Yield efficiency of the X-ray mutant svalöf's ‘pallas barley’

Summary Svalöf's Pallas barley is an X-ray mutant of the high-yielding Bonus barley variety. Both varieties were released by the Swedish Seed Association, Svalöf, and have gained wide distribution. Pallas barley arose as an erectoides mutant in 1947. After careful testing it was approved in 1958 as an original Swedish variety. It soon became widespread over great parts of western Europe owing to its high productivity, pronounced lodging resistance and high nitrogen utilization.Danish tests of barley varieties are recorded and published yearly. From 1958 when Pallas entered the Danish trials, and up to and including 1964, its parent Bonus barley was the official Danish standard variety. In 1965 it was replaced by Pallas barley. A careful biometrical comparison of the two varieties has been made with regard to grain yield, lodging resistance, straw height and strawproduction. In addition, the influence of increased lodging resistance on yield as well as the variance of yield and lodging have been analysed.

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Zusammenfassung Die Gerstensorte Svalöfs Pallas geht auf eine im Jahre 1947 entstandene röntgeninduzierte erectoides-Mutante der ertragreichen Sorte Bonus zurück. Beide Sorten wurden von dem Schwedischen Saatzuchtverein in Svalöf gezüchtet. Nach eingehender Prüfung wurde sie 1958 als schwedische Zuchtsorte zugelassen und erlangte durch ihre gute Ertragsfähigkeit, die wesentlich verbesserte Standfestigkeit und gute Stickstoffverwertung weite Verbreitung in großen Teilen Westeuropas.Die Ergebnisse der dänischen Gersten-Sortenprüfungen werden alljährlich veröffentlicht. 1958 wurde Pallas erstmalig in diese Versuche einbezogen, bei denen bis einschließlich 1964 ihre Ausgangssorte Bonus als offizielle Standardsorte diente. Sie wurde 1965 durch Pallas ersetzt. Beide Sorten wurden in bezug auf Ertragsfähigkeit, Standfestigkeit, Strohlänge und Strohertrag sorgfältigen biometrischen Prüfungen unterzogen, weiterhin wurden der Einfluß erhöhter Standfestigkeit auf die Ertragsfähigkeit untersucht sowie Ertrags- und Standfestigkeitsvarianzen geschätzt.

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This paper is cordially dedicated to Professor Hans Stubbe in appreciation of his pioneer research on induced mutations in cultivated plants.     

A root hairless barley mutant for elucidating genetic of root hairs and phosphorus uptake

This paper reports a new barley mutant missing root hairs. The mutant was spontaneously discovered among the population of wild type (Pallas, a spring barley cultivar), producing normal, 0.8 mm long root hairs. We have called the mutant bald root barley (brb). Root anatomical studies confirmed the lack of root hairs on mutant roots. Amplified Fragment Length Polymorphism (AFLP) analyses of the genomes of the mutant and Pallas supported that the brb mutant has its genetic background in Pallas. The segregation ratio of selfed F₂ plants, resulting from mutant and Pallas outcross, was 1:3 (–root hairs:+root hairs), suggesting a monogenic recessive mode of inheritance.In rhizosphere studies, Pallas absorbed nearly two times more phosphorus (P) than the mutant. Most of available inorganic P in the root hair zone (0.8 mm) of Pallas was depleted, as indicated by the uniform P depletion profile near its roots. The acid phosphatase (Apase) activity near the roots of Pallas was higher and Pallas mobilised more organic P in the rhizosphere than the mutant. The higher Apase activity near Pallas roots also suggests a link between root hair formation and rhizosphere Apase activity. Hence, root hairs are important for increasing plant P uptake of inorganic as well as mobilisation of organic P in soils.Laboratory, pot and field studies showed that barley cultivars with longer root hairs (1.10 mm), extracted more P from rhizosphere soil, absorbed more P in low-P field (Olsen P=14 mg P kg^–1 soil), and produced more shoot biomass than shorter root hair cultivars (0.63 mm). Especially in low-P soil, the differences in root hair length and P uptake among the cultivars were significantly larger. Based on the results, the perspectives of genetic analysis of root hairs and their importance in P uptake and field performance of cereals are discussed.     

