Ploidy level was determined for six species and 88 cultivars of the Rhododendron subgenus Tsutsusi. High-resolution flow cytometry of nuclear DNA was performed on macerated plant tissue. All plants analyzed were diploid (2n = 26) with the exception of `Euratom', `Euratom Orange', and `Red Wing', which were triploid (3n = 39), and `Casablanca Tetra', which was found to be a cytochimera: mixoploid (2n + 4n) in the LI and LII, but tetraploid in the LIII. The described method has proven to be useful in screening a large population of rhododendrons. Analysis of different organs and plant tissues was easily accomplished through flow cytometry, and has proven useful in determining the ploidy of different histogenic layers.
Three chrysanthemum (Dendranthema grandiflora) cultivars were cocultivated with 2 Agrobacterium tumefaciens strains in combination with 4 pBIN19 derived binary plasmids, all carrying the Nosnptll selection gene and 35Sgus(intron) reporter gene. All binary plasmids transferred DNA to chrysanthemum explants but only pMOG410 gave good stable expression of GUS. This plasmid differs from the other plasmids in 2 aspects: 1) It carries a restored nptll gene and 2) the selection gene is positioned at the left border side of the reporter gene. Cocultivation with AGLO(pMOG410) yielded up to 13 GUS positive shoots per 100 explants. The presence of the gus and nptll gene in recovered shoots was confirmed by PCR and Southern blot analysis.
Rhododendron simsii 'Hellmut Vogel' was regenerated using different types of explants, auxins and cytokinins. After a callus induction phase, with 2,4-dichlorophenoxyacetic acid or a-naphthaleneacetic acid, adventitious shoot regeneration was obtained on a medium supplemented with thidiazuron or zeatin. With thidiazuron shoots were small and a subsequent elongation step was required before rooting. An elongation step was not required when zeatin was used. The duration of the callus induction phase was negatively correlated with the regeneration capacity.
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