A DNA Fragment Mapped within the Submicroscopic Deletion of Ph1, a Chromosome Pairing Regulator Gene in Polyploid Wheat

Bread wheat is an allohexaploid consisting of three genetically related (homoeologous) genomes. The homoeologous chromosomes are capable of pairing but strict homologous pairing is observed at metaphase 1. The diploid-like pairing is regulated predominantly by Ph1, a gene mapped on long arm of chromosome 5B. We report direct evidence that a mutant of the gene (ph1b) arose from a submicroscopic deletion. A probe (XksuS1-5) detects the same missing fragment in two independent mutants ph1b and ph1c and a higher intensity fragment in a duplication of the Ph1 gene. It is likely that XksuS1-5 lies adjacent to Ph1 on the same chromosome fragment that is deleted in ph1b and ph1c. XksuS1-5 can be used to tag Ph1 gene to facilitate incorporation of genetic material from homoeologous genomes of the Triticeae. It may also be a useful marker in cloning Ph1 gene by chromosome walking.     

The parasporal inclusion of Bacillus thuringiensis subsp. shandongiensis: characterization and screening for insecticidal activity.

The parasporal body of Bacillus thuringiensis subsp. shandongiensis was characterized in terms of its structure, protein composition, and toxicological properties against several types of insects. The crystals of B. thuringiensis shandongiensis appear to consist of a major protein of 144 kDa present in an spherical inclusion, as determined by transmission electron microscopy, titration curve analysis, and SDS-PAGE of the solubilized crystals. A second protein of ca. 60 kDa is present in trace amounts and appears to be associated with a small bar-shaped inclusion. The 144-kDa protein has been characterized by isoelectric point determination, N-terminal amino acid sequence analysis, amino acid analysis, and immunological cross reactivity. Its N-terminal amino acid sequence differed from that of other B. thuringiensis crystal proteins. The 144-kDa protein was not immunologically related to the crystal proteins of two toxic serovars (B. thuringiensis israelensis and B. thuringiensis kurstaki HD-1) and one nontoxic serovar (B. thuringiensis indiana), as shown in immunoblots probed with antiserum raised against the 144-kDa B. thuringiensis shandongiensis protein, the B. thuringiensis israelensis crystal proteins, and the trypsin resistant fragment of B. thuringiensis kurstaki P1 proteins. In contrast to most B. thuringiensis serovars, B. thuringiensis shandongiensis crystals did not dissolve at pH 12. Solubilization was achieved in sodium bicarbonate at pH 8.3 and in the presence of 25 mM dithiothreitol.(ABSTRACT TRUNCATED AT 250 WORDS)     

Physical mapping of the MHC and grc by the pulse field electrophoresis

The development of the physical map of the major histocompatibility complex of the rat was undertaken using pulse field gel electrophoresis of fragments of genomic DNA from the BIL/2 (grc ⁺) and BIL/1 (grc ^–) strains obtained primarily from single and double digests with the enzymes Mlu I, Not I, and Sfi I and hybridized with a variety of mouse, rat, and human probes. Both strains are maintained by inbreeding the BIL heterozygote (forced heterozygosity; F31); hence, their differences lie almost entirely in the MHC-grc regions. The MHC-grc region was contained in five fragments of DNA comprising 3000–3200 kilobases (kb); thus, its size appears to be closer to that of the human MHC than to that of the mouse MHC. This didstance may be an underestimate of the size of the entire region, however, because the cluster of class I loci in the RT1.A region could not be defined in detail in this study. The most striking difference between the BIL/2 strain, which has normal growth and reproductive characteristics, and the BIL/1 strain, which has growth and reproductive defects and an enhanced susceptibility to chemical carcinogens, is a deletion of approximately 70 kb in the latter strain. The studies og grc ⁺ and grc ^– strain suggest that the phenotypic defects of the grc ^– stains may be due to the loss of genes that are normally present in this deleted region. Address correspondence and offprint request to: T. J. Gill III.     

