Relationship between powdery mildew infection, green leaf area and grain yield of barley

Barley plants (cv. Julia), grown in a greenhouse in large boxes of John Innes compost at a spacing equivalent to 300 plants/m², were exposed to powdery mildew from growth stage (G.S.) 2 (Feekes scale) until maturity. Treatment with ethirimol was used to provide five different epidemic patterns, viz. (i) no mildew, (ii) early mildew, (iii) late mildew, (iv) continuous mildew and (v) continuous mildew but delayed by seed dressing. None of the treatments affected the number of spikelet primordia differentiated on primary shoots, nor were the numbers and sizes of leaves affected. From graphs of mildew and green leaf area (GLA), plotted from G.S. 2 to G.S. 10.5, areas under the curves were determined. There was a strong negative correlation (r= -0.926) between areas under the mildew and GLA curves and it was also clear that reduction in GLA lagged behind disease. Grain yield was strongly correlated with total area under the GLA curve (r= 0.994 for primary shoots and 0.986 for tillers). For values of GLA at specific growth stages the best correlations were found at G.S. 9 (r= 0.967 for primary shoots and 0.992 for tillers). The correlation between total yield of primary shoots and area under the mildew curve was also high (r= 0.953) but for mildew estimates at specific times only those at G.S. 7 were significant (r= 0.980); at G.S. 10.5 there was no significant correlation. It may be postulated from the data that retranslocation of stored photosynthate, produced before anthesis, plays an important role in grain filling and that mildew post anthesis may have little effect on yield. The implications of the results are discussed in relation to methods of loss appraisal and strategies for mildew control.     

Effects of fungicide treatments and variety on development, grain growth and yield of spring barley

The effects of four fungicide treatments for the control of mildew on spring barley were assessed in three field experiments, one in each of the years 1981, 1982, 1983. The fungicide treatments (+/ - triadimenol seed treatment and +/ - triadimefon foliar spray applied during early booting) were chosen to control mildew, and hence affect yield determining processes, at different times in the life of the crop. Two of the experiments also tested different nitrogen amounts and the third tested four varieties differing in their degree of mildew resistance. Mildew appeared too late to affect the production and survival of spikelets and shoots, but reduced average grain weight by reducing the rate of grain growth. Grains in the upper part of the ear had a considerably lower growth rate and final weight than grains in central and basal positions but there was no evidence that the effects of mildew on grain size depended upon grain position within the ear. Mildew incidence increased with increasing nitrogen and varietal susceptibility but there were few significant interactions between these factors and fungicide treatment for grain yield. The degree of mildew control achieved by the seed treatment varied with barley variety. Use of the two successive fungicide treatments did not yield more barley than use of either alone. Amongst varieties, grain positions within the ear and fungicide treatments there was a close correlation between rate of grain growth and final grain weight. Duration of grain growth was not related to rate of grain growth or final grain weight but was inversely correlated with mean temperature during the period of rapid grain growth. The temperature sums during the period of rapid grain growth were similar for the three years and it is suggested that a more precise knowledge of the relationships between mildew incidence, varietal susceptibility and rate of grain growth may enable more accurate predictions to be made about likely yield responses to fungicide treatments.     

Size distribution and maturation of newly replicated DNA through the S and G2 phases of physarum polycephalum

The size distribution of newly made DNA and the dynamics of size maturation of progeny DNA molecules were studied in the synchronous S and G2 phases of Physarum polycephalum. Pulse labeling of DNA and analysis of the products on alkaline sucrose gradients showed that synthesis of primary replication units (which will also be referred to as “Okazaki” fragments) occurred throughout the S period. Pulse and pulse-chase experiments revealed a distinct pattern of size maturation. An apparently linear increase in molecular weight of progeny DNA molecules during the first hour of the S phase occurred at a rate of approximately 4–5 × 10⁵ daltons per min at 26°C, corresponding to the joining of 6–8 Okazaki fragments. The resulting 35–45S (1.1–2.2 × 10⁷ daltons) DNA molecules may correspond to the Physarum “replicon.” The further size increases of the newly made DNA appear to occur in steps, possibly reflecting a clustering of isochronous replicons along the chromatide. These observations are discussed with regard to mechanisms of DNA replication and size maturation.     

