Studies on Ergothioneine. IX. Ergothioneine Stimulates Mouse Spermatozoa and Fertilization in vitro

The effects of ergothioneine on spermatozoa and ova were investigated in vitro and in vivo. Spermatozoa were treated with ergothioneine in vitro , and injected into the uterine cavity of female mice immediately after the induction of superovulation. The ova were recovered 24 hr later and assessed for fertilization. Preincubation of spermatozoa with ergothioneine resulted in a significant increase in the fertilization rate. When ova were inseminated in the same manner in vitro with spermatozoa treated with 0.1 or 1.0 mM of ergothioneine, the penetration rate was significantly increased. These results suggest that ergothioneine is effective in inducing both capacitation and the acrosome reaction of mouse spermatozoa. Ergothioneine at concentrations of 0.1 and 1.0 mM in the preincubation medium was also effective in inducing the acrosome reaction of guinea pig spermatozoa. However, it had no significant effect on the development of 2-cell ova in vitro .     

<Emphasis Type="Italic">In vitro</Emphasis> Blastogenesis inhibited by <Emphasis Type="Italic">Erwinia carotovora</Emphasis><Emphasis Type="SmallCaps">L</Emphasis>-Asparaginase

Escherichia coliL-asparaginase, an antileukaemic agent in man¹, inhibits in vitro mitogen or antigen-induced blastogenesis in man^2,3 and in animals (M. Bennett, E. G. Mayhew and T. Han, unpublished data) and suppresses bone-marrow derived antibody precursor cells in the mouse⁴. We now report that another L-asparaginase preparation—from Erwinia carotovora—also possesses antileukaemic activity^5,6 and has a more pronounced immunosuppressive effect on in vitro blastogenesis than the E. coli enzyme.     

RESPONSIVENESS OF YEAST CELLS TO AUXIN, ANIMAL SEX HORMONES AND YEAST SEXUAL HORMONES IN RELATION TO SEX CONTROLLING GENES

Genetic control of the ability of yeast cells to respond to cell-expanding action of animal, plant and yeast hormones was studied. Mating type specificity in the ability to respond to yeast sexual hormones was changed by introduction of homothallism-controlling genes, D and HM. Auxin, a plant hormone can expand cells of most homothallic strains, but not those of heterothallic strains as far as tested. Animal sex hormones showed the action similar to that of yeast sexual hormones.     

The phylogeny of stiletto flies (Diptera: Therevidae)

The therevoid clade represents a group of four families (Apsilocephalidae, Evocoidae, Scenopinidae and Therevidae) of lower brachyceran Diptera in the superfamily Asiloidea. The largest of these families is that of the stiletto flies (Therevidae). A large‐scale (i.e. supermatrix) phylogeny of Therevidae is presented based on DNA sequence data from seven genetic loci (16S, 18S and 28S ribosomal DNA and four protein‐encoding genes: elongation factor 1‐alpha, triose phosphate isomerase, short‐wavelength rhodopsin and the CPSase region of carbamoyl‐phosphate synthase‐aspartate transcarbamoylase‐dihydroorotase). Results are presented from Bayesian phylogenetic analyses of approximately 8.7 kb of sequence data for 204 taxa representing all subfamilies and genus groups of Therevidae. Our results strongly support the sister‐group relationship between Therevidae and Scenopinidae, with Apsilocephalidae as sister to Evocoidae. Previous estimates of stiletto fly phylogeny based on morphology or DNA sequence data, or supertree analysis, have failed to find significant support for relationships among subfamilies. We report for the first time strong support for the placement of the subfamily Phycinae as sister to the remaining Therevidae, originating during the Mid Cretaceous. As in previous studies, the sister‐group relationship between the species‐rich subfamilies Agapophytinae and Therevinae is strongly supported. Agapophytinae are recovered as monophyletic, inclusive of the Taenogera group. Therevinae comprise the bulk of the species richness in the family and appear to be a relatively recent and rapid radiation originating in the southern hemisphere (Australia + Antarctica + South America) during the Late Cretaceous. Genus groups are defined for all subfamilies based on these results.     

Leaf Anatomy and Carbon Discrimination in NAD-malic Enzyme Panicum Species and their Hybrids Differing in Bundle Sheath Cell Ultrastructure

The relationship between leaf anatomy, ultrastructure and carbondiscrimination was investigated in leaves of two F₁hybrids (F₁-1and F₁-2) between two different types of the grassPanicum [anNAD-malic enzyme (ME) C₄species], which differ in bundle sheathultrastructure. The female parent was Kabulabula grass, whichhas centrifugal chloroplasts in bundle sheath cells and is designatedan NAD-ME(F) species, while the male parent was Makarikari grass,which has centripetal chloroplasts in the bundle sheath cellsand is designated an NAD-ME(P) species. Suberin lamellae arepresent in Kabulabula grass but are lacking in Makarikari grass.Both F₁hybrids had the same chromosome number (2n =36) as theparents but exhibited both univalent (about 45%) and bivalent(about 55%) chromosome pairing which was the major basis forthe identification of F₁hybrids. In F₁-1, elongated bundle sheathcell chloroplasts are arranged mainly in a centripetal position,similar to those in the male parent, Makarikari grass. In contrast,most of the bundle sheath cells in F₁-2 are packed with starch-containingchloroplasts, although in some cells chloroplasts tended tobe centripetally arranged. In both F₁hybrids, suberin lamellaewere found in the bundle sheath cell walls, similar to the femaleparent, Kabulabula grass. The ¹³C values of both F₁hybrids were-11.4 to -11.7, almost the same as those of Kabulabula grass(-11.4), but significantly higher than those of Makarikari grass(-12.7). These results indicate that the chloroplast orientationin the bundle sheath cells and the presence of suberin lamellaeare not obligatorily linked in their expression and suggestthat suberin lamellae may play an important role in discriminationagainst¹³C. Panicum ; NAD-malic enzyme species; hybrid; chloroplast position; ¹³C discrimination; suberin lamellae     

