Identification of two RAPD markers tightly linked with the Nicotiana debneyi gene for resistance to black root rot of tobacco

Linkage of randomly amplified polymorphic DNA (RAPD) markers with a single dominant gene for resistance to black root rot (Chalara elegans Nag Raj and Kendrick; Syn. Thielaviopsis basicola [Berk. and Broome] Ferraris) of tobacco (Nicotiana tabacum L.), which was transferred from N. debneyi Domin, was investigated in this study. There were 2594 repeatable RAPD fragments generated by 441 primers on DNAs of Delgold tobacco, a BC₅F₈ near isogenic line (NIL) carrying the resistance gene in a Delgold background, and PB19, the donor parent of the resistance gene. Only 7 of these primers produced eight RAPD markers polymorphic between Delgold and PB19, indicating there are few RAPD polymorphisms between them despite relatively dissimilar pedigrees. Five of the eight RAPD markers were not polymorphic between Delgold and the NIL. All of these markers proved to be unlinked with the resistance gene in F₂ linkage tests. Of the remaining three RAPD markers polymorphic between Delgold and the NIL, two were shown to be strongly linked with the resistance gene; one in coupling and the other in repulsion. Application of the two RAPDs in the elimination of linkage drag associated with the N. debneyi resistance gene and marker-assisted selection for the breeding of new tobacco cultivars with the resistance gene is discussed.     

Evolution of the response patterns to dietary carbohydrates and the developmental differentiation of gene expression of α-amylase in Drosophila

Intraspecific variation of -amylase activity in D. melanogaster and D. immigrans, which is distantly related to D. melanogaster, and interspecific variation of -amylase activity in 18 Drosophila species were examined. The amount of intraspecific variation of -amylase activities measured in terms of coefficient of variation in D. melanogaster and D. immigrans was one-half and one-tenth or less, respectively, of the interspecific variation in 18 Drosophila species. We also surveyed the response patterns of -amylase activity to dietary carbohydrates at the larval and adult stages. The levels of -amylase activity depended on both repression by dietary glucose (glucose repression) and induction by dietary starch (starch induction). In general, our data suggest that glucose repression was conserved among species at both stages while starch induction was mainly observed in larvae, although the degree of the response depended on species. In D. lebanonensis lebanonensis and D. serrata, larvae expressed electrophoretically different -amylase variants (isozymes) from those of adult flies. These results may suggest that the regulatory systems responsible both for the response to environment and developmental expression are different among species in Drosophila. Correspondence to: T. Yamazaki     

Über Konstitution und Erbgang eines neuen Delphinidinglycosids „Floridorin“ aus der Garteniris-Sorte cv. ‘Floridor’ (Cayeux 1929)

Zusammenfassung Bei der diploiden hohen Garten-Iris cv. Floridor (Cayeux 1929) wurde als bisher einziger Sorte ein neues Delphinidinglycosid Floridorin aufgefunden. Seine chemische Konstitution wurde als Delphinidin-3-Glukose-Rhamnose-p-Cumarsäure aufgeklärt. Es fand sich zusammen mit dem schon von uns aufgeklärten Anthozyan Tulipanin (Delphinidin-3-Glucose-Rhamnose). Das Hauptanthozyan der anderen Garten-Iris ist das von uns neuerdings nachuntersuchte Violanin. Die Untersuchungen wurden mit bereits von uns angegebenen neueren Methoden ausgeführt, wie der stufenweisen Hydrolyse und dem oxidativen Abbau. Das neue Floridorin zeigte bei den diploiden Garten-Iris einen monohybriden rezessiven Erbgang gegen Violanin. Die Blüten der Sorten, die Floridorin enthalten, sind schon mit dem Auge an einem charakteristischen taubenblauen Farbton zu erkennen.

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On the constitution and inheritance of a new delphinidine glycoside Floridorin from the cultivated iris variety cv. Floridor (Cayeux 1929)Studies on anthocyanins LI

Summary The diploid tall bearded garden Iris cv. Floridor (Cayeux 1929) proved to be thus far the only variety with a different anthocyanine, called Floridorin. Its chemical structure has been found to be delphinidine-3-glucose-rhamnose-p-coumaric-acid. It occurs together with tulipanine already analyzed by us as delphinidine-3-glucose-rhamnose. The main anthocyanine of thePogoniris garden varieties proved to be violanin the structure of which has been studied by us lately. The investigations were carried out by some newer methods, such as partial hydrolysis and oxidative degradation already published by us. The new Floridorin gives a monohybrid recessive Mendelian ratio with other diploid varieties ofPogoniris colored by violanine. The varieties colored by Floridorin show a characteristic greyish blue coloration which can be recognized with the naked eye.

