Search

Background: Strobilanthes cusia (Nees) Kuntze is a traditional medical plant distributed widely in south China. The indole compounds that originated from the plant are responsible for its pharmacological activities. However, the reason why indole ingredients are accumulated in this herb and how it is biosynthesized has remained largely unknown. Results: In this study, metabolic and transcriptional profiling measurement experiments of different S. cusia organs were carried out to understand the underlying molecular basis of indoles' biosynthetic logic. A metabolic investigation demonstrated that the indoles are primarily accumulated mainly in aerial parts, particularly in leaves. RNA-seq was employed to reveal the organ specific accumulation of indoles in different S. cusia organs. Meanwhile, a flavin-dependent monooxygenase gene (ScFMO1) was found in S. cusia, and it has capacity to produce indoxyl from indole by the fermentation assay. Finally, we assessed the outcomes of transient expression experiment in tobacco and confirmed that ScFMO1 localizes in cytoplasm. Conclusions: Our results suggest that ScFMO1 plays a key role in biosynthesis of indoles (Indigo, indirubin, indican, etc.), it will be useful for illuminating the molecular basis of the medicinal indoles' biosynthesis and developing strategies for improving their yields. [ABSTRACT FROM AUTHOR]

Ilex asprella is a plant from Aquifoliaceae. Its root is commonly used as folk medicinal materials in southern China. The chemical compositions of I. asprella are rich in pentacyclic triterpenoids, which show various biological activities and demonstrate a good prospect for drug development. The elucidation of biosynthesis mechanism of triterpenoids in I. asprella could lay important foundations for the production of these precious plant secondary metabolites by metabolic engineering. Our previous studies have revealed IaAO1 (a CYP716A210 homolog) responsible for the C-28 oxidation of α- and β-amyrin. Herein, we reported the identification of three more cytochrome P450 monooxygenase genes IaAO2 (a CYP716A212 homolog), IaAO4 (CYP714E88), IaAO5 (CYP93A220), and a cytochrome P450 reductase gene IaCPR by using Saccharomyces cerevisiae eukaryotic expression system and gas chromatography-mass spectrometry. Among them, the protein encoded by IaAO2 can catalyze the C-28 oxidation of α-amyrin and β-amyrin, IaAO4 can catalyze the C-23 oxidation of ursolic acid and oleanolic acid, while IaAO5 is responsible for the C-24 oxidation of β-amyrin. By introducing three genes IaAO1 , IaAO4 and IaCPR into S. cerevisiae. We constructed an engineered yeast strain that can produce C-23 hydroxyl ursane-type triterpenoid derivatives. This study contributes to a thorough understanding of triterpenoid biosynthesis of medicinal plants and provides important tools for further metabolic engineering. [ABSTRACT FROM AUTHOR]

BACKGROUND The seasonal long‐distance migration of pests amongst different seasonal habitats may encounter different degrees of insecticide selection pressure, but knowledge of the evolution of resistance and the underlying mechanisms remains sparse. Here, we show that the development of resistance of the oriental armyworm Mythimna separata (Walker), a notorious agricultural pest that migrates between northern and northeast China seasonally across the Bohai Gulf, is influenced by seasonal migration. RESULTS: There are two conspicuous migrations of M. separata, the northeastwards population in the second half of May and the returning population in the second half of August, between northern and northeast China per year, and the abundance values of migrants from April to October are significantly different. The resistance levels of seasonal migratory populations to λ‐cyhalothrin vary in different months, and the resistance levels and estimated frequency of the resistance allele of the first northeastward population (second half of May, May.‐2) in spring are significantly higher than those of the returning populations in autumn. Moreover, resistance decline was observed in migrating population Jul.‐2, with the resistance levels in the F1 progeny higher than those in their F2 progeny. Synergism tests indicate that cytochrome P450 monooxygenases (P450s) play a major role in resistance. Enzymatic assays show that P450 activity is significantly correlated with resistance levels in migratory populations, indicating that increased P450 activity is the main mechanism of resistance. The expression levels and correlation analysis of the relative expression of P450s with resistance levels show that three (CYP9A144, CYP9G40, and CYP6B79) out of 23 genes from CYP6 and CYP9 subfamilies are potentially involved in resistance to λ‐cyhalothrin. CONCLUSION: Our results show that the resistance of M. separata to λ‐cyhalothrin is unstable and likely to be metabolically driven by enhanced P450 activity mediated by the overexpression of multiple P450 genes. Notably, the development of resistance is probably affected by seasonal migration. [ABSTRACT FROM AUTHOR]

Choushenpilosulynes D–G (1-4): four new polyynes were isolated from the roots of Codonopsis pilosula (Campanulaceae) cultivated in Yunnan province, China. Their structures were identified by spectroscopic methods. Bioactive evaluation showed that choushenpilosulynes E (2) and F (3) demonstrated potent inhibitory effect on lipid formation induced by 100 μM oleic acid stimulation. In addition, choushenpilosulyne F (3) uncovered inhibitory activity against the expression of human 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) and squalene monooxygenase (SQLE) gene transcript in HepG2 cells. [ABSTRACT FROM AUTHOR]

