Open AccessApplication of Cleaved Amplified Polymorphic Sequence Method for Analysis of Cytoplasmic Genome among Aurantioideae Intergeneric Somatic Hybrids
Author(s) -
Samia Lotfy,
Luro François,
Françoise Carreel,
Yann Froelicher,
Delphine Rist,
Patrick Ollitrault
Publication year - 2003
Publication title -
journal of the american society for horticultural science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.408
H-Index - 80
eISSN - 2327-9788
pISSN - 0003-1062
DOI - 10.21273/jashs.128.2.0225
Subject(s) - biology , somatic fusion , protoplast , restriction fragment length polymorphism , genome , genetics , somatic cell , mitochondrial dna , southern blot , hybrid , microbiology and biotechnology , polymerase chain reaction , dna , gene , botany
Somatic hybridization allows the creation of new patterns of nuclear, mitochondrial and chloroplastic association. It is therefore necessary to master cytoplasmic molecular markers to determine the genetic origin of both organelles of plantlets obtained from protoplasts fusion. In the case of Citrus and related genera, only southern blot hybridization and restriction fragment-length polymorphism (RFLP) techniques were used for this task until now. Here, we describe the use in the Aurantioideae subfamily, of a simple and non labeling cleaved amplified polymorphic sequence (CAPS) technique, to determine the cytoplasmic genome origin of intergeneric somatic hybrids. Mitochondrial and chloroplastic universal primers previously selected for population genetic studies in Quercus by Demesure et al. (1995) are used with some modifications. The variability of cytoplasmic genome among somatic fusion partners is detected by coupling amplification and restriction reactions. Digested DNA fragments are analyzed by agarose gel electrophoresis (PCR-RFLP). This technique has been applied for the analysis of the cytoplasmic constitution of somatic hybrids arising from intergeneric, intersubtribal and intertribal combinations. Systematic transmission of the mitochondria from protoplasts isolated from embryogenic callus parents was confirmed.