Phytotron cultivation of early barley mutants

Conclusions The authors have tried to gather data which permit some information on the between and within locus reactions of induced early barley mutants to different photo- and thermoperiods. Eight mutant cases, showing rather drastic earliness in field cultivation and representing three different gene loci, were examined in phytotron experiments according to routine methods of cultivation. One of the mutants, mat-a ⁸, has been released as an original Swedish barley variety under the name of Svalöf's Mari. In a previous publication (Dormling et al., 1966) this mutant was compared to its parent variety, Svalöf's Bonus, under 30 different climatic conditions. In the present investigation three photoperiods (24, 16 and 8 hours of artificial light) were combined with three suitable thermoperiods (20-15°, 20-10° and 15-10°C).The results indicate that photoperiodic insensitivity, with regard to ear formation and heading capacity, as well as kernel production, is of rather frequent occurrence in connection with drastically early mutants in barley. Four out of eight induced mutants give a more or less pronounced insensitivity. Three of the four insensitive mutants represent locus a, one belongs to locus b. Of the two c-mutants none was insensitive; both were on the contrary pronounced long-day types.Photo- and thermoperiods interact in various ways. This is especially clear in the c-mutants just mentioned, which have a high generative productivity and efficiency at continuous light and high thermoperiods. They produce no grain but considerable vegetative matter at 8 hours of light, independently of thermoperiod, as well as at 16 hours of light with high temperatures. In fact, mutants of loci a and c differ strikingly with regard to their relations to the climatic conditions applied. The insensitive mutant b ¹³ is remarkably similar to the mutant a ¹², but its resemblance to the sensitive mutants b ⁷ and b ¹⁰ of the same locus is evidenced by its high average internode number.It ought to be pointed out here that the mutants of the three gene loci analysed in this study can be distinguished phenotypically with regard to morphological as well as physiological properties, in the field as well as in phytotron cultivation. The c-mutants are especially characteristic. However, there also seem to be clear differences in reaction between the allelic mutants of a locus. In fact, all eight mutants studied seem to react more or less differently.The insensitive mutant a ⁸, which has been released into practice, is also widely used in recombination work, and successful segregates have been isolated. The characteristics of a ⁸, which make the mutant valuable in practice, are also found in phytotron experimentation, specially with regard to earliness, generative efficiency and yield. Also the semidwarf habit and the insensitivity to changes in photo- and thermoperiods readily show up.This investigation has been generously supported by the Swedish Research Council of Forestry and Agriculture.     

Phosphorus (P) uptake and growth of a root hairless barley mutant (bald root barley, brb) and wild type in low- and high-P soils

The recently isolated root‐hairless mutant of barley (Hordeum vulgare L), bald root barley, brb offers a unique possibility to quantify the importance of root hairs in phosphorus (P) uptake from soil. In the present study the ability of brb and the wild‐type, barley genotype Pallas producing normal root hairs to deplete P in the rhizosphere soil was investigated and the theory of diffusion and mass flow applied to compare the predicted and measured depletion profiles of diffusible P. Pallas depleted twice as much P from the rhizosphere soil as brb. The P depletion profile of Pallas uniformly extended to 0.8 mm from the root surface, which was equal to the root hair length (RHL). The model based on the theory of diffusion and mass flow explained the observed P‐depletion profile of brb, and the P depletion outside the root‐hair zone of Pallas, suggesting that the model is valid only for P movement in rhizosphere soil outside the root‐hair zone. In low‐P soil (P in soil solution 3 µm ) brb did not survive after 30 d, whereas Pallas continued to grow, confirming the importance of root hairs in plant growth in a P‐limiting environment. In high‐P soil (P in soil solution 10 µm ) both brb and Pallas maintained their growth, and they were able to produce seeds. At the high‐P concentration, RHL of the Pallas was reduced from 0.80 ± 0.2 to 0.68 ± 0.14 mm. In low‐P soil, P‐uptake rate into the roots of Pallas was 4.0 × 10^?7 g mm^?1 d^?1 and that of brb was 1.9 × 10^?7 g mm^?1 d^?1, which agreed well with the double amount of P depleted from the rhizosphere soil of Pallas in comparison with that of brb. In high‐P soil, the P uptake rates into the roots of brb and Pallas were 3.3 and 5.5 × 10^?7 g mm^?1 d^?1, respectively. The results unequivocally confirmed that in a low‐P environment, root hairs are of immense importance in P acquisition and plants survival, but under high‐P conditions they may be dispensable. The characterization of phenotypes brb and Pallas and the ability to reproduce seeds offers a unique possibility of molecular mapping of QTLs and candidate genes conferring root‐hair formation and growth of barley.     

