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Genetic stability at nuclear and plastid DNA level in regenerated plants of Solanum species and hybrids

Aversano, Riccardo, Savarese, Salvatore, Maria De Nova, Jose, Frusciante, Luigi, Punzo, Maria, Carputo, Domenico
Euphytica 2009 v.165 no.2 pp. 353-361
Solanum cardiophyllum, Solanum commersonii, Solanum nigrum, Solanum phureja, Solanum tuberosum, potatoes, Solanum bulbocastanum, tissue culture, somaclonal variation, genetic markers, amplified fragment length polymorphism, chloroplast DNA, DNA, DNA fragmentation, genetic stability, Solanum stoloniferum, nuclear genome
In this work we detected the extent of variability at nuclear and cytoplasmic DNA level of regenerated plants belonging to Solanum genotypes with a different genetic background and somatic chromosome number. As for the nuclear characterization, a total of 66 (18.5%) polymorphic bands were scored using 13 ISSR primers on 45 randomly selected regenerants. Our results show that the regenerants obtained from clone cmm 1T and, at lower level, those from cph 1C are unstable under in vitro conditions or rather more prone to in vitro-induced stress leading to somaclonal variation than the other genotypes used. Two types of changes were observed: disappearance of parental ISSR fragments, termed “loss”; appearance of novel ISSR fragments, termed “gain”. The most frequent event occurring in the regenerants was the loss of fragments (41 bands). Regenerated plants were analyzed with seven plastid universal primers to determine the cytoplasmic composition at chloroplast level. All cpDNA primer pairs tested produced amplicons of the same size in all genotypes analyzed and no polymorphic fragments were observed with any universal primers used. Our results show that under in vitro culture conditions genotype affects the integrity of the genome. In addition, the absence of polymorphism at plastid level confirms the greater genetic stability of cytoplasmic DNA.