Effects of barley mildew on green leaf area and grain yield in field and greenhouse experiments

In 1980 the relationships between mildew severity, green leaf area (GLA) and grain yield of spring barley were examined using greenhouse-grown plants and plants grown in micro-plots in the field. Mildew, by causing premature senescence, reduced GLA and grain yield was strongly correlated with GLA integrated from growth stages 2–10.5 on the Feekes scale. Early mildew attack reduced all yield components (including grain size) even when fungicidal control had eliminated mildew by anthesis. Analyses of culms at anthesis and harvest supported the view that the smaller grain size associated with early mildew attack resulted from a deficiency in carbohydrate stored in culms before anthesis and available for retranslocation to the developing grain. Amounts of total soluble carbohydrate at anthesis and the amounts lost beween anthesis and harvest were both strongly correlated with GLA up to anthesis.     

The phylloplane microflora of ripening wheat and effect of late fungicide applications

Flag leaves and ears of spring wheat cv. Timmo (in 1980) and winter wheat cv. Maris Huntsman in 1981 and 1982 were colonised by a variety of micro-organisms whose numbers increased rapidly between anthesis and harvest. The predominant mycoflora were yeasts, yeast-like fungi and filamentous fungi which included Cladosporium spp., Verticillium lecanii, Alternaria alternata, Fusarium spp. and Epicoccum nigrum. Although similar species were isolated, their relative abundance on flag leaves and ears differed. The fungicide captafol was most effective as a protectant and significantly decreased populations of fungi on flag leaves and ears for 6 and 4 wk respectively, compared to untreated controls. Benomyl and Delsene M (carbendazim + maneb) were the most effective of the systemic sprays and formulations. In general, fungicides affected populations of yeasts, yeast-like fungi and Cladosporium spp. most while Alternaria was tolerant of all treatments. Yields of winter wheat were increased in two seasons by an average 0–2 t ha^-1 (2–4%) following a single late fungicide treatment at G.S. 50 or 60 and 0–41 t ha^-1 (5-1%) when this was combined with an early spray against foliar diseases (G.S. 38–40). Individual treatments increased yield by up to 12% with little difference between applications at G.S. 50 or 60. The yield benefit came mainly from increased 1000-grain weights. Germination of the treated grain was increased only slightly.     

Response of Galleria mellonella (Lepidoptera: Pyralidae) and Tenebrio molitor (Coleoptera: Tenebrionidae) to Spiroplasma citri inoculation

Injections into 4th instar larvae of Galleria mellonella using either in vitro or in vivo inoculum of the BR-6 isolate of Spiroplasma citri, propagated for one to nine passages, caused 5.7 to 24.7% mortality. Weight gain of the larvae injected at their 4th, 5th, and 6th instar was reduced in the first 4 days after inoculation but final pupal weight of the survivors was not significantly affected. Fourth instar larvae pupated within 10 days after the injection, but more larvae (6–13%) injected with 5th- to 9th-passage cultures pupated 5 or more days later than did larvae injected with 1st to 4th-passage cultures (0–3%). As many as one-third of the injected larvae developed into deformed pupae, with some external appendages missing or with a reduced and distorted thorax or abdomen with uneven tanning of the integument. Spiroplasma multiplied to dense concentrations (10⁸ to 10⁹/ml) in hemolymph smears from injected larvae incubated under oil. Larvae of Tenebrio molitor were not susceptible to S. citri by injection or feeding and G. mellonella were not susceptible by feeding. Transmissibility of S. citri by leafhopper vector to celery and periwinkle plants was retained after propagation for nine successive passages during 7 months in a nonhost insect such as Galleria.     

