An empirical comparison of selection methods for the improvement of biomass

Summary A single generation of upward truncation selection on families with 20% selected was carried out in each of five replicates using Tribolium castaneum as the test organism. The experiment involved eight lines: N — selected for offspring number; W — selected for pupal weight; B — selected for biomass; Q — quadratic index selected; L₂₁ — linear index selected with relative economic weights of 21 offspring number to pupal weight; L₁₁ — linear index selected with relative economic weights of 11 offspring number to pupal weight; L₁₂ — linear index selected with relative economic weights of 12 offspring number to pupal weight; C — an unselected control.Biomass (weight of offspring per family), offspring number, and pupal weight were measured. No differences in response to selection were found among the linear index lines and the pupal weight line with regard to any trait analysed. Generally, response to selection in the linear index lines and pupal weight line was small for offspring number and high for pupal weight. Selection pressure on offspring number in these lines seemed to be dependent on the correlation between offspring number and pupal weight. As a result, response to selection for biomass was poor in the linear index and pupal weight selected lines. In the case of the linear indices, poor response to selection for biomass appeared to be due to the violation of the assumption of additivity of the traits included in the definition of aggregate genotype.The responses in the quadratic index, biomass, and offspring number selected lines were equal with respect to selection for biomass. The response of the quadratic index selected line was less than the responses of the biomass and offspring number selected lines for offspring number, but the response in the quadratic index line was as large as that of any other line included in the experiment and greater than the biomass and offspring number selected lines where pupal weight was the criterion.Highly significant amounts of variation were found for all traits indicating that more replicates are needed for precise evaluation of selection systems.     

Experimental study of the efficiency of a reduced index in Tribolium

The relative efficiency of a reduced index (IR) relative to a full index (IF) was compared in an experiment with Tribolium. The selection objective included pupal length, adult weight, and egg mass. The reduced index was based on pupal length and adult weight, whereas the full index was based on these traits and also on egg mass. There were five generations of selection with four replicates, and a selected proportion of 20%. For each replicate, an unselected control was produced. Responses were significant in the IF and IR lines. Responses for the selection objective differed significantly between lines (p < 0.01). The efficiency of the IR line relative to the IF line was 0.52, similar to the expected efficiency of 0.51. The IF and IR lines did not differ significantly for pupal length nor adult weight, whereas the response for egg mass in the IF line was significantly different from the response in the IR line. Realized heritability was greater in the IF line (0.63 ± 0.05) than in the IR line (0.37 ± 0.16). The deleted trait (egg mass) has high heritability and genetic and phenotypic correlations nearly equal to zero with the other two traits included in the selection criterion. The results show the importance of including index traits with high genetic value that are independent of other traits, and they could be useful in breeding programs simultaneously considering production and reproduction traits with nearly zero correlations between them.     

Experimental comparison of methods for simultaneous selection of two correlated traits in Tribolium

Summary Two lines of Tribolium castaneum were selected in each of three replicates for egg laying between 7 and 11 days after adult emergency and for adult weight at 12 days, using theoretical (IT) and empirical (IP) index selection methods. Index coefficients were given empirically in the IP line and they were adjusted in the successive generations of selection according to the results obtained in the previous ones. Highly repeatable selection responses in all replicates occurred in both lines for the aggregate genotype (egg laying plus adult weight) and for each individual trait. The IP line tended to increase slightly more than the IT line for aggregate genotype and egg laying, while the highest response in adult weight was obtained with the IT method. The two methods gave consistently different responses in each replicate. The expected results were that IT selection should not exceed IP selection for the aggregate genotype and egg laying while theoretically the IT method should have been superior for increase adult weight. Theoretical expectations for adult weight have been fulfilled in practice. The IP method would be preferred in a practical sense because of its simplicity and freedom from need of parameter estimation.     

Successful oviposition and reproductive biology of Aprostocetus vaquitarum (Hymenoptera: Eulophidae): A predator of Diaprepes abbreviatus (Coleoptera: Curculionidae)

Aprostocetus vaquitarum (Wolcott) causes 78–91 percent mortality to eggs of Diaprepes abbreviatus (L.), under field conditions in southern Florida. In the laboratory, A. vaquitarum was reared on D. abbreviatus eggs at 25 °C, a photoperiod of 12:12 (L:D) and with abundant hosts, A. vaquitarum adult females lived around 15 days. Oviposition was significantly affected by the age of the host egg mass. Egg masses aged 0- to 3-day-old were accepted significantly better than those aged 4–6 days. The mean number of eggs deposited per female was around 53, with extreme values of 124 and 19 eggs per female. Using these data in combination with the sex ratio observed in the field (0.16) and the duration of the preimaginal stages, r_m (0.168–0142 day⁻¹), T (22.39–22.89 days), and R₀ (43.03–25.81 females per female) were calculated.     

