Triterpene methyl ethers of Chionochloa (Gramineae)

The leaf wax of twenty species of Chionochloa was examined for triterpene methyl ethers; twelve species gave positive yields. The known pentacyclic methyl ethers arundoin, miliacin, lupeol methyl ether and β-amyrin methyl ether were identified and the methyl ethers of the tetracyclic alcohols, cycloartenol and parkeol are reported as new natural products. Arundoin and miliacin occur in many species while the remaining compounds may be suitable chemotaxonomic markers.     

Association of a sperm-specific protein with the mitochondrial F1F0-ATPase in Heliothis: Implications for sterility of H. virescens × H. subflexa backcross hybrids

Evidence is presented that a mitochondrial protein that displays a species-specific net charge in the Heliothis spp complex is associated with subunits of the F₁-ATPase. The expression of this 63 kd polypeptide, p63, was restricted to sperm and its developmental pattern of synthesis and accumulation paralleled that of the putative β-subunit of the F₁-ATPase complex. Comparisons of the enzyme from fertile Heliothis virescens and sterile (H. virescens × H. subflexa) backcross hybrid males revealed two differences. First, the specific activity of the F₁-ATPase isolated from sterile males was half that of preparations from fertile males; and second, the p63 protein was bound less tightly to the complex in backcross sperm. The implications of these findings in relation to both the identification of the cause of backcross male sterility and prospects for future research are discussed.     

Biochemical genetics of hybrid sunfish: Differential survival of heterozygotes

Allelic segregation in reciprocal backcrosses involving the largemouth bass (Micropterus salmoides) and the F₁ hybrid (largemouth bass × smallmouth bass, M. dolomieui) was investigated to determine the extent of euheterosis and luxuriance. The frequencies of allelic isozymes encoded in the lactate dehydrogenase E, malate dehydrogenase B, and isocitrate dehydrogenase loci were determined for reciprocal backcross progeny subjected to different selection pressures. The progeny of the backcross (male F₁ × female largemouth bass) underwent a rapid loss of heterozygous individuals in a natural pond environment. When the offspring of this same mating were placed in artificial pools, where cannibalism is the main source of mortality, heterozygosity was advantageous. There was a marked correlation of increased heterozygosity at these enzyme loci with an increased growth rate. None of the above responses to selection was observed when the F₁ hybrid served as the maternal parent in the reciprocal backcross. A maternal factor in the egg cytoplasm may influence the expression of heterosis.     

Genetic control of “natural” killer lymphocytes in the mouse

Spleens from normal young mice contain lymphocytes that can kill certain in vitro grown Moloney lymphoma lines in a⁵¹Cr-release cytotoxicity test. A lymphoid cell without detectable T- or B-cell markers was previously shown to be responsible. Killing activity shows a marked dependence on the genotype of the donor mouse. When tested against a YAC line of strain A origin maintained in vitro spleens of A, A.CA, and A.SW mice had low activity, whereas CBA, C3H, C57L, and C57Bl spleens were highly active. In semisyngeneic F₁ crosses with strain A as one parent, reactivity resembled the opposite parental strain. Thus, (A×CBA)F₁, (A×C3H)F₁, (A×C57L)F₁, and (A×C57Bl)F₁ were reactive, whereas A×A.CA showed no significant activity. Analysis of the reactivity in (A×C57Bl)F₁×A backcross mice suggests that multiple genes are involved. Preliminary linkage analysis suggests at least oneH-2 linked factor. Another gene appears to be linked to theB (black) locus.     

An interstitial telomere array proximal to the distal telomere of mouse chromosome 13

