Effects of fungal culture filtrates of Verticillium lecanii (Lecanicillium spp.) hybrid strains on Heterodera glycines eggs and juveniles

Many nematode-antagonistic fungi produce secondary metabolites and enzymes that demonstrate toxicity against plant-parasitic nematodes. The objective of this study was to evaluate the effects of fungal culture filtrates of Verticillium lecanii hybrid strains on mature eggs, embryonated eggs (eggs fertilized but without development of juveniles), and second-stage juveniles (J2) of Heterodera glycines and to compare these effects with those of their parental strains. The fungal culture filtrates of certain hybrid strains inhibited egg hatch of mature eggs. Furthermore, the fungal culture filtrates of two hybrid strains, AaF23 and AaF42, exhibited high toxicity against embryonated eggs of H. glycines. However, most of the fungal culture filtrates of V. lecanii did not inactivate J2. These results suggested that enzymes or other active compounds produced by the fungal culture filtrates of V. lecanii exhibit activity against specific stages in the H. glycines life cycle. In addition, based on a visual assessment of the morphological changes in eggs caused by filtrates of each strain, there were differences between the hybrid strains and their respective parental strains with regard to the active substances produced by V. lecanii against the embryonated eggs. As a result of promoting recombination of whole genomes via protoplast fusion, several hybrid strains may have enhanced production of active substances that are different from those produced by their parental strains. It was concluded that natural substances produced by V. lecanii are one of the important factors involved in the suppression of H. glycines damage.     

<Emphasis Type="Italic">Arabidopsis</Emphasis> RETINOBLASTOMA-RELATED PROTEIN 1 is involved in G1 phase cell cycle arrest caused by sucrose starvation

Although sucrose availability is crucial for commitment to plant cell division during G1 phase by controlling the expression of D-type cyclins, it has remained unclear how these factors mediate entry into the cell cycle. Here we show that Arabidopsis RETINOBLASTOMA-RELATED PROTEIN 1 (AtRBR1) is involved in G1-phase cell cycle arrest caused by sucrose starvation. We generated estrogen-inducible AtRBR1 RNA interference (RNAi) Arabidopsis suspension MM2d cells, and found that downregulation of AtRBR1 leads to a higher frequency of arrest in G2 phase, instead of G1-phase arrest in the uninduced control, after sucrose starvation. Synchronization experiments confirmed that downregulation of AtRBR1 leads to a prolonged G2 phase and delayed activation of G2/M marker genes. Downregulation of AtRBR1 also stimulated the activation of E2F-regulated genes when these genes were repressed in the uninduced cells under the limited sucrose conditions. We conclude that AtRBR1 is a key effector for the ability of sucrose to modulate progression from G1 phase.     

Identification of human IgG1 variant with enhanced FcRn binding and without increased binding to rheumatoid factor autoantibody

Various studies have demonstrated that Fc engineering to enhance neonatal Fc receptor (FcRn) binding is effective for elongating half-life or increasing cellular uptake of IgG. A previous study has shown that a N434H mutation to enhance FcRn binding resulted in increased binding to rheumatoid factor (RF) autoantibody, which is not desirable for therapeutic use in autoimmune disease. In this study, we first showed that all the existing Fc variants with enhanced FcRn binding also show increased RF binding, and then identified specific mutations that could be introduced to those Fc variants to reduce the RF binding. Furthermore, we generated novel Fc variants that do not increase RF binding and show half-lives of 45 d in cynomolgus monkey, which is longer than those of previously reported Fc variants. In addition, we generated novel Fc variants with antigen sweeping activity that do not increase RF binding. We expect that these novel Fc variants will be useful as antibody therapeutics against autoimmune diseases.     

The plant vesicular transport engineering for production of useful recombinant proteins

The molecular breeding of plants that have been genetically engineered for improved disease resistance and stress tolerance has been undertaken with the goal of improving food production. More recently, it has been realized that transgenic plants can serve as bioreactors for the production of proteins or compounds with industrial or clinical uses. Several different recombinant enzymes and antibodies have been produced in this manner. To maximize the potential of industrial plants as a production system for proteins, efficient expression systems utilizing promoters that optimize transgene expression, 5′-untranslated region elements for efficient translation, and appropriate post-translational modifications and localization must be developed. This review summarizes successful examples of the production of recombinant enzymes, antibodies, and vaccines using signal peptides that direct vesicular localization in transgenic plants. We further discuss the modulation of recombinant protein localization to the endoplasmic reticulum, vacuolar system, or extracellular compartments by varying the signal peptide.     

Repeated Ubiquitin Genes in Trichophyton mentagrophytes

Using Trichophyton mentagrophytes cDNA as a template, we performed PCR amplification with the UB1S and UB1R primers for isolation of a partial fragment of T. mentagrophytes ubiquitin gene. We screened the T. mentagrophytes cDNA library prepared with Uni-ZAP phage vector by hybridization with the T. mentagrophytes ubiquitin fragment, UB1S-UB1R. The nucleotide and deduced amino acid sequences of T. mentagrophytes ubiquitin gene encoded two ubiquitin repeats in 162 bp to 624 bp. The deduced amino acid sequences of ubiquitin from T. mentagrophytes shared about 98% similarity with those of Saccharomyces cerevisiae, Neurospora crassa, and Cryptococcus neoformans. Received: 28 April 1999 / Accepted: 22 June 1999     

Microreview: Capsule-associated genes of Cryptococcus neoformans

Cryptococcosis, caused by Cryptococcus neoformans is a common systemic mycosis in man and animals, particularly immunocompromised patients. This pathogenic fungus produces a thick extracellular polysaccharide capsule. Four capsule-associated genes (CAP10, CAP59, CAP60, CAP64) were cloned and sequenced, and proved to be essential for capsule synthesis. However biochemical functions of CAP gene products have not been clarified yet. Recently, the relatedness of the polysaccharide capsule and four capsule-associated genes has partly been elucidated. Nucleotide sequence of four CAP gene fragments was analyzed for phylogenetic relationships, and they were in agreement with the conventional classification of varieties and serotypes within C. neoformans. Expression of four CAP genes and capsule size were examined using two media containing different amount of glucose, and the results indicated that CAP genes might play important roles in elaboration of extracellular polysaccharide capsule. Furthermore, analyses of CAP genes in various clinical samples would give the useful information to diagnose cryptococcosis in human and animals.     

Interspecies Transmission of Feline Immunodeficiency Virus from the Domestic Cat to the Tsushima Cat (Felis bengalensis euptilura) in the Wild

Feline immunodeficiency virus (FIV) was isolated from a wild-caught Tsushima cat (Felis bengalensis euptilura), an endangered Japanese nondomestic subspecies of leopard cat (F. bengalensis). Phylogenetic analysis of the env gene sequences indicated that the FIV from the Tsushima cat belonged to a cluster of subtype D FIVs from domestic cats. FIVs from both the Tsushima cat and the domestic cat showed similar levels of replication and cytopathicity in lymphoid cell lines derived from these two species. The results indicated the occurrence of interspecies transmission of FIV from the domestic cat to the Tsushima cat in the wild.