Effect of UVB radiation on the survival, feeding, and egg production of the brackish-water copepod, Sinocalanus tenellus, with notes on photoreactivation

We examined the effects of UVB radiation on hatching success of eggs, survival of various naupliar and copepodite stages, and feeding and egg production of adult females of the brackish-water copepod, Sinocalanus tenellus, by exposure to varying doses of UVB irradiance in the laboratory. Artificial UVB radiation resulted in an increased mortality of eggs, nauplii and copepodites with increasing UVB doses. UVB induced damage was stage-specific with eggs being most susceptible (LD₅₀= 4.1 kJ m^–2 ) and adult females being least susceptible (LD₅₀= 16.7 kJ m^–2). Effects on feeding and egg production of adult females were significantly evident at UVB doses higher than 11.0 kJ m^–2 and 7.0 kJ m^–2, respectively. We also examined the photorepair response of eggs and various developmental stages in simultaneous irradiation of UVB and enhanced PAR. With enhanced PAR there was a considerable recovery against UVB damage, being higher for younger animals than older ones. In nature, however, solar UVB radiation may rarely cause appreciable damage to S. tenellus population due to optically high attenuation properties of their habitat waters.     

The effect of isozyme selection on metric characters in an interspecific backcross of tomato — basis of an early screening procedure

Summary The extent of correlation was estimated between isozyme genotypes and the four widely segregating characters — leaf segment W/L ratio, stigma exsertion, fruit weight, and seed weight — in the first backcross of F₁ Lycopersicon esculentum x Solanum pennellii to the former parent. The inbred parents differ in their alleles at the 12 tested isozymic loci, which are known to mark a minimum of eight of the twelve tomato chromosomes. Based on the isozyme data, a mean heterozygosity value, ¯H, was calculated which estimates the proportion of pennillii alleles in each individual. Correlations between mean heterozygosity and observed levels of each quantitative trait were highly significant and positive or negative as expected from the relative parental values. Plants with the lowest mean heterozygosity — i.e., closest to the esculentum zymotype also had mean values closest to those of this parent amongst the whole backcross population for each of the quantitative traits.Bivariate and multiple regression analysis was used to evaluate the ability of isozymes vs diagnostic morphological characters to estimate the portion of recurrent parent genes carried in each backcross individual. The results suggest that isozyme data gives better estimates than single diagnostic morphological characters and approach the level obtained by combinations of three morphological traits. Since electrophoretic determinations are made on small seedlings, selection at that stage can effect great savings of space and effort by greatly deminishing the size of the population needed at maturity. As such, isozyme selection would precede morphological selection but not replace it, thus the predictive value of these biochemical markers as well as diagnostic morphological characters could be obtained.     

Inheritance of weight in Rhipicephalus appendiculatus ticks (Acari: Ixodidae) in the laboratory

A selection of the 10% lightest and 10% heaviest males and females of a population of individually weighed Rhipicephalus appendiculatus Neumann adults was made in two experiments. The offspring of homologous pairs were followed until the next adult stage (light x light, control x control and heavy x heavy). The engorged nymphal weights, unfed adult weights, engorged female weights of the parents, egg mass weights, egg weights, larval scutal lengths, engorged larval weights, unfed nymphal weights, engorged nymphal weights and adult weights of the progeny were determined. No significant differences could be demonstrated between the two lines for egg weight, larval scutal length, engorged larval weight and unfed nymphal weight. Significant differences were found between the egg masses, engorged nymphal weights and adult weights of the two lines. The heritability coefficients of body weight determined from adult to adult were 0.14 and 0.10, respectively, during the first and second experiments. Considering females and males separately, the coefficients were 0.10 and 0.18 during the frist experiment and 0.12 and 0.09 during the repeat experiment respectively.     