RFLP mapping of the barley homeotic mutant lax-a

The lax-a homeotic mutant of barley has flowers in which lodicules are replaced by stamens (giving five stamens per flower). RFLP mapping of an F₂ population from a Bonus lax-a ¹ x H. spontaneum cross showed that the mutation was on the short arm of chromosome 7(5H), closely linked to the centromere. An additional F₂ population was used to show that the lax-a mutation gave the five-stamen phenotype in all flowers of 6-rowed spikes and that hoods were elevated and reduced in size in lax-a/Hooded double-mutant plants.     

Colonization potential of bacteria in the rhizosphere

The effect of inoculum density on growth and steady-state populations of aPseudomonas sp., aMycoplana sp., and aCurtobacterium sp. in the rhizosphere if gnotobiotic barley plants was studied. Inoculation of sterile barley seedling at concentrations of about 1×10³, 1×10⁵ and 1×10⁷ viable cells (mg dry wt root)^–1 resulted in rapid colonization; maximum populations of about 5×10⁷ viable cells (mg dry wt root)^–1 developed in each case. We define this maximum population as the colonization potential. Measurement of growth of known rhizosphere bacteria might be a useful index of the amount of available carbon and energy lost by growing roots.     

Establishment of phosphate-solubilizing strains of Azotobacter chroococcum in the rhizosphere and their effect on wheat cultivars under green house conditions

A pot experiment was conducted in the green house to investigate the establishment of phosphate solubilizing strains of Azotobacter chroococcum, including soil isolates and their mutants, in the rhizosphere and their effect on growth parameters and root biomass of three genetically divergent wheat cultivars (Triticum aestivum L.). Five fertilizer treatments were performed: Control, 90 kg N ha^—1, 90 kg N + 60 kg P₂O₅ ha^—1, 120 kg N ha^—1 and 120 kg N + 60 kg P₂O₅ ha^—1. Phosphate solubilizing and phytohormone producing parent soil isolates and mutant strains of A. chroococcum were isolated and selected by an enrichment method. In vitro phosphate solubilization and growth hormone production by mutant strains was increased compared with soil isolates. Seed inoculation of wheat varieties with P solubilizing and phytohormone producing A. chroococcum showed better response compared with controls. Mutant strains of A. chroococcum showed higher increase in grain (12.6%) and straw (11.4%) yield over control and their survival (12—14%) in the rhizosphere as compared to their parent soil isolate (P4). Mutant strain M37 performed better in all three varieties in terms of increase in grain yield (14.0%) and root biomass (11.4%) over control.     

Comparative mapping of a gibberellic acid-insensitive dwarfing gene (Dwf2) on chromosome 4HS in barley

 We report the genetic mapping of Dwf2, a dominant gibberellic acid (GA₃)-insensitive dwarfing gene which has been previously described to cause a very short growth habit in barley (Hordeum vulgare) mutant ‘93/B694’. Using RFLP and microsatellite markers we performed segregation analysis in an F₂ population comprising 86 individuals developed from a cross of ‘93/B694’ (Dwf2) with ‘Bonus M2’ (dwf2). Dwf2 was mapped on the short arm of barley chromosome 4H proximal to microsatellite marker XhvOle (5.7 cM) and distal to RFLP marker Xmwg2299 (18.3 cM). The genetic localization of the Dwf2 gene at a homoeologous position to the multiallelic Rht-B1 and Rht-D1 loci in wheat suggests synteny of GA-insensitive dwarfing genes within the Triticeae. Moreover, the extremely prostrate growth habit exhibited in barley ‘93/B694’ (Dwf2) resembles that of wheat plants carrying the genes Rht-B1c (Rht3) or Rht-D1c (Rht10). Received: 1 July 1998 / Accepted: 17 September 1998     