Effects of 1,25-dihydroxyvitamin D3 on cell-cycle kinetics of T 47D human breast cancer cells

The replication of several human and animal cancer cell lines is regulated in vitro and in vivo by 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3], the hormonally active form of vitamin D3. We have examined the effects of concentrations of 1,25-(OH)2D3, which inhibit cellular replication, on the cell-cycle kinetics of a 1,25-(OH)2D3-responsive human breast cancer cell line, T 47D. After 6 or 7 days of treatment, a time period representing approximately five cell population doublings of control cultures, concentrations of 1,25-(OH)2D3 in the range 10(-9) M to 10(-6) M caused a time- and concentration-dependent decrease in cell numbers. Treatment of cells growing in charcoal-treated fetal calf serum with 10(-8) M 1,25-(OH)2D3 for 6 days reduced cell numbers to 49% +/- 9% (n = 9) of control, and this was associated with a marked increase in the proportion of cells in the G2 + M phase of the cell cycle from 9.7% +/- 0.5% (n = 11) to 19.6% +/- 2.3% (n = 9), significant by paired analysis (P less than 0.002). At higher concentrations of 1,25-(OH)2D3 (10(-7)-10(-6) M), there was a concentration-dependent decline in S phase and increases in both G0/G1 and G2 + M phase cells. Detailed analysis of the temporal changes in cell-cycle phase distribution following treatment with 2.5 X 10(-8) and 10(-7) M 1,25-(OH)2D3 showed an initial accumulation of cells in G0/G1 and depletion of S phase cells during the first 24 hr of treatment. This decline in S phase cells was not accompanied by a decline in % G2 + M indicating a transition delay in G2 or mitosis. At the lower dose these changes returned to control values at 48 hr and at later times were associated with a slight but consistent decline in G0/G1 phase and an increase in G2 + M. In contrast cells treated with 10(-7) M 1,25-(OH)2D3 had significantly elevated % G0/G1 cells at days 2 and 3, consistent with a transition delay through G1 phase. This was confirmed in stathmokinetic experiments which demonstrated an approximate sevenfold decrease in the rate of exit of cells from G0/G1 following 4 days of exposure to 10(-7) M 1,25-(OH)2D3. This accumulation of cells in G0/G1 was accompanied by a fall in % S phase cells.(ABSTRACT TRUNCATED AT 400 WORDS)     

Triterpenes and lignans from the leaves of Styrax tonkinensis

Two triterpenes (1 and 2) and eight lignans (3–10) were isolated from the ethyl acetate-soluble fraction of the leaves of Styrax tonkinensis (Pierre) Craib ex Hartw (Styracaceae). Their structures were established as ursolic acid (1), pomolic acid (2), 3,3′-bis(3,4-dihydro-6-methoxy-2H-1-benzopyran) (3), rac-(8α,8′β)-4,4′-dihydroxy-3,3′-dimethoxylignan-9,9′-diyldiacetate (4), (-)-secoisolariciresinol (5), (+)-pinoresinol (6), 4,4′-dihydroxy-3,3′-dimethoxy-9-ethoxy-9,9′-epoxylignan (7), (2S,3R, 4R)-4-[1-ethoxy-1-(4-hydroxy-3-methoxy)phenyl]methyl-2-(4-hydroxy-3-methoxy)phenyl-3-hydroxymethyl-tetrahydrofuran (8), (-)-neo-olivil-(9-O-9″)-seco-isolariciresinol (9) and isolariciresinol (10) based on MS, ¹H-and ¹³C-NMR spectral data. All these compounds (1–10) were firstly isolated from this plant, and compounds 2–5 and 7–9 were reported from the Styrax genus for the first time. Furthermore, the chemotaxonomic significance of the isolated compounds was discussed.     

