Περίληψη:
Different approaches to producing transgenic grapevines based on regeneration via embryogenesis were investigated. Embryogenic callus was initiated from anther tissue of Vitis vinifera cv. Sultana and three embryogenic
culture types (embryogenic callus, tissue type I; proliferating embryos, tissue type II; and a suspension) were
established. The three culture types were incolucaled with Agrobacterium tumefaciens harbouring a binary vector which contained a uidA reporter gene and either a hpt or nptII selectable marker gene or the cultures were
bombarded with microprojectiles carrying a uidA/nptII binary vector. Transgenic plants were produced only from
Agrobacterium transformation experiments. Transformed embryos were selected with kanamycin or hygromycin
antibiotics and recovered with the highest efficiency from inoculated type I cultures. Southern analysis of genomic
DNA extracted from ten transgenic plants showed that the number of T-DNA insertions in the genome ranged
from 1 to at least 4. Evidence for methylation of the T-DNA at cytosine and adenine residues in transgenic plants
was found by Southern analysis of DNA digested with two isoschizomer pairs of restriction endonucleases. No
evidence for genotype alterations or somatic meiosis was found when DNA from 80 somatic embryos and seven
plants regenerated from embryogenic culture were analysed at six sequence-tagged sites which are heterozygous
in cv. Sultana. Expression of the uidA gene in in vitro grown leaves of transgenic plants was most often high
and uniform but GUS staining was occasionally observed to be low and/or patchy. Transgenic plants and all plants
regenerated from embryogenic culture produced red veined, lobed leaves which are uncharacteristic of the accepted
ampelographic phenotype of Sultana. It is suggested that this phenotype may represent a juvenile growth stage.
Αναφορά NLM:
Franks T, Gang He D, Thomas M. Regeneration of transgenic shape Vitis vinifera L. Sultana plants: genotypic and phenotypic analysis. Mol Breed. 1998;4:321–33. https://doi.org/10.1023/A:1009673619456