Unveiling the role of bZIP transcription factors CREB and CEBP in detoxification metabolism of Nilaparvata lugens (Stål).

The basic leucine zipper (bZIP) superfamily is a crucial group of xenobiotics in insects. However, little is known about the function of CAAT enhancer binding proteins (CEBP) and cAMP response element binding protein (CREB) in Nilaparvata lugens. In the present study, NlCEBP and NlCREB were cloned and identified. Quantitative polymerase real-time chain reaction (qRT-PCR) analysis showed the expression of NlCEBP and NlCREB was significantly induced after chemical insecticides exposure. Silencing of NlCEBP and NlCREB increased the susceptibility of N. lugens to insecticides, and the detoxification enzyme activities were also significantly decreased. In addition, comparative transcriptome analysis revealed that 174 genes were significantly co-down-regulated after interfering with the two transcription factors. GO analysis showed that co-down-regulated genes are mostly related to energy transport and metabolic functions indicating the potential regulatory role of NlCEBP and NlCREB in detoxification metabolism. Our research shed lights on the functional roles of transcription factors NlCEBP and NlCREB in the detoxification metabolism of N. lugens, providing a theoretical basis for pest management and comprehensive control of this pest and increasing our understanding of insect toxicology.

Activator protein-1 mediated CYP6ER1 overexpression in the clothianidin resistance of Nilaparvata lugens (Stål).

BACKGROUND: Nilaparvata lugens, a destructive rice pest in Asia, has developed resistance to many insecticides, including the neonicotinoid clothianidin. CYP6ER1 plays an important role in N. lugens resistant to clothianidin, but only limited information on the transcriptional regulation of CYP6ER1 overexpression in clothianidin resistance is available. RESULTS: In this study, the transcription factor activator protein 1 (AP-1) was found to be overexpressed in a clothianidin-resistant strain of N. lugens and several field resistant populations. RNA interference-mediated silencing of NlAP-1 significantly decreased CYP6ER1 expression and increased the susceptibility of N. lugens to clothianidin. Additionally, NlAP-1 was highly expressed in egg and adult stages, and in midguts, and NlAP-1 was upregulated and induced to a greater extent in the clothianidin-resistant strain after exposure to clothianidin. Finally, dual-luciferase reporter assays confirmed the interaction between NlAP-1 and the two predicted binding sites in the CYP6ER1 promoter. CONCLUSION: NlAP-1 bound the -1388 to -1208-bp region of the CYP6ER1 promoter, enhancing its activity and then regulate the expression of CYP6ER1. These findings enhance our knowledge of the transcriptional regulation of the P450 genes that mediate insecticide resistance in insect pests. © 2021 Society of Chemical Industry.

Ionization basis for activation of enzymes soluble in ionic liquids.

BACKGROUND: The complex interactions between electrolytes and proteins have been studied for more than a century. However, understanding is not yet complete and does not provide a basis for predicting the activity of enzymes in ionic media. The use of ionic liquids (ILs) as reaction medium has opened up new opportunities for better understanding of the mechanism of enzymatic catalysis. Although a number of properties of ILs have been correlated with enzyme function, these relationships are not completely understood at a molecular level. METHODS: We propose that ILs must be able to promote ionization of protein ionizable groups in order to dissolve active enzymes. The biocompatible IL need to possess a functional group with large donor number and acceptor number in both cationic and anionic units, each of which is based on a high dielectric constant lead structure. We designed and synthesized two series of ILs and determined their ionizing-dissociating abilities and activities of lipases soluble in these new ILs. RESULTS: The results showed that the ionizing-dissociating abilities of ILs paralleled the catalytic activity trend of lipases dissolved in the ILs. The activities of lipases soluble in the newly designed ILs were comparable to those in water. CONCLUSIONS: We can conclude that ionizing-dissociating abilities of an IL can be used as a basis for predicting the activity of enzymes soluble in the IL. General significance Ionization basis for activation of enzymes gives a deeper understanding of the behavior of enzymes in non-aqueous media at a molecular level.

Phylogenetic shifts of bacterioplankton community composition along the Pearl Estuary: the potential impact of hypoxia and nutrients.

The significance of salinity in shaping bacterial communities dwelling in estuarine areas has been well documented. However, the influences of other environmental factors such as dissolved oxygen and nutrients in determining distribution patterns of both individual taxa and bacterial communities inhabited local estuarine regions remain elusive. Here, bacterioplankton community structures of surface and bottom waters from eight sites along the Pearl Estuary were characterized with 16S rRNA gene pyrosequencing. The results showed significant differences of bacterioplankton community between freshwater and saltwater sites, and further between surface and bottom waters of saltwater sites. Synechococcus dominated the surface water of saltwater sites while Oceanospirillales, SAR11 and SAR406 were prevalent in the bottom water. Betaproteobacteria was abundant in freshwater sites, with no significant difference between water layers. Occurrence of phylogenetic shifts in taxa affiliated to the same clade was also detected. Dissolved oxygen explained most of the bacterial community variation in the redundancy analysis targeting only freshwater sites, whereas nutrients and salinity explained most of the variation across all samples in the Pearl Estuary. Methylophilales (mainly PE2 clade) was positively correlated to dissolved oxygen, whereas Rhodocyclales (mainly R.12up clade) was negatively correlated. Moreover, high nutrient inputs to the freshwater area of the Pearl Estuary have shifted the bacterial communities toward copiotrophic groups, such as Sphingomonadales. The present study demonstrated that the overall nutrients and freshwater hypoxia play important roles in determining bacterioplankton compositions and provided insights into the potential ecological roles of specific taxa in estuarine environments.

