title: Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry.
abstract: Flow cytometry analyses were used to verify the ploidy stability of Quercus suber L. somatic embryogenesis process. Leaf explants of two adult cork oak trees (QsG0 and QsG5) of the North of Portugal were inoculated on MS medium with 2,4-D and zeatin. After 3 months, calluses with embryogenic structures were isolated and transferred to fresh MS medium without growth regulators and somatic embryo evolution was followed. Morphologically normal somatic embryos (with two cotyledons) and abnormal somatic embryos (with one or three cotyledons) were used in this assay. Flow cytometry combined with propidium iodide staining was employed to estimate DNA ploidy levels and nuclear DNA content of somatic embryos and leaves from mother plants. No significant differences (P< or =0.05) were detected among embryos, and between the embryos and the mother plants. Also, after conversion of these embryos, no significant morphological differences were observed among the somatic embryo-derived plants. These results and further studies using converted plantlet leaves and embryogenic callus tissue indicate that embryo cultures and converted plantlets were stable with regard to ploidy level. As no major somaclonal variation was detected our primary goal of "true-to-type" propagation of cork oak using somatic embryogenesis was assured at this level. The estimation of the 2C nuclear DNA content for this species is similar to the previously obtained value.
authors.affiliation: Department of Biology, University of Aveiro, 3810-193, Aveiro, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Loureiro
authors.forename: G
authors.initials: G
authors.lastname: Pinto
authors.forename: T
authors.initials: T
authors.lastname: Lopes
authors.forename: J
authors.initials: J
authors.lastname: Dolezel
authors.forename: C
authors.initials: C
authors.lastname: Santos
mesh_terms: D005434:Flow Cytometry; D010641:Phenotype; D011003:Ploidies; D029963:Quercus / Q000196:embryology* / Q000235:genetics*; D012639:Seeds / Q000254:growth & development*
references.citation: Biol Cell. 1993;78(1-2):41-51
references.pmid: 8220226
references.citation: Ann Bot. 2003 Jul;92(1):21-9
references.pmid: 12824068
references.citation: Ann Bot. 2002 Jun;89(6):741-53
references.pmid: 12102530
references.citation: Ann Bot. 2003 Aug;92(2):259-64
references.pmid: 12876189
references.citation: Plant Cell Rep. 2003 Jan;21(5):483-90
references.pmid: 12789452
references.citation: Theor Appl Genet. 2004 Aug;109(3):580-7
references.pmid: 15085264
references.citation: Cytometry. 1995 Dec 1;21(4):387-93
references.pmid: 8608738
references.citation: Ann Bot. 2002 Apr;89(4):385-9
references.pmid: 12096798
references.citation: Cytometry. 1995 Feb 1;19(2):103-6
references.pmid: 7743889
references.citation: Plant Cell Rep. 2004 Apr;22(9):632-9
references.pmid: 14615908
references.citation: Am J Bot. 1999 Oct;86(10):1373-81
references.pmid: 10523279
references.citation: Planta. 1998 Oct;206(2):215-24
references.pmid: 9737000
references.citation: Science. 1983 Jun 3;220(4601):1049-51
references.pmid: 17754551