The Effects of Selenium on Physiological Traits, Grain Selenium Content and Yield of Winter Wheat at Different Development Stages

The paper evaluated the effects of Se application time and rate on physiological traits, grain Se content, and yield of winter wheat by field experiment. Se application significantly increased grain Se content and yield, and the increased amount treated with 20 and 30 mg Se?L^?1 was the highest. At blooming–filling stage, Se application significantly increased grain Se content, but did not affect yield. Chlorophyll content was increased by Se application, and the increased amount at heading–blooming stage was higher than that in wheat leaves at the other stages. At four development stages, Se treatments (except for 10 mg Se?L^?1 at jointing–heading stage) significantly decreased the rate of superoxide (O₂ ^?) radical production. At heading–blooming (except for 50 mg Se?L^?1) and blooming–filling stages, hydrogen peroxide (H₂O₂) content was significantly decreased by Se treatments. The rate of O₂ ^? production and H₂O₂ content at 20 and 30 mg Se?L^?1 was the lowest. Se treatments (except for 10 mg Se?L^?1 at regreening–jointing and blooming–filling stages) also induced an evident decrease in malondialdehyde content. Proline content induced by Se treatments at jointing–heading and heading–blooming stages was higher than that in wheat leaves at regreening–jointing and blooming–filling stages. At four development stages, Se treatments all significantly increased glutathione peroxidase activity, and the treatments with 20 and 30 mg Se?L^?1 also evidently increased reduced glutathione content. These results suggested that Se application at different development stages increased antioxidant capacity of wheat, reduced oxidant stress to some extent, and the effects of Se treatments was the best if Se concentration ranged between 20 and 30 mg Se?L^?1. In addition, Se application time was more beneficial for Se accumulation and yield in wheat grain at heading–blooming stage.     

Interactions between fungicide, plant growth regulator, nitrogen fertiliser applications, foliar disease and yield of winter barley

Leaf size and foliar disease in winter barley increased with increasing total amounts of nitrogen applied to the crop: flag leaf areas increased at an average of 10% per 35 kg N ha^-1 Nitrogen top dressing applied in mid-March (G.S. 31) resulted in larger leaves, more foliar disease, more straw, delayed ear emergence, fewer grains ear^-1 and less grain yield than nitrogen applied in mid-April (G.S. 31). Application of chlormequat at G.S. 30 gave a variable response, but overall it increased fertile stems m-² and crop yield and decreased crop height but had no significant effect on straw yield. Fungicide treatments suppressed foliar disease and improved yield. Yield responses were greater when plant growth regulator and mid-March nitrogen had been applied at sites where more disease prevailed than with April-applied nitrogen. In one of the field experiments, on cv. Sonja, delaying the main nitrogen application until April, without fungicide treatment, gave a similar yield to that provided by nitrogen in March with two or three fungicide sprays.     

Temperature sex determination in the European sea bass,Dicentrarchus labrax (L., 1758) (Teleostei,Perciformes, Moronidae): critical sensitive ontogenetic phase

The temperature sex determination (TSD) mechanism in the European sea bass (Dicentrarchus labrax L.) was studied in respect to: a) the TSD sensitivity during the different developmental stages; and b) the intrapopulation correlation of sex determination with the growth rate up to the end of the TSD-sensitive period. At the stage of half-epiboly, eggs from the same batch were divided into four groups and subjected to different thermal treatments: a) 15 degrees C (G15 group) and b) 20 degrees C (G20 group) up to the middle of metamorphosis stage; c) 15 degrees C up to the end of yolk-sac larval stage and subsequently to 20 degrees C (G15-5 group); and d) 15 degrees C up to the end of the preflexion stage and then to 20 degrees C (G15-10 group). At the end of the treatments, size grading was applied and four additional populations were established from the upper (L) and lower (S) size portions of the G15 and G20 populations: G15L, G15S, G20L, and G20S. During the following growing phase, all populations were subjected to common rearing conditions. The sex ratios of each population were macroscopically determined at 190-210 mm mean total length. Female incidence was significantly affected (P < 0.05) by the different thermal treatments: 66.1% in the G15, 47.1% in the G15-10, 37.6% in the G15-5, and 18.1% in the G20 group. In addition, sex ratio was correlated with the growth rate of the fish up to the end of the TSD-sensitive period, with the larger fish presenting a significantly higher (P < 0.01) female incidence than the smaller fish in both thermal regimes tested: 73.1% in G15L vs. 57% in G15S, and 36.6% in G20L vs. 22.5% in G20S group. Results provide, for the first time, clear evidence that the sea bass is sensitive to TSD during all different ontogenetic stages up to metamorphosis, and that sex ratio is correlated with the growth rate of the fish well before the differentiation and maturation of the gonads.     

