Xiaohong Pan

Diversity of Microbial Community in Shihongtan Sandstone-Type Uranium Deposits, Xinjiang, China

Investigation of microbial communities in uranium deposits will be beneficial for understanding the indigenous microbial impacts on uranium-mineralization as well as developing appropriate remediation and long-term management strategies of uranium-contaminated repositories. In this study, microbial communities of Shihongtan uranium deposits, Xinjiang, China, were investigated using molecular biological techniques and traditional cultivation. PCR-DGGE and PCR-RFLP analyses suggested that there were a number of bacteria and archaea in Shihongtan uranium deposits. Phylogenetic analysis indicated that the bacterial communities were affiliated with Firmicutes, γ-Proteobacteria and Actinobacteria, while most of archaea showed close evolutionary relationship with Halobacteriaceae. Furthermore, a total of 27 bacterial strains were isolated from 8 core samples, and these strains were most closely related to Clostridium sp., Exiguobacterium sp., Enterobacter sp., Pseudomonas sp., Klebsiella sp., Aeromonas sp, Citrobacter sp., Tessaracoccus sp. and Jonesia sp., respectively. No archaea were cultured. We hope the current investigation of microbial communities in uranium deposits combining culture-dependent and -independent approaches, could not only help in understanding the microbial impacts on uranium-mineralization, but also help in identifying microbes that participate in uranium biomineralization for further study.

Physiological and biochemical response of Aedes aegypti tolerance to Bacillus thuringiensis

ABSTRACT Bacillus thuringiensis subsp. israelensis (Bti) represents the only eco-friendly bio-degradable insecticide for mosquito-borne disease control. Our research aims to identify if mosquito detoxification enzymes play an important role in Bti tolerance mechanisms in the dengue vector, Aedes aegypti. Several enzymes, such as amylase, cytochromes P450, Na+/K+-ATPase, acetylcholinesterase, protease and glutathione S-transferase (GST) were analysed and level of activity determined in Ae. aegypti larvae after Bti treatment. Bti exposure significantly increased the level of amylase (183.2%) as well as the activity of cytochromes P450 (177.5%), and Na+/K+-ATPase (142.9%). On the other hand, there was a decrease of 8.6% and 11.4% in acetylcholinesterase and GST activity, and no significant effect in the total level of protease activity. We suggest that the variation in amylase, cytochromes P450, Na+/K+-ATPase, acetylcholinesterase, protease and GST activity may be associated with the Bti insecticidal mechanism. This study provides the basis of detoxifying enzymes in Bti tolerance.

Microbial strategy for potential lead remediation: a review study.

The extensive exploitation and usage of lead compounds result in severe lead(II) pollution in water and soil environments, even in agricultural land, threatening the health of animals and humans via food chains. The recovery and remediation of lead(II) from water and soil environments have been intensively concerned in recent years. Compared with the traditional physic-chemistry treatment, microbial remediation strategy is a promising alternative to remediate lead(II)-contaminated environments due to its cost-effective and environmentally-friendly properties. Various microorganisms are capable of removing or immobilizing lead(II) from water and soil environments through bioaccumulation, precipitation or accelerated transformation of lead(II) into a very stable mineral, resulting in significant effects on lead(II) mobility and bioavailability. In the present review, we investigated a wide diversity of lead(II) bioremediation induced by different microbes and its multi-mechanisms. Moreover, we also discussed the progress and limitations, summarized the common rules of lead(II)-microbe interaction, and evaluated the environmental significance of microbes in lead biogeochemistry process. In addition, we further deliberated the feasibility and potential application of microbes in developing cost-effective, eco-friendly bioremediation or long-term management strategy for lead(II) contaminated repositories.

Cry11Aa Interacts with the ATP-Binding Protein from Culex quinquefasciatus To Improve the Toxicity

Cry11Aa displays high toxicity to the larvae of several mosquito species, including Aedes, Culex, and Anopheles. To study its binding characterization against Culex quinquefasciatus, Cry11Aa was purified and western blot results showed that Cry11Aa could bind successfully to the brush border membrane vesicles. To identify Cry11Aa-binding proteins in C. quinquefasciatus, a biotin-based protein pull-down experiment was performed and seven Cry11Aa-binding proteins were isolated from the midgut of C. quinquefasciatus larvae. Analysis of liquid chromatography-tandem mass spectrometry showed that one of the Cry11Aa-binding proteins is the ATP-binding domain 1 family member B. To investigate its binding property and effect on the toxicity of Cry11Aa, western blot, far-western blot, enzyme-linked immunosorbent assay, and bioassays of Cry11Aa in the presence and absence of the recombinant ATP-binding protein were performed. Our results showed that the ATP-binding protein interacted with Cry11Aa and increased the toxicity of Cry11Aa against C. quinquefasciatus. Our study suggests that midgut proteins other than the toxin receptors may modulate the toxicity of Cry toxins against mosquitoes.

