Ontogeny,function and evolution of extreme floral construction inMonimiaceae

In at least 4 genera of theMonimiaceae (Tambourissa, Wilkiea, Kibara, Hennecartia) extremely specialized flowers with a hyperstigma occur, i.e. a secretory zone in the narrow entrance of the floral cup. The mucilaginous secretion of the hyperstigma and of the carpels produces a transmitting medium for pollen tubes continuous from the mouth of the floral cup to the ovules. As to their floral morphology, the two extreme types,Hortonia andTambourissa, are connected gradually by various other genera. Possible evolutionary trends and systematic problems are outlined.     

Linkage disequilibrium ofMHC genes in the chicken

Two loci of the chicken MHC (theB complex) are expressed in erythrocytes,B-F andB-G. The former is homologous to the murineK andD loci and is also expressed in all white blood cells, while the latter is apparently restricted to RBC and is of unknown relationship toH-2 andHLA loci. A recombinant between two congenic, MHC-different strains, CB and CC, has permitted the production of antisera specific for theB-F andB-G alleles of these two strains, and these and other antisera have been used for typing of outbred populations of various chicken breeds.The cross-reactions found with haplotypes other than the donors' are extensive, sometimes even extreme, but it is possible to narrow the specificity of the typing sera by appropriate absorptions. With absorbed sera we have found a linkage disequilibrium which is almost certainly even stronger than that reported in studies of mammalianMHC loci. We have also made observations which suggest that the gametic association of a given set ofB-F andB-G alleles is probably not merely a matter of random crossing-over events.     

Theoretical study of the optimal shape of the front profile of the lens in the eye of the scallop,Pecten

The optimal shape of the front profile of the thick lens in the eye of the scallop,Pecten is theoretically, geometric optically investigated as a function of the refractive index of the lens and the retina, as well as of the geometrical parameters of the eye. The shape of the theoretical front surfaces is compared with that of the real, experimentally determined front face of the lens. The degree of correction of the lens for spherical aberration of the reflecting spherical mirror in thePecten eye is examined. The optimal shape of the front profile of the lens depends strongly on a set of parameters, such that a certain fine tuning is required among them to assure a full correction for spherical aberration. The extreme variability of the eye parameters and the shape of the front face of the lens in the scallop is inconsistent with this fine tuning requirement. The degree of correction of thePecten lens for spherical aberration might not be as good as it could be, a possible biooptical reason for which is discussed.     

Feeding Behavior and Survival of Glucose-Averse Blattella germanica (Orthoptera: Blattoidea: Blattellidae) Provided Glucose as a Sole Food Source

Glucose-averse (Glu/Glu) male German cockroaches, Blattella germanica, rejected brief exposures to glucose solutions despite food deprivation so extreme as to cause high mortality. In the 24-h period following 5 days of food deprivation, both Glu/Glu and wild-type (+/+) were given a continuous source of 1 M glucose solution. Although Glu/Glu visited the solution as frequently as +/+, feeding bouts were significantly shorter. These truncated feeding bouts were sufficient to extend Glu/Glu longevity to ca. 80 days, regardless of glucose concentration. When provided continuously, the glucose was completely rejected by Glu/Glu for the first 3 days. However, by day 9, glucose intake levels were similar to those of wild-type cockroaches. The initial feeding lag probably accounted for the lower long-term survival of the Glu/Glu vs the +/+ strain. Under the stress of food deprivation, glucose-averse B. germanica modify their feeding behavior toward glucose like many phytophagous insects provided non-host plants in no-choice situations.     

NADH oxidase ofThermus thermophilus HB8 overproduced fromEscherichia coli

Summary An NADH oxidase purified from the extreme thermophileThermus thermophilus HB8 is a monomeric flavoprotein with a 1 1 ratio of flavin-adenine dinucleotide (FAD) to the polypeptide chain. It catalyzes in vitro the oxidation of reduced NADH or NADPH with the formation of H₂O₂. The gene encoding the NADH oxidase fromT. thermophilus HB8 was cloned, and its nucleotide sequence was determined. The molecular mass of 22,749 Da, as deduced from thenox gene, agrees with that of the purified NADH oxidase fromT. thermophilus HB8, as estimated by mass spectrometry. Thenox gene does not contain a GX₄GK consensus sequence typical for nucleotide binding proteins. Thenox gene was overexpressed inEscherichia coli, and a protocol for the rapid purification of theE. coli-borneT. thermophilus NADH oxidase or its His₆-tagged analogue was developed by using thermal denaturation step and affinity chromatography.     