Abstract: Fufenozide, a novel non-steroidal ecdysone agonist, exhibits good efficacy and plays an important role in controlling lepidopteran pests in China. A high level of resistance to fufenozide (320.3-fold) in Plutella xylostella (L.) has been selected under laboratory conditions. In this study, the stability and biochemical basis of resistance to fufenozide were investigated. The resistance ratio to fufezonide declined substantially from 320.3- to 20.5-fold in the fufenozide resistant strain (JSR) when the selection pressure was removed for six consecutive generations. However, the susceptibility to fufezonide could not be fully restored after 28 generations without exposure to the compound. Increased relative fitness (0.87) was associated with the rapid reversal of resistance in the resistance decaying strain (JSRD). The enzymatic analyses showed reduced specific activities of monooxygenase (MO), glutathione S-transferases (GSTs), carboxylesterase and aryl-acylamidase in the JSRD strain, whereas the activity of chitinase increased. The current findings suggest the selected resistance to fufenozide in P. xylostella is not stable, and the four detoxifying enzymes including MO, GSTs, carboxylesterase, and aryl-acylamidase may be involved. For resistance management proposes, insecticide rotation should be a viable option to delay the occurrence of fufenozide resistance in the field. [Copyright &y& Elsevier]

The diversity and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in the sediment of the Pearl River Estuary were investigated by cloning and quantitative real-time polymerase chain reaction (qPCR). From one sediment sample S16, 36 AOA OTUs (3% cutoff) were obtained from three clone libraries constructed using three primer sets for amoA gene. Among the 36 OTUs, six were shared by all three clone libraries, two appeared in two clone libraries, and the other 28 were only recovered in one of the libraries. For AOB, only seven OTUs (based on 16S rRNA gene) and eight OTUs (based on amoA gene) were obtained, showing lower diversity than AOA. The qPCR results revealed that AOA amoA gene copy numbers ranged from 9.6 × 10 to 5.1 × 10 copies per gram of sediment and AOB amoA gene ranged from 9.5 × 10 to 6.2 × 10 copies per gram of sediment, indicating that the dominant ammonia-oxidizing microorganisms in the sediment of the Pearl River Estuary were AOA. The terminal restriction fragment length polymorphism results showed that the relative abundance of AOB species in the sediment samples of different salinity were significantly different, indicating that salinity might be a key factor shaping the AOB community composition. [ABSTRACT FROM AUTHOR]

1. Community structures of planktonic ammonia-oxidising archaea (AOA) and bacteria (AOB) were investigated for five high-altitude Tibetan lakes, which could be classified as freshwater, oligosaline or mesosaline, to develop a general view of the AOA and AOB in lakes on the Tibetan Plateau. 2. Based on PCR screening of the ammonia monooxygenase α-subunit ( amoA) gene, AOA were present in 14 out of 17 samples, whereas AOB were detected in only four samples. Phylogenetic analyses indicated that the AOB communities were dominated by a unique monophylogenetic lineage within Nitrosomonas, which may represent a novel cluster of AOB. AOA, on the other hand, were distinct among lakes with different salinities. 3. Multivariate statistical analyses indicated a heterogeneous distribution of the AOA communities among lakes largely caused by lake salinity, whereas the uniform chemical properties within lakes and their geographical isolation may favour relatively homogeneous AOA communities within lakes. 4. Our results suggest a wide occurrence of AOA in Tibetan lakes and provide the first evidence of salinity-related differentiation of AOA community composition as well as potential geographical isolation of AOA in inland aquatic environments. [ABSTRACT FROM AUTHOR]

Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), has become the economically most important insect defoliator of potatoes, Solanum tuberosum L., in northern Xinjiang Uygur autonomous region in China. Currently, control of Colorado potato beetle relies mainly on chemical insecticides. And this may result in insecticide resistance. In this study, LDm values were measured by a topical bioassay for 14 conventional insecticides in seven local populations from Urumqi, Changji, Tacheng, Nilka, Gongliu, Qapqal, and Tekes counties (cities). The Tekes field population was the most susceptible population and was selected as a reference strain. Compared with the Tekes strain, the Changji, Qapqal, Nilka, Tacheng, and Congliu populations exhibited moderate to very high levels of resistance to cyhalothrin. The Qapqal and Changji populations showed a moderate and a very high resistance to deltarnethrin, respectively. And the Changji population developed a high resistance against a-cypermethrin. Moreover, the Qapqal population had a moderate resistance to carbofuran, and the Urumqi population reached high level of resistance to endosulfan. Possible resistance mechanisms of the Changji and Qapqal populations were determined using three enzyme inhibitors. Triphenyl phosphate (TPP), diethylmeleate, and piperonyl butoxide (PBO) had little synergism to cyhalothrin in the two populations. In contrast, PBO and TPP exhibited some synergistic effects to carbofuran in the Qapqal population, indicating the involvement of monooxygenases and esterases in conferring carbofuran resistance. It seems that additional mechanisms, such as target site insensitivity, should play an important role in Colorado potato beetle resistances to cyhalothrin and carbofuran in northern Xinjiang local populations. [ABSTRACT FROM AUTHOR]

The article presents a study which investigates the cross-resistance and biochemical mechanism of thiamethoxam resistance in B-biotype Bemisia tabaci in China. A thiamethoxam-resistant B. tabaci strain (TH-R) was selected in the laboratory to examine the risks and mechanism of resistance in B-biotype B. tabaci. The study shows that B-biotype B. tabaci can develop resistance to thiamethoxam, in which cytochrome P450 carboxylesterase and monooxygenase are responsible for the resistance.