The effects of different thymidine concentrations on DNA replication in pea-root cells synchronized by a protracted 5-fluorodeoxyuridine treatment

Single-cell and DNA fiber autoradiography, cytophotometry and velocity sedimentation in alkaline sucrose gradients were used to analyse DNA replication and nascent replicon maturation in 5-fluorodeoxyuridine (FUdR)-synchronized cells of Pisum sativum. The replicon size was not significantly changed by the protracted FUdR treatment. When the synchronized cells were released from the inhibitor, labeled with [³H]TdR for 30 min, and chased in medium containing 1 × 10⁻⁶ M or lower concentrations of cold thymidine, DNA replication stopped after approx. 25% of the genome had replicated, and the nascent strands failed to grow above 9–12 × 10⁶ D single-stranded (ss) DNA. When the cells were chased in medium with 1 × 10⁻⁵ M cold thymidine, the DNA content of the labeled cells steadily increased with time and the size of the nascent molecules grew continuously until replicon size was achieved; then they were accumulated at replicon size until the cells arrived in late S or G2. When the FUdR-synchronized cells were chased in medium containing 1 × 10⁻⁴ M cold thymidine, the size of the nascent strands increased continuously with time, indicating that some neighbouring nascent replicons were joined as soon as they completed their replication. These observations led us to postulate that in FUdR-synchronized cells the rates of chain elongation, cell progression through the S phase and nascent replicon maturation are controlled by thymidine availability.     

Molecular cytogenetic identification of a wheat–<Emphasis Type="Italic">Aegilops geniculata</Emphasis> Roth 7M<Superscript>g</Superscript> disomic addition line with powdery mildew resistance

Aegilops geniculata Roth is an important germplasm resource for the transfer of beneficial genes into common wheat (Triticum aestivum L.). A new disomic addition line NA0973-5-4-1-2-9-1 was developed from the BC₁F₆ progeny of the cross wheat cv. Chinese Spring (CS)/Ae. geniculata SY159//CS. We characterized this new line by morphological and cytogenetic identification, analysis of functional molecular markers, genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), and disease resistance evaluation. Cytological observations suggested that NA0973-5-4-1-2-9-1 contained 44 chromosomes and formed 22 bivalents at meiotic metaphase I. The GISH investigations showed that the line contained 42 wheat chromosomes and a pair of Ae. geniculata chromosomes. EST-STS multiple loci markers and PLUG (PCR-based landmark unique gene) markers confirmed that the introduced Ae. geniculata chromosomes belonged to homoeologous group 7. FISH identification suggested that NA0973-5-4-1-2-9-1 possessed an additional pair of 7M^g chromosomes, and at the same time, there were structural differences in a pair of 6D chromosomes between NA0973-5-4-1-2-9-1 and TA7661 (CS-AEGEN DA 7M^g). After inoculation with powdery mildew (Blumeria graminis f. sp. tritici, Bgt) isolates E09, NA0973-5-4-1-2-9-1 exhibited a powdery mildew resistance infection type different from that of TA7661, and we conclude that the powdery mildew resistance of NA0973-5-4-1-2-9-1 originated from its parent Ae. geniculata SY159. Therefore, NA0973-5-4-1-2-9-1 can be used as a donor source for introducing novel disease resistance genes into wheat during breeding programs with the assistance of molecular and cytogenetic markers.     