Anticonvulsant properties of spirohydantoins derived from optical isomers of camphor

Natural camphor exists as thed(+) form but thel(–) form has been synthesized. Replacement of the keto group on carbon 2 of each form with a hydantoin moiety led to only one spirohydantoin derivative. Bothd andl derivatives were synthesized. Both forms and their racemic mixture were tested in vivo for toxicity and behavioral effects in mice. A dose of 100 mg/kg of thed form was not toxic: mice showed normal grooming and exploratory behavior; thel form induced hunched posture, body jerks and myoclonic manifestations followed by quiescence. Thedl form showed intermediate effects. Challenge with the convulsant pentylenetetrazol (Metrazol) 2 hr after treatment with placebo or the camphor spirohydantoins produced seizure manifestations in all controls, in half of the subjects pretreated with thed-camphor derivative, in none of those pretreated with thel derivative and an intermediate response in those pretreated with racemic mixture. Thus, a spirohydantoin moiety added to camphor conferred strong anticonvulsive properties on thel form and modest ones on thed form; thed form did not seem to antagonize thel form.     

Temperature sensitivity as a general phenomenon in a collection of chlorophyll-deficient mutants of sweetclover (Melilotus alba)

A collection of chlorophyll (Chl)-deficient mutants of sweetclover (Melilotus alba) with defects in eight nuclear loci were grown at 17 or 26° C. Plants grown at either temperature were examined for Chl content, Chla/b ratio, expression of the light-harvesting complex II (LHC-II) apoproteins, and protochlorophyllide (Pchlide) biosynthetic capacity. Except for thech4 mutant, the parental strain and all mutants accumulate more Chl when grown at 26° C than at 17° C. Thech5 mutants, lacking Chl b under any growth condition, and thech12 mutant showed little temperature-dependent phenotypic plasticity, whereas this was a marked phenomenon in the other mutants. Thech10 andch11 mutants demonstrated extreme temperature sensitivity with regard to the production of Chlb and the Chlb-binding LHC-II apoproteins. When excised trifoliolates were supplemented with exogenously supplied -aminolevulinic acid, only thech4 mutant was markedly impaired in the ability to produce Pchlide. These data indicate that temperature-sensitive phenotypic plasticity is a common phenomenon of chlorophyll-deficient mutants and substantiate that only a minority of Chl-deficient mutants is impaired in the biosynthesis of Chl.This research was supported by Grants GM84-CRCR-1-1479 (J.C.O.) and 89-00641 (J.M.) of the United States Department of Agriculture and by National Science Foundation Grant DMB87-03100 (J.M.). This is paper No. 8971, Nebraska Agricultural Research Division.     

Evolutionary trends of the hairy-faceSaguinus in terms of the dental and cranial morphology

Shape variations in the dentition and the cranium were analyzed for sevenSaguinus forms of the hairy-face tamarin by applying the factor analysis method. The results obtained for the dental and cranial measurements were almost consistent with each other. The magnitude of the difference in shape factors between theS. nigricollis group and theS. midas group is appreciably larger than that between the former group and theS. mystax group. If the ancestral geographic centre of origin is postulated as being within the region which is inhabited by the livingS. nigricollis group, the morphological distances between any pairs of groups correlate well with the geographic distances between them. Concerning the dental and cranial morphologies, the physical changes in the three species group probably took place in two directions; that is, from theS. nigricollis group to theS. mystax group, and from theS. nigricollis group to theS. midas group. The forms belonging to each species group are more closely related to each other, with the exception ofS. imperator in theS. mystax group. The uniqueness ofS. imperator was clearly demonstrated by factor analysis and distance analysis. In theS. mystax group, although still hypothetical,S. imperator may have been related only through the basic ancestral stock toS. labiatus andS. mystax.     

Vegetative diversification and radiation in subtribeDendrobiinae (Orchidaceae): Evidence from chloroplast DNA phylogeny and anatomical characters

The data derived from a chloroplast DNA restriction site analysis of subtribeDendrobiinae (Orchidaceae) indicate that extreme vegetative diversification is concentrated in two limited parts of this group. Overlaying the vegetative character states onto the chloroplast DNA cladogram suggests that several xeromorphic, vegetative characters evolved in the lines leading to the above-mentioned clades. Several anatomical characters are also associated with xeromorphy. These vegetative and anatomical characters facilitated the establishment of this group in various dry habitats. On the other hand, the modifications of size and number of parenchymatous cells substantially contributed to the vegetative diversification. This fact implies that a simple structural adjustment can result in a major modification of growth habits in theDendrobiinae.     

