Cytological evidence of male heterogamety in sex determination of Telmatoscopus albipunctatus (Diptera: Psychodidae)

Telmatoscopus albipunctatus is polymorphic for several polytene chromosome bands. In examining the inheritance of a polymorphic heterochromatin-like band in chromosome IV we verified that it is inherited like a sex-linked factor. There are two types of chromosome IV in regard to this band: one bears a very thick heterochromatin-like band (H⁺), and the other bears a thinner corresponding band (H^–). Three kinds of combinations are found in our stocks: H⁺H⁺, H⁺H^– and H^–H^–. All three combinations can be found in females; however, in males, only the combinations H⁺H^– and H^–H^– are found. Through specific crosses, it was concluded that the sex determining factor is located in chromosome IV in close vicinity to these bands.     

Nucleolar relationships in some Australian Chironomus species

Members of a group of Australian Chironomus species in the pseudothummi complex show wide variation in number and location of nucleolar organizing regions (NORs). The structure of these regions has been examined by phase contrast microscopy and silver banding of salivary gland polytene chromosomes. Presence of nucleoli was also checked on other types of chromosomes in some species. The contribution of the silver banding technique to nucleolar studies in these chironomid chromosomes is discussed. Nucleoli often seem to emerge from groups of (up to 9) bands. Further studies are necessary to confirm the presence of rRNA cistrons in all of these bands. Banding differences, in particular absence of bands from homologous regions of some species which have smaller nucleoli or lack particular nucleoli, have been found. In the case of Ch. tepperi, however, little banding difference is apparent in the 16B region between the N(IV)⁺ and N(IV)^– chromosomes, although in situ hybridization (Eigenbrod 1978) shows a deletion of rRNA cistrons in the N(IV)^– stock. Differences in heterochromatin amount have also been observed at different NORs. A scheme for the evolution of nucleolar-producing regions in this Chironomus group in terms of these and other known chromosomal changes is presented and discussed.     

Effects of planted tree fallows on soil nitrogen dynamics,above-ground and root biomass,N2-fixation and subsequent maize crop productivity in Kenya

The effects of planted fallows of Sesbania sesban (L.) Merr. and Calliandra calothyrsus (Meissner) on soil inorganic nitrogen dynamics and two subsequent maize crops were evaluated under field conditions in the highlands of eastern Kenya. Continuous unfertilised maize, maize/bean rotation and natural regrowth of vegetation (weed fallow) were used as control treatments. The proportion of symbiotic N₂-fixation was estimated by measuring both leaf ¹⁵N enrichment and whole-plant ¹⁵N enrichment by the ¹⁵N dilution technique for Sesbania and Calliandra, using Eucalyptus saligna (Sm.) and Grevillea robusta (A. Cunn) as reference species. Above- and below-ground biomass and N contents were examined in Sesbania, Calliandra, Eucalyptus and Grevillea 22 months after planting. Both the content of inorganic N in the topsoil and the quantity of N mineralised during rainy seasons were higher after the Sesbania fallows than after the other treatments. Compared to the continuous unfertilised maize treatment, both residual crop yields were significantly higher when mineral N (one application of 60 kg N ha^–1) was added. Furthermore, the second crop following the Sesbania fallow was significantly higher than the continuous maize crop. The above-ground biomass of the trees at final harvest were 31.5, 24.5, 32.5 and 43.5 Mg ha^–1 for the Sesbania, Calliandra, Grevillea and Eucalyptus, respectively. For the total below-ground biomass the values for these same tree species were 11.1, 15.5, 17.7, and 19.1 Mg ha^–1, respectively, of which coarse roots (>2 mm), including tap roots, amounted to 70–90%. About 70–90% of the N in Sesbania, and 50–70% in Calliandra, was derived from N₂-fixation. Estimates based on leaf ¹⁵N enrichment and whole-plant ¹⁵N enrichment were strongly correlated. The N added by N₂-fixation amounted to 280–360 kg N ha^–1 for Sesbania and 120–170 kg N ha^–1 for Calliandra, resulting in a positive N balance after two maize cropping seasons of 170–250 kg N ha^–1 and 90–140 kg N ha^–1, for Sesbania and Calliandra, respectively. All the other treatments gave negative N balances after two cropping seasons. We conclude that Sesbania sesban is a tree species well suited for short duration fallows due to its fast growth, high nutrient content, high litter quality and its ability to fix large amounts of N₂ from the atmosphere.     