Properties of a 72-kilodalton mosquitocidal protein from Bacillus thuringiensis subsp. morrisoni PG-14 expressed in B. thuringiensis subsp. kurstaki by using the shuttle vector pHT3101.

The mosquitocidal properties of Bacillus thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni PG-14 are attributable to protein inclusions grouped together within a parasporal body. In both of these strains, the mosquitocidal activity resides in proteins with molecular masses of 27, 72, 128, and 135 kDa. In an attempt to determine the toxicity of each protein, the shuttle vector pHT3101 was used to express the cryIVD gene (encoding the 72-kDa CryIVD protein) from B. thuringiensis subsp. morrisoni in an acrystalliferous mutant of B. thuringiensis subsp. kurstaki. With this system, parasporal inclusions of the 72-kDa protein were obtained that were comparable in size, shape, and toxicity to those produced by parental B. thuringiensis subsp. morrisoni. The inclusions were bar shaped, measured 500 by 300 by 150 nm, and were easily visible with phase-contrast microscopy by 16 h of cell growth. A 50% lethal concentration of 64 ng/ml for these inclusions was determined in bioassays against fourth instars of Culex quinquefasciatus, which was similar to the 50% lethal concentration of 55 ng/ml obtained for the 72-kDa inclusion from B. thuringiensis subsp. israelensis. In contrast, expression of the cryIVD gene in Escherichia coli was very low and only detectable by immunoblot analysis. These results demonstrate that the pHT3101-B. thuringiensis expression system can be used to express the CryIVD protein in quantities and with properties comparable to that obtained with the natural host. This system may prove useful for the expression of other B. thuringiensis proteins and, in particular, for reconstitution experiments with inclusions produced by the mosquitocidal subspecies of B. thuringiensis.     

Extracellular development, in vitro, of the erythrocytic cycle of Plasmodium falciparum

One way to explore the nature of the dependence o f intracellular parasites on their host cell is to replace the living host cell with a non-living environment that supports development o f the parasite. Bill Trager, Jonathan Williams and Gokal Gill describe their methods for obtaining extracellulor development of erythrocytic stages of Plasmodium falciparum.     

Cytogenetic studies in Cochlearia L. (Cruciferae). The chromosomal constitution of C. danica L.

Genome analysis has been used to investigate the relationship of C. danica L. to the diploid and tetraploid species in the genus. The results of this analysis suggest that C. danica has the genomic constitution A¹ ₇ A¹ ₇ A² ₇ A² ₇ T₇ T₇. Both A₇ genomes in C. danica are segmentally differentiated from the A₇ genome in C. groenlandica L. and from the similar A₆ genomes in C. pyrenaica DC. and C. officinalis L. but, in hybrids, are capable of some pairing with these genomes. A² ₇ is more distantly related to A₇ and A₆ than is A¹ ₇ and in A₇ some of the differentiation has taken place by reciprocal translocations. A¹ ₇ and A² ₇ still retain enough homology for pairing between them. The T₇ genome has no homology with any of the A genomes but may be derived from an ancestral T₆ genome.     

Deviation from homeoviscous adaptation in Escherichia coli membranes.

The process by which an organism changes the composition of its membranal fatty acids in response to growth temperature, so as to maintain optimal membrane functioning, is known as homeoviscous adaptation (HA). One expression of HA is the constancy of the fluorescence polarization (P) of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene (DPH) in membranes of cells grown at various temperatures. The P of DPH in the membranes of Escherichia coli was shown by us to be inversely proportional to bacterial growth rate on different carbon sources. This result, implying failure of HA, is now complemented by measurements of DPH lifetimes, which indicate that the dominant variables contributing to the drop in P are (a) the order parameter of the membrane, which goes down, and (b) the fluidity, which may slightly increase. These are then the changes induced by enhanced growth rate. Two additional effects, cell membrane permeability and sensitivity to thermal shock, determined by the diffusion of o-nitrophenylgalactoside (ONPG) and by exposure to 52 degrees C, respectively, are reported to increase with growth rate. We can now conclude that there is a deviation from the principle of HA in E. coli grown at various rates, brought about by controlling the growth media at constant temperatures.     

