The identification and metabolism of GA9 and its metabolites in seed coats and shoots of pea,line G2

[³H]gibberellin A₉ was applied to shoots or seed parts of G2 pea to produce radiolabeled metabolites. These were used as markers during purification for the recovery of endogenous GA₉ and its naturally occurring metabolites. GA₉ and its metabolites were purified by HPLC, derivatized and examined by GC-MS. Endogenous GA₉, GA₂₀, GA₂₉ and GA₅₁ were identified in pea shoots and seed coats. GA₅₁-catabolite and GA₂₉-catabolite were also detected in seed coats. GA₇₀ was detected in seed coats following the application of 1 g of GA₉. Applied [³H]GA₉ was metabolized through both the 13-hydroxylation and 2-hydroxylation pathways. Labeled metabolites were tentatively identified on the basis of co-chromatography on HPLC with endogenous compounds identified by GC-MS. In shoots [³H]GA₅₁ and [³H]GA₅₁-catabolite were the predominant metabolites after 6 hrs, but by 24 hrs there was little of these metabolites remaining, while [³H]GA₂₉-catabolite and an unidentified metabolite predominated. In seed coats [³H]GA₅₁ was the initial product, later followed by [³H]GA₅₁-catabolite and an unidentified metabolite (different from that in shoots), with lesser amounts of [³H]GA₂₀, [³H]GA₂₉ and [³H]GA₂₉-catabolite. [³H]GA₇₀ was a very minor product in both cases. [³H]GA₉ was not metabolized by pea cotyledons.Edited by T.J. Gianfagna.Author for correspondence     

Fatal association between dieldrin-resistant and susceptible Australian sheep blowflies, Lucilia cuprina.

A novel phenomenon of interactions between genotypes of the dieldrin-resistance (Rdl) locus of the Australian sheep blowfly (Lucilia cuprina) is described. Susceptible adult flies exposed to dieldrin-resistant (Rdl/Rdl or Rdl/S) adults, raised from larvae grown on media containing sublethal concentrations of dieldrin, display mortality related to the concentration on which the resistant flies developed. The resistant flies excrete quantities of dieldrin that are toxic to susceptible flies. These observations provide an additional mechanism to those previously identified for the rapid evolution of resistance to dieldrin by L. cuprina.     

Vacuolar H(+)-translocating pyrophosphatases: a new category of ion translocase.

The membrane surrounding the central vacuole of plant cells contains an H(+)-translocating ATPase (H(+)-ATPase) and an H(+)-translocating inorganic pyrophosphatase (H(+)-PPase). Both enzymes are abundant and ubiquitous in plants but the H(+)-PPase is unusual in its exclusive use of inorganic pyrophosphate (PPi) as an energy source. The lack of sequence identity between the vacuolar H(+)-PPase and any other characterized ion pump implies a different evolutionary origin for this translocase. The existence of the vacuolar H(+)-PPase, in conjunction with increasing recognition of PPi as a key metabolite in plant systems, necessitates reconsideration of ATP as the primary energy source for membrane transport in plant cells.     

Adenine photodimerization in deoxyadenylate sequences: elucidation of the mechanism through structural studies of a major d(ApA) photoproduct.

The mechanism of the photodimerization of adjacent adenine bases on the same strand of DNA has been elucidated by determining the structure of one of the two major photoproducts that are formed by UV irradiation of the deoxydinucleoside monophosphate d(ApA). The photoproduct, denoted d(ApA)*, corresponds to a species of adenine photodimer first described by P?rschke (P?rschke, D. (1973) J.Am.Chem.Soc. 95, 8440-8446). From a detailed examination of its chemical and spectroscopic properties, including comparisons with the model compound N-cyano-N1-(1-methylimidazol-5-yl)formamidine, it is deduced that d(ApA)* contains a deoxyadenosine unit covalently linked through its C(8) position to C(4) of an imidazole N(1) deoxyribonucleoside moiety bearing an N-cyanoformamidino substituent at C(5). On treatment with acid, d(ApA)* is degraded with high specificity to 8-(5-amino-imidazol-4-yl)adenine whose identity has been confirmed by independent chemical synthesis. It is concluded that the primary event in adenine photodimerization entails photoaddition of the N(7)-C(8) double bond of the 5'-adenine across the C(6) and C(5) positions of the 3'-adenine. The azetidine species thus generated acts as a common precursor to both types of d(ApA) photoproduct which are formed from it by competing modes of azetidine ring fission.     

Acclimatory responses in enzymatic activity and metabolite concentrations in the tropical scorpion Heterometrus fulvipes

1. 1.|Changes in tissue metabolite concentrations and enzyme activities in the pedipalpal (PM) and heart (HM) muscles of the tropical scorpion Heterometrus fulvipes show that the metabolism in PM and HM is fundamentally reorganized following low (18°C) and high (38°C) temperature acclimation.

2. 2.|Changes in metabolite concentrations show that metabolite biosynthesis showed increases after cold acclimation but decreases after warm acclimation.

3. 3.|Similarly, changes in enzyme activities show a preponderance of glycolysis and HMP shunt activity after cold acclimation, while after warm acclimation glycogenolysis, oxidative metabolism and gluconeogenesis predominated.

