In previous studies, the regeneration rates of Miscanthus × giganteus J.M.Greef, Deuter ex Hodk., Renvoize from callus tissue cultured on semi-solid media significantly declined after 4 mo of culture, which presents problems with germplasm conservation and use as an alternative propagation system. Due to the species’ lignocellulosic nature, it was hypothesized that the accumulation of phenolic compounds in the callus may be responsible for inhibiting regeneration. The current study aimed to optimize regeneration of M. × giganteus callus by culturing the callus tissue in the presence of 2-aminoindan-2-phosphonic acid (AIP), a competitive inhibitor of phenylalanine ammonia lyase (PAL), to reduce the biosynthesis of phenolics. Embryogenic callus was cultured on media supplemented with 9.0- or 11.3-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0-, 1-, 10-, 100-, or 1000-μM AIP. Every 28 d for 7 mo, the callus tissue was visually classified based on morphology and regeneration rate. Over the duration of the study, regeneration of shoots was consistently highest in callus cultured on 11.3-μM 2,4-D supplemented with 10- and 100-μM AIP (13–58.3%), and in vitro plantlet development from callus cultured on all concentrations of AIP demonstrated tillering and rooting. Total soluble phenolic content of the callus decreased in a dose-dependent manner from 2242.34-μg g−1 dry weight in the control to 1569.71-μg g−1 dry weight in AIP-treated callus. These data indicate that inhibiting PAL in M. × giganteus cultures increased the percentage of calluses exhibiting regeneration over time.
Mutation of the gene drop-dead (drd) causes adult Drosophila to die within 2 weeks of eclosion and is associated with reduced rates of defecation and increased volumes of crop contents. In the current study, we demonstrate that flies carrying the strong allele drdlwf display a reduction in the transfer of ingested food from the crop to the midgut, as measured both as a change in the steady-state distribution of food within the gut and also in the rates of crop emptying and midgut filling following a single meal. Mutant flies have abnormal triglyceride (TG) and glycogen stores over the first 4 days post-eclosion, consistent with their inability to move food into the midgut for digestion and nutrient absorption. However, the lifespan of mutants was dependent upon food presence and quality, suggesting that at least some individual flies were able to digest some food. Finally, spontaneous motility of the crop was abnormal in drdlwf flies, with the crops of mutant flies contracting significantly more rapidly than those of heterozygous controls. We therefore hypothesize that mutation of drd causes a structural or regulatory defect that inhibits the entry of food into the midgut. 相似文献
Gαq directly activates p63RhoGEF and closely related catalytic domains found in Trio and Kalirin, thereby linking Gq-coupled receptors to the activation of RhoA. Although the crystal structure of Gαq in complex with the catalytic domains of p63RhoGEF is available, the molecular mechanism of activation has not yet been defined. In this study, we show that membrane translocation does not appear to play a role in Gαq-mediated activation of p63RhoGEF, as it does in some other RhoGEFs. Gαq instead must act allosterically. We next identify specific structural elements in the PH domain that inhibit basal nucleotide exchange activity, and provide evidence that Gαq overcomes this inhibition by altering the conformation of the α6–αN linker that joins the DH and PH domains, a region that forms direct contacts with RhoA. We also identify residues in Gαq that are important for the activation of p63RhoGEF and that contribute to Gα subfamily selectivity, including a critical residue in the Gαq C-terminal helix, and demonstrate the importance of these residues for RhoA activation in living cells. 相似文献
Hfq is a global regulatory RNA‐binding protein. We have identified and characterized an atypical Hfq required for gene regulation and infectivity in the Lyme disease spirochete Borrelia burgdorferi. Sequence analyses of the putative B. burgdorferi Hfq protein revealed only a modest level of similarity with the Hfq from Escherichia coli, although a few key residues are retained and the predicted tertiary structure is similar. Several lines of evidence suggest that the B. burgdorferi bb0268 gene encodes a functional Hfq homologue. First, the hfqBb gene (bb0268) restores the efficient translation of an rpoS::lacZ fusion in an E. coli hfq null mutant. Second, the Hfq from B. burgdorferi binds to the small RNA DsrABb and the rpoS mRNA. Third, a B. burgdorferi hfq null mutant was generated and has a pleiotropic phenotype that includes increased cell length and decreased growth rate, as found in hfq mutants in other bacteria. The hfqBb mutant phenotype is complemented in trans with the hfq gene from either B. burgdorferi or, surprisingly, E. coli. This is the first example of a heterologous bacterial gene complementing a B. burgdorferi mutant. The alternative sigma factor RpoS and the outer membrane lipoprotein OspC, which are induced by increased temperature and required for mammalian infection, are not upregulated in the hfq mutant. Consequently, the hfq mutant is not infectious by needle inoculation in the murine model. These data suggest that Hfq plays a key role in the regulation of pathogenicity factors in B. burgdorferi and we hypothesize that the spirochete has a complex Hfq‐dependent sRNA network. 相似文献
Proteins belonging to the Hsp70 class of molecular chaperones are highly conserved and ubiquitous, performing an essential role in the maintenance of cellular homeostasis in almost all known organisms. Trypanosoma brucei, Trypanosoma cruzi and Leishmania major are human parasites collectively known as the Tritryps. The Tritryps undergo extensive morphological changes during their life cycles, largely triggered by the marked differences between conditions in their insect vector and human host. Hsp70s are synthesised in response to these marked changes in environment and are proposed to be required for these parasites to successfully transition between differentiation stages while remaining viable and infective. While the Tritryps Hsp70 complement consists of homologues of all the major eukaryotic Hsp70s, there are a number of novel members, and some unique structural features. This review critically evaluates the current knowledge on the Tritryps Hsp70 proteins with an emphasis on T. brucei, and highlights some novel and previously unstudied aspects of these multifaceted molecular chaperones. 相似文献
Summary Calling characteristics of field crickets (Teleogryllus oceanicus) differ between Pacific populations parasitized and unparasitized by a phonotactic fly (Ormia ochracea). Although we inferred that these song differences were due to natural selection by the fly, other environmental differences among sampling localities may also influence the cricket song. To evaluate the contribution of parasitoid pressure to variation in song structure, we analysed calls of crickets from five areas arrayed along a gradient of prevalence of parasitization. A novel use of canonical correlation analysis allowed us to test simultaneously the robustness of alternative hypotheses and their predictions. There is strong inference that selection pressures by phonotactic parasitoid flies have shaped song characteristics of field crickets in the Hawaiian Islands. Not all song components appear to have been equally affected by parasitoid selection and approximately 80–90% of total song variation among individuals is associated with other, unmeasured ecological and environmental attributes.That songs evolve is not disputed, but that they evolved in certain directions or that song structure is due to the action of certain forces, or that differences are due to random evolutionary events, or that they are the result of selection, or that they arise mainly in association with speciation — these are matters for speculation, debate, and analysis.Otte (1992, P. 25) 相似文献