Species-specific diversity among simian immunodeficiency viruses from African green monkeys

The prevalence, natural history, and genetic characteristics of simian immunodeficiency virus (SIV) infections in most feral African monkey species are presently unknown, yet this information is essential to elucidate their origin and relationship to other simian and human immunodeficiency viruses. In this study, a combination of classical and molecular approaches were used to identify and characterize SIV isolates from West African green monkeys (Cercopithecus sabaeus) (SIVagm isolates). Four SIVagm viruses from wild-caught West African green monkeys were isolated and analyzed biologically and molecularly. Amplification, cloning, and sequencing of a 279-bp polymerase fragment directly from uncultured peripheral blood mononuclear cells was facilitated by the use of nested polymerase chain reaction. The results indicated that West African green monkeys are naturally infected with SIVs which are closely related to East African SIVagm isolates. However, structural, antigenic, and genetic differences were observed which strongly suggest that the West African green monkey viruses comprise a phylogenetically distinct subgroup of SIVagm. These findings support our previous hypothesis that SIVagm viruses may have evolved and diverged coincident with the evolution and divergence of their African green monkey host. In addition, this study describes a polymerase chain reaction-based approach that allows the identification and molecular analysis of divergent SIV strains directly from primary monkey tissue. This approach, which does not depend on virus isolation methods, should facilitate future studies aimed at elucidating the origins and natural history of SIVs in feral African green monkey populations.     

A plastome mutation affects processing of both chloroplast and nuclear DNA-encoded plastid proteins

Summary A bovine tRNA gene cluster has been characterized and the sequences of four tDNAs determined. Two of the tDNAs could encode tRNA^Ser _IGA, one tDNA^Ser _UGA, and the fourth tRNA^Gln _CUG. The three serine tDNAs representing the UCN codon isoacceptor family are almost identical. However, the sequence of the tDNA^Ser _TGA differs from a previously sequenced bovine tDNA^Ser _TGA at 12 positions (ca. 14%). This finding suggests that in the bovine genome, two subfamilies of genes might encode tRNA^Ser _UGA. It also raises the possibility that new genes for a specific UCN isoacceptor might arise from the genes of a different isoacceptor, and could explain previously observed differences between species in the anticodons of coevolving pairs of tRNAs^Ser _UCN. The gene cluster also contains complete and partial copies, and fragments, of the BCS (bovine consensus sequence) SINE (short interspersed nuclear element) family, six examples of which were sequenced. Some of these elements occur in close proximity to two of the serine tDNAs.     

Action Spectrum for Resetting the Circadian Phototaxis Rhythm in the CW15 Strain of Chlamydomonas: II. Illuminated Cells

The action spectrum for resetting the phase of the circadian clock in Chlamydomonas reinhardtii is different depending upon whether the light stimuli are presented to cells that were in darkness versus dim illumination before stimulation. In this report, we show that phase resetting of illuminated cells appears to be mediated by components of the photosynthetic apparatus. This conclusion is based upon the action spectrum for phase-shifting illuminated cells (which looks like that for photosynthesis) and upon the fact that inhibitors of photosynthetic electron transport also inhibit the light-induced phase shift of illuminated cells. Both of these characteristics differ from that of cells taken from darkness. We, therefore, believe that at least two resetting pathways for this circadian clock exist and that both of these pathways are ecologically significant.     

Regulation of Phytochrome Message Abundance in Root Caps of Maize : Spatial, Environmental, and Genetic Specificity

In many cultivars of maize (Zea mays L.) red light affects root development via the photomorphogenetic pigment phytochrome. The site of perception for the light is the root cap. In the maize cultivar Merit, we investigated phytochrome-mediated events in the cap. We established that the message encoded by the phyA1 gene was most abundant in dark-grown tissue and was asymmetrically distributed in the root cap, with greatest expression in the cells which make up the central columella core of the cap. Phytochrome message was negatively autoregulated in a specific region within the root cap. This autoregulation was sensitive to very-low-fluence red light, and thus was characterized as a phytochrome-mediated, very-low-fluence event. The kinetics of message reaccumulation in the dark were also examined and compared to the kinetics of the light requirement for root gravitropism in this cultivar. Similarly, the degree of autoregulation present in two other maize cultivars with different light requirements for gravitropic sensitivity was investigated. It appears that the Merit cultivar expresses a condition of hypersensitivity to phytochromemediated light regulation in root tissues. We conclude that phytochrome regulates many activities within the cap, but the degree to which these activities share common phytochrome-mediated steps in not known.     