Lambda clones of mouse DNA from BALB/c and C57BL/10, each containing an array of telomere hexamers, were localized by FISH to a region close to the telomere of Chr 13. Amplification of mouse genomic DNA with primers flanking SSRs within the cloned DNA showed several alleles, which were used to type eight sets of RI strains. The two lambda clones contained allelic versions of the interstitial telomere array, Tel-rs4, which is 495 bp in C57BL/10 and which includes a variety of sequence changes from the consensus telomere hexamer. Comparison of the segregation of the amplification products of the SSRs with the segregation of other loci in an interspecies backcross (C57BL/6JEi × SPRET/Ei) F₁× SPRET/Ei shows recombination suppression, possibly associated with ribosomal DNA sequences present on distal Chr 13 in Mus spretus, when compared with recombination in an interstrain backcross, (C57BL/6J × DBA/J) F₁× C57BL/6J, and with the MIT F₂ intercross. Analysis of recombination in females using a second interstrain backcross, (ICR/Ha × C57BL/6Ha) F₁× C57BL/6Ha, also indicates recombination suppression when compared with recombination in males of the same strains, using backcross C57BL/6Ha × (ICR/Ha × C57BL/6Ha) F₁. Thus, more than one cause may contribute to recombination suppression in this region. The combined order of the loci typed was D13Mit37–D13Mit30–D13Mit148–(D13Rp1, 2, 3, 4, Tel-rs4)–D13Mit53–D13Mit196–D13Mit77–(D13Mit78, 35). Data from crosses where apparently normal frequencies of recombination occur suggest that the telomere array is about 6 map units proximal to the most distal loci on Chr 13. This distance is consistent with evidence from markers identified in two YAC clones obtained from the region. Received: 24 September 1996/Accepted: 20 January 1997     

Fine mapping of Ath6, a quantitative trait locus for atherosclerosis in mice

Ath6 is a novel quantitative trait locus associated with differences in susceptibility to atherosclerosis between C57BL/6J (B6) and C57BLKS/J (BKS) inbred mouse strains. Combining data from an intercross and a backcross (1593 meioses) between mice from B6 and BKS strains and from The Jackson Laboratory interspecific backcross panels, (C57BL/6J ×Mus spretus) F₁× C57BL/6J and (C57BL/6J × SPRET/Ei) F₁× SPRET/Ei, we constructed a consensus genetic map and narrowed Ath6 to a 1.07 ± 0.26 cM interval between the anonymous DNA marker D12Pgn4 and the gene Nmyc1. This region is near the proximal end of murine Chromosome (Chr) 12, which is homologous to the human chromosomal region 2p24-p25. Marker order in the Ath6 region was concordant among the two crosses and The Jackson Laboratory interspecific backcross panels. This high resolution map rules out candidate genes encoding apolipoprotein B, syndecan 1, and Adam17. The two Ath6 crosses have a combined potential resolution of 0.06 cM. Received: 12 September 2000 / Accepted: 22 February 2001     

Murine responses to (Tyr-Glu-Ala-Gly)n: II. H-2 and non-H-2 gene effects

Mice of the H-2^b haplotype responded to the sequential polymer poly(Tyr-Glu-Ala-Gly) in the in vitro T-cell proliferative assay, irrespective of whether they were homozygous or heterozygous at the H-2^b locus. The antibody responses of the H-2^b congenic mice to this polymer were variable, with A.BY and BALB.B showing responses better than those of C57BL/6 and C57BL/10 strains. The antibody responses of the F₁ progeny of (responder × nonresponder) strains of mice to this polymer are generally lower than the responder parents. F₁ mice with C57BL/10 background were the poorest responders. Studies with F₂ mice and backcross progenies of selective breeding of high and low antibody responder (C57BL/6 × BALB/c) F₁ to high responder C57BL/6 mice indicated that both non-H-2 genes and H-2 gene dosage effects influenced the magnitude of the humoral antibody responses. Animals having low responder non-H-2 background and only half the dosage of the responder immune response genes has greatly diminished antibody responses.     

Enzymatic removal of a cephalosporin methyl ester blocking group

Over 7000 microorganisms were screened to find an enzyme source for the hydrolysis of a C₄ methyl ester blocking group on 7-aminodesacetoxycephalosporanic acid (7-ADCA). Only one culture, Streptomyces capillispira Mertz and Higgens nov. sp., produced an enzyme that catalysed the reaction. Enzyme synthesis in a defined mineral salts medium was repressed by NH₃ and amino acids. Under optimum fermentation conditions, the maximum rate of substrate hydrolysis was 6 × 10^?10 mol min^?1 mg^?1 cell. The enzyme was recovered from the mycelia and partially purified by gel filtration. Kinetic studies by pH-stat titration indicated that the pH optimum was 7.5–8.5, the temperature optimum was 25–30°C, and the substrate K_m value was 2.3 mg ml^?1. The reaction products, 7-ADCA and methanol, were weak competitive inhibitors of the enzyme with K₁ values of 6.63 and 0.188 mg ml^?1, respectively. The enzyme also hydrolysed cefaclor and cephalexin methyl esters but did not hydrolyse cephalosporin ethyl esters. With further improvements in enzyme yields and stability, enzymatic deblocking of cephalosporins could provide an alternative to chemical deblocking processes.     