Ammonia-nitrogen excretion in Daphnia pulex

Ammonia-nitrogen excretion rates were measured in natural summer and cultured populations of Daphnia pulex from Silver Lake, Clay County, Minnesota, USA during 1973. The mean rate of ammonia-nitrogen excretion for the summer populations was 0.20 µg N animal^–1 day^–1 or 5.11 µg N mg^–1 dry body weight day^–1 (N = 80) measured at 15°, 20°, and 25°C. These rates appear to be temperature and weight dependent, but they are probably affected by factors other than temperature and dry body weight. Ammonia-nitrogen excretion rates of Daphnia pulex cultured on Chlamydomonas reinhardi yielded the following relationship with temperature: Log₁₀E = (0.061) T 1.773, where E is µg N animal^–1 day^–1 and T is temperature °C. The ammonia-nitrogen excretion on a mg^–1 dry body weight day^–1 basis was related to temperature according to the following similar expression Log₁₀E = (0.043) T + 0.153, where E is µg N mg^–1 dry body weight day^–1, and T is temperature °C. The length-weight relationship of Daphnia pulex for the summer populations (N = 1583) was log₁₀W = (0.526) Log₁₀L + 1.357, where W is weight in µg and L is length in mm.     

Cycling of nitrogen in modern agricultural systems

Summary Agro-ecosystems have developed from mixed- and multiple-cropping systems with relatively closed N cycles to intensively managed monocultures with large N inputs in the form of commercial fertilizers. Cultivation of increasingly larger areas of land has resulted in substantial losses of soil organic matter and N. Also, the move from slash and burn agriculture to intensively ploughed systems has resulted in losses through increased erosion.The use of N fertilizers has increased rapidly toca. 60 Tg N yr^–1 (1980/81), which is equivalent to at least 40% of the N fixed biologically in all terrestrial systems and 36% more than is fixed in all croplands. On a global scale, the major losses of N from agro-ecosystems are estimated to be: harvest, 30 Tg; leaching, 2 Tg; erosion, 2–20 Tg; denitrification 1–44 Tg; and ammonia volatilization, 13–23 Tg. However, the data base is very crude and several estimates may be wrong by as much as one order of magnitude.Additions of N fertilizers have both direct and indirect effects on soil microorganisms. The possible importance of such effects is briefly discussed and a specific example is given on long-term effects on soil microbial biomass and nitrification rates in 27-year-old cropping systems with different N additions: (i) 0 kg N ha^–1 yr^–1, (ii) 80 kg N ha^–1 yr^–1, (iii) farmyard manureca. 80 kg N ha^–1 yr^–1.Few detailed N budgets exist for agro-ecosystems, despite its major importance as a limiting plant nutrient and the large losses of N from such systems. In conclusion, preliminary nitrogen budgets for four cropping systems (barley receiving 0 or 120 kg N ha^–1 yr^–1; meadow fescue ley with 200 kg N ha^–1 yr^–1 and a lucerne ley) are presented, with special attention to N flow through the soil organisms.Keynote address     

Position of Igl-1, md,and Bst loci on chromosome 16 of the mouse

The experiments described here delineate the position of the chromosome 16 markers Igl-1(immunoglobulin ₁ light chain), md (mahoganoid), and Bst (belly spot and tail), and suggest their location relative to the endogenous proviral locus Akv-2, which is linked within 5.9 centimorgans to Igl-1 (Epstein et al. 1984). The data from an intercross and a three-point backcross detailed herein show the order of these three genes and distances between them to be: centromere-md –10.4 ± 1.6 - Igl-1 –15.6 ± 2.6 - Bst. Using a recombinant chromosome recovered in the intercross, we have constructed a stock homozygous for md and Igl-1 ^b(KpnI^–), that will aid in mapping other genes on chromosome 16.In previous publications, alleles at the structural locus have been referred to as + and –. To conform to standard mouse nomenclature, we propose here the allele symbols a for + and b for –. We refer to genes encoding immunoglobulin molecules (i. e., v, c) in lower case letters, while the gene products are capitalized     