     

Genetics of two tissue esterase polymorphisms (Est-4 and Est-5) in the rabbit

Two polymorphic esterase systems were found after electrophoresis of rabbit tissue homogenates. Each of these systems is controlled by an autosomal locus with two alleles. Est-4 determines the absence (Est-4^a) or presence (Est-4^b) of two bands of esterase activity with intermediate anodal mobility and broad substrate specificity. This polymorphism was found to be present in liver, small intestine, and spleen but not in kidney, heart, and testis. Est-5 is coding for cathodally migrating esterases which differ in mobility (Est-5^a and Est-5^b). This polymorphism was found only in kidney and testis homogenates. Est-5 esterases are more active against -naphthyl acetate than against -naphthyl acetate and have no activity against -naphthyl butyrate. Linkage analysis indicated that Est-4 is localized on rabbit LG VI as part of a cluster of esterase loci, whereas Est-5 segregates independently. Rabbits from two inbred and nine partly inbred strains were tested for these polymorphisms.This investigation was supported in part by Public Health Service Research Grant RR-00251 from the Division of Research Resources and by funds from the University of Utrecht. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Schmelzverhalten und kalorische Eigenschaften einer Mischung von Rinderserumalbumin,Kochsalz und Wasser

Zusammenfassung Es wurde die mittlere EnthalpiedifferenzH _m von Rinderserumalbumin (RSA)-Salz-Wassergemischen im Temperaturbereich von –47 °C bis 0°C bestimmt.H _m enthält neben der Energie zur Erwärmung auch noch die Schmelzwärme der Mischung. Aus dem Verlauf vonH _m als Funktion des Lösungsmittels (0,5 molale NaCl-Lösung) ergibt sich für den Anteil des kalorisch gebundenen Lösungsmittels ein Wert von 0,54 g Lösungsmittel pro g RSA. Dieser Anteil ist unabhängig von der mittleren Ladung des RSA-Moleküls.Aus einer Analyse der Meßergebnisse nach der Theorie der Mischungen folgt, daß die Partialenthalpie des RSA in verdünnten Mischungen negativ ist. Die Partialenthalpie des Lösungsmittels ist dagegen in diesen verdünnten Gemischen unverändert gleich der Enthalpiedifferenz des Lösungsmittels selbst. Die Messungen wurden mit Hilfe eines Differential-Scanning-Kalorimeters der Fa. Perkin-Elmer durchgeführt.

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Melting behaviour and caloric properties of a mixture of bovine serum albumin, sodium chloride and water

Summary In the temperature range from –47 °C to 0 °C the enthalpy differencesH _m of bovine serum albumin (BSA)-salt-water mixtures were measured.H _m includes the energy to increase the temperatures as well as the melting heat of the mixture. The plot ofH _m as a function of the solvent content (0,5 molal NaCl solution) gives 0,54 g solvent per g dry BSA for the caloric bound solvent. This value is independent of the average charge of the BSA molecule.The theory of mixtures applied to this measurements shows that the partial enthalpy difference of BSA in dilute mixtures is negative while that of the solvent in this mixtures is equal to the enthalpy difference of the pure solvent.The enthalpy differences were measured with a Differential-Scanning-Calorimeter (Perkin-Elmer).

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Herrn Prof. Dr. Dr. h. c. mult. Boris Rajewsky zum 80. Geburtstag gewidmet.     

