Development of <Emphasis Type="Italic">mazF</Emphasis>-based markerless genome editing system and metabolic pathway engineering in <Emphasis Type="Italic">Candida tropicalis</Emphasis> for producing long-chain dicarboxylic acids

Candida tropicalis can grow with alkanes or plant oils as the sole carbon source, and its industrial application thus has great potential. However, the choice of a suitable genetic operating system can effectively increase the speed of metabolic engineering. MazF functions as an mRNA interferase that preferentially cleaves single-stranded mRNAs at ACA sequences to inhibit protein synthesis, leading to cell growth arrest. Here, we constructed a suicide plasmid named pPICPJ-mazF that uses the mazF gene of Escherichia coli as a counterselectable marker for the markerless editing of C. tropicalis genes to increase the rate of conversion of oils into long-chain dicarboxylic acids. To reduce the β-oxidation of fatty acids, the carnitine acetyltransferase gene (CART) was deleted using the gene editing system, and the yield of long-chain acids from the strain was increased to 8.27 g/L. By two homologous single exchanges, the promoters of both the cytochrome P450 gene and the NADPH–cytochrome P450 reductase gene were subsequently replaced by the constitutively expressed promoter pGAP, and the production of long-chain dicarboxylic acids by the generated strain (C. tropicalis PJPP1702) reached 11.39 g/L. The results of fed-batch fermentation showed that the yield of long-chain acids from the strain was further increased to 32.84 g/L, which was 11.4 times higher than that from the original strain. The results also showed that the pPICPJ-mazF-based markerless editing system may be more suited for completing the genetic editing of C. tropicalis.     

Gut microbiota composition is associated with environmental landscape in honey bees

There is growing recognition that the gut microbial community regulates a wide variety of important functions in its animal hosts, including host health. However, the complex interactions between gut microbes and environment are still unclear. Honey bees are ecologically and economically important pollinators that host a core gut microbial community that is thought to be constant across populations. Here, we examined whether the composition of the gut microbial community of honey bees is affected by the environmental landscape the bees are exposed to. We placed honey bee colonies reared under identical conditions in two main landscape types for 6 weeks: either oilseed rape farmland or agricultural farmland distant to fields of flowering oilseed rape. The gut bacterial communities of adult bees from the colonies were then characterized and compared based on amplicon sequencing of the 16S rRNA gene. While previous studies have delineated a characteristic core set of bacteria inhabiting the honey bee gut, our results suggest that the broad environment that bees are exposed to has some influence on the relative abundance of some members of that microbial community. This includes known dominant taxa thought to have functions in nutrition and health. Our results provide evidence for an influence of landscape exposure on honey bee microbial community and highlight the potential effect of exposure to different environmental parameters, such as forage type and neonicotinoid pesticides, on key honey bee gut bacteria. This work emphasizes the complexity of the relationship between the host, its gut bacteria, and the environment and identifies target microbial taxa for functional analyses.     

Determinant landscape‐scale factors on pond odonate assemblages

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Different fermentation strategies by Schizochytrium mangrovei strain pq6 to produce feedstock for exploitation of squalene and omega‐3 fatty acids

Schizochytrium mangrovei strain PQ 6 was investigated for coproduction of docosahexaenoic acid (C22: 6ω‐3, DHA ) and squalene using a 30‐L bioreactor with a working volume of 15 L under various batch and fed‐batch fermentation process regimes. The fed‐batch process was a more efficient cultivation strategy for achieving higher biomass production rich in DHA and squalene. The final biomass, total lipid, unsaponifiable lipid content, and DHA productivity were 105.25 g · L^?1, 43.40% of dry cell weight, 8.58% total lipid, and 61.66 mg · g^?1 · L^?1, respectively, after a 96 h fed‐batch fermentation. The squalene content was highest at 48 h after feeding glucose (98.07 mg · g^?1 of lipid). Differences in lipid accumulation during fermentation were correlated with changes in ultrastructure using transmission electron microscopy and Nile Red staining of cells. The results may be of relevance to industrial‐scale coproduction of DHA and squalene in heterotrophic marine microalgae such as Schizochytrium .     

Two new species of Phyllonema (Rivulariaceae,Cyanobacteria) with an emendation of the genus

Two untapered, heterocytous species were observed and collected from the intertidal and supratidal zones of the Mexican coastline of the Pacific Ocean near Oaxaca and from the Gulf of Mexico. These populations were highly similar in morphology to the freshwater taxon Petalonema incrustans in the Scytonemataceae. However, 16S rRNA sequence data and phylogenetic analysis indicated that they were sister taxa to the epiphyllic, Brazilian species Phyllonema aveceniicola in the Rivulariaceae, described from culture material. While genetic identity between the two new species was high, they differed significantly in morphology, 16S rRNA gene sequence identity, and sequence and structure of the 16S–23S ITS region. Their morphology differed markedly from the generitype of the previously monotypic Phyllonema, which has tapered, heteropolar, single‐false branched trichomes with very thin or absent sheath. The two new species, Phyllonema ansata and Phyllonema tangolundensis, described from both culture and environmental material, have untapered, isopolar, geminately false branched trichomes with thick, lamellated sheaths, differences so significant that the species would not be placed in Phyllonema without molecular corroboration. The morphological differences are so significant that a formal emendation of the genus is required. These taxa provide a challenge to algal taxonomy because the morphological differences are such that one would logically conclude that they represent different genera, but the phylogenetic evidence for including them all in the same genus is conclusive. This conclusion is counter to the current trend in algal taxonomy in which taxa with minor morphological differences have been repeatedly placed in separate genera based primarily upon DNA sequence evidence.     