Ribosomal genes not integrated into chromosomal DNA in Drosophila melanogaster

DNA obtained by a gentle lysis procedure from adult Drosophila melanogaster was analyzed by sucrose gradient sedimentation. The major portion of the DNA has an estimated weight of at least 5–10×10⁹ daltons. All of the ribosomal genes are present in this high molecular weight DNA in adult males with one nucleolus organizer or in adult females with two nucleolus organizers as shown by hybridizing fractions of the gradient with ribosomal RNA. In female adults with one nucleolus organizer instead of the usual two, 68% of the ribosomal genes are found in high molecular weight DNA and 32% are found in DNA of smaller size (3×10⁸ daltons). We propose that these latter genes are not integrated into the DNA of the chromosome.     

Decomposition and nutrient release by green manures in a tropical hillside agroecosystem

The decomposition and nutrient release of 12 plant materials were assessed in a 20-week litterbag field study in hillsides from Cauca, Colombia. Leaves of Tithonia diversifolia (TTH) and Indigofera constricta (IND) decomposed quickly (k=0.035±0.002 d^–1), while those of Cratylia argentea (CRA) and the stems evaluated decomposed slowly (k=0.007±0.002 d^–1). Potassium presented the highest release rates (k>0.085 d^–1). Rates of N and P release were high for all leaf materials evaluated (k>0.028 d^–1) with the exception of CRA (N and P), TTH and IND (P). While Mg release rates ranged from 0.013 to 0.122 d^–1, Ca release was generally slower (k=0.008–0.041 d^–1). Initial quality parameters that best correlated with decomposition (P>0.001) were neutral detergent fibre, NDF (r=–0.96) and in vitro dry matter digestibility, IVDMD (r=0.87). It is argued that NDF or IVDMD could be useful lab-based tests during screening of plant materials as green manures. Significant correlations (P>0.05) were also found for initial quality parameters and nutrient release, being most important the lignin/N ratio (r=–0.71) and (lignin+polyphenol)/N ratios (r=–0.70) for N release, the C/N (r=0.70) and N/P ratios (r=–0.66) for P release, the hemicellulose content (r=–0.75) for K release, the Ca content (r=0.82) for Ca release, and the C/P ratio (r=0.65) for Mg release. After 20 weeks, the leaves of Mucuna deerengianum released the highest amounts of N and P (144.5 and 11.4 kg ha^–1, respectively), while TTH released the highest amounts of K, Ca and Mg (129.3, 112.6 and 25.9 kg ha^–1, respectively). These results show the potential of some plant materials studied as sources of nutrients in tropical hillside agroecosystems.     

Comparison of soil nitrogen mineralization and nitrification in a mixed grassland and forested ecosystem in central Taiwan

We used the buried bag incubation method to study temporal patterns of net N mineralization and net nitrification in soils at Ta-Ta-Chia forest in central Taiwan. The site included a grassland zone, (dominant vegetation consists of Yushania niitakayamensis and Miscanthus transmorrisonensis Hayata) and a forest zone (Tsuga chinensis var. formosana and Yushania niitakamensis). In the grassland, soil concentration NH₄ ⁺ in the organic horizon (0.1–0.2 m) ranged from 1.0 to 12.4 mg N kg^–1 soil and that of NO₃ ^– varied from 0.2 to 2.1 mg N kg^–1 soil. In the forest zone, NH₄ ⁺ concentration was between 2.8 and 25.0 mg N kg^–1 soil and NO₃ ^–varied from 0.2 to 1.3 mg N kg^–1 soil. There were lower soil NH₄ ⁺ concentrations during the summer than other seasons. Net N mineralization was higher during the summer while net nitrification rates did not show a distinct seasonal pattern. In the grassland, net N mineralization and net nitrification rates were between –0.1 and 0.24 and from –0.04 to 0.04 mg N kg^–1 soil day^–1, respectively. In the forest zone, net N mineralization rates were between –0.03 and 0.45 mg N kg^–1 soil day^–1 and net nitrification rates were between –0.01 and 0.03 mg N kg^–1 soil day^–1. These differences likely result from differing vegetation communities (C₃ versus C₄ plant type) and soil characteristics.     