Chromosomal homologies betweenDrosophila lebanonensis andD. melanogaster determined by in situ hybridization

Twelve biotin-labelled recombinant DNA probes were hybridized to polytene chromosomes ofDrosophila melanogaster andD. lebanonesis. Probes were chosen in order to cover the whole chromosomal complement. Six probes correspond to known genes fromD. melanogaster (RpII215, H3–H4, MHC, hsp28/23, hsp83, hsp70), four probes are clones isolated from aD. subobscura library (Xdh, DsubS3, DsubG3, DsubG4) and the remaining two probes correspond to the Adh gene ofD. lebanonensis and to one sequence (262), not yet characterized, from the same species. The chromosomal homologies obtained from the in situ hybridization results allow us to determine that Muller's C and D chromosomal elements are fused in the karyotype ofD. lebanonensis and constitute the large metacentric chromosome. Single pericentric inversions in theE andB elements have generated the medium and small metacentric chromosomes, respectively. No great changes are detected in Muller'sA element, which remains acrocentric. The changes detected in the karyotypic evolution ofD. lebanonensis are frequently observed inDrosophila evolution, as deduced from chromosomal homologies of severalDrosophila species. The results are also consistent with Muller's proposal that chromosomal elements have been conserved during the evolution ofDrosophila.     

Untersuchungen zur Regulation der Bacteriochlorophyll-Synthese bei Rhodospirillum rubrum

Zusammenfassung Wenn Rhodospirillum rubrum aus aeroben Dunkelkulturen in ein synthetisches Medium übertragen und anaerob im Licht bebrütet wird, beginnt die Bacteriochlorophyllbildung bereits nach 1 Std, das Wachstum erst nach 5–8 Std Bebrütung. — Puromycin (10 g/ml) und Chloramphenicol (20 g/ml) hemmen in diesen Kulturen Protein- und Pigmentsynthese vollständig. Eine Hemmung wird auch beobachtet, wenn die Antibiotica erst mehrere Stunden nach Beginn der Lichtbebrütung zugesetzt werden. Synthese von Thylakoidprotein und Bacteriochlorophyll scheinen regulatorisch gekoppelt zu sein.Actinomycin C (D) (40 g/ml) und Mitomycin (1 g/ml) hemmen die Bacteriochlorophyllbildung enbenfalls. Die Thylakoidbildung ist vom Vorhandensein einer funktionsfähigen DNS und RNS abhängig.Die spezifische Aktivität der -Aminolaevulinsäuresynthetase nimmt in anaeroben Lichtkulturen gegenüber aeroben Dunkelkulturen um etwa das Vierfache zu. Sie wird durch die verwendeten antibiotica nicht beeinflußt. Die Biosynthese des Enzyms wird durch Mitomycin und Puromycin, nicht aber durch Actinomycin gehemmt.

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Summary If dark-aerobically grown Rhodospirillum rubrum is transferred to anaerobic conditions in the light, the synthesis of photosynthetic pigments starts after 1 or 2 hours. The growth of the culture begins in the synthetic medium not before 5 hours incubation. In these cultures Puromycin (10 g/ml) and Chloramphenicol (20 g/ml) inhibit synthesis of protein and bacteriochlorophyll both. An inhibition is also observed when the antibiotics are added some hours after the beginning of anaerobic light incubation.The synthesis of chromatophore protein and bacteriochlorophyll are likely connected by gen-regulation.Actinomycin C (D) (40 g/ml) and Mitomycin C (1 g/ml) inhibit the bacteriochlorophyll synthesis likewise. The effect of actinomycin is increased by preincubation with the antibiotic in the dark. Mitomycin C stops synthesis of bacteriochlorophyll and protein even if it added after preincubation in the light.The level of -aminolevulinic acid-synthetase increased fourfold in anaerobic light-cultures compared to dark-aerobically grown cells. The activity of the enzyme is not influenced by the antibiotics. But the rate of biosynthesis is inhibited by Puromycin and Mitomycin, but not by Actinomycin.

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Abkürzungen im Text ALS -Aminolaevulinsäure - B-Chlorophyll Bacteriochlorophyll - DNS Desoxyribonucleinsäure - RNS Ribonucleinsäure     

Novel 2D and 3D multiple-quantum bi-directional HCNCH experiments for the correlation of ribose and base protons/carbons in 13C/15N labeled RNA

