Anemone

Abstract: Relative amounts of DNA were determined by Feulgen cytophotometry in 22 diploid species of Ranunculaceae (n=7, 8, 9) representing six genera, and exhibiting large differences in chromosome size, but no marked differences in karyotype pattern. Chemical determination of absolute amounts of DNA for six of these species, allowed conversion of all the photometric data into absolute units of DNA. The mean DNA content per nucleus varied from.13×10−11gm in Aquilegia to 5.25×10−11gm in species of Anemone in the section Homalocarpus. The DNA values obtained appeared to be “quantized”, and data for the majority of species fitted a non-geometrical series with the observed relative terms: 1—8—12—16—20—24—40. The magnitude of these variations in DNA content, the preservation of the karyotype and the tendency towards a simple numerical progression in DNA values, lead us to prefer an interpretation of the evolution of DNA content in terms of differential polynemy to one postulating changes in size of genetic units in an unchanging number of strands per chromosome.

Abstract: Chloroplast DNA cleavage sites for 10 restriction enzymes were mapped for 46 species representing all sections of Anemone, four closely related genera (Clematis, Pulsatilla, Hepatica, and Knowltonia), and three more distantly related outgroups (Caltha, Ranunculus, and Adonis). Comparison of the maps revealed that the chloroplast genomes of Anemone and related genera have sustained an unusual number and variety of rearrangements. A single inversion of a 42-kb segment was found in the large single-copy region of Adonis aestivalis. Two types of rearrangements were found in the chloroplast genome of Clematis, Anemone, Pulsatilla, Hepatica, and Knowltonia: An approximately 4-kb expansion of the inverted repeat and four inversions within the large single-copy region. These rearrangements support the monophyletic status of these genera, clearly separating them from Caltha, Ranunculus, and Adonis. Two further inversions were found in two Clematis species and three Anemone species. While appearing to support a monophyletic grouping for these taxa, these two inversions conflict with data from both chloroplast restriction sites and morphology and are better interpreted as having occurred twice independently. These are the first two documented cases of homoplastic inversions in chloroplast DNA. Finally, the second intron of the chloroplast rps12 gene was shown to have been lost in the common ancestor of the same three Anemone species that feature the two homoplastic inversions.

Abstract: To understand the flexibility of symbiotic associations in coral reefs, we investigated the specificity of the Aiptasia (cf. insignis)-Symbiodinium association in the laboratory by rendering the anemones aposymbiotic and inoculating them with different isolates of SYMBIODINIUM: Infective algal symbionts were monitored over 3 months by re-isolation and identification using denaturing-gradient gel electrophoresis and sequence comparison of their amplified 18S rRNA hypervariable V1 + V2 gene region. Despite similarity in their external morphology, the algal isolates differed in their infectivity towards the host. Within days of single-isolate inoculation, aposymbiotic anemones formed associations with fresh or cultured isolates (clade B) from the anemones Aiptasia sp. or A. tagetes, respectively. They associated to a limited extent with cultured isolates (clade A) from the tridacnids Tridacna crocea or Hippopus hippopus, and not at all with a cultured isolate (clade C) from the stony coral Montipora verrucosa, nor with a free-living isolate (clade A) from subtidal sands. Aposymbiotic anemones inoculated with a mixture of all isolates had only the anemone taxon as their detectable symbionts. Re-inoculation of induced symbioses with a mixture of all isolates and incubation with wild anemones showed that the initial induced symbioses with the anemone taxon were stable. Anemones originally infected with tridacnid isolates either additionally acquired the anemone taxon or had the former outgrown by the latter. These results demonstrate the presence of a host-symbiont recognition mechanism, and possibly competition among potential algal symbionts in the Aiptasia-Symbiodinium association. Here we present a method that may be useful in monitoring the algal population dynamics in symbiotic corals in the field, along with an efficient method of rendering Aiptasia aposymbiotic for further laboratory investigation of Aiptasia-Symbiodinium symbioses.

Abstract: Embryogenesis in anther cultures of Anemone Canadensis L., Anemone hupehensis Lemoine and Nicotiana tabacum L. was shown to be inhibited by abscisic acid added to the medium. However, this inhibition was reduced in the presence of activated charcoal (AC). The presence of AC in the culture medium strongly promoted embryogenesis in anther cultures of Anemone Canadensis compared with other media combinations. Treatment of the agar medium with AC, which was removed before inoculation of the anthers, also stimulated embryogenesis, but treatment of the water constituent did not. The number of embryos produced in anther cultures of Anemone Canadensis and Nicotiana tabacum was shown to be positively related to the length of lime of incubation on medium supplemented with AC. In the case of Anemone Canadensis the stimulating effect of AC was most pronounced when the first visible embryos had emerged. The presence of anther-derived embryos from Anemone Canadensis in anther cultures of Anemone Canadensis and Nicotiana tabacum was shown to inhibit embryogenesis. It was also demonstrated that embryos from anther cultures of Anemone canadensis, Papaver setigerum DC and Clematis viticella L. produced phenolic substances, and that the concentration of these substances was higher in culture medium lacking AC. Treatment of such medium with AC could reduce the concentration of phenolic substances by more than 80%.