Biosynthesis of abscisic acid in fungi: identification of a sesquiterpene cyclase as the key enzyme in Botrytis cinerea

While abscisic acid (ABA) is known as a hormone produced by plants through the carotenoid pathway, a small number of phytopathogenic fungi are also able to produce this sesquiterpene but they use a distinct pathway that starts with the cyclization of farnesyl diphosphate (FPP) into 2Z,4E‐α‐ionylideneethane which is then subjected to several oxidation steps. To identify the sesquiterpene cyclase (STC) responsible for the biosynthesis of ABA in fungi, we conducted a genomic approach in Botrytis cinerea. The genome of the ABA‐overproducing strain ATCC58025 was fully sequenced and five STC‐coding genes were identified. Among them, Bcstc5 exhibits an expression profile concomitant with ABA production. Gene inactivation, complementation and chemical analysis demonstrated that BcStc5/BcAba5 is the key enzyme responsible for the key step of ABA biosynthesis in fungi. Unlike what is observed for most of the fungal secondary metabolism genes, the key enzyme‐coding gene Bcstc5/Bcaba5 is not clustered with the other biosynthetic genes, i.e., Bcaba1 to Bcaba4 that are responsible for the oxidative transformation of 2Z,4E‐α‐ionylideneethane. Finally, our study revealed that the presence of the Bcaba genes among Botrytis species is rare and that the majority of them do not possess the ability to produce ABA.     

Water availability,fundamental niches and realized niches: A case study from the Cape flora

The ability of plants to survive drought or waterlogging constitutes an important niche parameter, which might be particularly significant in explaining species coexistence in the species‐rich and seasonally dry Cape Floristic Region of South Africa. However, the degree of physiological adaptation and specialization to these eco‐hydrological parameters (the fundamental niche) cannot be readily inferred from correlative studies based on species distributions and spatial variation in environmental parameters (the realized niche). We used an ex situ greenhouse experiment to compare the fundamental hydrological niches (different mean annual precipitation, rainfall seasonality and soil drainage) of six eco‐hydrologically divergent African Restionaceae species. Juvenile plants were subjected to six different watering treatments, ranging from no watering to waterlogging, to determine drought and waterlogging susceptibility and optimal growth conditions. We used the rate of biomass accumulation and survival rate as response measures. We found that species from dry and mesic (but well‐drained) habitats had optimal or near‐optimal growth at benign conditions (under which most restio species grow well). All species performed worse when droughted and died when not watered. Species from dry habitats tended to perform better (assessed in growth) than species from wet habitats under droughting. Species from wet habitats performed best when waterlogged, whereas species from dry habitats performed very poorly when waterlogged – thus showing that realized and fundamental niches covaried at the wet end of the hydrological gradient. We conclude that eco‐hydrological parameters are part of the fundamental niche, and fundamental and realized species niches are approximately correlated along them. The distribution of wet habitat species appears not to reflect their drought tolerance, suggesting that it may not be predicted by bioclimatic variables, but rather by soil drainage characteristics.     

Response of maize stemborers and associated parasitoids to the spread of grasses in the rainforest zone of Kisangani,DR Congo: effect on stemborers biological control

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Struktur und quantitatives Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums der Mäuseleber

Zusammenfassung In normalen Leberzellen der Maus wurde das quantitative Verhalten des perimitochondrialen granulären endoplasmatischen Retikulums untersucht. 74% der Mitochondrien zeigen Beziehungen zum granulären endoplasmatischen Retikulum. Die einzelnen Mitochondrien werden zu 52%19,5 von den Membranen des granulären endoplasmatischen Retikulums bedeckt. Je Mikrometer Membranstrecke sind auf der mitochondriennahen Seite des granulären endoplasmatischen Retikulums 21 und auf der mitochondrienfernen Seite 20 Ribosomen zu finden, was der Zahl im übrigen granulären endoplasmatischen Retikulum entspricht. Die Befunde stellen die Grundlage für Untersuchungen des perimitochondrialen granulären endoplasmatischen Retikulums unter pathologischen Bedingungen dar.

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Structure and quantitative behaviour of the perimitochondrial granular endoplasmic reticulum in the liver cells of the mouse

Summary The perimitochondrial granular endoplasmic reticulum in normal mouse liver cells has been investigated quantitatively. 74% of the mitochondria are in association with the granular endoplasmic reticulum. The individual mitochondrion is covered by the membranes of the granulated E. R. in 52%19.5. The outer surface of the endoplasmic membrane, facing the mitochondrion, is occupied by 21 ribosomes per m; the corresponding surface of the membrane facing the free cytoplasm is occupied by 20 ribosomes per m. These data are in agreement with those of that fraction of the E. R., which is not in association with mitochondria. These findings represent a basis for investigations of the perimitochondrial endoplasmic reticulum under pathological conditions.