Transcriptomic Analysis of Aedes aegypti in Response to Mosquitocidal Bacillus thuringiensis LLP29 Toxin

Globally, Aedes aegypti is one of the most dangerous mosquitoes that plays a crucial role as a vector for human diseases, such as yellow fever, dengue, and chikungunya. To identify (1) transcriptomic basis of midgut (2) key genes that are involved in the toxicity process by a comparative transcriptomic analysis between the control and Bacillus thuringiensis (Bt) toxin (LLP29 proteins)-treated groups. Next-generation sequencing technology was used to sequence the midgut transcriptome of A. aegypti. A total of 17130 unigenes, including 574 new unigenes, were identified containing 16358 (95.49%) unigenes that were functionally annotated. According to differentially expressed gene (DEG) analysis, 557 DEGs were annotated, including 226 upregulated and 231 downregulated unigenes in the Bt toxin-treated group. A total of 442 DEGs were functionally annotated; among these, 33 were specific to multidrug resistance, 6 were immune-system-related (Lectin, Defensin, Lysozyme), 28 were related to putative proteases, 7 were lipase-related, 8 were related to phosphatases, and 30 were related to other transporters. In addition, the relative expression of 28 DEGs was further confirmed through quantitative real time polymerase chain reaction. The results provide a transcriptomic basis for the identification and functional authentication of DEGs in A. aegypti.

Nematicidal Activity of Cry1Ea11 from Bacillus thuringiensis BRC-XQ12 Against the Pine Wood Nematode (Bursaphelenchus xylophilus)

The nematicidal activity of 92 Bacillus thuringiensis strains against the pine wood nematode Bursaphelenchus xylophilus, one of the worlds top 10 plant-parasitic nematodes, was determined. The insecticidal crystal proteins (ICPs) from Bacillus thuringiensis BRC-XQ12 were the most toxic to Bursaphelenchus xylophilus, with a lethal concentration 50 (LC50) of 32.13 μg/ml. Because the ICPs expressed by Bacillus thuringiensis BRC-XQ12 were closest to Cry1Ea6 and B. thuringiensis BRC-XQ12 contained four kinds of cry1 subgenes (cry1Aa, cry1Cb, cry1Ea, and cry1Ia), Cry1Ea was most likely to be the key active component against the nematode. The 3,516-bp cry1Ea11 gene from BRC-XQ12, as designated by the B. thuringiensis δ-endotoxin nomenclature committee, was expressed in Escherichia coli. Purified Cry1Ea11 showed an LC50 of 32.53 and 23.23 μg/ml at 24 and 48 h, with corresponding virulence equations of Y = 32.15X + 1.38 (R2 = 0.9951) and Y = 34.29X + 3.16 (R2 = 0.9792), respectively. In order to detect the pathway of B. thuringiensis Cry1Ea11 into Bursaphelenchus xylophilus, the nematode was fed with NHS-rhodamine-labeled GST-Cry1Ea11. The results of confocal laser-scanning microscopy showed that the 159-kDa GST-Cry1Ea11 could be detected in the stylet and the esophageal lumen of the pine wood nematode, indicating that GST-Cry1Ea11 could enter into the nematode through the stylet. As far as we know, no Cry1 proteins have been shown to have activity against plant-parasitic nematodes before. These results demonstrate that Cry1Ea11 is a promising nematicidal protein for controlling pine wilt disease rendered by B. xylophilus, further dramatically broadening the spectrum of Bacillus thuringiensis ICPs.