Population studies of Laminaria hyperborea from its northern range of distribution in Norway

Eight populations of Laminaria hyperborea from wave-exposed localities in Finnmark (70–71°N) were compared with two populations in Vega (65°N). Standing stock in shallow water (3–5 m) was within the same range in the two areas (6–16 kg fr. wt. m^–2). Both the highest and lowest value of standing stock were registered in Finnmark. Degree of wave-exposure was determined by the numbers of sectors exposed to open sea at each locality, with each sector given a relative wind force value. The most wave-exposed locality in Vega had a slightly higher annual biomass production as a function of plant age compared with the populations from Finnmark within a similar range of wave exposure. Of the two localities in Vega the most wave-exposed one had a higher annual biomass production per plant than the more sheltered one. The most wave-exposed locality examined was situated in Finnmark and exhibited the lowest annual biomass production per plant. Recruits (one-year-old plants) were found at all the localities except at one locality in Finnmark. The plants reached a higher age in Finnmark (13–18 years) than in Vega (8–9 years).     

Effects of nutrient (N,P, C) enrichment,grazing and depth upon littoral periphyton of a softwater lake

Three field experiments were performed in Lake Lacawac, PA to determine the importance of potentially limiting nutrients relative to other factors (grazing, depth) in structuring shallow water algal periphyton communities. All three experiments measured periphyton growth (as chlorophyll-a, AFDM or biovolumes of the algal taxa) on artificial clay flower pot substrates which released specified nutrients to their outer surfaces.Control of standing crop by nutrient supply rate vs. grazing was examined in Expt. I. Substrates releasing excess N and P, together with one of 4 levels of C (as bicarbonate) were placed either inside or outside exclosures designed to reduce grazer densities. Chlorophyll-a rose from 1.1–25.6 µg.cm^–2, and some dominant taxa (e.g., Oedogonium, Nostoc, Anacystis) were replaced by others (e.g., Scenedesmus, Cryptomonas) as bicarbonate supply increased. Reductions in invertebrate density did not significantly affect chlorophyll-a at any of the nutrient levels.Reasons for the species shift were further evaluated in Expt. II, using a minielectrode to measure the elevation of pH within the periphyton mat through photosynthetic utilization of bicarbonate. The pH adjacent to pots diffusing N, P and large quantities of bicarbonate, and supporting high chlorophyll-a densities of 32 µg cm^–2, averaged 10.0 compared to 6.3 in the water column. Pots diffusing only N and P supported 0.7 µg chlorophyll-a cm^–2 and elevated pH to 8.2. We suspect that bicarbonate addition favored efficient bicarbonate users (e.g., Scenedesmus), while inhibiting other taxa (e.g., Oedogonium) because of the attendant high pH.Expt. III was designed to test effects of depth (0.1 m vs. 0.5 m) and N (NH₄ ⁺ vs. NO₃ ^–) upon the growth response to bicarbonate observed in Expts. I and II. Similar standing crop and species composition were noted on pots at 0.1 m vs. 0.5 m. Enrichment with NH₄ ⁺ vs. NO₃ ^– also appeared to have little effect upon the periphyton community.Shallow water periphyton communities in Lake Lacawac, when supplied with sufficient N and P, appear to show a distinctive response to increasing bicarbonate concentration and pH which is robust to moderate variation in grazer densities, distance from the water surface, and the form of N enrichment.     

Nitrogen concentration of cauliflower organs as determined by organ size,N supply,and radiation environment

