Use of EDTA to prevent spontaneous cell-to-cell transformation provides a more accurate estimation of gene transfer frequencies

Summary Spontaneous cell-to-cell transformation between naturally competent bacteria on selective media resulted in an overestimation of the transferability of genetic information. EDTA effectively prevented transformation on selective media whereas DNaseI did not reliably inhibit cell-to-cell transformation. An improved method to estimate gene transfer frequencies is described.     

Regeneration of Triticum aestivum apical explants after microinjection of germ line progenitor cells with DNA

A highly efficient method of regenerating fertile, phenotypically normal plants from shoot apex cultures of T. aestivum was developed. The hypodermal layer (L2) of the vegetative apex containing germ line precursor cells could be located with bright field microscopy and targeted for microinjection. Fluorescently labelled dextrans were used as markers to develop a microinjection procedure which did not disrupt nuclear or cytoplasmic structure. This procedure was used to inject plasmid DNA into L2 cells. Capillary microinjection did not shear the plasmid DNA and delivery of DNA was confirmed by polymerase chain reaction analysis of DNA isolated from injected apices. The significance of these findings in relation to the development of cereal transformation systems will be discussed.     

Na+ and K+ binding by glycerinated muscle fibers with reciprocal cation concentrations in the medium

The binding of Na+ and K+ by glycerinated muscle fibres was observed at reserve concentrations of NaCl in the medium. Under external concentrations of Na+ of K+ up to 0.4-0.5 mM, a constant fraction (0.15-0.25 mmoles/kg dry weight of the fibres) bound by glycerinated fibres was revealed. With the increase of NaCl or KCl concentration in the medium up to 10 mM the concentration of bound cations increased too. The parameters of Na+ and K+ sorption by glycerinated models were calculated. The values of Na+ and K+ binding limits were 4.4 and 1.8 mmole/kg dry weight of the fibres and those of affinity, 3.2 and 4.1 kcal/mol, respectively. The binding of one cation took place in conditions when its concentration was 10,000-20,000 fold less than that of the other cation. This points to the fact that Na+ and K+ binding is highly specific and is carried out by different centres. It is suggested that myosin ATPase is a substratum binding Na+ and K+ in glycerinated muscle fibres at reverse ratio concentrations of these cations in the medium.     

Patterns of Reproductive Isolation in Toads

Understanding the general features of speciation is an important goal in evolutionary biology, and despite significant progress, several unresolved questions remain. We analyzed an extensive comparative dataset consisting of more than 1900 crosses between 92 species of toads to infer patterns of reproductive isolation. This unique dataset provides an opportunity to examine the strength of reproductive isolation, the development and sex ratios of hybrid offspring, patterns of fertility and infertility, and polyploidization in hybrids all in the context of genetic divergence between parental species. We found that the strength of intrinsic postzygotic isolation increases with genetic divergence, but relatively high levels of divergence are necessary before reproductive isolation is complete in toads. Fertilization rates were not correlated to genetic divergence, but hatching success, the number of larvae produced, and the percentage of tadpoles reaching metamorphosis were all inversely related with genetic divergence. Hybrids between species with lower levels of divergence developed to metamorphosis, while hybrids with higher levels of divergence stopped developing in gastrula and larval stages. Sex ratios of hybrid offspring were biased towards males in 70% of crosses and biased towards females in 30% of crosses. Hybrid females from crosses between closely related species were completely fertile, while approximately half (53%) of hybrid males were sterile, with sterility predicted by genetic divergence. The degree of abnormal ploidy in hybrids was positively related to genetic divergence between parental species, but surprisingly, polyploidization had no effect on patterns of asymmetrical inviability. We discuss explanations for these patterns, including the role of Haldane''s rule in toads and anurans in general, and suggest mechanisms generating patterns of reproductive isolation in anurans.     

Scanning electron microscope study of Pseudomonas putida colonies.

Pseudomonas putida colonies were examined by scanning electron microscope. A variety of cell morphologies, multicellular arrangements, and extracellular materials were observed in the fixed material. Different regions of a single colony showed characteristic organizations of these architectural elements. In some cases, the detailed microstructure of the fixed colony surfaces observed by scanning electron microscopy could be correlated with macroscopic patterns visualized by histochemical staining and surface relief photography of live colonies. Extracellular materials were seen to extend onto the agar surface beyond the boundaries of the cell mass, and the final structures of these materials, after fixation and desiccation, were colony specific. The significance of these features of colony microstructure for formulating hypotheses about the control of colony morphogenesis is discussed.