50 I. 50 II. 52 III. 53 IV. 66 V. 71 VI. 72 VII. 74 75 References 75 SUMMARY: Cryptospores, recovered from Ordovician through Devonian rocks, differ from trilete spores in possessing distinctive ...configurations (i.e. hilate monads, dyads, and permanent tetrads). Their affinities are contentious, but knowledge of their relationships is essential to understanding the nature of the earliest land flora. This review brings together evidence about the source plants, mostly obtained from spores extracted from minute, fragmented, yet exceptionally anatomically preserved fossils. We coin the term ‘cryptophytes’ for plants that produced the cryptospores and show them to have been simple terrestrial organisms of short stature (i.e. millimetres high). Two lineages are currently recognized. Partitatheca shows a combination of characters (e.g. spo‐rophyte bifurcation, stomata, and dyads) unknown in plants today. Lenticulatheca encompasses discoidal sporangia containing monads formed from dyads with ultrastructure closer to that of higher plants, as exemplified by Cooksonia. Other emerging groupings are less well characterized, and their precise affinities to living clades remain unclear. Some may be stem group embryophytes or tracheophytes. Others are more closely related to the bryophytes, but they are not bryophytes as defined by extant representatives. Cryptophytes encompass a pool of diversity from which modern bryophytes and vascular plants emerged, but were competitively replaced by early tracheophytes. Sporogenesis always produced either dyads or tetrads, indicating strict genetic control. The long‐held consensus that tetrads were the archetypal condition in land plants is challenged.
Full text
Available for:
BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NMLJ, NUK, OILJ, PNG, SAZU, SBCE, SBMB, UL, UM, UPUK
Spores of Bacillus species can remain in their dormant and resistant states for years, but exposure to agents such as specific nutrients can cause spores' return to life within minutes in the process ...of germination. This process requires a number of spore-specific proteins, most of which are in or associated with the inner spore membrane (IM). These proteins include the (i) germinant receptors (GRs) that respond to nutrient germinants, (ii) GerD protein, which is essential for GR-dependent germination, (iii) SpoVA proteins that form a channel in spores' IM through which the spore core's huge depot of dipicolinic acid is released during germination, and (iv) cortex-lytic enzymes (CLEs) that degrade the large peptidoglycan cortex layer, allowing the spore core to take up much water and swell, thus completing spore germination. While much has been learned about nutrient germination, major questions remain unanswered, including the following. (i) How do nutrient germinants penetrate through spores' outer layers to access GRs in the IM? (ii) What happens during the highly variable and often long lag period between the exposure of spores to nutrient germinants and the commitment of spores to germinate? (iii) What do GRs and GerD do, and how do these proteins interact? (iv) What is the structure of the SpoVA channel in spores' IM, and how is this channel gated? (v) What is the precise state of the spore IM, which has a number of novel properties even though its lipid composition is very similar to that of growing cells? (vi) How is CLE activity regulated such that these enzymes act only when germination has been initiated? (vii) And finally, how does the germination of spores of clostridia compare with that of spores of bacilli?
Some Bacillus species are causative agents of food spoilage and a wide array of diseases. Due to their ability to form highly heat-resistant spores, it is of great interest to develop more effective ...inactivation strategies whereby these spores could be inactivated. Therefore, this work assessed inactivation of thermal and ultrasound treatments against Bacillus subtilis spores. The study further investigated the thermosonication (thermal and ultrasound, TS) -induced inactivation to the spores through a combination of morphology observation and internal factor analyses. The results of TS indicated that the TS combination synergistically inactivated spores by the maximum log reduction of 2.43 ± 0.08 at 80 °C and 20 W/ml and caused severe cell damage. The visual images revealed that the destructive mode of action of TS had multitarget sites, including coat, cortex, and inner membrane. Three distinct sub-populations were detected by Flow cytometry (FCM), and an unknown step with some physical compromise of the spore's inner membrane and partially hydrolyzed cortex involving the three steps model of inactivation was suggested. The combination of DPA (pyridine-2,6 dicarboxylic acid) content and the relative viabilities of the fractions suggested that during the TS treatment DPA release took place largely after spore death. The dead spores that retained DPA germinated relatively normally, but outgrow poorly, indicating that some key enzymes of intermediary metabolism has been damaged by TS treatment. Such understanding of the lethal action of TS may lead to the development of novel strategies involving spore destruction.
