Alternative biofuel production in non-natural hosts

Global energy and environmental concerns have stimulated increased efforts in synthesizing petroleum-derived products from renewable resources. Biological production of metabolites for fuel is increasingly becoming a feasible, renewable, environmentally sound alternative. However, many of these chemicals are not highly produced in any known native organism. Here we review the current progress of modifying microorganisms with heterogeneous elements for the production of biofuels. This strategy has been extensively employed in a variety of hosts for the development of production of various alcohols, fatty acids, alkenes and alkanes.     

Molecular data and ecological niche modelling reveal a highly dynamic evolutionary history of the East Asian Tertiary relict Cercidiphyllum (Cercidiphyllaceae)

? East Asia's temperate deciduous forests served as sanctuary for Tertiary relict trees, but their ages and response to past climate change remain largely unknown. To address this issue, we elucidated the evolutionary and population demographic history of Cercdiphyllum, comprising species in China/Japan (Cercdiphyllum?japonicum) and central Japan (Cercdiphyllum magnificum). ? Fifty-three populations were genotyped using chloroplast and ribosomal DNA sequences and microsatellite loci to assess molecular structure and diversity in relation to past (Last Glacial Maximum) and present distributions based on ecological niche modelling. ? Late Tertiary climate cooling was reflected in a relatively recent speciation event, dated at the Mio-/Pliocene boundary. During glacials, the warm-temperate C.?japonicum experienced massive habitat losses in some areas (north-central China/north Japan) but increases in others (southwest/-east China, East China Sea landbridge, south Japan). In China, the Sichuan Basin and/or the middle-Yangtze were source areas of postglacial northward recolonization; in Japan, this may have been facilitated through introgressive hybridization with the cool-temperate C.?magnificum. ? Our findings challenge the notion of relative evolutionary and demographic stability of Tertiary relict trees, and may serve as a guideline for assessing the impact of Neogene climate change on the evolution and distribution of East Asian temperate plants.     

Climate oscillation during the Quaternary associated with landscape heterogeneity promoted allopatric lineage divergence of a temperate tree Kalopanax septemlobus (Araliaceae) in East Asia

We investigated the biogeographic history of Kalopanax septemlobus, one of the most widespread temperate tree species in East Asia, using a combined phylogeographic and palaeodistribution modelling approach. Range-wide genetic differentiation at nuclear microsatellites (G'(ST) = 0.709; 2205 samples genotyped at five loci) and chloroplast DNA (G(ST) = 0.697; 576 samples sequenced for 2055 bp at three fragments) was high. A major phylogeographic break in Central China corresponded with those of other temperate species and the spatial delineation of the two temperate forest subkingdoms of East Asia, consistent with the forests having been isolated within both East and West China for multiple glacial-interglacial cycles. Evidence for multiple glacial refugia was found in most of its current range in China, South Japan and the southernmost part of the Korean Peninsula. In contrast, lineage admixture and absence of private alleles and haplotypes in Hokkaido and the northern Korean Peninsula support a postglacial origin of northernmost populations. Although palaeodistribution modelling predicted suitable climate across a land-bridge extending from South Japan to East China during the Last Glacial Maximum, the genetic differentiation of regional populations indicated a limited role of the exposed sea floor as a dispersal corridor at that time. Overall, this study provides evidence that differential impacts of Quaternary climate oscillation associated with landscape heterogeneity have shaped the genetic structure of a wide-ranging temperate tree in East Asia.     

Major components of the KARRIKIN INSENSITIVE2-dependent signaling pathway are conserved in the liverwort Marchantia polymorpha

KARRIKIN INSENSITIVE2 (KAI2) was first identified as a receptor of karrikins, smoke-derived germination stimulants. KAI2 is also considered a receptor of an unidentified endogenous molecule called the KAI2 ligand. Upon KAI2 activation, signals are transmitted through the degradation of D53/SMXL proteins via MAX2-dependent ubiquitination. Although components in the KAI2-dependent signaling pathway, namely MpKAI2A and MpKAI2B, MpMAX2, and MpSMXL, exist in the genome of the liverwort Marchantia polymorpha, their functions remain unknown. Here, we show that early thallus growth is retarded and gemma dormancy in the dark is suppressed in Mpkai2a and Mpmax2 loss-of-function mutants. These defects are counteracted in Mpkai2a Mpsmxl and Mpmax2 Mpsmxl double mutants indicating that MpKAI2A, MpMAX2, and MpSMXL act in the same genetic pathway. Introduction of MpSMXL^d53, in which a domain required for degradation is mutated, into wild-type plants mimicks Mpkai2a and Mpmax2 plants. In addition, the detection of citrine fluorescence in Nicotiana benthamiana cells transiently expressing a SMXL-Citrine fusion protein requires treatment with MG132, a proteasome inhibitor. These findings imply that MpSMXL is subjected to degradation, and that the degradation of MpSMXL is crucial for MpKAI2A-dependent signaling in M. polymorpha. Therefore, we claim that the basic mechanisms in the KAI2-dependent signaling pathway are conserved in M. polymorpha.

Functions of genes in the KARRIKIN INSENSITIVE2-dependent signaling pathway are conserved in the liverwort Marchantia polymorpha and control early development of the thallus.     