Simultaneous selection for grain yield and protein percentage in backcross populations fromAvena sterilis ×A. sativa matings by using the independent culling levels procedure

Summary The method of independent culling levels was applied for simultaneous improvement in grain yield, protein percentage, and protein yield in a population of oat lines from the BC₀–BC₅ of the interspecific matingAvena sativa ×A. sterilis. Grain yield and protein percentage were subjected to selection with various combination of culling levels to give samples with 2% of the lines from the original population. — Intense selection for grain yield resulted in samples with high grain and protein yields but low protein percentage. Intense selection for protein percentage resulted in samples with high protein percentage but low grain and protein yields. The sample selected for protein yield only showed no significant difference from the recurrent parent for any trait. The recommended regime for improving protein percentage and grain and protein yields simultaneously was one that initially saved approximately 25–50% of the original population on the basis of protein percentage, and then provided intense selection for grain yield on the lines that remained. At least three backcrosses were necessary to obtain lines with high protein percentage and acceptable agronomic traits.Journal Paper No. J-11439 of the Iowa Agric. and Home Econ. Exp. Stn., Ames, IA 50011. Project No. 2447     

Modelling continuous culture of Rhodospirillum rubrum in photobioreactor under light limited conditions

Rhodospirillum rubrum was grown continuously and photoheterotrophically under light limitation using a cylindrical photobioreactor in which the steady state biomass concentration was varied between 0.4 to 4 kg m^–3 at a constant radiant incident flux of 100 W m^–2. Kinetic and stoichiometric models for the growth are proposed. The biomass productivities, acetate consumption rate and the CO₂ production rate can be quantitatively predicted to a high level of accuracy by the proposed model calculations. Nomenclature: C _X, biomass concentration (kg m^–3) D, dilution rate (h^–1) Ea, mean mass absorption coefficient (m² kg^–1) I , total available radiant light energy (W m^–2) K, half saturation constant for light (W m^–2) R _W, boundary radius defining the working illuminated volume (m) r _X, local biomass volumetric rate (kg m^–3 h^–1) <r _X>, mean volumetric growth rate (kg m^–3 h^–1) V _W, illuminated working volume in the PBR (m^–3). Greek letters: , working illuminated fraction (–) _M, maximum quantum yield (–) bar, mean energetic yield (kg J^–1).     

The kinetics of P515 in relation to the lipid composition of the thylakoid membrane

Flash-induced P515 absorbance changes have been studied in dark-adapted chloroplasts isolated from spinach plants grown under two different light intensities. The slow component (reaction 2), normally present in the P515 response of chloroplasts isolated from plants grown at an intensity of 60 W · m^–2, was largely reduced in chloroplasts isolated from plants grown at an intensity of 6 W · m^–2. This reduction of the slow component in the P515 response appeared to be coincident with an alteration in the lipid composition of the thylakoid membrane. Mainly the ratio monogalactosyldiacylglycerol to digalactosyldiacylglycerol appeared to be altered. In thylakoids from plants grown at 6 W · m^–2, the ratio was approximately 35% lower than that of plants grown at 60 W · m^–2. The amount of both cytochromeb ₅₆₃ and cytochromef was largely reduced in chloroplasts isolated from plants grown at low light intensity. These results may indicate a possible correlation between structural organization of the thylakoid membrane and the kinetics of the flash-induced P515 response.     

Genetic studies of self-fertility in rye (Secale cereale L.). 2. The search for isozyme marker genes linked to self-incompatibility loci

The segregation of several isozyme marker genes has been studied in F₂ inbred families from hybrids between self-sterile and five self-fertile inbred lines (nos. 2, 3, 4, 5, and 8) as well as from interline hybrids. Self-pollination of F₁ hybrids between self-sterile forms and lines 5 and 8 gave an F₂ segregation ratio of 1 heterozygote:1 homozygote for the gene Prx7 (chromosome 1R) against the allele from the line. This is interpreted as a result of tight linkage of the Prx7 gene with the S1 gene in chromosome 1R (recombination at a level of 0–1%). The self-pollination of such hybrids with lines 2,3 and 4 gave normal segregation for the Prx7 gene (1:2:1). This means that these lines carry a self-fertility allele which is not on chromosome 1R. Interline hybrids 5×2, 5×3 and 5×4 had self-fertility alleles for the two S genes and in inbred F₂ progenies gave the expected deviating segregation for the Prx7 gene in a ratio of 2:3:1. The segregation of interline hybrid 5×8 was normal, 1:2:1, as expected. Highly-deviating segregation in an inbred F₂ family of a hybrid with line 5 has also been obtained for another gene from chromosome 1R — Pgi2 (recombination with the S1 locus of 16.7%). By using the same method it has been estimated that line 4 has a self-fertility allele of the S2 locus from chromosome 2R and that the genes -Glu and Est4/11 are linked with it (recombination 16.7% and 17.5–20% respectively). Lines 2 and 3 have a self-fertility allele of the S5 locus from chromosome 5R which is linked with the Est5-7 gene complex (recombination at a level of 28.8–36.0%).     

