New Global Insights on the Regulation of the Biphasic Life Cycle and Virulence Via ClpP-Dependent Proteolysis in Legionella pneumophila

Legionella pneumophila, an environmental bacterium that parasitizes protozoa, causes Legionnaires’ disease in humans that is characterized by severe pneumonia. This bacterium adopts a distinct biphasic life cycle consisting of a nonvirulent replicative phase and a virulent transmissive phase in response to different environmental conditions. Hence, the timely and fine-tuned expression of growth and virulence factors in a life cycle–dependent manner is crucial for survival and replication. Here, we report that the completion of the biphasic life cycle and bacterial pathogenesis is greatly dependent on the protein homeostasis regulated by caseinolytic protease P (ClpP)-dependent proteolysis. We characterized the ClpP-dependent dynamic profiles of the regulatory and substrate proteins during the biphasic life cycle of L. pneumophila using proteomic approaches and discovered that ClpP-dependent proteolysis specifically and conditionally degraded the substrate proteins, thereby directly playing a regulatory role or indirectly controlling cellular events via the regulatory proteins. We further observed that ClpP-dependent proteolysis is required to monitor the abundance of fatty acid biosynthesis–related protein Lpg0102/Lpg0361/Lpg0362 and SpoT for the normal regulation of L. pneumophila differentiation. We also found that the control of the biphasic life cycle and bacterial virulence is independent. Furthermore, the ClpP-dependent proteolysis of Dot/Icm (defect in organelle trafficking/intracellular multiplication) type IVB secretion system and effector proteins at a specific phase of the life cycle is essential for bacterial pathogenesis. Therefore, our findings provide novel insights on ClpP-dependent proteolysis, which spans a broad physiological spectrum involving key metabolic pathways that regulate the transition of the biphasic life cycle and bacterial virulence of L. pneumophila, facilitating adaptation to aquatic and intracellular niches.     

Feeding Preferences in Greylag Geese and the Effect of Activated Charcoal

ABSTRACT Greylag geese (Anser anser) can cause serious damage to agricultural fields near wetlands that are attractive for resting and nesting but not for feeding. Alternative plantings or spraying fields may prevent goose damage. We randomly designed 64 plots in spring 2004 and prepared plantings of white clover (Trifolium repens), white clover with perennial ryegrass (Lolium perenne; mixture), fertilized perennial ryegrass (grass), or unfertilized perennial ryegrass. We measured goose-dropping densities in plots as a measure of feeding preference in autumn 2004 (7 weeks), spring 2005 (6 weeks), and autumn 2005 (7 weeks) following removal of a protective fence and vegetation sampling for content analysis in 2004. We also sprayed activated charcoal (20 kg/ha) in a suspension on 32 plots (8/planting) to deter geese in autumn 2004 only. In a second experiment we examined pairs of greylag geese in cages for preferences between grass treated with or without activated charcoal. Charcoal did not deter geese in either experiment. However, dropping density averaged highest for clover (1.01/m²), followed by the mixture (0.65/m²), then fertilized (0.23/m²) and unfertilized grass (0.16/m²). Preferences were consistent in all 3 experimental periods. Fertilized grass reached 31.8 cm in height on average in spring, whereas clover measured 15.4 cm. Crude protein and water-soluble carbohydrate content (g/kg dry matter) was 294 and 49, respectively, in white clover and 183 and 139, respectively, in fertilized grass. We found a positive partial correlation independent of vegetation type between dropping densities and crude protein and a negative correlation with water-soluble carbohydrate content. Thus, to prevent grazing damage to agricultural fields, we recommend planting white clover, strongly preferred by feeding geese, in areas (fallow agricultural or nonagricultural) adjacent to their habitat and not in agricultural fields under production.     

Assessment of the changes of 2,4-dichlorophenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal

The rate of adsorption of 2,4-dichlorophenoxyacetic acid (2,4-D) by activated charcoal (AC) from liquid and semi-solid tissue culture media was determined using 2-[¹⁴C]-2,4-D. In liquid medium 99.5% of the added 2,4-D (10^-4 M) was adsorbed by AC (2.5 gl^-1) within 5 days of preparation of the medium. Higher 2,4-D levels of reduced AC concentrations increased the level of available 2,4-D in the medium and extended the period necessary for the level of 2,4-D in the medium to become stabilized. In semi-solid medium the rate of adsorption of 2,4-D by AC was considerably reduced. A stable ratio of gel/2,4-D:AC/2,4-D was only reached after 10 to 20 days, depending on the 2,4-D concentration used. Low pH levels and maintenance of the medium at higher temperatures (20–30°C) accelerated the adsorption of 2,4-D by AC. In vitro tissue cultures of coconut palm showed marked differences in their growth response according to the age of the medium used and the associated variations in 2,4-D concentrations.Abbreviations AC activated charcoal - BAP 6-benzylamino-purine - 2,4-D 2,4-dichlorophenoxyacetic acid     

