In vitro propagation systems were developed for Carnegiea gigantea (Engelm.) Britt & Rose, Pachycereus pringlei (Berger) Britt & Rose and Stenocereus thurberi (Engelm.) Buxb, three North American species of columnar cacti. In vitro germinated seedlings were used as a source of explants. Multiple shoot formation from areoles was achieved for three types of explants (apical, lateral, and transverse) cultured on Murashige and Skoog (MS) basal media supplemented with 3% sucrose, 10 g·L-1 agar and various treatments with growth regulators. The highest shoot production efficiency for C. gigantea was obtained on transverse explants cultured on a medium with 2 mg·μmL-1 (8.87 μm) BA, where 5.3 shoots per explant were obtained. In P. pringlei and S. thurberi the best response was obtained using transverse explants on medium with 1 mg·L-1 (4.44 μm) BA (3.8 and 4.3 shoots per explant, respectively). Rooting of the in vitro generated shoots was achieved most efficiently on MS basal media with 3% sucrose, 10 g·L-1 agar and 1 mg·L-1 (4.9 μm) indole-3-butyric acid. Rooting frequencies were 92%, 88%, and 96% for C. gigantea, P. pringlei and S. thurberi, respectively, and the frequency of survival of the plants once transferred to soil was 86% on average. Chemical name used: benzyladenine (BA).
Susceptibility of Mexican cacti to Agrobacterium rhizogenes was evaluated in 65 species of 22 genera. Stem discs taken from in vitro cultured plants were inoculated with Agrobacterium rhizogenes A4 agropine-type strain that contains the wild RiA4 plasmid and the binary vector pESC4 with the nptII and gus genes. Hairy roots were produced directly from wounds, or starting from calli generated on the wounded surface, in 34 of the evaluated species. The frequency of hairy roots formation, the number of roots per explant and its growth rates were variable among the tested species. In the 31 remaining species the production of transformed roots was not observed under the conditions used in these experiments. Histochemical detection of β-glucuronidase (GUS) activity demonstrated the expression of this foreign gene in the hairy roots. PCR analyses demonstrated the presence of the rolB and nptII genes in the DNA of the transformed roots. The patterns of alkaloid-like compounds obtained by thin layer chromatography in some of the tested species were qualitatively similar between the transformed and non-transformed roots.
In vitro propagation systems by means of areole activation were developed for Echinocereus knippelianus, Echinocereus schmollii, Mammillaria carmenae, M. carmenae fo. rubrisprina, M. herrerae, M. theresae, Melocactus curvispinus, Escontria chiotilla and Polaskia chichipe. In vitro germinated seedlings were used as source of explants. Multiple shoot formation from areoles was achieved on MS basal medium supplemented with 3% sucrose, 10 g L-1 agar and 6-benzylaminopurine (BA) or 6-(, -dimethylallylamino)purine (2iP). Efficiencies ranged from 6.0 shoots per explant in M. carmenae fo. rubrisprina to 13.5 shoots per explant in Echinocereus schmollii. Rooting of the in vitro generated shoots was achieved in MS basal medium, or MS basal medium supplemented with indoleacetic acid, indolebutyric acid or activated charcoal. Finally, 49-98% of these plants survived.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.