Establishment of the evergreen broad-leaved tree species Castanopsis cuspidata in an abandoned secondary forest in western Japan

Recently, populations of Castanopsis cuspidata have often expanded into secondary forests in western Japan. To determine the establishment processes of this species, we analyzed its spatial distribution in a secondary forest dominated by Quercus variabilis and Quercus serrata that is located adjacent to a stand dominated by C. cuspidata. Saplings, defined as ≥30 cm stem length (SL) and <5 cm diameter at breast height (DBH), were abundant and their size distribution was inversely J-shaped, indicating continuous recruitment. Although seedlings (SL < 30 cm) and small saplings (30 ≤ SL < 50 cm) of C. cuspidata were aggregated near flowering trees of this species, some were found ≥40 m from the nearest adults, suggesting that there is animal-aided dispersal of acorns. The distribution of larger-sized individuals (≥100 cm SL) of C. cuspidata was unrelated to distance from the nearest flowering C. cuspidata or dominant Quercus species (≥5 cm DBH), but was associated with dead Pinus densiflora trees, which were abundant at the site. Thus, the establishment of C. cuspidata in this forest is controlled mainly by two factors, viz. patterns of acorn dispersal by animals, and forest disturbance regime (i.e., deaths of pine trees).     

Effect of glucosamine and related compounds on the degranulation of mast cells and ear swelling induced by dinitrofluorobenzene in mice

AimsGlucosamine has been used safely to relieve osteoarthritis in humans, but the precise mechanism underlying its efficacy is still unclear. In this study, we investigated the direct effects of glucosamine and related compounds on mast cell mediated inflammation using cultured mast cells and an animal model.Main methodsDinitrophenyl (DNP)-IgE-sensitized rat basophilic leukemia RBL-2H3 cells were treated with glucosamine-HCl (GlcN-HCl), N-acetylglucosamine (GlcNAc), chitin oligomer or chitosan oligomer. Cells were stimulated by DNP-BSA to induce degranulation and released β-hexosaminedase was determined colorimetrically to measure the degree of degranulation. Dinitrofluorobenzene (DNFB) sensitized BALB/c mice were administrated orally with 1 or 0.1 mg GlcN-HCl or GlcNAc for 6 days. One hour after the final administration, mice were challenged by DNFB to induce ear swelling.Key findingsGlcN-HCl significantly inhibited the antigen-induced degranulation of RBL-2H3 cells at higher than 0.01 mg/mL for 24 h-treatment while GlcNAc, a chitin oligomer and a chitosan oligomer had no effect. GlcN-HCl also suppressed intracellular calcium mobilization. GlcN-HCl and GlcNAc significantly suppressed the antigen-induced up-regulation of TNF-α and IL-6 mRNA. Ear swelling and histamine levels of plasma and ear in DNFB-treated mice were significantly suppressed by oral administration of GlcN-HCl or GlcNAc (0.1 and 1 mg) for 6 days.SignificanceOur results strongly suggest that GlcN-HCl and GlcNAc have anti-inflammatory effects in vivo by suppressing the activation of mast cells.     

Myeloid Differentiation Factor 88 (MyD88)-Deficiency Increases Risk of Diabetes in Mice

Background

Multiple lines of evidence suggest innate immune response pathways to be involved in the development of obesity-associated diabetes although the molecular mechanism underling the disease is unknown. Recent observations suggest that saturated fatty acids can act as a ligand for toll-like receptor (TLR) 4, which is thought to mediate obesity-associated insulin resistance. Myeloid differentiation factor 88 (MyD88) is an adapter protein for TLR/IL-1 receptor signaling, which is involved in the activation of inflammatory pathways. To evaluate molecular mechanisms linking obesity-associated diabetes down-stream of TLR4, we investigated physiological role of MyD88 in high-fat diet (HFD)-induced obesity.

Methodology/Principal Findings

In the present study, we found MyD88-deficient mice fed a HFD had increased circulating levels of insulin, leptin and cholesterol, as well as liver dysfunction (increased induction of ALT levels, increased activation of JNK and cleavage of PARP), which were linked to the onset of severe diabetes. On the other hand, TNF-α would not be involved in HFD-induced diabetes in MyD88-deficient mice, because TNF-α level was attenuated in MyD88-deficient mice fed with HFD.

Conclusions/Significance

The present finding of an unexpected role for MyD88 in preventing diabetes may provide a potential novel target/strategy for treating metabolic syndrome.     

Immortalized dendritic cell line with efficient cross-priming ability established from transgenic mice harboring the temperature-sensitive SV40 large T-antigen gene

The uptake of an antigen and its presentation to specific T cells by dendritic cells (DCs) is a primary event in initiation of humoral and cellular immune responses as well as the induction of cytotoxic T cells (CTLs). DCs are induced by culturing bone marrow cells in the presence of GM-CSF. However, the resulting DCs are short-lived and the culture usually contains CD11c-negative non-DC cells, which adversely affects reproducibility and makes interpretation of the experimental results difficult. Therefore, it would be useful if DCs could be readily immortalized with their functions being retained. In this study we established a novel, immortalized murine DC line with antigen-presenting capacity in vitro as well as an augmenting effect on humoral and cellular immune responses in vivo, utilizing bone marrow cells from transgenic mice harboring the temperature-sensitive SV40 large T-antigen gene. In the presence of GM-CSF, the resulting DC line, termed SVDC, could be continuously subcultured for more than 12 months. When pulsed with OVA alone or OVA-IgG immune complexes via Fcgamma receptors, SVDC augmented OVA-specific T cell proliferation efficiently in vitro, and elicited OVA-specific IgG production in vivo on the adoptive transfer of pulsed SVDC into naive mice. Interestingly, SVDC exhibited significantly high cross-priming ability compared to DCs in a short-term culture, thus leading to their extremely high effectiveness in inducing anti-tumor immunity in vivo. Thus, SVDC is useful for the detailed characterization of antigen presentation, and for research on the various therapeutic benefits of DC vaccination to elicit specific immune responses in immunodeficiencies, infectious diseases and cancer.