Mitogen‐activated protein kinase phosphatase 1 reduces the replication efficiency of Bamboo mosaic virus in Nicotiana benthamiana

In plants, the mitogen‐activated protein kinase (MAPK) cascades are the central signaling pathways of the complicated defense network triggered by the perception of pathogen‐associated molecular patterns to repel pathogens. The Arabidopsis thaliana MAPK phosphatase 1 (AtMKP1) negatively regulates the activation of MAPKs. Recently, the AtMKP1 homolog of Nicotiana benthamiana (NbMKP1) was found in association with the Bamboo mosaic virus (BaMV) replication complex. This study aimed to investigate the role of NbMKP1 in BaMV multiplication in N. benthamiana. Silencing of NbMKP1 increased accumulations of the BaMV‐encoded proteins and the viral genomic RNA, although the same condition reduced the infectivity of Pseudomonas syringae pv. tomato DC3000 in N. benthamiana. On the other hand, overexpression of NbMKP1 decreased the BaMV coat protein accumulation in a phosphatase activity‐dependent manner in protoplasts. NbMKP1 also negatively affected the in vitro RNA polymerase activity of the BaMV replication complex. Collectively, the activity of NbMKP1 seems to reduce BaMV multiplication, inconsistent with the negatively regulatory role of MKP1 in MAPK cascades in terms of warding off fungal and bacterial invasion. In addition, silencing of NbMKP1 increased the accumulation of Foxtail mosaic virus but decreased Potato virus X. The discrepant effects exerted by NbMKP1 on different pathogens foresee the difficulty to develop plants with broad‐spectrum resistance through genetically manipulating a single player in MAPK cascades.     

Properties of antibacterial bioboard from bamboo macromolecule by hot press

Bamboo macromolecules were pretreated with bamboo vinegar, which has an antibacterial property, and processed into antibacterial bioboard (ABB) by hot pressing. The ABB was then analyzed by conducting Fourier-transform infrared spectroscopy, thermogravimetric analysis and differential thermal analysis. Results showed that ABB samples had average density of 1.0 g/cm³, which is appropriate for application. The physical and mechanical properties were best for the ABB sample pretreated with bamboo vinegar and hot pressed at 165 °C for 10 min. Fourier-transform infrared spectroscopy revealed that the optimum conditions for hot pressing were a temperature of 165 °C, duration of 10 min, and the addition of bamboo vinegar. Thermogravimetric analysis/differential thermal analysis curves indicated that the thermal degradation of the ABB was less than that of bamboo, revealing that hot pressing increased the thermal stability of ABB samples. Analysis revealed that pretreatment with bamboo vinegar improved the antibacterial property of the ABB.     

In vitro biotechnological advancements in Malabar nut (Adhatoda vasica Nees): Achievements,status and prospects

Adhatoda vasica Nees, belonging to family Acanthaceae, is a well-known medicinal plant. It is endorsed for its pyrroloquinazoline alkaloids and its derivatives, such as vasicine and vasicinone. Germinating A. vasica seeds is a tedious task; on that account, vegetative propagation is the preferred method for its multiplication. For rapid and large-scale multiplication, germplasm conservation as well as secondary metabolites production, in vitro culture of A. vasica was preferred over conventional propagation by several researchers; however, some major applications of this tissue culture technique are still awaiting to undergo extensive research. The present review, for the first time, illustrates all the major achievements associated with in vitro regeneration of A. vasica, reported till date and highlights the future prospects.     

Recent advances in polysaccharide bio-based flocculants

Natural polysaccharides, derived from biomass feedstocks, marine resources, and microorganisms, have been attracting considerable attention as benign and environmentally friendly substitutes for synthetic polymeric products. Besides many other applications, these biopolymers are rapidly emerging as viable alternatives to harmful synthetic flocculating agents for the removal of contaminants from water and wastewater. In recent years, a great deal of effort has been devoted to improve the production and performance of polysaccharide bio-based flocculants. In this review, current trends in preparation and chemical modification of polysaccharide bio-based flocculants and their flocculation performance are discussed. Aspects including mechanisms of flocculation, biosynthesis, classification, purification and characterization, chemical modification, the effect of physicochemical factors on flocculating activity, and recent applications of polysaccharide bio-based flocculants are summarized and presented.     

采用竹管蜂巢繁育苜蓿切叶蜂

对5种繁育苜蓿切叶蜂的蜂巢材料进行了筛选测定,结果表明,竹管的繁蜂效果较好,子代蜂的雌雄性比为1：1,而且竹管内的蜂茧均未被寄生蜂寄生。