4. 4.|Higher metabolite concentrations and enzyme activities both before and after thermal acclimation in HM reflect its greater compensatory abilities.

Evidence for the existence of cytoskeleton-bound polysomes in plants.

When conventional, high ionic strength buffers were used for the isolation of polysomes from pea plants, less than 20% were retained in the detergent-insoluble pellet. Reducing Tris, K+ and Mg++ to 10 mM increased retention to 70%, and when a new, microfilament-stabilizing buffer was used, retention increased to 80%. Conditions which favoured polysome pelleting at lower g forces permitted the retention of actin in the pellet. The data are consistent with the hypothesis that higher plants, like animals, contain cytoskeleton-(actin)-bound polysomes.     

The survival and growth of embryonic proprioceptive neurons is promoted by a factor present in skeletal muscle

To date, the neurotrophic factor requirements of developing sensory neurons have been studied using heterogeneous populations of neurons that innervate a wide variety of different sensory structures. To ascertain the particular neurotrophic factor requirements of different kinds of sensory neurons and to determine whether these requirements are related to the type of sensory receptors innervated, it is necessary to study homogeneous preparations of functionally distinct sensory neurons. For this reason I have studied the influence of a soluble extract of skeletal muscle on the survival and growth of proprioceptive neurons isolated from the trigeminal mesencephalic nucleus (TMN) of the embryonic chick. Explants of the TMN and dissociated glia-free cultures of TMN neurons were established from chick embryos of 10 to 18 days incubation (E10 to E18). Skeletal muscle extract prepared from E18 chick pectoral muscle and enriched for neurotrophic activity by ammonium sulfate fractionation promoted marked neurite outgrowth from explants and substantial survival in dissociated cultures established during the period of natural neuronal death in the TMN. In these latter cultures 70 to 80% of the neurons survived and grew in the presence of the extract compared with less than 2% in control cultures. At later ages, following the period of natural neuronal death, these effects were less marked. The neurotrophic activity of extracts prepared from muscle of different ages increased steadily from E10 to E20 (the oldest muscle studied). The active factor is heat labile, trypsin sensitive, and non-dialyzable, it is neither functionally nor immunochemically related to NGF and it has negligible neurotrophic effect on the predominantly cutaneous sensory neuron population of the trigeminal ganglion. These findings demonstrate that skeletal muscle contains a neurotrophic factor which supports the survival and growth of proprioceptive neurons and suggest that this factor has some specificity among functionally distinct kinds of sensory neurons.     

The influence of invertebrates on the breakdown ofPotamogeton pectinatus L. in a coastal marina (Zandvlei,South Africa)

The influence of invertebrates upon the decomposition ofPotamogeton pectinatus L. in a coastal Marina system was examined over 112 days using litter bags. Invertebrate inclusion bags (2 mm mesh, 5 mm holes) registered a dry mass loss of 1% d^–1, while exclusion litter bags (80 µm mesh) produced a 0.4% mass loss d^–1 (a 2.5 fold difference). Losses of ash and N from inclusion bags were greater than those from exclusion bags (p < 0.05). There was a three fold difference between the two treatments in the time taken for litter to breakdown to half the initial stock: T_1/2 for inclusion bags = 43 d, exclusion bags = 130 d. In both treatments, minerals showed an expected rapid loss, due to leaching, with a subsequent slow increase relative to the dry material remaining. A total of nine invertebrate taxa was recorded from inclusion bags, with a peak biomass of 64 mg g^–1 dry massPotamogeton bag^–1 reached at 64 days after immersion. Grazing amphipods,Melita zeylanica Stebbing andAustrochiltonia subtenuis (Barnard), numerically dominated the litter bag fauna, whileM. zeylanica and nymphs of the zygopteran predatorIschnura senegalensis (Rambur) formed most of the biomass. Scanning Electron Microscopy indicated heavy grazing of micro-organisms by invertebrates, with major qualitative differences occurring 112 days after immersion. Invertebrates significantly accelerated the rate of litter breakdown through their feeding activities, assisting fragmentation and thus contributing to plant losses and also by increasing the surface area for microbial colonisation and attack.     

Movements of some indigenous riverine fish in Sri Lanka

The migratory movements of reproductive potamodromus fish established in man-made tropical lakes in the dry zone of Sri Lanka were determined. It was shown that the nine indigenous riverine species do not spawn in these lakes but move into the upstream channels when sexually mature. The pattern of upstream movement was found to be species-specific although most species showed a diurnal periodicity. It is suggested that the development of totally lacustrine forms of riverine species is very unlikely to occur in the man-made lakes of Sri Lanka because of their relatively short duration time.     

Biosynthesis of porphyrins and corrins. 1. 1H and 13C NMR spectra of (hydroxymethyl)bilane and uroporphyrinogens I and III

High-field NMR spectroscopic methods have been applied to study the reactions catalyzed by porphobilinogen (PBG) deaminase and uroporphyrinogen III (uro'gen III) cosynthase, which are the enzymes responsible for the formation of the porphyrin macrocycle. The action of these enzymes in the conversion of PBG, [2,11-13C]PBG, and [3,5-13C]PBG to uro'gens I and III has been followed by 1H and 13C NMR, and assignments are presented. The principal intermediate that accumulated was the correspondingly labeled (hydroxymethyl)bilane (HMB), the assignments for which are also presented.