Defective retinal depolarizing bipolar cells in regulators of G protein signaling (RGS) 7 and 11 double null mice

Two members of the R7 subfamily of regulators of G protein signaling, RGS7 and RGS11, are present at dendritic tips of retinal depolarizing bipolar cells (DBCs). Their involvement in the mGluR6/Gα(o)/TRPM1 pathway that mediates DBC light responses has been implicated. However, previous genetic studies employed an RGS7 mutant mouse that is hypomorphic, and hence the exact role of RGS7 in DBCs remains unclear. We have made a true RGS7-null mouse line with exons 6-8 deleted. The RGS7(-/-) mouse is viable and fertile but smaller in body size. Electroretinogram (ERG) b-wave implicit time in young RGS7(-/-) mice is prolonged at eye opening, but the phenotype disappears at 2 months of age. Expression levels of RGS6 and RGS11 are unchanged in RGS7(-/-) retina, but the Gβ5S level is significantly reduced. By characterizing a complete RGS7 and RGS11 double knock-out (711dKO) mouse line, we found that Gβ5S expression in the retinal outer plexiform layer is eliminated, as is the ERG b-wave. Ultrastructural defects akin to those of Gβ5(-/-) mice are evident in 711dKO mice. In retinas of mice lacking RGS6, RGS7, and RGS11, Gβ5S is undetectable, whereas levels of the photoreceptor-specific Gβ5L remain unchanged. Whereas RGS6 alone sustains a significant amount of Gβ5S expression in retina, the DBC-related defects in Gβ5(-/-) mice are caused solely by a combined loss of RGS7 and RGS11. Our data support the notion that the role of Gβ5 in the retina, and likely in the entire nervous system, is mediated exclusively by R7 RGS proteins.     

Preventative and Curative Applications of Verticillium lecanii for Biological Control of Cucumber Powdery Mildew

The effect of timing of the application of the mycoparasite , Verticillium lecanii, on cucumber powdery mildew , Sphaerotheca fuliginea, was studied in a rooted cucumber leaf bioassay . The mycoparasite was applied at different times before and after mildew inoculation . At near - to maximum humidity ( 95% relative humidity) , early preventative (9 and 5 days before mildew inoculation) and early curative control treatments (2 days after mildew inoculation) gave considerable reduction in mildewed leaf areas , while late curative treatments resulted in greater mildewed leaf area but ultimately a reduced amount of healthy mildewed leaf area ( 20 % ) . Appropriate timing of biocontrol treatment application of V. lecanii is important to achieve good control .     

Optimization of the Salmonella/mammalian microsome assay for urine mutagenesis by experimental designs

Assessing urine mutagenicity with the Salmonella mutagenicity test is often limited by the volumes of the samples. Optimization of the assay was performed with factorial and Doehlert designs. Two fractional factorial designs 2^3-1 (3 factors, 4 experiments) were used to estimate the main effects of the percent S9 in the mix, the time of liquid incubation, the inoculum size and the growth conditions. A Doehlert design (3 factors, 13 experiments) was used to study the main effects and the interactions of the NADP, G6P and S9 in the mix. The positive markers were benzo[a]pyrene (BaP, 0.3 μg/plate) and a pool of smokers' urine (SU, 1.25 ml equivalent/plate). The response was limited to the induction factor (IF, number of induced revertants/number of spontaneous revertants) with Salmonella typhimurium TA98. The optimal conditions for BaP were: a 60 min period of liquid incubation and a volume of 0.1 ml (approx. 10⁸ cells/plate) of an overnight culture grown in 50 ml of Nutrient Broth No. 2 from a 250 ml flask. The S9 mix (0.1 ml, final volume) included 1.5% of S9, 1.0 mM NADP and 4.4 mM G6P. The maximal IF was 15.79. The optimal conditions for SU were: a 60 min period of liquid incubation and a volume of 0.1 ml (approx. 10⁸ cells/plate) of an overnight culture grown in 7 ml of Nutrient Broth No. 2 from a 20 × 180 mm tube. The S9 mix (0.1 ml, final volume) included: 4% S9, 4.2 mM NADP and 5.2 mM G6P. The maximal I7F was 10.95. These optimal conditions did not modify the spontaneous frequencies of the tester strains: TA97a, TA98, TA100 and TA102. The dose-response curves of mutagenic urine samples were found to be non-linear. This micromethod required 8-fold less urine sample and 12.5-fold less liver homogenate as compared to the standard plate incorporation assay and was from 6.2- to 11.8-fold more sensitive to evaluate urine mutagenicity. The sensitivity of this technique was found to be limited to individuals smoking more than approx. 5 cigarettes/day by the standard extraction-concentration procedure.     