Display omitted
•Synergetic inactivation of ultrasound and heat was conducted.•Electron microscopy revealed appearance and ultrastructure changes of Bacillus. subtilis spores.•Flow cytometry distinguished different physiological states of stressed B. subtilis spores.•Internal factor analyses elucidated the initial events in the killing of spores of B. subtilis by thermosonication.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Aims
Determine how supercritical CO2 (scCO2) plus peracetic acid (PAA) inactivates Bacillus subtilis spores, factors important in spore resistance to scCO2‐PAA, and if spores inactivated by scCO2‐PAA ...are truly dead.
Methods and Results
Spores of wild‐type B. subtilis and isogenic mutants lacking spore protective proteins were treated with scCO2‐PAA in liquid or dry at 35°C. Wild‐type wet spores (aqueous suspension) were more susceptible than dry spores. Treated spores were examined for viability (and were truly dead), dipicolinic acid (DPA), mutations, permeability to nucleic acid stains, germination under different conditions, energy metabolism and outgrowth. ScCO2‐PAA‐inactivated spores retained DPA, and survivors had no notable DNA damage. However, DPA was released from inactivated spores at a normally innocuous temperature (85°C), and colony formation from treated spores was salt sensitive. The inactivated spores germinated but did not outgrow, and these germinated spores had altered plasma membrane permeability and defective energy metabolism. Wet or dry coat‐defective spores had increased scCO2‐PAA sensitivity, and dry spores but not wet spores lacking DNA protective proteins were more scCO2‐PAA sensitive.
Conclusions
These findings suggest that scCO2‐PAA inactivates spores by damaging spores’ inner membrane. The spore coat provided scCO2‐PAA resistance for both wet and dry spores. DNA protective proteins provided scCO2‐PAA resistance only for dry spores.
Significance and Impact of the Study
These results provide information on mechanisms of spore inactivation of and resistance to scCO2‐PAA, an agent with increasing use in sterilization applications.
Full text
Available for:
BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Common sterilization techniques for labile and sensitive materials have far-reaching applications in medical, pharmaceutical, and industrial fields. Heat inactivation, chemical treatment, and ...radiation are established methods to inactivate microorganisms, but pose a threat to humans and the environment and can damage susceptible materials or products. Recent studies have demonstrated that cold low-pressure plasma (LPP) treatment is an efficient alternative to common sterilization methods, as LPP's levels of radicals, ions, (V)UV-radiation, and exposure to an electromagnetic field can be modulated using different process gases, such as oxygen, nitrogen, argon, or synthetic (ambient) air. To further investigate the effects of LPP, spores of the Gram-positive model organism
were tested for their LPP susceptibility including wild-type spores and isogenic spores lacking DNA-repair mechanisms such as non-homologous end-joining (NHEJ) or abasic endonucleases, and protective proteins like α/β-type small acid-soluble spore proteins (SASP), coat proteins, and catalase. These studies aimed to learn how spores resist LPP damage by examining the roles of key spore proteins and DNA-repair mechanisms. As expected, LPP treatment decreased spore survival, and survival after potential DNA damage generated by LPP involved efficient DNA repair following spore germination, spore DNA protection by α/β-type SASP, and catalase breakdown of hydrogen peroxide that can generate oxygen radicals. Depending on the LPP composition and treatment time, LPP treatment offers another method to efficiently inactivate spore-forming bacteria.IMPORTANCESurface-associated contamination by endospore-forming bacteria poses a major challenge in sterilization, since the omnipresence of these highly resistant spores throughout nature makes contamination unavoidable, especially in unprocessed foods. Common bactericidal agents such as heat, UV and γ radiation, and toxic chemicals such as strong oxidizers: (i) are often not sufficient to completely inactivate spores; (ii) can pose risks to the applicant; or (iii) can cause unintended damage to the materials to be sterilized. Cold low-pressure plasma (LPP) has been proposed as an additional method for spore eradication. However, efficient use of LPP in decontamination requires understanding of spores' mechanisms of resistance to and protection against LPP.