Chromosome mapping in barley by means of telotrisomic analysis

Summary A total of 37 genetic markers located in chromosomes 2, 3, 4 and 5 were associated with specific arms by means of telotrisomic analysis in five telotrisomics (Triplo 2 L, 2 S, 3 S, 4 S, 5 L) of barley (Hordeum vulgare L.). The genes v, gp (= gp 2), li, gs 5, tr and msg2 showed a trisomic ratio with Triplo 2 L indicating that these genes were on the long arm of chromosome 2. A disomic ratio was obtained for genes wst 4, gs 5, and v with Triplo 2 S, confirming that these genes were on the long arm of chromosome 2(2 L). A disomic ratio was observed for genes e, f(= lg), sk, and gs6 with Triplo 2 L. Two genes, f(= lg) and gs6 showed a trisomic ratio with Triplo 2S. These results indicated that genes e, f(= lg), sk, and gs 6 are on the short arm of chromosome 2 (2S). Since only one telocentric chromosome was available for chromosome 3, 4 and 5, most of the well-mapped marker genes were tested with those telocentric chromosomes. The genes cu 2, uz, wst, als, gs 2, zb,f2, and cer-zn ³⁴⁸ showed trisomic ratio with the telocentric for chromosome 3. These genes were located on the short arm of chromosome 3 (Robertson 1971). This indicated that the telocentric chromosome is for the short arm of chromosome 3(3 S). A disomic ratio was obtained for genes yst, x _c, al, yst2, a _n, ari-a ⁶ and x _s, indicating that these genes are on the long arm of chromosome 3. Two genes, f9 and K, showed trisomic ratio with the telocentric chromosome for 4, while genes gl(= gl2), br2, yh, lg 3, lg 4 and lk 5 showed disomic ratios. This indicated that the telocentric chromosome is for the short arm of chromosome 4. Two genes, fs 2 and g, were studied with Triplo 5 L. Both showed trisomic ratio, indicating that fs 2 and g are located on Triplo 5 L. The centromere position (C) on chromosome 2, 3 and 4 was thus located as (the left side of C is the short arm and the right is the long arm): chromosome 2: f — sk — gs6 — e — C — gs5 — msg2 — wst4 — v — gp — li — tr; chromosome 3: f2 — cer-zn ³⁴⁸ — uz — gs2 — als — cu2 — wst — zb — C — yst — x _c — al — yst2 — a _n — ari-a ⁶ — x _s; chromosome 4: f9 — K — C — lg4 — lg ³ — gl2 — br2 — lk5 — yh. The centromere position on chromosome 5 was not precisely located.Contribution from the Department of Agronomy, Published with the approval of the director of the Colorado State University Experiment Station as Scientific Series Paper No. 2606. This research was supported in part by by NSF Grant GB 4482X and GB 30 493 to T. Tsuchiya and Colorado State University Experiment Station Hatch Project     

Gull contributions of phosphorus and nitrogen to a Cape Cod kettle pond

Nutrient excretion rates and the annual contribution of P from the feces of the gulls Larus argentatus and L. marinus (and of N from L. argentatus) to the nutrient budget of Gull Pond (Wellfleet), a soft water seepage lake, have been estimated. Intensive year-round gull counts by species were combined with determinations of defecation rate and the nutrient content of feces to quantitatively assess the P loading rates associated with regular gull use of this coastal pond on a seasonal and annual basis. Total P loading from gulls was estimated to be 52 kg yr^–1, with 17 kg from L. argentatus and 35 kg from L. marinus, resulting from about 5.0 × 10⁶ h yr^–1 and 1.7 × 10⁶ h yr^–1 of pond use. This compares with P loading estimates of 67 kg yr^–1 from upgradient septic systems, 2 kg yr^–1 from precipitation and 3 kg yr^–1 from unpolluted ground water. Fifty-six percent of annual gull P loading was associated with migratory activity in late fall. Estimated annual N loading by L. argentatus was 14 kg TKN, 206 g NO₃-N, and 1.85 g g NH₃-N.     

A molecular genetic linkage map of mouse chromosome 18, includingspm,Grl-1, Fim-2/c-fms,andMbp

Restriction endonuclease fragment length variations (RFLV) were detected in mice with DNA probes for myelin basic protein (Mbp), glucocorticoid receptor-1 (Grl-1), and Friend MuLV integration site-2 (Fim-2). RFLV of theMbp gene were found inSacI restriction patterns, RFLV of theGrl-1 gene were found inEcoRV patterns, and RFLV of theFim-2 were found inBglII patterns. A three-point backcross was carried out by the backcross mating (C57BL/KsJ-spm/spm × MOL-MIT)F₁ males × C57BL/KsJ-spm/spm; spm is an autosomal recessive gene causing sphingomyelinosis. From the results,spm, Grl-1, Fim-2, andMbp loci were mapped on chromosome 18, and the following order of genes is proposed, with distances between genes in parentheses: centromere—spm—(7.8 cM)—Grl-1—(7.8 cM)—Fim-2—(39.1 cM)—Mbp—telomere. All laboratory strains and two European subspecies (Mus mus domesticus andM. m. brevirostris) carry theGrl-1 ^a ,Fim-2 ^a , andMbp ^a alleles. In contrast, another wild subspecies from Europe (M. m. musculus) and some Asian subspecies (M. m. molossinus, Chinese mice of wild origin, andM. m. yamashinai) carry theGrl-1 ^b ,Fim-2 ^b , andMbp ^b alleles. Onlycastaneus strains carry the intermediate combination of theGrl-1 ^b ,Fim-2 ^a , andMbp ^b alleles.     