Leaf Longevity,Construction, and Maintenance Costs of Three Mangrove Species Under Field Conditions

This study assessed the effect of leaf age on construction cost (CC) in the mangrove species Avicennia germinans, Laguncularia racemosa, and Rhizophora mangle growing in their natural habitat. Leaf osmolality values were species-specific, the highest in A. germinans (1 693 mmol kg^–1) and the lowest in L. racemosa (1 270 mmol kg^–1). In the three species, contents of chlorophyll (a+b) (Chl_a+b) and nitrogen (N) per unit of leaf area were maximal in adult leaves and tended to decline with age. Leaf mass to leaf area ratio (LMA) and ash content increased during leaf ageing. Similarly, as leaves aged, a significant increase in leaf construction cost per leaf area (CC_a) was observed, while per leaf mass (CC_m) it remained almost constant, suggesting a sustained production of leaf compounds as leaves became older. CC was positively correlated with LMA and heat of combustion (Hc) per leaf area, suggesting differences among species in the quantity and composition of expensive compounds. Leaf half lifetime (t_0.5) showed contrasting values in the three mangrove species (60, 111, and 160 d in L. racemosa, R. mangle, and A. germinans, respectively). Overall, L. racemosa was the species with less expensive leaves to construct while leaves of A. germinans and R. mangle had the highest CC_m and CC_a, respectively. Leaf longevity was positively correlated with the ratio between CC and maximum photosynthetic rate (P _max), clearly showing the existence of a balance between leaf costs and benefits.     

Physiological energetics of a midge,Chironomus riparius Meigen (Insecta,Diptera): normoxic heat output over the whole life cycle and response of larva to hypoxia and anoxia

The rate of metabolism of laboratory reared Chironomus riparius was monitored by direct calorimetry over the entire life cycle from egg to adult stage. The metabolic response of the fourth instar larva to decreasing oxygen concentrations and anoxia was also measured. Normoxic measurements were carried out at 20°C and the hypoxic-anoxic experiments at 10°C. In larvae with body sizes ranging from 0.0028 to 0.645 mg ash-free dry mass (afdm), the rate of heat dissipation was related to body mass by a power function, with a mass exponent of 0.71±0.02 corresponding to an exponent of -0.29 for the relationship between mass-specific metabolic rate and body mass. However, the allometric equations applicable to larvae would not predict the metabolic rates of eggs, pupae and adults. Single egg batches used in the experiments consisted of 354±90 eggs, the individual egg with a mass of 0.99±0.01 g (mean±SD). The mass-specific rate of heat dissipation of the egg (13.7±1.8 W mg^-1 afdm) was considerably lower than that of the first and second instar larvae (44–53 W mg^-1) but equal to that of fourth instar larvae (13.1±3.9 W mg^-1). Heat dissipation by a pupa shortly before adult emergence was high (14.8±1.8 W mg^-1), probably due to high metabolism during metamorphosis. Emergence of the adult in the calorimeter was indicated by a short but intense burst of heat. The newly emerged imago had a ca. 20–35% higher metabolic rate than the pupa. In response to reduced O₂ partial pressure the fourth instar larva of C. riparius displayed metabolic regulation. In continuously declining oxygen partial pressure, the fourth instar larva maintained its aerobic energy metabolism (4.2 W mg^-1) with only a small decrease down to 0.8 kPa, corresponding to an oxygen concentration of 0.42 mg O₂l^-1 H₂O. Below this critical oxygen concentration (P_c), the rate of heat dissipation decreased rapidly down to the anoxic level which was only 14–17% of the normoxic level. The high relative reduction of metabolic rate under anoxia gives a wrong impression of short-term tolerance of C. riparius to anoxia. The absolute energetic costs of C. riparius associated with anaerobic energy metabolism (0.64±0.11 W mg^-1) are almost 6 times higher than those of more anoxia tolerant invertebrates such as sphaeriid bivalves.     

Synthesis and evaluation of Triticum durum — T. monococcum amphiploids

Summary Nine Triticum durum — T. monococcum amphiploids (AABBA^mA^m) were synthesized by chromosome doubling of sterile triploid F₁ hybrids involving nine T. durum (AABB) cultivars and a T. monococcum (A^mA^m) line. The triploid F₁ hybrids had a range of 4–7 bivalents and 7–13 univalents per PMC. The synthetic amphiploids, however, showed a high degree of preferential pairing of chromosomes of the A genomes of diploid and tetraploid wheats. The amphiploids were meiotically stable and fully fertile. Superiority of four amphiploids for tiller number per plant, 100-grain weight, protein content and resistance to Karnal bunt demonstrated that these could either be commercially exploited as such after overcoming certain inherent defects or used to introgress desirable genes into durum and bread wheat cultivars. Methods for improvement of these amphiploids are discussed.     