Data from field experiments carried out in three consecutive years under contrasting N supply and radiation environment altered by artificial shading were used to identify (a) the relationship between N concentration and organ size under conditions of unrestricted N supply and (b) critical levels of soil nitrate (Nmin_crit), where nitrogen concentration of cauliflower organs begin to decline because of N limitations. The decline of N concentrations in cauliflower was analysed at different levels of morphological aggregation, i.e., the whole shoot level, the organ level (leaves, stem, and curd), and within different leaf groups within the canopy. Nmin_crit values (0–60 cm soil depth) for total nitrogen concentration of cauliflower organs leaves, stem and curd were estimated at 85, 93 and 28 kg N ha^–1, respectively. Within the canopy, Nmin_crit values for total N of leaves increased from the top to the bottom from 44 to 188 kg N ha^–1. Nmin_crit values for protein N in leaves from different layers of the canopy were much lower at around 30 kg N ha^–1, without a gradient within the canopy. It is discussed that these differences in Nmin_crit values are most likely a consequence of N redistribution associated with nitrogen deficiency. The decline of average shoot nitrogen concentrations, [Nm] (%N DM), with shoot dry matter, W _sh, (t ha^–1) under conditions of optimal N supply was [Nm]= 4.84 (±0.071) W _sh ^{–0.089(± 0.011)}, r ²=0.67 (±S.E.). The reduction of radiation intensity by artificial shading (60% of control) had no significant influence on total nitrogen concentrations of leaves and only a small influence on protein nitrogen concentrations in lower layers of the canopy. The leaf nitrate nitrogen fraction of nitrogen, f _nitr (–), within the canopy decreased linearly with increased average incident irradiance in different canopy layers (I _av, W PAR m^–2) (f _Nitr. = 0.2456(±0.0188) – 0.0023(±0.0004)I _av, r ² = 0.67.     

The genomic relationship between Glycine max (L.) Merr. and G. soja Sieb. and Zucc. as revealed by pachytene chromosome analysis

Summary This study was conducted with the objective of determining the genomic relationship between cultivated soybean (Glycine max) and wild soybean (G. soja) of the subgenus Soja, genus Glycine. Observations on cross-ability rate, hybrid viability, meiotic chromosome pairing, and pollen fertility in F ₁ hybrids of G. max × G. soja and reciprocals elucidated that both species hybridized readily and set mature putative hybrid pods, generated vigorous F₁ plants, had a majority of sporocytes that showed 18II + 1IV chromosome association at diakinesis and metaphase I, and had a pollen fertility that ranged from 49.2% to 53.3%. A quadrivalent was often associated with the nucleolus, suggesting that one of the chromosomes involved in the interchange is a satellited chromosome. Thus, G. max and G. soja genetic stocks used in this study have been differentiated by a reciprocal translocation. Pachytene analysis of F₁ hybrids helped construct chromosome maps based on chromosome length and euchromatin and heterochromatin distribution. Chromosomes were numbered in descending order of 1–20. Pachytene chromosomes in soybean showed heterochromatin distribution on either side of the centromeres. Pachytene analysis revealed small structural differences for chromosomes 6 and 11 which were not detected at diakinesis and metaphase I. This study suggests that G. max and G. soja carry similar genomes and validates the previously assigned genome symbol GG.Research supported in part by the Illinois Agricultural Experiment Station and U.S. Department of Agriculture Competitive Research Grant (85-CRCR-1-1616)     

A chromosome rearrangement in Neurospora that produces segmental aneuploid progeny containing only part of the nucleolus organizer

In translocation T(ILVL)OY321 of Neurospora crassa a distal portion of the nucleolus organizer chromosome, including ribosomal DNA sequences and the nucleolus satellite, is interchanged with a long terminal segment of IL. When OY321 is crossed by Normal sequence, one-fourth of the meiotic products are segmental aneuploids that contain two copies of the long IL segment and that are deficient for the distal portion of the organizer. Each such product forms a nucleolus and is viable. The complementary aneuploid products are deficient for the IL segment and are therefore inviable. — In crosses of OY321xOY321, each product is capable of making two nucleoli; nucleoli formed by the separated nucleolus organizer parts usually fuse, but most 8-spored asci contain some nuclei in which two separate nucleoli can be seen. One nucleolus is then terminal on its chromosome while the second is interstitial and somewhat smaller. — In crosses of OY321 x Normal, half of the meiotic products are capable of making two nucleoli. However, only about 15% of 8-spored asci have one or more nuclei containing separate nucleoli. At pachytene and later in prophase I, the single fusion nucleolus is associated with three bivalent chromosome segments. Each nucleus of every ascus contains at least one nucleolus, even in asci where some nuclei display two nucleoli. — Crosses of Aneuploid x Normal are usually semibarren, producing a reduced number of ascospores, some of which are inviable. Some aneuploid cultures become fully fertile by reverting to a quasinormal sequence lacking a satellite. In some crosses of Aneuploid x Normal, individual asci may show at prophase I either complete loss, partial loss, or pycnosis of the translocated IL segment. This observation of pycnosis suggests chromosome inactivation. — Growth from aneuploid ascospores is initially slow, but can accelerate to the wild-type rate.     