Genetic regulation of alcohol dehydrogenase C2 in the mouse. Developmental consequences of the temporal locus (Adh-3t) and positioning of Adh-3 on chromosome 3

The tissue specificity of a proposed cis-acting temporal locus (Adh-3t), which regulates alcohol dehydrogenase C₂ (ADH-C₂) activity in mouse reproductive tissue extracts, has been examined in C5 7BL/6J, SM/J, F₁ (SM/J × C5 7BL/6J) mice as well as in progeny of an (F₁ [SM/J × C5 7BL/6J] × C5 7BL/6J) back-cross. Electrophoretic variants for ADH-C₂, previously used to localize the gene (Adh-3) encoding this enzyme on chromosome 3, enabled the relative parental contributions to ADH-C₂ phenotype in F¹ and backcross mouse tissues to be determined. These analyses demonstrated that (1) stomach, kidney, lung, adrenals, seminal vesicles, epididymis, uterus, and ovary ADH-C₂ is encoded by a single locus (Adh-3); Adh-3t is differentially active in various tissues, eg, lung exhibits no apparent activity whereas the temporal locus is fully active in seminal vesicles; (3) Adh-3t is probably differentically active in different cells of some tissues, eg, adrenals. Specific activity profiles of stomach and epididymal ADH-C₂ during the neonatal development of C5 7BL/6J, SM/J, and F₁ (SM/J × C5 7BL/6J) male mice supported the proposal for a cis-acting temporal locus for this enzyme. Genetic analyses examining segregation of Adh-3 and Adh-3t among backcross progeny suggested that these are distinct but closely linked loci, since one recombinant among 256 progeny was observed. Linkage data of Adh-3 with Va (varitint-waddler) and de (droopy ear) was also obtained, which suggested that Adh-3 is localized on chromosome 3 between Va and de.     

A gene locus affecting tolerance to BGG in mice.

A genetic analysis was made of the ease of tolerance induction to bovine γ-globulin (BGG) in DBA/2, BALB/c, F₁ and backcross generation mice. Like parental DBA/2 mice, the F₁ generation of BALB/c × DBA/2 becomes tolerant when treated with 2 mg BGG. A backcross of this F₁ to DBA/2 parents produced mice that all became tolerant to this dose of BGG. A backcross of F₁ mice to BALB/c parents produced 50% offspring tolerized by the same dose of BGG and 50% resistant to tolerance induction.The data suggest a single autosomal locus affecting tolerance induction. Data presented elsewhere suggest that the locus affects macrophage function. We propose that this locus be called tolerance (symbol Tol-l) and the two alleles be (Tol-l^a (DBA/2 type) and Tol-l^b (BALB/c type) with Tol-l^a being dominant.     

A multiplexed set of microsatellite markers for discriminating <Emphasis Type="Italic">Acacia mangium</Emphasis>, <Emphasis Type="Italic">A. auriculiformis</Emphasis>, and their hybrid

In order to assist breeding and gene pool conservation in tropical Acacias, we aimed to develop a set of multipurpose SSR markers for use in both Acacia mangium and A. auriculiformis. A total of 51 SSR markers (developed in A. mangium and natural A. mangium x A. auriculiformis hybrid) were tested. A final set of 16 well-performing SSR markers were identified, six of which were species diagnostic. The markers were optimized for assay in four multiplex mixes and used to genotype range-wide samples of A. mangium, A. auriculiformis, and putative F₁ hybrids. Simulation analysis was used to investigate the power of the markers for identifying the pure species and their F₁, F₂, and backcross hybrids. The six species diagnostic markers were particularly powerful for detecting F₁ hybrids from pure species but could also discriminate the pure species from F₂ and backcross progenies in most cases (97 %). STRUCTURE analysis using all 16 markers was likewise able to distinguish these cross types and pure species sets. Both sets of markers had difficulties in distinguishing F₂ and backcross progenies. However, identifying F₁ from pure species is the current primary concern in countries where these species are planted. The SSR marker set also has direct application in DNA profiling (probability of identity?=?4.1?×?10^?13), breeding system analysis, and population genetics.     

