High frequency Agrobacterium-mediated transformation and plant regeneration via direct shoot formation from leaf explants in Beta vulgaris and Beta maritima.
We have developed a new procedure for Agrobacterium-mediated transformation of plants in the genus Beta using shoot-base as the material for Agrobacterium infection. The frequency of regeneration from shoot bases was analyzed in seven accessions of sugarbeet ( Beta vulgaris) and two accessions of B. maritima to select materials suitable for obtaining transformed plants. The frequency of transformation of the chosen accessions using Agrobacterium strain LBA4404 and selection on 150-mg/l kanamycin was found to be higher than that in previously published methods. Genomic DNA analysis and beta-glucuronidase reporter assays showed that the transgene was inherited and expressed in subsequent generations. In our method, shoot bases are prepared by a simple procedure, and transformation does not involve the callus phase, thus minimizing the occurrence of somaclonal variations.
Hisano
H
H
Graduate School of Agriculture, Hokkaido University, 060-8589, Sapporo, Japan.
Kimoto
Y
Y
Hayakawa
H
H
Takeichi
J
J
Domae
T
T
Hashimoto
R
R
Abe
J
J
Asano
S
S
Kanazawa
A
A
Shimamoto
Y
Y
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001483:Base Sequence; D027461:Beta vulgaris / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D019663:Chenopodiaceae / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002934:Cinnamates / Q000494:pharmacology; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D005822:Genetic Vectors; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D018520:Plant Shoots / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000502:physiology*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
D002934:Cinnamates; D017931:DNA Primers; D018744:DNA, Plant; D006921:Hygromycin B; C026273:hygromycin A