Feeding rate of slimy sculpin and burbot on young lake charr in laboratory reefs

Synopsis Predation and contaminants are two possible factors in the poor recruitment of young lake charr Salvelinus namaycush in the Great Lakes. We measured the feeding rate of slimy sculpins Cottus cognatus and burbot Lota lota on young lake charr (uncontaminated young from eggs of a hatchery brood stock and contaminated young from eggs of Lake Michigan lake charr) in laboratory test chambers with a cobble substrate. The median daily consumption rate of sculpins for all tests was 2 lake charr eggs (N = 22 tests; 95% confidence interval, 0–13) and 2 lake charr free embryos (N = 31 tests; 95% confidence interval, 0–10). Feeding rate did not differ between hatchery and contaminated prey. Slimy sculpins continued to feed on lake charr when another prey organism, the deepwater amphipod Pontoporeia hoyi, was present. Feeding by burbot on free embryos (4–36 d^–1) increased as the mobility of young increased, but burbot consumed about 10% of their body weight weekly in free-swimming young (140–380 d^–1). Predation on lake charr eggs by sculpins could be considerable over the 100 to 140 d incubation period, and burbot could eat large numbers of free-swimming lake charr as the young fish left the reef. Predation pressure on young lake charr may inhibit rehabilitation of self-sustaining populations of lake charr on some reefs unless a critical egg density has been reached.     

Astragaloside IV and polysaccharide production by hairy roots of Astragalus membranaceus in bioreactors

Hairy roots of Astragalus membranaceus were grown in bioreactors up to 30 l for 20 d. Cultures from a 30 l airlift bioreactor gave 11.5 g l dry wt with 1.4 mg g^–1 astragaloside IV, similar to cultures from 250 ml and 1 l flasks, but greater than yields from a 10 l bioreactor (dry wt 9.4 g l^–1, astragaloside IV 0.9 mg g^–1). Polysaccharide yields were similar amongst the different bioreactors (range 25–32 mg g^–1). The active constituent content of the cells approached that of plant extracts, indicating that large scale hairy root cultures of A. membranaceus has the potential to provide an alternative to plant crops without compromising yield or pharmacological potential.     

Genetic polymorphism of human urine deoxyribonuclease I

Summary A genetic polymorphism of human urine deoxyribonuclease I (DNase I) has been detected by the technique of polyacrylamide gel isoelectric focusing (IEF-PAGE) followed by immunoblotting with anti-DNase I antibody. Family studies showed that the three common phenotypes —DNASE1 1, 1–2, and 2 — and the other four rare phenotypes — DNASE1 1–3, 2–3, 2–4, and 3–4 — represent homozygosity or heterozygosity for four autosomal codominant alleles, DNASE1 ^* 1, ^* 2, ^* 3, and ^* 4. The frequencies of the DNASE1 ^* 1, DNASE1 ^* 2, DNASE1 ^* 3, and DNASE1 ^* 4 alleles in a studied Japanese population were 0.5453, 0.4396, 0.0117, and 0.0034, respectively.     

Using species-specific paleotemperature equations with foraminifera: a case study in the Southern California Bight

Species-specific paleotemperature equations were used to reconstruct a record of temperature from foraminiferal δ¹⁸O values over the last 25 kyr in the Southern California Bight. The equations yield similar temperatures for the δ¹⁸O values of Globigerina bulloides and Neogloboquadrina pachyderma. In contrast, applying a single paleotemperature equation to G. bulloides and N. pachyderma δ¹⁸O yields different temperatures, which has been used to suggest that these species record the surface-to-thermocline temperature gradient. In Santa Barbara Basin, an isotopically distinct morphotype of G. bulloides dominates during glacial intervals and yields temperatures that appear too cold when using a paleotemperature equation calibrated for the morphotype common today. When a more appropriate paleotemperature equation is used for glacial G. bulloides, we obtain more realistic glacial temperatures. Glacial–interglacial temperature differences (G–I ΔT) calculated in the present study indicate significant cooling (8–10°C) throughout the Southern California Bight during the last glacial maximum (LGM). The magnitude of glacial cooling varies from 8°C near the middle of the Southern California Bight (Tanner Basin and San Nicolas Basin) to 9°C in the north (Santa Barbara Basin) and 9.5–10°C in the south (Velero Basin and No Name Basin). Our temperature calculations agree well with previous estimates based on the modern analog technique. In contrast, studies using N. pachyderma coiling ratios, U^k′₃₇ indices, and transfer functions estimate considerably warmer LGM temperatures and smaller G–I ΔT.     