Incorporation of hygromycin resistance in Brassica nigra and its transfer to B. napus through asymmetric protoplast fusion

Summary With the idea to develop a selection system for asymmetric somatic hybrids between oilseed rape (Brassica napus) and black mustard (B. nigra), the marker gene hygromycin resistance was introduced in this last species by protoplast transformation with the disarmed Agrobacterium tumefaciens strain C58 pGV 3850 HPT. The B. nigra lines used for transformation had been previously selected for resistance to two important rape pathogens (Phoma lingam, Plasmodiophora brassicae). Asymmetric somatic hybrids were obtained through fusion of X-ray irradiated (mitotically inactivated) B. nigra protoplasts from transformed lines as donor with intact protoplasts of B. napus, using the hygromycin resistance as selection marker for fusion products. The somatic hybrids hitherto obtained expressed both hygromycin phosphotransferase and nopaline synthase genes. Previous experience with other plant species had demonstrated that besides the T-DNA, other genes of the donor genome can be co-transferred. In this way, the produced hybrids constitute a valuable material for studying the possibility to transfer agronomically relevant characters — in our case, diseases resistances — through asymmetric protoplast fusion.     

Identification and characterization of common bean (Phaseolus vulgaris L.) lines well nodulated in the presence of high nitrate

Common bean,Phaseolus vulgaris L., is known to be ‘inefficient’ in nodulation and N₂ fixation although it responds to applied nitrogen. An experiment was conducted to identify and to characterize bean cultivars nodulating in the presence of a high level of nitrogen. Sixteen cultivars and a check for inefficient nodulation, OAC Seaforth, were inoculated and grown for 40 days in replicated pots supplied with zero, 3.5 and 10.5 mM combined nitrogen as NO ₃ ⁻ and NH ₄ ⁺ . Seven traits relating to nodulation and N₂ fixation were all significantly affected by N level (N), cultivar (Cv) and N × Cv interactions (except for root dry weight), indicating that cultivars responded differently to the N treatments. Total dry weight (W) and shoot to root ratio (S/R) increased with the increased N levels. Nodule dry weight (Wn), visual nodulation score (Nv) and nodulation index (Nx) decreased as the N increased. Percent N and N content per plant increased with the increased N level. Plant weight (W) was positively correlated with Wn, Nv and N content and negatively correlated with %N. Nodulation score was positively associated with Wn and plant N content. Genotypes superior in nodulation and N₂ fixation in the presence of N were identified. Cultivars Italian Barlotti, California Light Red Kidney, Kentucky Wonder A and Pueblo 152 were selected for further testing and use in improving the nitrate tolerant nodulating characteristic of bean.     

SEVERE INBREEDING DEPRESSION AND RAPID FITNESS REBOUND IN THE BUTTERFLY BICYCLUS ANYNANA (SATYRIDAE)

We established inbred laboratory lines of the satyrid Bicyclus anynana with one, three and 10 pairs of butterflies, which were subsequently allowed to increase freely to a maximum size of 300 butterflies. Minimally inbred control lines were established with 300 randomly selected virgin butterflies of equal sex ratio. We measured fecundity, egg weight, egg hatching, adult emergence, adult size, and the proportion of crippled adults in generations F₂, F₃, F₅, and F₇ (the latter two for the one pair bottleneck lines only). The most striking result was an unexpectedly large decrease in egg hatching with increase in inbreeding (25% per 10% increase in inbreeding). Such a level of inbreeding depression has not been reported previously for any insect. The distribution of egg hatching rate for individual clutches within inbred lines was markedly skewed, with a large fraction of clutches producing no eggs at all. This is interpreted as a relatively lower ratio of detrimental to lethal (or sterile) mutation loads than is found in Drosophila, the only insects for which mutation loads have been well characterized. Possible explanations for this severe inbreeding depression include a relatively high rate of mutation to recessive alleles with substantial damaging effects and infrequent episodes of inbreeding in nature. In the experiments, average egg hatching rate recovered rapidly between F₂ and F₇ in three of the six one-pair lines. We discuss the implications of these results for survival of populations through extreme bottlenecks in nature and in captivity.     