Intercellular communication and the control of growth: XII. Alteration of junctional permeability by simian virus 40. roles of the large and smallT antigens

Summary We studied the action of temperature-sensitive mutant simian virus 40—a transformation-inducing DNA virus—on the junctional permeability to mono-, di- and triglutamate in rat embryo-, pancreas islet (epithelia)-, and 10T1/2 cell cultures. Junctional permeability was reduced (reversibly) in the transformed state. To dissect the genetics of this alteration, we used two kinds of mutant virus DNA. One kind had a temperature-sensitive mutation on theA gene, rendering the largeT antigen (the gene product) thermolabile (T ⁺ T ^–). The other had a deletion on theF gene, in addition, abolishing (permanently) the expression of the littlet antigen (t ^–). The junctional alteration occurred in the conditionT ⁺ t ⁺, but not in the conditionsT ^– t ⁺,T ⁺ t ^– orT ^– t ^–. Both antigens, thus, are necessary for this junctional alteration—a genetic requirement identical to that for decontrol of growth (but distinct from that of the cytoskeletal alteration).     

Genetic mapping,distribution, and properties of an aconitase isozyme in Anopheles albimanus (Diptera:Culicidae)

Electrophoretic analysis of the developmental stages and tissues of Anopheles albimanus showed that qualitatively similar allozymes of aconitase (Acon-2) occur at all stages, and the enzyme is widespread in every larval and adult tissues. Relative heat stabilities of the allozymes were investigated by electrophoresis of heated aqueous extracts and by heating the enzyme in situ in acrylamide gels after electrophoretic separation in Tris-citrate and Tris-maleate buffer systems. The pupal aconitase in the crude extract is more stable to heat than the larval and adult enzyme. The presence of citrate ions in the gel increased the stability of aconitase to heat. Studies of substrate specificities indicated that cis-aconitic acid is the best substrate but citric acid can also serve as a substrate. Zymograms developed with isocitric acid as a substrate showed no aconitase electromorphs and produced only isocitrate dehydrogenase bands. Aconitase has a pH optimum of 8.0 and this enzyme is completely inhibited if treated in situ with ethylenediaminetetra-acetic acid (EDTA), p-chloromercuribenzoate (PCMB), and urea at concentrations higher than 5mm, 5×10^–5 m, and 2 m, respectively. Acon-2¹⁰⁰ and Acon-2¹⁰⁵ do not respond differently to the above treatments. Genetic crosses involving a holandric translocation, pericentric inversions, visible mutants, and allozyme markers were analyzed to map the aconitase (Acon-2) locus on the left arm of chromosome 3. The gene sequence (and map distances) on 3L is centromere—esterase-8 (Est-8)—2—esterase-4 (Est-4)—25—esterase-2 (Est-2)—9—Acon-2—5—phosphoglucomutase (Pgm)—7—esterase-6 (Est-6).     

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.     

Blood parameters of two hill stream cobitids in relation to their habitat

The blood of two fresh water cobitids — Botia lohachata, an exclusive water breathing form and Lepidocephalus guntea a dual breather, — showed a comparatively higher range of Hb (16.0–19.0 g%), Hct (50.0–61.1%) and number of RBC (2.71–6.7 millions/mm³) than many other water and air breathing fishes. Significant sexual difference exists in these characteristics (P > 0.05).The impact of life in oxygen depleted water, also inhabited by L. guntea as a result of its air breathing habit, is well reflected in its greater RBC size (11.86 × 8.66 µm) and their larger surface area (83.96 µm²) compared to that of Botia (53.16 µm²). The smaller size (9.92 × 6.45 µm) and consequently greater number of erythrocytes (4.67 millions/mm³) in Botia, are related to its active mode of life in the swiftly flowing water of hilly rivers. Though the higher nucleo-cytoplasmic ratio of 1 : 5.2 in Botia and 1 : 6.9 in Lepidocephalus suggest a slower red cell metabolism, the greater number of erythrocytes seems to have compensated for their active mode of life.     