Reproductive relationships between annual species ofCalendula (Compositae)

Breeding experiments were carried out inCalendula species. In the annuals, which are selfers, rarely some outcrossing was observed only in the most peripheral flowers. In experimental crosses fruit was produced in all combinations. Fertile F₁ and F₂ hybrids could be grown from crosses between parents with similar chromosome numbers:C. palaestina ×C. pachysperma and crosses of different morphological forms ofC. arvensis. In crosses of species with different chromosome numbers at least partly fertile F₁ hybrids were obtained fromC. tripterocarpa ×C. stellata andC. tripterocarpa ×C. arvensis and crosses of the latter withC. palaestina. Fertile F₂ plants were grown from the combination ofC. arvensis ×C. tripterocarpa. Considering this information and previously obtained data, a scheme is proposed for explaining speciation in the genusCalendula.     

Genetic mapping of the whirler mutation

The whirler (wi) mutation on mouse Chromosome (Chr) 4 results in an autosomal recessive neuroepithelial deafness and vestibular dysfunction exhibited as a characteristic shaker-waltzer behavior (deafness, circling, and head-bobbing). We have constructed a genetic linkage map across the wi region in both an interspecific [(wi/wi× CAST/Ei)F₁×wi/wi] backcross (n = 817) and an intraspecific [(wi/wi× CBA/Ca)F₁×wi/wi)] backcross (n = 335). In the interspecific backcross, wi was found to be non-recombinant with Orm1, 0.12 cM distal of D4Mit87 and Ambp, and 0.12 cM proximal of CD301. In the intraspecific backcross, wi was found to be non-recombinant with Orm1 and D4Mit244, 0.3 cM distal of Mup1, and 0.6 cM proximal of Tnc. We also report a family from the interspecific backcross that shows evidence of multiple recombinations across the region of mouse Chr 4 around the wi locus. These rearrangements appear specific to both the region and the family. Received: 10 July 1998 / Accepted: 19 January 1999     

Genetic analysis and molecular mapping of seedling survival drought tolerance gene in lentil (<Emphasis Type="Italic">Lens culinaris</Emphasis> Medikus)

Lentil populations were developed from crosses between ‘JL-3’ (sensitive to drought stress) and ‘PDL-1’ and ‘FLIP-96-51’ (tolerant to drought stress), to study the inheritance of drought tolerance and to identify the markers associated with it. The parental types, F₁, F₂, F₃, and backcross (BC) generations were screened for drought tolerance using seedling survivability and drought scores. The F₁ hybrids responded similar to the drought-tolerant parent, indicating dominance of seedling drought tolerance over sensitivity. Segregation for seedling survival drought tolerance versus sensitivity in F₂ generation was in complete agreement with monogenic 3:1 ratio. The F₃ families and backcross data additionally confirmed monogenic tolerance based on seedling survival under drought. Out of 51 SSR markers screened, thirteen markers were polymorphic between the parental types. Seven markers among them were found to be associated with seedling survival drought tolerance through bulk segregant analysis. Association of these markers with seedling survival drought tolerance was further confirmed through their screening on 10 drought-tolerant and drought-sensitive genotypes. These seven markers were screened in F₂ mapping population (JL-3 × PDL-1) of 101 individuals to map their position in relation to the gene for seedling survival drought tolerance. Linkage analysis mapped the seven markers within a map distance of 133.2 cM. A single major gene Sdt was identified with a LOD value of 19.9 and phenotypic variation (R ²) of 69.7 %. The Sdt locus was obtained in the marker interval of PLC_105–PBA_LC_1480 spanning 24.9 cM with the closest marker PLC_105 at a distance of 9.0 cM on the obtained linkage group. This is the first report on genetic control and linkage of SSR markers for drought tolerance in lentil. These linked markers can be used in molecular breeding programmes for introgression of seedling survival drought tolerance gene in high-yielding cultivars.     