Multiple-quantum 2D and 3D bi-directional HCNCH experiments are presented for the correlation of base and ribose protons/carbons in ¹³C/¹⁵N labeled HIV-1 TAR RNA. In both 2D and 3D experiments, the magnetization of H1 is transferred to H6/H8 and H1 through H1-C1-N1/9-C6/8-H6/8 and H1-C1-N1/9-C1-H1 pathways, and the magnetization of H6/8 is transferred to H1 and H6/8 through H6/8-C6/8-N1/9-C1-H1 and H6/8-C6/8-N1/9-C6/8-H6/8 pathways. Chemical shifts of four different nuclei (H1, C1, C6/8 and H6/8) are sampled in the 2D experiment. The correlation of base and ribose protons/carbons is established by the rectangular arrangement of crossover and out-and-back peaks in the proton/carbon correlated spectrum. The rectangular connections can be further resolved using the nitrogen dimension in a ¹H/¹³C/¹⁵N 3D experiment. Furthermore, by taking advantage of the well separated chemical shifts of N1 (pyrimidine) and N9 (purine), the 2D spectrum can be simplified into two sub-spectra based on their base type. Both experiments were tested on a ¹³C/¹⁵N labeled 27-mer HIV-1 TAR RNA containing a UUCG hairpin loop.     

Quantitative measurement of cell-bubble attachment in bubble columns using foam flotation techniques

A simple and convenient system for quantitatively measuring the number of adsorbed animal cells per unit of bubble surface area (, unit: cells/cm2) was developed. The system was successfully applied to recombinant Chinese hamster ovary (r-CHO) suspension cultures to investigate the dynamic cell-bubble attachment in a bubble column. In serum-free medium, values increased with bubble rising height (H) and cell concentration (C) and then became constant (about 1750 cells/cm2) when H and C were sufficiently high. In medium containing protective additives, the trends of values with H were similar to that in serum-free medium. Compared with serum-free medium, polyvinyl-pyrrolidone (PVP) increased the values to 1941 cell/cm2 whereas other tested additives decreased the values of in some different degree.     

Glycoconjugates of the intestinal epithelium of the domestic fowl (Gallus domesticus): a lectin histochemistry study

Summary A lectin histochemical study was performed on formalin-fixed paraffin-embedded tissues of duodenum, jejunum, ileum, caecum and colon from six fasted and six non-fasted 8-week-old chickens (Gallus domesticus). The purpose of this study was to identifyin situ the pattern of carbohydrate residues present on the luminal surface of the intestinal epithelium. Ten biotinylated lectins with different sugar specificities were used as probes, and avidin—biotin—peroxidase complex (ABC) was used as a visualant. The most significant finding was the binding pattern ofLens culinaris agglutinin to various segments of the intestines. The luminal surface of the small intestinal epithelium did not stain with this lectin. In the colon the luminal surface was lightly stained, while the caecal luminal surface was intensely stained. Throughout the intestine the luminal surface stained withCanavalia ensiformis agglutinin,Ricinus communis agglutinin-I and wheatgerm agglutinin, but it did not stain withDolichos biflorus agglutinin. These findings indicated that, throughout the intestine, the luminal surface contains glycoconjugates with eitherN- orO-linked glycoprotein, or both, with terminal non-reducing -galactosyl and sialyl residues. Furthermore, the caecal surface is rich inN-linked glycoproteins with an -(16)-linked fucosyl residue near the glycosidic linkage. The potential significance of these observations and the role of glycoconjugates in host—parasite interaction (i.e.Eimeria sp. versusGallus domesticus) are discussed.     

Two-dimensional 1H NMR study of two cyclobutane type photodimers of thymidylyl-(3′→5′)-thymidine

2D NMR spectroscopy and J coupling constant analysis are applied to resolve the structure of two photoproducts of thymidylyl-(35)-thymidine. These products are cyclobutane type thymine dimers possessing the cis-syn (the predominant one) and trans-syn geometry. The cis-syn is formed in an ANTI-ANTI conformation about the N-glycosyl linkages and resembles the normal base-stacked configuration. The glycosidic conformation in solution of the 5 terminal fragment differs from the crystal in which the less common SYN conformation is observed. In this isomer only the sugar pucker of the 3 terminal fragment is changed substantially with respect to the dinucleotide. The trans-syn isomer is formed in a SYN-ANTI glycosidic conformation. In this isomer the sugar puckers of both deoxyribose rings are affected and a preference for a pure 2-endo conformation is observed.Abbreviations dTpdT 2-deoxythymidylyl-(35)-2-deoxythymidine - dTp[]dT cyclobutane type photodimers of dTpdT - dTp- and dTp[]- their 5' terminal fragments (fragment A) - -pdT and-[]pdT their 3 terminal fragments (fragment B) - RP-HPLC reversed-phase high-performance liquid chromatography - COSY two-dimensional correlated spectroscopy - 2D NOE two-dimensional nuclear Overhauser spectroscopy     