Some properties of shikimate: NADP oxidoreductase produced in sweet potato root tissue after slicing

The activity of shikimate: NADP oxidoreductase [EC 1. 1. 1.25] in sweet potato root tissue increased soon after slicing.Enzyme preparations obtained from both sliced tissue and fromfresh tissue probably contained a single enzyme component, andthey showed identical chromatographical behaviour. K_m values of the enzyme for NADP and shikimate were 1.0x10^–4Mand 1.3 x 10^–3M, respectively. Enzyme activity was potentlyinhibited by SH-inhibitors such as p-chloromercuribenzoate andoxidized glutathione. Enzyme activity was affected neither by mononucleotides suchas ATP, ADP and AMP, divalent cations, Mg⁺⁺, Ca⁺⁺ and Mn⁺⁺,nor by metabolites such as tryptophan, phenylalanine, tyrosineand t-cinnamic acid which are involved in aromatic compoundsyntheses. The enzyme rapidly lost its activity. This inactivation reactionshowed a time course consisting of two steps of the first-orderreaction. The inactivated enzyme preparation was not reactivatedby thiol compounds such as cysteine, 2-mercaptoethanol and glutathione,although these reagents, to a certain extent, protected theenzyme from inactivation. The results suggest that denaturationof the enzyme protein was involved in inactivation of the enzyme. ¹Part 74 of the phytopathological chemistry of sweet potatowith black rot and injury. ²Present address: Department of Biology, Faculty of Science,Tokyo Metropolitan University, Setagaya-ku, Tokyo. (Received August 5, 1968; )     

Frost tolerance in excised leaves of the common bugle (Ajuga reptans L.) correlates positively with the concentrations of raffinose family oligosaccharides (RFOs)

Mass increases in raffinose family oligosaccharides (RFOs, α 1,6-galactosyl extensions of sucrose) are well documented in the generative tissues of many plants upon cold acclimation, and they (i.e. mainly the two shortest RFO members, raffinose and stachyose) have been suggested as frost stress protectants. Our focus here was on the longer RFO members as they commonly occur in the frost-hardy evergreen labiate Ajuga reptans in its natural habitat, and accumulate to their highest concentrations in winter when the plant is faced with sub-zero temperatures. We examined the effects of RFO concentration and chain length on frost tolerance using excised leaves which accumulate long-chain RFOs under both cold and warm conditions, thereby uncoupling the acclimation temperature from RFO production. We demonstrated that frost tolerance in excised A. reptans leaves correlates positively with long-chain RFO accumulation under both acclimation temperatures. After 24 d post-excision in the warm, the leaves had increased their RFO concentrations (mainly long-chain RFOs) 22-fold to 78 mg g⁻¹ fresh weight, and decreased their EL₅₀ values (temperature at which 50% leakage occurred) from −10.5 to −24.5 °C, suggesting a protective role for these oligosaccharides in the natural frost tolerance of A. reptans .     

Does stand structure influence susceptibility of eucalypt floodplain forests to dieback?

Forest dieback is a worldwide problem that is likely to increase with climate change and increasing human demands for resources. Eucalyptus camaldulensis forests are an acute example of forest dieback, with 70% of the Victorian Murray River floodplain in some state of dieback. If we are to halt dieback in these floodplain forests, we need to understand what makes stands susceptible to dieback. Forest diebacks are often related to stand structure, with dieback more severe in senescent or high‐density stands. We determined whether certain stand structures make these forests more susceptible to dieback. We undertook an extensive survey of 176 stands across 100 000 ha of forest, covering the range of stand condition on this floodplain. Large and small trees (20‐, 40‐, 80‐ and 120‐cm diameter) showed a similar reduction in the probability of being alive with decreasing stand condition. A slight improvement in stand condition was found at higher densities and basal areas, which may reflect the higher productivity or younger age of these stands. Stand condition was moderately, positively correlated with longitude, with stand condition being higher in the east of the Murray River floodplain where flooding frequencies are currently higher. This suggests that dieback of these floodplain forests would be more effectively mitigated by increased water availability through flooding than by altering stand structure.     

Content of Adenosine Phosphate Compounds in a Long-Day Duckweed, Lemna gibba G3, under Different Light and Nutritional Conditions

The levels of ATP ADP and AMP as well as the energy charge were examined in the long-day duckweed. Lemna gibba G3, under different light and nutritional conditions. ATP and ADP content, but not AMP, decreased slightly when the plant was cultured with a medium depleted of sucrose or was exposed to continuous darkness. The energy charge was not affected either by changes in the light conditions or by the depletion of sucrose from the medium, although increase of fresh weight changed drastically under these conditions. The levels of ADP and AMP and the value of the energy charge remained nearly constant throughout a 1-day period when the plant was exposed to short-day, continuous light and continuous dark. ATP content decreased gradually during the light period of the short day and thereafter remained constant during the dark period. When the plant was exposed to continuous light. ATP content decreased until the 8th hour after beginning of the continuous light, only to recover rapidly to its original level during the phase coinciding with the preceding dark period of the short day. Under continuous dark conditions the ATP content remained constant throughout the day. It was concluded that the diurnal rhythm of physiological activities previously reported cannot be related to the energy charge.