Comparison and Mechanism of the UV-Resistant Mosquitocidal Bt Mutant LLP29-M19

Abstract Bacillus thuringiensis (Bt) is one of the most widely used and studied biopesticides. However, it is vulnerable to the influence of ultraviolet (UV) radiation, causing shorter persistence under field conditions. To obtain a high-active and effective Bt new product, the main objective of this study is to obtain a highly UV-resistant Bt mutant from the mosquitocidal Bt LLP29 through UV exposure. After 19 rounds of UV exposure, a Bt mutant named LLP29-M19 was obtained, showing resistance to UV radiation for up to 67 min. The mosquitocidal fatality rate of LLP29-M19 was 95%, which was slightly higher than that of LLP29 (90%). Comparative characterization showed that there were no substantial differences in morphology between LLP29-M19 and the original strain, LLP29. However, some changes were detected in physiological and biochemical characteristic reactions, including fructose, glucose, and xylose metabolism. Furthermore, although both LLP29-M19 and LLP29 showed negative zeta potentials, the surface charge of LLP29 was −28.1 mV and that of LLP29-M19 was −42.8 mV. The size distribution of LLP29-M19 was also slightly larger than that of LLP29. Fourier transform infrared analysis indicated that amide functional groups might be involved in the resistance mechanism of LLP29-M19. Quantitative analysis using inductive coupled plasma emission spectrometry showed that some elements increased greatly in LLP29-M19, such as K. All of these results will be highly valuable for better understanding the mechanism of Bt resistance. Explanations regarding the resistance mechanism of this novel Bt mutant may lead to the development of new biopesticides with high mosquitocidal activity and persistence.

Flowerlike Mg(OH)2 Cross-Nanosheets for Controlling Cry1Ac Protein Loss: Evaluation of Insecticidal Activity and Biosecurity

Bacillus thuringiensis (Bt) can produce Cry proteins during the sporulation phase, and Cry protein is effective against lepidopteran, coleopteran, and dipteran insects and nematodes. However, Cry protein tends to be discharged into soil and nontarget plants through rainwater runoff, leading to reduced effective period toward target insects. In the present study, nano-Mg(OH)2 (magnesium hydroxide nanoparticles, MHNPs) were synthesized to control the loss of Cry1Ac protein and deliver protein to Helicoverpa armigera (Lepidoptera: Noctuidae). The results showed that Cry1Ac protein could be loaded onto MHNPs through electrostatic adsorption, and both MHNPs and Cry protein were stable during the adsorption process. Meanwhile, the Cry1Ac-loaded MHNPs could remain on the surface of cotton leaves, resulting in enhanced adhesion of Cry1Ac protein by 59.50% and increased pest mortality by 75.00%. Additionally, MHNPs could be slowly decomposed by acid medium and MHNPs showed no obvious influence on cotton, Bt, Escherichia coli, and H. armigera. Therefore, MHNPs could serve as an efficient nanocarrier for delivery of Cry1Ac protein and be used as a potential adjuvant for biopesticide in agricultural applications.

The Synergistic Antibacterial Mechanism of Gentamicin-Loaded CaCO3 Nanoparticles

In the present study, we used CaCO3 nanoparticles (CCNPs) as carriers to assess the physicochemical characteristics and antibacterial effect of gentamicin sulfate (GS)-loaded CCNPs (CGPs). The results indicated that CCNPs had relatively regular chain-like structure, and the size of the crystallites was around 62.5 nm. FT-IR analysis indicated that the GS could effectively load onto CCNPs. Meanwhile, the dosage of CCNPs would affect the drug loading and entrapment efficiency of GS. CCNPs could prolong the release of GS, and the complete release of GS from CCNPs was extended up to 24 h. Additionally, CCNPs could obviously increase the antibacterial effect of GS. The zeta potential analysis and microscopic investigations indicated that the adsorbed CCNPs could increase the damage level of bacterial cell wall and enhance the permeability of cell membranes, leading to increased bacterial death.

The adsorption features between insecticidal crystal protein and nano-Mg(OH)2

Nano-Mg(OH)2, with low biological toxicity, is an ideal nano-carrier for insecticidal protein to improve the bioactivity. In this work, the adsorption features of insecticidal protein by nano-Mg(OH)2 have been studied. The adsorption capacity could reach as high as 136 mg g−1, and the adsorption isotherm had been fitted with Langmuir and Freundlich models. Moreover, the adsorption kinetics followed a pseudo-first or -second order rate model, and the adsorption was spontaneous and an exothermic process. However, high temperatures are not suitable for adsorption, which implies that the temperature would be a critical factor during the adsorption process. In addition, FT-IR confirmed that the protein was adsorbed on the nano-Mg(OH)2, zeta potential analysis suggested that insecticidal protein was loaded onto the nano-Mg(OH)2 not by electrostatic adsorption but maybe by intermolecular forces, and circular dichroism spectroscopy of Cry11Aa protein before and after loading with nano-Mg(OH)2 was changed. The study applied the adsorption information between Cry11Aa and nano-Mg(OH)2, which would be useful in the practical application of nano-Mg(OH)2 as a nano-carrier.