Tree pre-planting at Rutland Water

The aesthetic problems posed by reservoir margins have been recognised for many years. They stem from a complex pattern of action and interaction of several environmental variables — water manipulation patterns, wave action, substrate characteristics, grazing and other factors.These factors usually result in reservoir margins being not only visually unattractive and physically unstable, but also of low ecological diversity.The Natural Environment Research Council and Water Research Centre have, since 1968, together funded a research programme to investigate the use of plant species to combat these problems.In isolated instances in the UK, woody species in particular have succeeded in colonising margins naturally, to very good visual effect (e.g. at Lake Vyrnwy). Biotechnical treatments — using artificially planted shrubs and trees — have been successfully used in large Central European reservoirs to combat soil erosion by waves.Following trials of various woody species on the margins of established reservoirs, 10 species/cultivars were pre-planted above and below the future top water level (TWL) of Rutland Water in 1973, 5 years before reservoir filling was completed. Pre-planting theoretically allows better survival by trees of subsequent partial inundation. The precise siting of pre-planted trees in the vertical dimension should normally be done using modelled predictions of reservoir summer median water level (a variable important to tree survival). The complex factors affecting the operation of Rutland Water made precise prediction impossible; the experimental plot eventually extended from 500 mm above TWL, to 2 000 mm below TWL, in 500 mm steps.252 trees were involved in the plot and their growth has been monitored for the past 7 years. The species being predominantly hydrophytic, drought was a problem in the years prior to reservoir filling. Mulching with grass-cuttings proved to be helpful in this respect. Subsequently, prolonged periods of high water level have been deleterious to tree survival in the lower areas of the plot. Once abstraction/recharging patterns become more cyclic, it is anticipated that tree growth will stabilise.Department of Applied Biology, University of Cambridge     

Epistatic,additive and dominance variation in a triple test cross of bread wheat (Triticum aestivum L.)

Summary Triple test cross progenies resulting from the crossing of three testers (Kloka, UP 368 and an f₁ intermediate between them) and 24 varieties of bread wheat have been studied for plant height (cm), peduncle length (cm), ear length (cm), number of spikelets per spike and harvest index (ratio between economic and total yield). Epistasis was not significant for any of the characters studied. The testers were inadequate for plant height and for peduncle length although the testers varied considerably for these traits. Additive variance played a significant role in the inheritance of all the characters except number of spikelets per spike. The dominance variance was important for plant height, ear length and harvest index. The degree of dominance was in the over-dominance range for plant height. Complete dominance was operative for ear length, number of spikelets per spike and harvest index whereas for peduncle length only partial dominance was observed. The possibility of the isolation of the recombinants with high harvest index has been stressed.     

Recovery growth following periods of restricted rations and starvation in rainbow trout Salmo gairdneri Richardson

Fingerling rainbow trout (6.9 to 11.6 cm fork length) maintained at 12° C were either: fed restricted rations (3% of dry body weight/day) for 16 weeks, or starved for 3 weeks (14.5% weight loss), or for 13 weeks (32.5% weight loss). Whole body wet weights and dry weights (as % of wet weight), and wet and dry weights of tissues (heart, spleen, liver, skin, gonad, gut, visceral fat, and 'carcase') resulting from these treatments were compared to corresponding weights of an 'initial sample'. Visceral fat was completely utilized during both short and long-term starvation and gut was significantly reduced in the extended starvation group.
During subsequent recovery growth on full rations, following food deprivation, fish grew at approximately the same rate as fully-fed controls, which had not suffered food deprivation, as regards body wet weight and condition ( K ), but tended to surpass controls in % dry weight.
Heart, liver, gonad, gut and visceral fat also increased during recovery growth in a manner suggestive of overcompensation in response to former food deprivation. Gut, skin and carcase dry weights were less in the control, and in the 14.5% weight loss groups following recovery growth than in those in 3% rations and in the severely starved group, indicating that the effects of slow growth resulting from limited rations resemble those of severe starvation more than those of limited starvation.