Shifts in archaeaplankton community structure along ecological gradients of Pearl Estuary.

The significance of archaea in regulating biogeochemical processes has led to an interest in their community compositions. Using 454 pyrosequencing, the present study examined the archaeal communities along a subtropical estuary, Pearl Estuary, China. Marine Group I Thaumarchaeota (MG-I) were predominant in freshwater sites and one novel subgroup of MG-I, that is MG-Iν, was proposed. In addition, the previously defined MG-Iα II was grouped into two clusters (MG-Iα II-1, II-2). MG-Iα II-1 and MG-Iλ II were both freshwater-specific, with MG-Iα II-1 being prevalent in the oxic water and MG-Iλ II in the hypoxic water. Salinity, dissolved oxygen, nutrients and pH were the most important determinants that shaped the differential distribution of MG-I subgroups along Pearl Estuary. Marine Group II Euryarchaeota (MG-II) dominated the saltwater sites, but their abundance was higher in surface waters. The phylogenetic patterns of MG-I subgroups and their habitat preferences provide insight into their phylogeographic relationships. These results highlight the diversification of various ecotypes of archaea, especially of MG-I, under distinct environmental factors in Pearl Estuary, which are of great value for further exploring their ecological functions.

Glycerol carbonate: a novel biosolvent with strong ionizing and dissociating powers.

The activity of biocatalysts in nonaqueous solvents is related to the interaction of organic solvents with cells or enzymes. The behavior of proteins is strongly dependent on the protonation state of their ionizable groups, which ionization constants are greatly affected by the solvent. Due to the weak ionizing and dissociating powers of common organic solvents, the charge of the protein will change significantly when the protein is transferred from water to common organic solvents, resulting in protein denaturation. In this work, glycerol carbonate (GC) was synthesized, which ionizing and dissociating abilities were very close to those of water. Transesterification activities of Candida antarctica lipase B (CALB) in GC were comparable to those in water and remained constant during 4-week storage. Bacillus subtilis and Saccharomyecs cerevisiae were cultured in liquid media containing GC with test tubes. In the medium containing low GC concentration, Bacillus subtilis and Saccharomyecs cerevisiae grew well as in a medium containing no organic solvent, but, in the medium containing high GC concentration, the growth of Bacillus subtilis and Saccharomyecs cerevisiae was suppressed. The results suggested that GC is a potential biosolvent, which has great significance to biocatalysis in nonaqueous solvents.

Design of biosolvents through hydroxyl functionalization of compounds with high dielectric constant.

We proposed basic principles for biosolvent design on the viewpoint of ionization. Two classes of biosolvents, based on cyclic carbonate moiety and amide moiety, were designed through hydroxyl functionalization of highly dielectric compound. The newly designed compounds, glycerol carbonate (GC) and N-hydroxymethyl formamide (HOF), were synthesized for the development of soluble enzymatic systems and characterized by (13)C NMR and (1)H NMR. All the characterization data were consistent with the expected structures. Using conductance measurements, the pK (a) values of trichloroacetic acid in GC and HOF were determined as 0.80 and 0.85 at 25.0 °C, which was very close to that in water (pK (a) = 0.70), suggesting that the ionizing and dissociating abilities of GC and HOF are similar to those of water. The effects of various reaction parameters on activity and stability of Candida antarctica lipase B and lipase from Pseudomonas cepacia were investigated using the transesterification of ethyl butyrate with n-butanol as a model reaction. The activities of lipases in GC and HOF were comparable to those in water, indicating that the newly designed compounds were biocompactible. Biosolvent design is a promising and versatile method for developing new biosolvents.

Lipases are soluble and active in glycerol carbonate as a novel biosolvent.

Glycerol carbonate was synthesized as biosolvent for the development of soluble enzymatic system. The effects of various reaction parameters on activity and stability of lipases were investigated using the transesterification of ethyl butyrate with n-butanol as a model reaction. Enzymatic activity in glycerol carbonate was compared with that in water and in conventional organic solvents with different ionizing and dissociating abilities. The pK(a) value of trichloroacetic acid and transesterification activities of Candida antarctica lipase B and Candida rugosa lipase in glycerol carbonate are similar to those in water, indicating that ionizing and dissociating powers are capable of satisfactorily predicting the biocompatibility of organic solvents for soluble enzymatic systems.

Enhanced stability of charged dendrimer-encapsulated Pd nanoparticles in ionic liquids.

Highly stable dendrimer-encapsulated Pd nanoparticles in ionic liquids were prepared for the first time by using charged PAMAM dendrimers as templates, which could maintain hydrogenation efficiency for up to at least 12 recycles.