The current classification of the phylum
(new name,
) features eight distinct classes, six of which include known spore-forming bacteria. In Bacillus subtilis, sporulation involves up to 500 genes, ...many of which do not have orthologs in other bacilli and/or clostridia. Previous studies identified about 60 sporulation genes of B. subtilis that were shared by all spore-forming members of the
. These genes are referred to as the sporulation core or signature, although many of these are also found in genomes of nonsporeformers. Using an expanded set of 180 firmicute genomes from 160 genera, including 76 spore-forming species, we investigated the conservation of the sporulation genes, in particular seeking to identify lineages that lack some of the genes from the conserved sporulation core. The results of this analysis confirmed that many small acid-soluble spore proteins (SASPs), spore coat proteins, and germination proteins, which were previously characterized in bacilli, are missing in spore-forming members of
and other classes of
. A particularly dramatic loss of sporulation genes was observed in the spore-forming members of the families
and
. Fifteen species from diverse lineages were found to carry
(
-interrupting) elements of different sizes that all encoded SpoIVCA-like recombinases but did not share any other genes. Phylogenetic trees built from concatenated alignments of sporulation proteins and ribosomal proteins showed similar topology, indicating an early origin and subsequent vertical inheritance of the sporulation genes.
Many members of the phylum
(
) are capable of producing endospores, which enhance the survival of important Gram-positive pathogens that cause such diseases as anthrax, botulism, colitis, gas gangrene, and tetanus. We show that the core set of sporulation genes, defined previously through genome comparisons of several bacilli and clostridia, is conserved in a wide variety of sporeformers from several distinct lineages of
. We also detected widespread loss of sporulation genes in many organisms, particularly within the families
and
Members of these families, such as Lysinibacillus sphaericus and Clostridium innocuum, could be excellent model organisms for studying sporulation mechanisms, such as engulfment, formation of the spore coat, and spore germination.
Upon starvation, rod-shaped Myxococcus xanthus bacteria form mounds and then differentiate into round, stress-resistant spores. Little is known about the regulation of late-acting operons important ...for spore formation. C-signaling has been proposed to activate FruA, which binds DNA cooperatively with MrpC to stimulate transcription of developmental genes. We report that this model can explain regulation of the fadIJ operon involved in spore metabolism, but not that of the spore coat biogenesis operons exoA-I, exoL-P, and nfsA-H. Rather, a mutation in fruA increased the transcript levels from these operons early in development, suggesting negative regulation by FruA, and a mutation in mrpC affected transcript levels from each operon differently. FruA bound to all four promoter regions in vitro, but strikingly each promoter region was unique in terms of whether or not MrpC and/or the DNA-binding domain of Nla6 bound, and in terms of cooperative binding. Furthermore, the DevI component of a CRISPR-Cas system is a negative regulator of all four operons, based on transcript measurements. Our results demonstrate complex regulation of sporulation genes by three transcription factors and a CRISPR-Cas component, which we propose produces spores suited to withstand starvation and environmental insults.
Full text
Available for:
BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
The propensity for aerobic bacilli and allied genera to produce highly heat-resistant spores and thermally stable spoilage enzymes are major bacteriological issues faced by the dairy industry. Most ...of the enzymes are able to survive any heat treatment applied during the manufacture of milk powders and have the potential to remain active in milk powders and other dairy products during storage, and may explain some of the sensory and functionality defects reported in dairy products. Despite many reports on the occurrence of spore-forming bacteria in dairy products, knowledge about food quality related properties of many aerobic sporeformers is still scarce. Therefore, the aim of this study was to determine thermal resistance and spoilage potential of a large pool of mesophilic and thermophilic sporeformers, representing 738 isolates and 31 different RAPD groups, recently isolated from Chinese milk powders. Spore formers producing highly heat resistant spores (surviving 125°C for 30min) included 2 thermophiles (Geobacillus thermoleovorans group and Geobacillus stearothermophilus) and one mesophilic species (Brevibacillus brevis). Paenibacillus macerans showed the highest proteolytic activity followed by members of the Bacillus cereus group, Br. brevis, Bacillus subtilis, G. thermoleovorans group and Virgibacillus proomii. The highest lipase producing strains belonged to Bacillus licheniformis. Phospholipase activity was only shown by members of the B. cereus group and Brevibacillus parabrevis. Ten strains showed positive β-galactosidase activity, while, 4 strains showed positive haemolytic activity. B. licheniformis strains, despite belonging to one RAPD group or sub-group showed markedly different phenotypic characters which support the previous findings of heterogeneity in RAPD-based B. licheniformis groups. The results of this study will broaden the knowledge about the spoilage potential and thermal resistance of many strains of dairy origin.