A single site-specific trans-opened 7,8,9,10-tetrahydrobenzo[a]pyrene 7,8-diol 9,10-epoxide N2-deoxyguanosine adduct induces mutations at multiple sites in DNA

Site-specific mutagenicity of trans-opened adducts at the exocyclic N(2)-amino group of guanine by the (+)-(7R,8S,9S,10R)- and (-)-(7S,8R,9R,10S)-enantiomers of a benzo[a]pyrene 7,8-diol 9,10-epoxide (7-hydroxyl and epoxide oxygen are trans, BPDE-2) has been determined in Chinese hamster V79 cells and their repair-deficient counterpart, V-H1 cells. Four vectors containing single 10S-BPDE-dG or 10R-BPDE-dG adducts positioned at G(0) or G(-1) in the analyzed 5'-ACTG(0)G(-1)GA sequence of the non-transcribed strand were separately transfected into the cells. Mutations at each of the seven nucleotides were analyzed by a novel primer extension assay using a mixture of one dNTP complementary to the mutated nucleotide and three other ddNTPs and were optimized to quantify levels of a mutation as low as 1%. Only G --> T mutations were detected at the adducted sites; the 10S adduct derived from the highly carcinogenic (+)-diol epoxide was 40-50 and 75-140% more mutagenic than the 10R adduct in V79 and V-H1 cells, respectively. Importantly, the 10S adducts, but not the 10R adducts, induced separate non-targeted mutations at sites 5' to the G(-1) and G(0) lesions (G(0) --> T and C --> T, respectively) in both cell lines. Neither the T 5' to G(0) nor sites 3' to the lesions showed mutations. Non-targeted mutations may enhance overall mutagenicity of the 10S-BPDE-dG lesion and contribute to the much higher carcinogenicity and mutagenicity of (+)-BPDE-2 compared with its (-)-enantiomer. Our study reports a definitive demonstration of mutations distal to a site-specific polycyclic aromatic hydrocarbon adduct.     

In the higher plant Pisum sativum maturation of nascent DNA is blocked by cycloheximide, but only after 4-8 replicons are joined.

Velocity sedimentation in alkaline sucrose gradients, single cell autoradiography and cytophotometry were used to determine if protein synthesis is required for the maturation of nascent replicons to chromosomal-sized molecules in cultured pea-root cells. The results obtained showed that cycloheximide at 5 and 10 microgram/ml, added either before or during labeling with tritiated thymidine, blocked maturation of nascent DNA at an intermediate size of 72-140 X 10(6) daltons single-stranded DNA. To reach this size, nascent replicons - which are 18 X 10(6) daltons single-stranded DNA each - were replicated and groups of 4-8 replicons were joined even though protein synthesis was reduced to 15% of the control. Further maturation of the nascent molecules to chromosomal size, however, was prevented and this resulted in the accumulation of nascent molecules in the 72-140 X 10(6) daltons range. The experiments also showed that the joining of nascent replicons is not an absolute function of late S or G2 phase of the cell cycle, since cells treated with cycloheximide and blocked in mid-S phase had nascent DNA of a size corresponding to 4-8 joined replicons. Finally, the results support the hypothesis that at least one step in the process of nascent DNA maturation may require replication, during late-S phase, of DNA segments that are interspersed within replicon-clusters that replicate early in the S phase.     

