Analysis of genetic variability in various tissue culture-derived lemon plant populations using RAPD and flow cytometry

Abstract

Five populations of lemon plants [Citrus limon (L.) Burm] obtained from undeveloped ovules through diVerent tissue culture procedures were examined for the presence of somaclonal and irradiationinduced genetic variation. Tested groups were: (1) nucellar seedlings; (2) organogenic, regenerated via adventitious buds from nucellar seedling internodes;(3) embryogenic population, regenerated from nonirradiated nucellar callus via somatic embryogenesis; (4) embryogenic population, regenerated from irradiated nucellar callus via somatic embryogenesis; and (5) protoplast-derived, regenerated via somatic embryogenesis. Genomic DNA samples from 360 plants (72 from each group) were screened for polymorphism among RAPD Wngerprints ampliWed by 10 decamer primers. Among all tested plants, genetic variation was detected only within the group of plants recovered from irradiated embryogenic calli. Out of 72 plants from that group, three had RAPD Wngerprints diVerent from the rest of the population, and fourth plant was found to be cytochimeric, consisting of diploid and tetraploid cells as revealed by Xow cytometry. In all other populations of regenerated plants, we did not come across any plants with changed ploidy level.

Current Importance and Potential Use of Low Doses of Gamma Radiation in Forest Species

“…Irradiation treatments performed at in vitro culture has been also employed to increase genetic variability and mutants as a potential source of new commercial cultivars (Rasheed et al, 2003;Orbovic´ et al, 2008). So, tissue culture techniques offer a wide choice of explants (initial plant material) for gamma radiation treatment (cells, tissues, somatic embryos and organs).…”

Genetic and Epigenetic Variations in Barley Calli Cultures

“…explants source, ploidy level, donor genotype, medium composition, and environmental conditions are some of the factors causing somaclonal variation (20). Molecular markers such as RAPD (4,13) are utilized to detect somaclonal variation at nucleotide sequence level. it involves amplification of genomic DNA with short and random primers at low annealing temperatures (22).…”

Haploids in Conifer Species: Characterization and Chromosomal Integrity of a Maritime Pine Cell Line

“…Thus, random amplified polymorphic DNA (RAPD) markers have been used in peach [31] cedar [32], lemon [33] and chestnut [34], whereas amplified fragment length polymorphism (AFLP) has been used in kiwifruit [35], grape [36] and apple [37]. In the last few years, microsatellites have been widely used to analyze many plant species, including arabian coffee [38], cork oak [39] and trembling aspen [40].…”