Abstract: A Helicoverpa armigera population was collected from Shandong province, China. After 15 generations of selection in the laboratory, the H. armigera strain developed more than 20-fold resistance to spinosad. At LD50 level, no significant cross-resistance was found between spinosad and chlorpyrifos, methomyl, avermectin and chlorfenapyr except for fenvalerate with a low cross-resistance of 2.4-fold. However, LD99 values of fenvalerate against the parental and resistant strains were not different significantly. After inhibitors were used, spinosad resistance could be partially suppressed by piperonylbutoxide (PBO) and triphenylphosphate (TPP), but not by diethylmaleate (DEM). Activities of p-nitroanisole O-demethylase (ODM) developed to 8.26-fold compared with the parental strain, but no obvious changes were found in activities of carboxyl esterase (CarE) and glutathione-S-transferase (GST). The results indicated that resistance to spinosad in the cotton bollworm might be associated with an increase in cytochrome P450 monooxygenase. [Copyright &y& Elsevier]

Abstract: An extremely thermophilic alkane-degrading bacterium, strain MH-1, was isolated from the deep subterranean petroleum reservoir in Shengli oil field, PR China. Based on its physiological characteristics and analysis of its 16S rRNA gene sequence, strain MH-1 was identified as Geobacillus stearothermophilus. Strain MH-1 was able to grow at temperatures ranging from 50 to 72°C and effectively degraded hexadecane as the sole carbon source at 70°C. Strain MH-1 degraded alkanes with different length chains (C12–C31) in crude oil, but it preferentially degraded middle chain-alkanes. The alkane hydroxylase system of G. stearothermophilus MH-1 was characterized. It contained at least three alkane monooxygenase gene homologs (alkB-geo1, alkB-geo4, and alkB-geo6). The AlkB-geo6 cluster, which had a high sequence similarity to the alkB2 cluster of Rhodococcus strains NRRL B-16531 and Q15, contained alkB-geo6, two rubredoxins and a putative transcriptional regulatory protein. [Copyright &y& Elsevier]

The abundance and diversity of amoA gene, a gene putatively encoding ammonia monooxygenase subunit A, was evaluated in oxic lake water and anoxic sediments from two adjacent sites of Qinghai Lake, China. An integrated approach was employed including geochemistry, clone library construction, and reverse-transcription quantitative polymerase chain reaction. Whereas ammonia-oxidizing archaea (AOA) were more abundant than ammonia-oxidizing bacteria (AOB) in oxic lake water, the opposite was true in anoxic sediments. The AOB community was the same in the water and sediments and consisted of Nitrosomonas- and Nitrosospira-like sequences, suggesting that the sediment-associated AOB may be derived from the water column, but metabolically inactive. In contrast, the AOA community was distinct between the water column and the sediments, and clone sequences were related to those from aquatic environments and soils, respectively, suggesting that AOA in the sediments may be native and metabolically active. Our data have important implications for a better understanding of the potential role of ammonia oxidizers in lake ecosystems. [ABSTRACT FROM AUTHOR]

Polypogon fugax is a common winter weed in China and other Asia countries. We have previously found a P. fugax biotype (R) resistant to acetyl co-enzyme A carboxylase (ACCase) herbicides also cannot be effectively controlled by some acetolactate synthase (ALS) herbicides. This study evaluated the level of resistance to four ALS herbicides (metsulfuron-methyl, chlorsulfuron, monosulfuron, pyribambenz isopropyl) in the R biotype and the associated resistance mechanism. The R biotype exhibited moderate level of resistance to metsulfuron-methyl (6.0-fold) compared with the sensitive biotype (S). Sequence analysis of ALS gene revealed that two ALS genes existed in P. fugax. However, no substitution associated with ALS resistance mechanism were found in ALS genes between the S and R biotypes. The activity of ALS enzyme isolated from the R biotype was inherently higher and less sensitive to metsulfuron-methyl than the S biotype. Glutathione S-transferases (GST) activity was also less sensitive to metsulfuron-methyl in the R than as the S biotypes. Malathion, a cytochrome P450 (CYP) monooxygenase inhibitor, had much greater synergistic effect with metsulfuron-methyl on the R than as the S plants, reducing the ED50 value (herbicide dose to inhibit growth by 50%) of metsulfuron-methyl by 23- and 6-fold, respectively, suggesting that CYP mediated enhanced metabolism might contribute to the resistance to ALS herbicides. These results suggest that metsulfuron-methyl resistance in the R biotype was associated with the up-regulated ALS enzymatic activity and the GST and CYP-mediated enhanced herbicide metabolism. [ABSTRACT FROM AUTHOR]