Effects of powdery mildew on cucumber yields, and its chemical control

Effects of Sphaerotheca fuliginea were deduced from experiments in which cucumber foliage, exposed to naturally occurring inocula, was fungicidally sprayed. Yields increased as the incidence of mildew decreased but the relation was sometimes affected by differing amounts of phytotoxicity. The fungitoxicity and phytotoxicity of a range of chemicals applied as high-volume sprays, fumigants or soil drenches, were tested. Non-phytotoxic concentrations of some fungicides adequately protected cucumber foliage from subsequent attack but were often insufficient to eradicate established infections, the increased amounts needed for this being phytotoxic. Powdery mildew was controlled more effectively when drazoxolon and quinomethionate were applied as sprays with 0·1 and 0·02% a.i. respectively, than when used as fumigants at 2–4 and 1·3 g/28·3 m³(= 1000 ft³). Sprays of drazoxolon (0·1% a.i.) increased yields from 7·0 to 11·2 kg/ plant during 8 weeks picking and in another experiment weights of fruit were increased by applying quinomethionate (0·02% a.i.) from 10·8 to 52·2 kg/plot of four plants. In the former experiment appreciable amounts of phytotoxicity and infection were tolerated before yields decreased but, in the latter, yields were inversely proportional to numbers of dead leaves which were directly related to the incidence of mildew. Spraying with quinomethionate, or drazoxolon plus tetradifon (0·012% a.i.) increased numbers of female flowers from 59·5 in the mildewed controls to 93·0 and 142·8 per plant respectively, and of these 17·6, 32·8 and 16·2 % subsequently produced marketable cucumbers. In addition to decreasing yields, severe S. fuliginea infestations were associated with increased numbers of misshapen cucumbers. Increasing sulphur concentrations from 0·5 to 1·0g/28·3m³during nightly fumigations significantly decreased mildew incidence and increased (a) yields from 35·8 to 52·9 kg during 5 weeks picking, and (b) the proportion of high-quality cucumbers from 49 to 63 %. Similar trends occurred in another trial where sulphur concentrations were increased from 0·75 to 1·0 g/28·3 m³, but comparisons with fortnightly sprays of 0·1% drazoxolon suggest that these sulphur concentrations caused some damage.     

20-Hydroxyecdysone accelerates the flow of cells into the G1 phase and the S phase in a male accessory gland of the mealworm pupa (Tenebrio molitor)

The cells of the bean-shaped accessory glands of mealworms proliferate through the first 7 days of the 9-day pupal stage. Immediately after larval-pupal ecdysis, 25-27% of the cells were in the G1 phase, 60-65% were in the G2 phase, and the balance were in S phase. Over the first 4 days of normal development, the S fraction gradually increased, to reach its highest level in the mid-pupa at the time of the major ecdysteroid peak (Delbecque et al., 1978). Thereafter, the S fraction declined until over 95% of the cells had accumulated in G2 on Day 8. When 0-day pupal glands were explanted into Landureau's S-20 medium for 6 days, the G1 fraction remained fairly constant (25-30%) while S and the G2 fractions fluctuated. On the first day in vitro, the G2 fraction declined and the S fraction rose. On the second day in basal media, the S fraction fell and G2 rose correspondingly until 70% of the cells reached G2 when cycling stopped on the third day. With addition of 20-hydroxyecdysone to 0-day cultures, the S fraction increased quite sharply. It remained large for all 6 days of the experiment in the continuing presence of hormone. A 1-day pulse of hormone produced a transient increase in S. We blocked cell cycling with hydroxyurea in a stathmokinetic experiment and showed that 20-hydroxyecdysone accelerated the flow of cells from the G2 phase to the G1 phase by 2.5-fold. An increase in the G1 fraction was detected within 10 hr of hormone administration and the effect was dose-dependent with an ED50 of 5 X 10(-7) M for 20-hydroxyecdysone. We conclude that 20-hydroxyecdysone acts at a control point in the G2 phase. Incubation of the glands with 20-hydroxyecdysone for only 30-60 min followed by washout stimulated the flow from G2 to G1 and the effect persisted after transfer of the tissues to hormone-free media. Dose-dependent stimulation also occurred with ponasterone A (ED50 3 X 10(-9] but not with cholesterol.