IgG-specific helper activity of t lymphocytes from mice lacking theIr-IgG gene

Cooperative interactions between T and B cells from the congenic inbred mouse strains B10.A(2R) and B10.A(4R) in antibody responses controlled byIr genes have been studied. Within theI region of the MHC, these strains share only theI-A subregion. TheIr gene controlling responsiveness to IgA maps in theI-A subregion, both strains being responders to IgA. T cells from 2R mice collaborate effectively with B cells from 2R or 4R mice for antihapten antibody responses to DNP-IgA. TheIr gene controlling responses to IgG maps in theI-B subregion, and 2R mice are nonresponders for this antigen. Nevertheless, 2R T cells primed with IgG can help responder (4R) B cells -but not syngeneic nonresponder (2R) B cells -in responding to DNP-IgG. These results indicate that mice lacking theIr-IgG gene nonetheless may develop helper T lymphocytes specific for myeloma proteins. In addition, they indicate that cells from congenic mice sharing only theA subregion of theI region can collaborate efficiently.     

Genetic mapping of AFLP/AMF-derived DNA markers in the vicinity of the self-incompatibility locus in<Emphasis Type="Italic"> Ipomoea trifida</Emphasis>

Sporophytic self-incompatibility of diploid Ipomoea trifida is controlled by a single multiallelic locus, the S-locus. To make a fine linkage map around the S-locus, AFLP (amplified restriction fragment length polymorphism) and AMF (AFLP-based mRNA fingerprinting) analyses were performed using bulked genomic DNA and mRNA, respectively, from several plants of each S-haplotype in a segregating population. Putative S-haplotype-specific fragments were obtained and subjected to RFLP analysis of genomic DNA to confirm genetic linkage to the S-locus. Eight DNA markers co-segregating with the S-haplotype were identified and mapped in close proximity to the S-locus. One of them, AAM-68, was the most tightly linked to the S-locus, because no recombinants were detected in the 873 plants of the segregating population analyzed. The S-locus region was defined to be within 1.25 cM in the linkage map. These markers are useful for positional cloning of the S-locus genes in Ipomoea.     

New classification of liverworts based on molecular and morphological data

Complete sequences for the 18S-rRNA gene of 22 bryophytes (12 completely new) were determined and used to construct phylogenetic trees. The evaluation of sequence data according to the maximum parsimony principle (PAUP 3.1.1) and the neighbor-joining method (MEGA) results in similar phylogenetic trees in which theBryopsida appear as a sister group to theJungermanniopsida, and both together as a sister group to theMarchantiopsida. Among theMarchantiopsida, theSphaerocarpales diverge early as a separate clade. TheMetzgeriales andJungermanniales are monophyletic. They belong to one clade and cannot be separated by either method of evaluation.     

TCRVΒ17 gene polymorphism in wild populations of IndianMus musculus

The Indian house mouse populations, in addition to extreme polymorphism for nuclear genes encoding for various isozymes and mitochondrial DNA also show a high level of variation for the TCRVΒ17 gene. RFLP analysis has revealed six variant forms of the gene which are classified on the basis of size of the restriction fragment. These forms are not present in the laboratory inbred strains. The new alleles are present at different frequencies in nine Indian localities. Gene expression study showed that all the alleles are functional and expressed, though at varying levels. No case of clonal deletion was observed in these animals. Variable expression levels of theVΒ17 gene in these natural populations of mice might be under the control of the MHC haplotype.     

