Use of M281 and M671 markers for detection of male fertility in garlic Allium sativum L.

Relevance. Male sterility in garlic (Allium sativum L.) restricts sexual reproduction and slows breeding.One proposed molecular mechanism is impaired degradation of the callose wall surrounding microspores, associated with reduced activity of the AsaNRF1 gene. The study aimed to characterize polymorphism in the AsaNRF1 promoter region in garlic varieties and breeding accessions and to evaluate M281 and M671 markers for preliminary selection of genotypes with different reproductive potential.Material and methods. A collection of Allium sativum L. comprising 3 varieties and 57 breeding accessions was examined. Total DNA was isolated from young leaves of two-week-old plants using the Sorb-GMO-B kit. Insertions of 281 and 671 bp were detected by PCR with the published M281 marker and an M671 primer pair designed by the authors. Amplification included 35 cycles with annealing at 58 °C; products were separated in a 1% agarose gel. Each accession was analyzed in two analytical replicates.Results. M281 was detected in 26 accessions (43.3%), whereas M671 was detected in 23 (38.3%). Both insertions occurred in 17 genotypes (28.3%); M281 alone in 9 (15.0%), and M671 alone in 6 (10.0%). Neither insertion was detected in 28 accessions (46.7%). The variety Imperator had the M281-/M671+ genotype, confirming incomplete overlap between marker-defined groups.Combined use of M281 and M671 improves preliminary molecular screening; however, reliable identification of fertile garlic forms requires integration of marker data with morphological and cytological assessment of reproductive organs.