Evolution of thedec-1 eggshell locus inDrosophila. I. Restriction site mapping and limited sequence comparison in themelanogaster species subgroup

Summary We have analyzed 18 kb of DNA in and upstream of thedefective chorion-1 (dec-1) locus of the eight known species of themelanogaster species subgroup ofDrosophila. The restriction maps ofD. simulans, D. mauritiana, D. sechellia, D. erecta, andD. orena are shown to have basically the restriction map ofD. melanogaster, whereas the maps ofD. teissieri andD. yakuba were more difficult to align. However, the basic amount of DNA and sequence arrangement appear to have been conserved in these species. A small deletion of varying length (65–200 bp) is found in a repeated sequence of the central transcribed region ofD. melanogaster, D. simulans, andD. erecta. Restriction site mapping indicated that thedec-1 gene is highly conserved in themelanogaster species subgroup. However, sequence comparison revealed that the amount of nucleotide and amino acid substitution in the repeated region is much larger than in the 5 translated region. The 5 flanking region showed noticeable restriction site polymorphisms between species. Based on calculations from the restriction maps a dendrogram was derived that supports earlier published phylogenetic relationships within themelanogaster species subgroup except that theerecta-orena pair is placed closer to themelanogaster complex than toD. teissieri andD. yakuba.     

Genetics of esterases in Drosophila. IV. Slow-migrating S-esterase in Drosophila of the virilis group

A slow-migrating -esterase (S-esterase) is described which has been detected in Drosophila montana, Drosophila imeretensis, and some stocks of Drosophila virilis when mixtures of - and -naphthyl acetate are used as substrates in histochemical reactions after electrophoresis. Sexual dimorphism for S-esterase has been demonstrated. This esterase is contained in male genitalia only, predominantly in the ejaculatory bulb (waxy plug). It appears 3–4 days after emergence of flies. In hybrids between S⁺ and S⁰ species, the activity of the slow esterase is either decreased or inhibited. An autonomous synthesis of the S-esterase in the ejaculatory bulb was established by transplantation of imaginal genital discs into larvae of different Drosophila stocks. Based on analysis of physicochemical and immunochemical properties, S-esterase is suggested to be an independent fraction of esterase, possibly dimeric, which does not cross-react with -esterase antiserum.     

Esterases in histochemistry and ultrahistochemistry

Synopsis The histochemical identification of individual esterases is a problem that has not yet been overcome. Inhibitors and different substrates reveal different patterns of distribution. 8-hydroxyquinoline acetate is a useful substrate in ultrahistochemistry. There is evidence of a relationship between esterase distribution and function.ACTH adrenocorticotropic hormone - 5Bri–O-2 5-bromoindoxyl acetate - 5Br–4ClI–O-2 5-bromo-4-chloro indoxyl acetate - cAMP cyclic adenosine monophosphate - DFP di-isopropyl-fluorophosphate - hCG human chorion gonadotropin - HS-2/4 thiol acetate/butyrate - I-O-2/4 indoxyl acetate/butyrate - N-O-2/3/4 -naphthyl acetate/propionate/butyrate - N-O-2 -naphthyl acetate - N-S-2/9 -naphthyl thiolacetate/nonanoate - NAS-O-2 naphthol AS acetate - NASD-O-2 naphthol AS-D acetate - 4NP-O-2/3 p-nitrophenyl acetate/propionate - 4NP-S-2 p-nitrophenyl thiol acetate - P-O-2 phenyl acetate - Q-O-2/4 8-hydroxyquinoline acetate/butyrate - Q-S-2/4 8-mercaptoquinoline acetate/butyrate - TBA-S-2/9 -thiolbenzanilide acetate/nonanoate - TSH thyroid-stimulating hormone     

Isoenzymes and histochemistry of non-specific esterases in normal and cancerous skin

