Somatic embryogenesis in perennial statice Limonium bellidifolium,Plumbaginaceae

Cotyledon explants from perennial statice Limonium bellidifolium (Statice caspia Willd.) were cultured on Murashige and Skoog's basal medium (MS) supplemented with various levels of 2,4-D, kinetin and sucrose. Embryogenic calluses developed over a period of 10 days with the highest response at 4.5 M (1 mg l^–1) 2,4-D, 0.5 M (0.1 mg l^–1) kinetin and 117 mM sucrose. Following induction, the calluses were transferred to MS media supplemented with 88 or 117 mM sucrose and 0 or 0.5 M kinetin. Somatic embryos at the globular, heart-shaped, torpedo, and cotyledonary stages developed. Fully germinated plantlets developed with the best response in medium supplemented with 117 mM sucrose and 0.5 M kinetin. Direct somatic embryogenesis without a callus phase was observed with some of the cotyledon explants. Induction, maturation and germination of somatic embryos on the optimized media were equally effective using cotyledon, hypocotyl and root explants. Serial sections of L. bellidifolium cotyledon explants cultured for two weeks indicated that pro-embryogenic masses originated from parenchyma cells below the epidermis. Further histological observations of embryogenic calluses confirmed the initiation and development of globular and heart-shaped embryos and repetitive somatic embryogenesis. Ultrastructural observations indicated that the embryogenic cells were less vacuolate with abundant organelles compared to the cells of the explant. This is the first report of somatic embryogenesis in the Plumbaginaceae.     

Universal restriction endonucleases: designing novel cleavage specificities by combining adapter oligodeoxynucleotide and enzyme moieties

Class IIS restriction endonucleases cleave double-stranded (ds) DNA at precise distances from their recognition sequences. A method is proposed which utilizes this separation between the recognition site and the cut site to allow a class IIS enzyme, e.g., FokI, to cleave practically any predetermined sequence by combining the enzyme with a properly designed oligodeoxynucleotide adapter. Such an adapter is constructed from the constant recognition site domain (a hairpin containing the ds sequence, e.g., GGATG CCTAC for FokI) and a variable, single-stranded (ss) domain complementary to the ss sequence to be cleaved (at 9 and 13 nucleotides on the paired strands from the recognition sequence in the example of FokI). The ss sequence designated to be cleaved could be provided by ss phage DNA (e.g., M13), gapped ds plasmids, or supercoiled ds plasmids that were alkali denatured and rapidly neutralized. Combination of all three components, namely the class IIS enzyme, the ss DNA target sequence, and the complementing adapter, would result in target DNA cleavage at the specific predetermined site. The target ss DNA could be converted to the precisely cleaved ds DNA by DNA polymerase, utilizing the adapter oligodeoxynucleotide as primer. This novel procedure represents the first example of changing enzyme specificity by synthetic design. A practically unlimited assortment of new restriction specificities could be produced. The method should have many specific and general applications when its numerous ramifications are exploited.     

Plant growth regulators and the orchid cut-flower industry

Research involving plant growth regulators (PGRs) and orchids in areas of orchid growth and development are reviewed. For all areas covered in the review — seed germination and seedling growth, lateral shoot production, root production, flower initiation and development, postharvest physiology, and photosynthate partitioning — it was concluded that further studies would assist in clarifying potential uses for PGRs in the orchid cut-flower industry. It is suggested that extra PGR research on orchids is justified at the present time because of favourable prospects facing the orchid cut-flower industry.     

Cut5 is a component of the UV-responsive DNA damage checkpoint in fission yeast

A checkpoint responding to DNA damage in G2 results in a delay in the onset of mitosis through inhibition of p34^cdc2 kinase activity via maintenance of inhibitory tyrosine phosphorylation. Genetic analyses of this checkpoint in fission yeast have identified single alleles of several genes, suggesting these screens are not yet saturating, and hence further genes await identification. To fully understand the complexity of this checkpoint it will be necessary to define all the genes involved. To this end we screened for new mutants defective in the ability to delay mitosis in the presence of DNA-damaging agents. Twenty-four mutants were isolated that were defective in UV-C and MMS-induced checkpoint delay. Amongst these mutants was an allele of cut5 that was also defective in the checkpoint responses. We show here, contrary to previous reports, that the UV-C induced checkpoint response is defective in cut5 mutants. Therefore, like all other checkpoint mutants, cut5 is required for G2 checkpoint arrest following DNA damage, regardless of the nature of the lesions involved. Received: 24 July 1998 / Accepted: 14 September 1998     

Organogenesis and somatic embryogenesis in callus cultures ofRosa hybrida andRosa chinensis minima