Identification and chromosomal locations of a family of cytochrome P-450 genes for pisatin detoxification in the fungus Nectria haematococca

Summary The ability to detoxify the phytoalexin, pisatin, an antimicrobial compound produced by pea (Pisum sativum L.), is one requirement for pathogenicity of the fungus Nectria haematococca on this plant. Detoxification is mediated by a cytochrome P-450, pisatin demethylase, encoded by any one of six Pda genes, which differ with respect to the inducibility and level of pisatin demethylase activity they confer, and which are associated with different levels of virulence on pea. A previously cloned Pda gene (PdaT9) was used in this study to characterize further the known genes and to identify additional members of the Pda family in this fungus by Southern analysis. DNA from all isolates which demethylate pisatin (Pda⁺ isolates) hybridized to PdaT9, while only one Pda^– isolate possessed DNA homologous to the probe. Hybridization intensity and, in some cases, restriction fragment size, were correlated with enzyme inducibility. XhoI/BamHI restricted DNA from reference strains with a single active Pda allele had only one fragment with homology to PdaT9; no homology attributable to alleles associated with the Pda^– phenotype was found. Homology to this probe was also limited to one or two restriction fragments in most of the 31 field isolates examined. Some unusual progeny from laboratory crosses that failed to inherit demethylase activity also lost the single restriction fragment homologous to PdaT9. At the chromosome level, N. haematococca is highly variable, each isolate having a unique electrophoretic karyotype. In most instances, PdaT9 hybridized to one or two chromosomes containing 1.6–2 million bases of DNA, while many Pda- isolates lacked chromosomes in this size class. The results from this study of the Pda family support the hypothesis that deletion of large amounts of genomic DNA is one mechanism that reduces the frequency of Pda genes in N. haematococca, while simultaneously increasing its karyotypic variation.     

Effect of insulin on exocrine pancreatic secretion in healthy and diabetic anaesthetised rats

This investigation characterised the effects of exogenous insulin on exocrine pancreatic secretion in anaesthetised healthy and diabetic rats. Animals were rendered diabetic by a single injection of streptozotocin (STZ, 60 mg kg^–1 I.P.). Age-matched controls were injected citrate buffer. Rats were tested for hyperglycaemia 4 days after STZ injection and 7–8 weeks later when they were used for the experiments. Following anaesthesia (1 g kg^–1 urethane I.P.), laparotomy was performed and the pancreatic duct cannulated for collection of pure pancreatic juice. Basal pancreatic juice flow rate in diabetic rats was significantly (p < 0.001) increased whereas protein and amylase outputs were significantly (p < 0.001) decreased compared to control rats. Insulin (1 IU, I.P.) produced in healthy rats significant increases in pancreatic flow rate, amylase secretion and protein output compared to basal (p < 0.05). Insulin action also included a reduction in blood glucose (152.7 ± 16.9 mg dl^–1, n= 6, prior to insulin and 42.0 ± 8.4 mg dl^–1, n= 4, 100 min later). In fact, flow rate and glycaemia showed a strong negative correlation (p < 0.01, Pearson). Pretreatment with atropine (0.2 mg kg^–1, I.V.) abolished the effects of insulin on secretory parameters despite a similar reduction in glycaemia; in this series of experiments the correlation between flow rate and blood glucose was lost. In diabetic rats, insulin (4 IU, I.P.) did not modify exocrine pancreatic secretion. There was a fall in blood glucose (467.6 ± 14.0 mg dl^–1, n= 10, prior to insulin and 386.6 ± 43.6 mg dl^–1, n= 7, 120 min later). Rats, however, did not become hypoglycaemic. Similar results were observed in diabetic atropinized rats. The results of this study indicate that the effects of insulin on exocrine pancreatic secretion in anaesthetised healthy rats are mediated by hypoglycaemia-evoked vagal cholinergic activation. (Mol Cell Biochem 261: 105–110, 2004)