Hydrogen accumulation by H2-uptake negative strains of Rhizobium

Summary Hydrogen evolution from root nodules has been reported to decrease the efficiency of the nitrogen fixing system. Mutants ofRhizobium meliloti andRhizobium leguminosarum were selected which were deficient in H₂-uptake capacity (Hup^–). The relative efficiency of the nitrogen fixation for both species assessed with C₂H₂ reduction was 0.66.The hydrogen production was monitored using a simple root incubation method. As such, hydrogen production up to 3.83 and 15.57 ml.day^–1.g^–1 plant dry weight were recorded forPisum sativum — Rhizobium leguminosarum 4.20 Hup^– andMedicago sativa — Rhizobium meliloti 1.5 Hup^– respectively. In a closed container (250 ml), hydrogen concentrations up to 20% (v/v) could be reached in the root phase ofMedicago sativa in a time period of 320 hours.     

The semidwarf gene,sd-1, of rice (Oryza sativa L.). II. Molecular mapping and marker-assisted selection

To establish the location of the semidwarf gene, sd-1, the anthocyanin activator (A), purple node (Pn), purple auricle (Pau), and the isozyme locus, EstI-2, in relation to DNA markers on the molecular linkage map of rice, 20 RFLP markers, previously mapped to the central region of chromosome 1 (McCouch et al. 1988), were mapped onto an F₂ population derived from the cross Taichung 65 (A,Pn,Pau)/Taichung 65 (sd-1). sd-1 and EstI-2 were determined to be linked most tightly to RFLP markers RG 109 and RG 220, which cosegregated with each other. The distance between these RFLP markers and sd-1 was estimated to be 0.8 cM, based on an observed recombination value of 0.8%. The order of genes and markers in this region of chromosome 1 was determined to be sd-1 — (EstI-2 — RG220 — RG109) — RG381 — A — Pn — Pau. To test the efficacy of selection for sd-1 based on these linked markers, 50-day-old F₂ seedlings derived from another cross, Milyang 23/Gihobyeo, were analyzed for marker genotype. At this age, the semidwarf character could not be clearly detected based on phenotype. In addition, plant height was normally distributed in this population, making it difficult to unambiguously identify plants carrying sd-1. Thirteen seedlings homozygous for the sd-1-associated allele at EstI-2, RG220 and RG109, and 13 seedlings homozygous for the Sd-1-associated allele at all three marker loci were selected for further genetic analysis. At 20 days after heading, the culm lengths of these 26 plants were measured and the expected phenotype was confirmed in every case. These 26 plants were then selfed for four generations and F₆ lines were again evaluated to determine whether any recombination among the three molecular markers, or between these markers and the sd-1 gene, could be detected. No recombinants were identified, confirming the tight linkage of these loci and the usefulness of genotypic selection for this recessive semidwarf character prior to the time when it can be evaluated based on phenotype.     

Pyrimidine biosynthesis in Serratia marcescens: Polypeptide interactions of three nonsequential enzymes

Orotidine-5-monophosphate pyrophosphorylase (OMPppase, E.C. 2.4.2.10) and orotidylate decarboxylase (OMPdecase, E.C. 4.1.1.23) were purified from Serratia marcescens HY. These enzymes required physical association for maximal catalytic activities and formed a fragile complex with dihydroorotase (DHOase, E.C. 3.5.2.3.). OMPppase reversibly lost 50% of its activity upon separation from DHOase. The kinetic characteristics of OMPppase were modified by this separation. In the presence of DHOase, the K _ms for PRPP and orotate were stoichiometric: 2.3×10^–6 m and 2.6×10^–6 m, respectively. Following separation, the K _ms were significantly different: 1.3 × 10^–6 m for PRPP and 4.1×10^–6 m for orotate. OMPppase and OMPdecase could be reversibly separated by acrylamide gel electrophoresis, but the separation was accompanied by a loss of catalytic efficiency for both enzymes. DHOase readily associated into multiple molecular forms and could not be purified. The DHOase-OMPppase-OMPdecase interactions demonstrate that a weakly aggregated, multifunctional enzyme complex participates in the biosynthesis of pyrimidine nucleotides in S. marcescens. This unique association of nonsequential biosynthetic enzymes may represent a larger complex which provides a channeling or regulatory unit.This work was supported by grants from the National Science Foundation (NSF GB 5811) and the Office of Naval Research (Nonr 4413). One of us (J.W.) was a National Science Foundation Graduate Fellow.