A high-resolution genetic map and a diagnostic RFLP marker for the Mlg resistance locus to powdery mildew in barley

The semi-dominantly acting Mlg resistance locus in barley confers race-specific resistance to the obligate biotrophic fungus Erysiphe graminis f.sp. hordei. A high-resolution genetic map was constructed at Mlg based on a cross between the near-isogenic barley lines Pallas BC₅ Mlg and Pallas mlg. A total of 2000 F₂ progeny were inspected by cleaved amplified polymorphic sequence (CAPS) analysis, defining a 4.47 cM interval encompassing the resistance locus. Pathogen challenge of the segregants with multiple powdery mildew isolates uncovered a novel resistance specificity in Pallas BC₅ Mlg. Probes from within 4.0 cM of Mlg were mapped in rice, revealing orthologues on five different rice chromosomes and suggesting multiple breaks of chromosomal collinearity in this region between the two grass species. The most tightly Mlg-linked RFLP marker, MWG032, was shown to reliably detect the presence of the resistance allele in a collection of 30 European barley cultivars. Received: 23 March 2000 / Accepted: 20 April 2000     

Characteristics of a photorespiratory mutant of barley (Hordeum vulgare L.) deficient in phosphogly collate phosphatase

A barley mutant RPr84/90 has been isolated by selecting for plants which grow poorly in natural air, but normally in air enriched to 0.8% CO₂. After 5 minutes of photosynthesis in air containing¹⁴CO₂ this mutant incorporated 26% of the¹⁴C carbon into phosphoglycollate, a compound not normally labelled in wild type (cv. Maris Mink) leaves.The activity of phosphoglycollate phosphatase (EC 3.1.1.18) was 1.2 nkat mg^–1 protein at 30°C in RPr 84/90 compared to 19.2 nkat mg^–1 protein in the wild-type leaves. Phosphoglycollate phosphatase activity was not detected after protein separation by electrophoresis of leaf extracts from the mutant on polyacrylamide gels; on linear 5% acrylamide gels three bands with enzyme activity were separated from extracts of wild type plants. Gradient gel electrophoresis followed by activity staining showed two bands in Maris Mink tracks of MW 86,000 and 96,000, but no bands in 84/90. This is the first report of isozymes of phosphoglycollate phosphatase in barley which were absent in the mutant extracts. Our results confirm an earlier report of isozymes of this phosphatase in Phaseolus vulgaris [18].The photosynthetic rate of RPr 84/90 in 1% O₂, 350 l CO₂ l^–1 was 9–12 mg CO₂ dm^–2 h^–1 at 20°C, whereas the wild-type rate was 27–29 mg CO₂ dm^–2 h^–1 at 20°C. In 21% O₂, 350 l CO₂ l^–1 the rate was 2–3 mg CO₂ dm^–2 h^–1 in the mutant and 20 mg CO₂ dm^–2 h^–1 in the wild type.Genetic analysis has shown that the mutation segregates as a single recessive nuclear gene.     

Use of microbial peptide inhibitors for characterization of the substrate specificity of thermitase,a thermostable serine protease fromThermoactinomyces vulgaris

Seven microbial peptide inhibitors—chymostatin, antipain, elastatinal, leupeptin, pepstatin, bestatin, and phosphoramidon—were tested for their efficiency to inhibit thermitase, a thermostable serine protease fromThermoactinomyces vulgaris. Chymostatin and antipain were the most effective inhibitors, with K_i values of 7×10^–8 M and 2×10^–7 M, respectively. Except for leupeptin, all inhibitors resist hydrolysis by thermitase. Leupeptin, however, is cleaved by thermitase between the two leucylresidues. Further, a close relationship in specificity between thermitase and subtilisin BPN and their distinct discrimination from elastase specificity was demonstrated by using these inhibitors.     

Second-element turn-on of gene expression in an IS1 insertion mutant

Summary To learn more about the ways in which genes silenced by insertion mutations can be reactivated, we have undertaken a systematic investigation of Gal⁺ revertants of the polar mutant galOP-306::IS1 in Escherichia coli K12. The selective conditions used excluded reversion to wild type by precise excision of IS1. In this system (which resisded on a multi-copy plasmid) reversion to the Gal⁺ phenotype occurred with a frequency of about 10^-7 per cell and per generation. Analysis of the revertants revealed that — with the single exception of the previously published chromosomal mutant sis1 — alterations in the structure of IS1 lead to reactivation of gal operon expression. These events fall into four classes: (I) insertion of IS2 at position 327 in IS1, insertion of IS2 at position 687 in IS1, (III) insertion of a hitherto undetected mobile element, IS150, at position 387, (IV) a 16-bp deletion encompassing IS1 coordinates 553–568. Of some 200 independent reversion events studied, all but one were of types I–III i.e. they involved the intervention of a second mobile element.