•Heat resistance and spoilage potential of a wide range of mesophilic and thermophilic sporeformers have been determined•Spores of Geobacillus thermoleovorans group and Geobacillus stearothermophilus survived the heat treatment of 125°C for 30min•Paenibacillus macerans isolates are most proteolytic among sporeformers investigated•Members of the Bacillus cereus group are able to produce phospholipases, haemolysins, proteases and lipases•The phenotypic similarity within the same RAPD groups of Bacillus licheniformis is questioned
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
9.
physical state of water in bacterial spores Sunde, Erik P; Setlow, Peter; Hederstedt, Lars ...
Proceedings of the National Academy of Sciences - PNAS,
11/2009, Volume:
106, Issue:
46
Journal Article
Peer reviewed
Open access
The bacterial spore, the hardiest known life form, can survive in a metabolically dormant state for many years and can withstand high temperatures, radiation, and toxic chemicals. The molecular basis ...of spore dormancy and resistance is not understood, but the physical state of water in the different spore compartments is thought to play a key role. To characterize this water in situ, we recorded the water ²H and ¹⁷O spin relaxation rates in D₂O-exchanged Bacillus subtilis spores over a wide frequency range. The data indicate high water mobility throughout the spore, comparable with binary protein-water systems at similar hydration levels. Even in the dense core, the average water rotational correlation time is only 50 ps. Spore dormancy therefore cannot be explained by glass-like quenching of molecular diffusion but may be linked to dehydration-induced conformational changes in key enzymes. The data demonstrate that most spore proteins are rotationally immobilized, which may contribute to heat resistance by preventing heat-denatured proteins from aggregating irreversibly. We also find that the water permeability of the inner membrane is at least 2 orders of magnitude lower than for model membranes, consistent with the reported high degree of lipid immobilization in this membrane and with its proposed role in spore resistance to chemicals that damage DNA. The quantitative results reported here on water mobility and transport provide important clues about the mechanism of spore dormancy and resistance, with relevance to food preservation, disease prevention, and astrobiology.
Full text
Available for:
BFBNIB, NMLJ, NUK, PNG, SAZU, UL, UM, UPUK
2Duf, named after the presence of a transmembrane (TM) Duf421 domain and a small Duf1657 domain in its sequence, is likely located in the inner membrane (IM) of spores in some
species carrying a ...transposon with an operon termed
. These spores are known for their extreme resistance to wet heat, and 2Duf is believed to be the primary contributor to this trait. In this study, we found that the absence of YetF or YdfS, both Duf421 domain-containing proteins and found only in wild-type (wt)
spores with YetF more abundant, leads to decreased resistance to wet heat and agents that can damage spore core components. The IM phospholipid compositions and core water and calcium-dipicolinic acid levels of YetF-deficient spores are similar to those of wt spores, but the deficiency could be restored by ectopic insertion of
, and overexpression of YetF increased wt spore resistance to wet heat. In addition,
and
spores have decreased germination rates as individuals and populations with germinant receptor-dependent germinants and increased sensitivity to wet heat during germination, potentially due to damage to IM proteins. These data are consistent with a model in which YetF, YdfS and their homologs modify IM structure to reduce IM permeability and stabilize IM proteins against wet heat damage. Multiple
homologs are also present in other spore forming
and
and even some asporogenous
, but fewer in asporogenous species. The crystal structure of a YetF tetramer lacking the TM helices has been reported and features two distinct globular subdomains in each monomer. Sequence alignment and structure prediction suggest this fold is likely shared by other Duf421-containing proteins, including 2Duf. We have also identified naturally occurring
homologs in some
and
species and in wt
spores, but not in wt
. Notably, the genomic organization around the
gene in most of these species is similar to that in
, suggesting that one of these species was the source of the genes on this operon in the extremely wet heat resistant spore formers.
PDF