Synopsis Non-specific esterases in normal and carcinomatous skin of the mouse have been investigated electrophoretically and histochemically. Three esterase bands were obtained on electrophoresis from homogenates of normal skin; homogenates of carcinomas showed an accumulation of esterase-Ia and esterase-Ib.^* However, using several ester substrates, substrate-specific patterns were demonstrated in the electrophoresis separations and histochemically in tissue sections. On the electrophoresis separations, -naphthyl acetate, -naphthyl acetate, 6-bromo-2-naphthyl acetate, naphthol AS acetate, naphthol AS-D acetate and naphthol AS-LC acetate gave rise to similar patterns, but with -naphthyl propionate as subsmate, more esterase-Ib was indicated and with 5-bromo-indoxyl acetate a distinctive preponderance. Peripheral or uniformly distributed staining was found histochemically in tumour epithelium using -naphthyl acetate, -naphthyl propionate and -naphthyl acetate, whereas with the substrates of naphthol AS acetate, naphthol AS-D acetate and indoxyl acetate an intermediate pattern of staining related to keratinization was obtained.     

Small enzymes with esterase activities from two thermophilic fungi, Emericella nidulans and Talaromyces emersonii

Extracellular esterase activities in Emericella nidulans and Talaromyces emersonii are attributed to small enzymes with molecular weights less than 10 kDa (microenzymes). A 1.6 kDa esterase accounted for most of the esterase activity observed in both organisms and one of them also contained a 4.1 kDa microenzyme with weaker esterase activity. These esterases were growth-associated and active towards fluorescein dibutyrate and -naphthyl acetate as well as tributyrin.     

Social mutilation in the Ponerine ant <Emphasis Type="Italic">Diacamma</Emphasis>: cues originate in the victims

Summary. In the queenless ponerine ant genus Diacamma, all workers eclose with a pair of innervated thoracic appendages termed gemmae. The gamergate (= mated egg laying worker) maintains reproductive monopoly by mutilating the gemmae of all eclosing individuals. Such mutilation leads to irreversible behavioural and neurological changes such that the individual lacking gemmae becomes incapable of appropriate sexual calling and mating. In one population related to Diacamma ceylonense from India, Diacamma sp. from Nilgiri (hereafter referred to as nilgiri), gamergates do not mutilate their nestmates and yet maintain reproductive monopoly. To understand what triggers mutilation, we exchanged cocoons between the mutilating D. ceylonense colonies and the non mutilating nilgiri colonies. nilgiri callows were not mutilated even in D. ceylonense colonies while D. ceylonense callows were mutilated even in nilgiri colonies, suggesting that the cues for mutilation originate in the victims (callows), presumably in the gemmae themselves. This finding should facilitate understanding the proximate mechanism and evolutionary significance of mutilation of gemmae as a method of resolution of reproductive conflicts in the genus Diacamma.Received 27 August 2003; revised 26 April 2004; accepted 27 April 2004.     

A comparative study of embryonic gene expression inDrosophila andEphestia

Summary The ontogeny of allozyme patterns has been studied in embryos ofDrosophilamelanogaster, which are doubly heterozygous for alleles specifying the slow and fast forms of alcohol dehydrogenase (ADH) and -glycerophosphate dehydrogenase (GPDH). The ontogeny of esterase-2 was studied in embryos and young larvae of the flour mothEphestia kühniella, which are heterozygous for two of the three existing esterase-2 alleles. In freshly laidDrosophila eggs only the maternal enzyme forms are present and during the first 15 hours of development the staining of these forms becomes progressively fainter. After 16 and 17 h, the paternal and hybrid bands of ADH and GPDH respectively become obvious. Before hatching, the intensity distribution in the three-banded pattern of reciprocal hybrids is asymmetric in favour of the persisting maternal enzyme form. InEphestia embryos, however, there is no persistence of the maternal esterases. In all reciprocal heterozygotes a three-banded pattern suddenly appears 96 h after egg deposition, indicating synchronous activation of both parental alleles. The relative intensity distribution in the hybrid patterns approaches that of the mature larvae stepwise and in an allele-specific manner. This result and the fact that the various heterozygous types exhibit unequal total activities suggest that the Esterase-2 alleles have different activities, which are fixed late in embryogenesis.