Pre-incubation of somatic tissues of the cut rose Carefree Beauty and miniature roses Red Sunblaze and Baby Katie in 10, 100, or 200 M 2,4-D induced the development of highly rhizogenic callus. Upon transfer of rhizogenic callus to a regeneration medium containing 23 M TDZ and 3 M GA₃, a low frequency of shoot organogenesis (3.3%) and somatic embryogenesis (6.6%) was observed. Incubation of leaf and stem internodes in 11, 27, 54, 81, or 108 M NAA followed by transfer of explants to the regeneration medium resulted in up to a 25% increase in shoot organogenesis from callus-derived internodal explants of Red Sunblaze, but no somatic embryogenesis was observed. The influence of glucose versus sucrose in the regeneration medium on leaf explants of Carefree Beauty and Baby Katie pre-incubated in 100 M 2,4-D revealed a difference in genotypic response to shoot organogenesis and somatic embryogenesis. For Carefree Beauty, a concentration of 111 mM glucose induced higher frequencies of both organogenic (33%) and embryogenic calluses (25%) than either 59 mM or 117 mM sucrose. For Baby Katie, no significant difference was found for number of organogenic calluses induced on 59 mM sucrose and 111 mM glucose.Abbreviations ABA Abscisic acid - BA Benzyladenine - 2,4-D 2,4-Dichlorophenoxyacetic acid - GA₃ Gibberellic acid - IAA Indoleacetic acid - MS Murashige & Skoog (1962) - NAA -Naphthaleneacetic acid - TDZ Thidiazuron     

Biological consequences of stream routing through a final-cut Strip mine pit: Benthic macroinvertebrates

Benthic macroinvertebrates were collected from the Plachek Pit, a final-cut coal strip mine pit at the Big Horn Mine near Sheridan, Wyoming, USA. Goose Creek was routed through the pit to allow mining under the original creek bed. The benthic community in the pit was dominated by the worm Limnodrilus hoffmeisteri and various midge genera, including Chironomus sp., Procladius sp. and Tanypus stellatus. Insects other than chironomids were minor components of the benthic fauna, and were collected only in the upstream portions of the pit; they were concluded to have originated from drift deposition. The pit functions as a sink for drifting macroinvertebrates. Based on community composition and supplemental water quality information it was concluded that the Plachek Pit best exemplified an organically-enriched, lake-like ecosystem. Overall, rerouting of Goose Creek through the Plachek Pit was not viewed as deleterious to Goose Creek, but rather benefited the creek by serving as a sink for inputs from upstream municipal and agricultural effluents.     

Testing meat-eating in early hominids: an analysis of butchery marks on defleshed carcases

The diet of early hominids responsible for the bone accumulations at Plio-Pleistocene sites is still a controversial issue. Meateating and bone marrow consumption are often presented either as complementary or as opposing strategies of carcass exploitation. The occurrence of cut marks on fossil bones at early sites is a potential source of information that has not been consistently used as evidence of what products hominids obtained from carcasses. Some authors interpret them as the result of manipulating meat-bearing bones, whereas others believe that they can also be the result of extracting marginal scraps of flesh that have survived carnivores’ initial consumption of carcasses. In this study, a referential framework concerning the latter process is presented and it is concluded, according to the data drawn from the FLK “Zinj” site and the results obtained in the experiment, that hominids processed meat-bearing bones (in which flesh was abundant) and not defleshed carcasses from felid kills. This work constitutes a reference that can also be used for later Pleistocene sites and adds a further dimension to the hunting-versus-scavenging debate.     

Micropropagation of Alpinia purpurata from inflorescence buds

Alpinia purpurata K Schum inflorescence buds inoculated on Murashige and Skoog medium (MS) containing 10 M 6-benzyladenine with 5 M naphthaleneacetic acid gave rise to multiple shoot formation with a mean increase of 15 to 20 new shoots each 4 weeks. Plants could be stored for more than 6 months in flasks containing deionized water with 5 g 1^-1 sucrose with minimal vegetative growth.Abbreviations IAA indole-3-acetic acid - NAA naphthaleneacetic acid - BA 6-benzyladenine - AS Adenine sulphate - MS Murashige and Skoog medium     

Invertase and sucrose synthase in flowers

Sucrose and reducing sugar concentrations in petals of cut carnation flowers, whose life was prolonged up to 7 days by bathing stalks in sucrose solutions, were respectively 3-fold and 2-fold higher than those bathed in water. Reducing sugar concentrations were about 7-fold higher than sucrose concentrations. A study of invertase and sucrose synthase activities in flower petals of carnation and four other species of flowers revealed that both enzymes may be involved in hydrolysis of translocated sucrose. Invertase activity, while being up to 20-fold higher than sucrose synthase activity in some species was approximately comparable in others. More detailed studies on invertase from petals of 3 flower species demonstrated the presence of only the acid form of the enzyme with a K_m value for sucrose of about 2.5 mM.