Fission yeast Cdc37 is required for multiple cell cycle functions

The identification of a Schizosaccharomyces pombe homologue of the cdc37 gene is described. The gene product is most similar to the budding yeast homologue, but shows similarity to metazoan Cdc37 proteins, with a region of high similarity at the extreme N-terminus. Gene transplacement experiments in diploid cells followed by tetrad dissection show that the gene is essential. Depletion of the gene product after switching off expression of cdc37 from the regulatable nmt81 promoter results in cessation of growth and division. The cells arrest heterogeneously, with a significant proportion showing mitotic defects; paradoxically, a proportion of the cells show a short-cell phenotype consistent with an advanced cell cycle.Communicated by D. Y. Thomas     

The PSO4 gene is responsible for an error-prone recombinational DNA repair pathway in Saccharomyces cerevisiae

Summary The induction of mitotic gene conversion and crossing-over inSaccharomyces cerevisiae diploid cells homozygous for thepso4-1 mutation was examined in comparison to the corresponding wild-type strain. Thepso4-1 mutant strain was found to be completely blocked in mitotic recombination induced by photoaddition of mono- and bifunctional psoralen derivatives as well as by mono- (HN1) and bifunctional (HN2) nitrogen mustards or 254 nm UV radiation in both stationary and exponential phases of growth. Concerning the lethal effect, diploids homozygous for thepso4-1 mutation are more sensitive to all agents tested in any growth phase. However, this effect is more pronounced in the G2 phase of the cell cycle. These results imply that the ploidy effect and the resistance of budding cells are under the control of thePSO4 gene. On the other hand, thepso4-1 mutant is mutationally defective for all agents used. Therefore, thepso4-1 mutant has a generalized block in both recombination and mutation ability. This indicates that thePSO4 gene is involved in an error-prone repair pathway which relies on a recombinational mechanism, strongly suggesting an analogy between thepso4-1 mutation and theRecA orLexA mutation ofEscherichia coli.     

Reduced transmitter release conferred by mutations in theslowpoke-encoded Ca2+-activated K+ channel gene ofDrosophila

Potassium channels control the repolarization of nerve terminals and thus play important roles in the control of synaptic transmission. Here we describe the effects of mutations in theslowpoke gene, which is the structural gene for a calcium activated potassium channel, on transmitter release at the neuromuscular junction inDrosophila melanogaster. Surprisingly, we find that theslowpoke mutant exhibits reduced transmitter release compared to normal. Similarly, theslowpoke mutation significantly suppresses the increased transmitter release conferred either by a mutation inShaker or by application of 4-aminopyridine, which blocks theShaker-encoded potassium channel at theDrosophila nerve terminal. Furthermore, theslowpoke mutation suppresses the striking increase in transmitter release that occurs following application of 4-aminopyridine to theether a go-go mutant. This suppression is most likely the result of a reduction of Ca²⁺ influx into the nerve terminal in theslowpoke mutant. We hypothesize that the effects of theslowpoke mutation are indirect, perhaps resulting from increased Ca²⁺ channel inactivation, decreased Na⁺ or Ca²⁺ channel localization or gene expression, or by increases in the expression or activity of potassium channels distinct fromslowpoke.     

Restimulation in secondary MLC by a non-D-locus determinant within the MHC

By testing a family in which one offspring had inherited a chromosome including a recombination between theHLA-B andD loci, we sought to obtain some insight into whether determinants other than those atHLA-D are capable of restimulating in a secondary MLC. Results obtained in the primary MLC indicated that the recombinant child did not express products of theHLA-D region normally associated with this haplotype. When sensitization in a primary MLC was to the entire haplotype, everyone who had the sensitizing haplotype restimulated strongly and specifically. In addition, however, weak to moderate stimulation was obtained when the cells of the recombinant child were used to restimulate. Presumably these cells possessed determinants coded for within theHLA-A toB chromosomal segments of the sensitizing haplotype, but not those coded for by theHLA-D locus. Our results indicate that a simple structure atHLA-D is not the sole factor in the secondary MLC. Either the products of loci outside theHLA-D locus can also restimulate or the recombination occurred within theD locus, suggesting that theD region contains more than one locus coding for restimulating determinants.     

Expression ofEscherichia coli recA andada genes inSaccharomyces cerevisiœ using a vector with geneticin resistance

Construction ofE. coli-yeast shuttle plasmids containing theneo selection gene is described. The protein-coding regions of theE. coli ada orrecA genes under the control of theADH1 promoter and terminator were ligated into theSphI unique site of pNF2 to produce pMSada and pMSrecA, respectively. The plasmids were used for transformation of the haploid and diploidpso4-1 strains ofS. cerevisiœ and their corresponding wild types. Transformants were obtained by selection for geneticin (G418) resistance. Crude protein samples were extracted from the individual transformants. Both the RecA and Ada proteins were present in all strains containing therecA andada genes on plasmids, respectively. Thus the geneticin selection system was successfully used for the preparation of model, yeast strains.