Expected utility maximization and selection of stable plant cultivars

Summary In most plant breeding programs, selection of the best commercially suitable cultivars for a target group of environments is based on information obtained from evaluation trials cultivated in a sample of environments. Information on the performance of cultivars collected in a sample of environments can only be approximate and, consequently, selection of the best cultivar involves choosing among cultivars that respond uncertainly in many environments. The agronomic and/or economic value of a cultivar across environments may be considered the general or overall utility of the cultivar. Data from a sample of environments therefore provides only an estimate of any cultivar's overall utility, with the overall goal of selection among all cultivars being the maximization of the expected utility. Within this frame-work, expected utility maximization, an approach to decision making that has been well developed in the disciplines of economics and statistics, can assist the plant breeder in making such decisions. This research was initiated (1) to determine how expected utility maximization might be used to develop indices that are useful for selecting broadly adapted plant cultivars, and (2) to determine how the breeder's preferences might affect choice of the best cultivar. The data used in this research were from USDA Regional Soybean Tests. The results indicated that expected utility maximization, which explicitly incorporates into the selection rule the plant breeder's preferences regarding stability, can be a useful aid in the selection of stable plant cultivars.     

Hematological and blood chemistry profiles of American bison grazing on Konza Prairie of Kansas

Normal hematological and blood chemistry parameters were measured in 45 American bison (Bison bison) that were divided into three age groups for comparison. There was a statistically significant (P less than 0.05) increase with advancing age in mean corpuscular volume, mean corpuscular hemoglobin, absolute neutrophil and eosinophil counts, total protein, globulin, creatinine, and blood urea nitrogen. There was a statistically significant (P less than 0.05) decrease with advancing age in levels of sorbital dehydrogenase, alkaline phosphatase, glucose, sodium, calcium and phosphorus.     

Survival of early preimplantation porcine embryos after co-culture with cells producing an avian retrovirus

To determine the relative survival of porcine embryos after co-culture with cells producing an avian retrovirus, four-cell stage embryos were obtained from sows following synchronization with altrenogest and superovulation with gonadotropins. These embryos were randomly assigned to the following treatments: no manipulation (zona-intact); zona removed with acidified Tyrode's solution (zona-free); and zona removed followed by co-culture with D-17 canine cells producing an avian retrovirus vector derived from spleen necrosis virus (zona-free + co-culture). The survival rates of four-cell stage embryos to morulae or early blastocysts during a 48-h culture period were 93.3, 80.0 and 57.7% in zona-intact, zona-free and zona-free + co-culture groups, respectively. Following embryo transfer, the development of embryos to fetuses at six weeks of gestation was 37.5, 30.0 and 11.7% in zona-intact, zona-free and zona-free + co-culture groups. These results indicate that early preimplantation porcine embryos can develop to apparently normal fetuses following co-culture with cells producing a retrovirus, and the feasibility of this method for retrovirus-mediated gene transfer in pigs was demonstrated.     

Insecticidal toxins from Bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with B. thuringiensis subsp. kurstaki CryIA(c) toxins

Genes encoding insecticidal crystal proteins were cloned from three strains of Bacillus thuringiensis subsp. kenyae and two strains of B. thuringiensis subsp. kurstaki. Characterization of the B. thuringiensis subsp. kenyae toxin genes showed that they are most closely related to cryIA(c) from B. thuringiensis subsp. kurstaki. The cloned genes were introduced into Bacillus host strains, and the spectra of insecticidal activities of each Cry protein were determined for six pest lepidopteran insects. CryIA(c) proteins from B. thuringiensis subsp. kenyae are as active as CryIA(c) proteins from B. thuringiensis subsp. kurstaki against Trichoplusia ni, Lymantria dispar, Heliothis zea, and H. virescens but are significantly less active against Plutella xylostella and, in some cases, Ostrinia nubilalis. The sequence of a cryIA(c) gene from B. thuringiensis subsp. kenyae was determined (GenBank M35524) and compared with that of cryIA(c) from B. thuringiensis subsp. kurstaki. The two genes are more than 99% identical and show seven amino acid differences among the predicted sequences of 1,177 amino acids.     

Transposon Tn916 mutagenesis in Clostridium botulinum.

The study of toxinogenesis and other properties in Clostridium botulinum is limited by the absence of genetic methods that enable construction of defined mutants. In this study, tetracycline-resistant transposon Tn916 in Enterococcus faecalis was conjugatively transferred in filter matings to group I Clostridium botulinum strains Hall A and 113B. The Tn916 transfer frequencies to C. botulinum ranged from 10(-8) to 10(-5) Tcr transconjugant per recipient depending on the donor strain. Southern blot analyses of EcoRI or HindIII chromosomal digests extracted from randomly selected Tcr transconjugants showed that the transposon inserted at different sites in the recipient chromosome, and the copy number of Tn916 varied from one to three. Tn916 insertion gave several different auxotrophic mutants. This approach should be useful for the study of genes important in growth, survival, and toxinogenesis in C. botulinum.