Linkage of diabetes insipidus and agouti genes in the rat

Linkage of the hooded (h), agouti (A), and diabetes insipidus (di) genes was found in (ACI×DI)F₁×DI backcross rats. The genetic map distance A-di for females and for males was 19±5 and 28±5 cM, respectively. However, this difference was not significant. The combined data showed the map distance to be 25±4 cM. The three-point cross showed the following corrected distances and order of genes: h-42±4-A-25±4-di. However, the linkage of h and A, although significant (x²=9.03, P<0.001), is only tentative and must be confirmed by additional studies.     

Structural polymorphism of murine factor B controlled by a locus closely linked to the H-2 complex and demonstration of multiple alleles

New alleles of murine factor B (Bf) protein were demonstrated. When ethylenediaminetetraacetic acid (EDTA)-plasmas from inbred and wild mice were analyzed by isoelectro-focusing (IEF) and immunofixation, murine Bf proteins were visualized as distinct protein bands in all mice tested. Four variants of murine Bf could be demonstrated in a large number of tested mice: Bf 1 (isoelectro-focusing point (P.I.) range of 5.8–6.1) exemplified by B10 and B10.BR, Bf 2 (P.I. range of 5.8–6.0) exemplified by B10.MOL (OHM), Bf 3 (P.I. range of 5.6–5.9) exemplified by B10.MOL (TEN2) and Mus musculus (Mus m.) subspecies Chc, Bf4 (P.I. range of 6.0–6.3) exemplified by Mus m. subspecies Shh. The genetic linkage between S locus and Bf locus was studied with two backcross progenies — [B 10.BR × (B10.BR × Mus m. subspecies Chc)F₁] and [B 10.BR × (B10.BR × Mus m. subspecies Shh)F₁]. Totally, 256 backcross progenies were typed for Bf type and for Ss type (plasma level of the fourth complement protein regulated by S locus). The results indicated that murine Bf was controlled by a single codominant locus located close to the H-2 complex because no mouse showing recombination between Bf locus and S locus was found.     

Control of the antibody response of C57BL/6 Lyt-congenic strains to the Lyt-3.1 alloantigen by a gene linked to theLyt-2 locus

Congenic anti-Lyt-3.1 sera have recently been produced by immunizing B6-Lyt-2^a mice with thymocytes from either B6-Lyt-2^a, Lyt-3^a or B6-Lyt-2^a, Lyt-3^a, H-2^k mice (Boos et al. 1978). Surprisingly, mice of the congenic strain B6 failed to produce either anti-Lyt-2.1 or anti-Lyt-3.1 cytotoxic antibodies after identical immunizations. To determine the genetic basis for the difference in response to Lyt-3.1, (B6 × B6-Lyt-2^a)F_a mice and progeny of the backcross, (B6 × B6-Lyt-2^a)F₁ × B6-Lyt-2^a, were immunized with B6-Lyt-2^a, Lyt-3^a, H-2^k thymocytes. In addition, thymic biopsies of backcross progeny were performed and thymocytes tested for the Lyt-2.2 antigenic specificity. Results indicate that gene(s) governing the immune response to Lyt-3.1 is (are) linked to theLyt-2 locus, and that the responder allele (linked toLyt-2 ^a ) shows very poor penetrance in Lyt-2^a/Lyt-2^b mice.     

Hybridization of Australian Haemonchus placei (place, 1893), Haemonchus contortus cayugensis (Das & Whitlock, 1960) and Haemonchus contortus (Rudolphi, 1803) from Louisiana

