However, it’s cultivation is difficult being climate sensitive crop. Likewise very less information is available on genomic studies of N.sativa. Simple Sequence Repeats (SSRs) are co‑dominant nuclear markers in plant genetics studies. The present study was performed to explore the microsatellite loci (SSR) related to quinone and phenol biosynthesis. Seventy-five accessions of N. sativa were collected from different regions including Pakistan, Jordan, Ukraine, and U.S. Idaho. Genomic DNA was extracted from leaf samples using the common CTAB method. Primers were designed from the GPPS gene- mainly involved in the biosynthesis of quinone, phenols, and monoterpenes via Geranyl Diphosphate Synthase (GPPS) pathway. 100 primer pairs were successfully amplified from the 135 designed primers. The amplified products were electrophoresed using Metaphor Agarose (Lonza, USA). A significant number SSRs revealed polymorphism. A total of 705 repeat motifs (di nucleotide, trinucleotide, and tetra nucleotide) were detected based on DNA sequencing. Data were analyzed through Cluster analysis, Principal component analysis using SPSS version 20, and XLSTAT. Gene ALX, sequences were further analyzed using MEGA X software and Gramene SSR locator tool. The identified molecular markers may provide a valuable tool for molecular breeding, mapping, and genetic improvement of thymoquinone biosynthesis.
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