Journal Article

Plant somatic hybrid cytoplasmic DNA characterization by single-strand conformation polymorphism

Óscar Olivares-Fuster, María Hernández-Garrido, José Guerri and Luis Navarro

in Tree Physiology

Volume 27, issue 6, pages 785-792
Published in print June 2007 | ISSN: 0829-318X
Published online June 2007 | e-ISSN: 1758-4469 | DOI:
Plant somatic hybrid cytoplasmic DNA characterization by single-strand conformation polymorphism

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Unlike maternal inheritance in sexual hybridization, plant somatic hybridization allows transfer, mixing and recombination of cytoplasmic genomes. In addition to the use of somatic hybridization in plant breeding programs, application of this unique tool should lead to a better understanding of the roles played by the chloroplastic and mitochondrial genomes in determining agronomically important traits. The nucleotide sequences of cytoplasmic genomes are much more conserved than those of nuclear genomes. Cytoplasmic DNA composition in somatic hybrids is commonly elucidated either by length polymorphism analysis of restricted genome regions amplified with universal primers (PCR-RF) or by hybridization of total DNA using universal cytoplasmic probes. In this study, we demonstrate that single-stranded conformational polymorphism (SSCP) analysis is a powerful, quick and easy alternative method for cytoplasmic DNA characterization of somatic hybrids, especially for mitochondrial DNA. The technique allows detection of polymorphisms based on both size and sequence of amplified targets. Twenty-two species of the subfamily Aurantioideae were analyzed with eight universal primers (four from chloroplastic and four from mitochondrial regions). Differences in chloroplastic DNA composition were scored in 98% of all possible two-parent combinations, and different mitochondrial DNA profiles were found in 87% of them. Analysis by SSCP was also successfully used to characterize somatic hybrids and cybrids obtained by fusion of Citrus sinensis (L.) Osb. and C. excelsa Wester protoplasts.

Keywords: chloroplastic DNA; citrus; mitochondrial DNA; plant protoplast fusion

Journal Article.  0 words. 

Subjects: Plant Sciences and Forestry

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