There were distinct ecotypie differences in the ability to develop to third stage larvae at a constant 11 or 13°C for two weeks. H. conforms cayugensis could develop at both 11 and 13°C; H. contortus from Louisiana could develop at 13°C but not 11°C and H. placei could not complete development at either temperature. Eggs produced from the first cross between ecotypes had the same cold tolerance as their maternal parent. F₁ eggs were intermediate between parental ecotypes, while F₂ eggs, when fertile, developed at the same temperature as the most cold-tolerant parent in the first cross. The H. placei knobbed vulvar morph type was dominant to H. contortus cayugensis smooth morph type and recessive to the H. contortus (Louisiana) smooth morph type. Knobbed morph type of H. contortus (Louisiana) was dominant to smooth of H. contortus cayugensis. Dominance of morph type was enhanced by a matroclinous effect in the between species matings but not in the within species matings. No fertile eggs were produced by the F1 of the mating between female H. placei × male H. contortus (Louisiana) or by the F₂ of the reciprocal mating. Reciprocal matings between H. placei × H. contortus cayugensis produced an F₁ and F₂ that had reduced fertility but were not completely sterile. No decrease in fertility was observed in the F₂ and F₁ from the H. contortus cayugensis × H. contortus(Louisiana)matings. Cytological studies revealed several kinds of meiotic disturbances in the between species F₁ and F₂. The most frequent were aneuploidy, failure of chromosome pairing, and pairing between non-homologous chromosomes. Males were more seriously affected than females by meiotic disturbances. The H. placei X chromosome appeared to be selected against by the hybrid genetic background. Within species hybrids showed no meiotic disturbances and the percentage of male offspring from these crosses was the same as for female offspring.     

Use of single nucleotide polymorphisms identifies backcrossing and species misidentifications among three San Francisco estuary osmerids

Two threatened osmerid species native to the San Francisco Estuary (SFE)—Delta Smelt (Hypomesus transpacificus) and Longfin Smelt (Spirinchus thaleichthys)—are subject to broad human influence, including significant habitat alteration and the presence of the introduced osmerid, Wakasagi (Hypomesus nipponensis). The identification of these closely related species and their hybrids is difficult in field collected specimens which are subject to damage through handling and may be difficult to identify morphologically, especially when young. In addition, it is known that these three species hybridize, but the extent and effect of hybridization is difficult to quantify and monitor. We developed assays for 24 species-specific single nucleotide polymorphisms (SNPs) that identify whether a sample is a pure species (Delta Smelt, Longfin Smelt, or Wakasagi), a first generation (F₁) hybrid, or a backcross. We used this SNP panel to genetically identify wild osmerids collected in Yolo Bypass from 2010 to 2016 and detected nine Delta Smelt × Wakasagi F₁ hybrids and two Wakasagi × (Delta Smelt × Wakasagi) backcross hybrids; all assayed hybrids had Wakasagi as the maternal parent. The backcrossing into Wakasagi suggests that hybridization may only occur in one direction and thus preclude introgression to Delta Smelt. We also found substantial morphological field misidentifications (32.7%) in the Yolo Bypass samples resulting in more Wakasagi and fewer Delta Smelt than previously recorded when based on morphology. The SNP panel described in this study constitutes a valuable resource for monitoring hybridization in the SFE and assigning species identifications with accuracy and efficiency.     

Development of a ploidy series from a single interspecific Trifolium repens L.Ā . nigrescens Viv. F1 hybrid

 The objective of the current research was to generate a ploidy series of backcross progenies from a single triploid (2n=3x=24) Trifolium repens×T. nigrescens F₁ hybrid (3x H-6909-5). The 3x H-6909-5 plant was highly sterile and produced no seeds from approximately 3000 reciprocal backcrosses to both parental species. Chromosome doubling by an in vitro colchicine method resulted in a marked increase in fertility. Pollen stainability was increased from 9.9% in 3x H-6909-5 to an average of 89.2% (range 87.7–90.9%) in the three chromosome-doubled 6x H-6909-5 plants. Subsequent backcrosses of 6x H-6909-5 and interbreeding of backcross derivatives resulted in an array of fertile hybrids at 4x, 5x and 7x levels and some aneuploids. The occurrence of 7x BC₁F₁ progeny from the T. repens×6x H-6909-5 (4x×6x) cross is the first unequivocal evidence of functional female 2n gametes in white clover. Meiotic pairing in F₁ and BC₁F₁ progeny indicated the presence of allosyndetic pairing, suggesting that genetic exchange between the two species is possible. Received: 17 October 1996 / Accepted: 8 November 1996