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Ethnopharmacological relevance: Present survey was conducted to explore ethnomedicinal uses and cultural importance of wild edible fruits species by the inhabitants of Lesser Himalayas-Pakistan. Materials and methods: Information was obtained through informed consent semi-structured interviews, questionnaires, market survey, focus group conversation, unceremonious dialogue and village walks with key informants. Cultural significance of each species was calculated based on use report by participants at each study site. Results: A total of 35 wild edible fruits belonging to 21 genera and 17 families were used for the treatment of various ailments and consumed. Rosaceae was found dominating family with (8 spp.), followed by Moraceae (6 spp.), Rhamnaceae (5 spp.), Palmae and Vitaceae (2 spp. each) and remaining families were represented by one species each. Fruits (48%) were found highly utilized plant parts, followed by leaves (34%), bark, flowers and seeds (4% each), branches, latex and roots (2% each). Water was used as a medium for preparation while milk, ghee, oil, egg and butter are used for application. Modes of preparation were fall into seven categories like fresh parts eaten raw (38%), powder (24%), decoction (20%), extract (12 %), paste (4%), juice and latex (2% each). Based on cultural important index (CI) Morus nigra was found most significant species within top ten fruit plants followed by Morus alba, Olea ferruginea, Berberis lycium, Pyrus pashia, Ficus carica, Ficus palmata, Ziziphus mauritiana, Diospyros lotus and Ziziphus nummularia. Conclusions: Traditional uses of wild edible plant depend mainly on socio-economic factors rather than climatic conditions or wealth of flora. Use reports and citation demonstrated that there is a common cultural heritage regarding the gathered food plants. Further investigation is required for Antioxidant study, essential and toxic components, pharmacological applications; dietary requirements and biotechnological techniques to improve yields.
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Plant retinoblastoma-related (RBR) proteins are primarily considered as key regulators of G(1)/S phase transition, with functional roles in a variety of cellular events during plant growth and organ development. Polyclonal antibody against the C-terminal region of the Arabidopsis RBR1 protein also specifically recognizes the alfalfa 115 kDa MsRBR protein, as shown by the antigen competition assay. The MsRBR protein was detected in all cell cycle phases, with a moderate increase in samples representing G(2)/M cells. Antibody against the human phospho-pRb peptide (Ser807/811) cross-reacted with the same 115 kDa MsRBR protein and with the in vitro phosphorylated MsRBR protein C-terminal fragment. Phospho-MsRBR protein was low in G(1) cells. Its amount increased upon entry into the S phase and remained high during the G(2)/M phases. Roscovitine treatment abolished the activity of alfalfa MsCDKA1;1 and MsCDKB2;1, and the phospho-MsRBR protein level was significantly decreased in the treated cells. Colchicine block increased the detected levels of both forms of MsRBR protein. Reduced levels of the MsRBR protein in cells at stationary phase or grown in hormone-free medium can be a sign of the division-dependent presence of plant RBR proteins. Immunolocalization of the phospho-MsRBR protein indicated spots of variable number and size in the labelled interphase nuclei and high signal intensity of nuclear granules in prophase. Structures similar to phospho-MsRBR proteins cannot be recognized in later mitotic phases. Based on the presented western blot and immunolocalization data, the possible involvement of RBR proteins in G(2)/M phase regulation in plant cells is discussed.
Associations of salicylate-like phenolic glycosides (PGs) with biological activity have been reported in Salix and Populus trees, but only for a few compounds, and in relation to a limited number of herbivores. By considering the full diversity of PGs, we may improve our ability to recognize genotypes or chemotype groups and enhance our understanding of their ecological function. Here, we present a fast and efficient general method for salicylate determination in leaves of Eurasian aspen that uses ultra-high performance liquid chromatography-electrospray ionization/time-of-flight mass spectrometry (UHPLC-ESI/TOFMS). The time required for the liquid chromatography separations was 13.5 min per sample, compared to around 60 min per sample for most HPLC protocols. In leaf samples from identical P. tremula genotypes with diverse propagation and treatment histories, we identified nine PGs. We found the compound-specific mass chromatograms to be more informative than the UV-visible chromatograms for compound identification and when quantitating samples with large variability in PG content. Signature compounds previously reported for P. tremoloides (tremulacin, tremuloidin, salicin, and salicortin) always were present, and five PGs (2'-O-cinnamoyl-salicortin, 2'-O-acetyl-salicortin, 2'-O-acetyl-salicin, acetyl-tremulacin, and salicyloyl-salicin) were detected for the first time in P. tremula. By using information about the formic acid adduct that appeared for PGs in the LTQ-Orbitrap MS environment, novel compounds like acetyl-tremulacin could be tentatively identified without the use of standards. The novel PGs were consistently either present in genotypes regardless of propagation and damage treatment or were not detectable. In some genotypes, concentrations of 2'-O-acetyl-salicortin and 2'-O-cinnamoyl-salicortin were similar to levels of biologically active PGs in other Salicaceous trees. Our study suggests that we may expect a wide variation in PG content in aspen populations which is of interest both for studies of interactions with herbivores and for mapping population structure.
The identity and scope of chloroplast and mitochondrial proteases in higher plants has only started to become apparent in recent years. Biochemical and molecular studies suggested the existence of Clp, FtsH, and DegP proteases in chloroplasts, and a Lon protease in mitochondria, although currently the full extent of their role in organellar biogenesis and function remains poorly understood. Rapidly accumulating DNA sequence data, especially from Arabidopsis, has revealed that these proteolytic enzymes are found in plant cells in multiple isomeric forms. As a consequence, a systematic approach was taken to catalog all these isomers, to predict their intracellular location and putative processing sites, and to propose a standard nomenclature to avoid confusion and facilitate scientific communication. For the Clp protease most of the ClpP isomers are found in chloroplasts, whereas one is mitochondrial. Of the ATPase subunits, the one ClpD and two ClpC isomers are located in chloroplasts, whereas both ClpX isomers are present in mitochondria. Isomers of the Lon protease are predicted in both compartments, as are the different forms of FtsH protease. DegP, the least characterized protease in plant cells, has the most number of isomers and they are predicted to localize in several cell compartments. These predictions, along with the proposed nomenclature, will serve as a framework for future studies of all four families of proteases and their individual isomers.
Ecological significance of trees growing in urban and peri-urban settings is likely to increase in future land-use regimes, calling for better understanding of their role as potential reservoirs or stepping stones for associated biodiversity. We studied the diversity of fungal endophytes in woody tissues of asymptomatic even aged pedunculate oak trees, growing as amenity trees in a peri-urban setting. The trees were classified into three groups according to their phenotypic vitality (high, medium, and low). Endophytes were cultured on potato dextrose media from surface sterilized twigs and DNA sequencing was performed to reveal the taxonomic identity of the morphotypes. In xylem tissues, the frequency and diversity of endophytes was highest in oak trees showing reduced vitality. This difference was not found for bark samples, in which the endophyte infections were more frequent and communities more diverse than in xylem. In general, most taxa were shared across the samples with few morphotypes being recovered in unique samples. Leaf phenolic profiles were found to accurately classify the trees according to their phenotypic vitality. Our results confirm that xylem is more selective substrate for endophytes than bark and that endophyte assemblages in xylem are correlated to the degree of host vitality. Thus, high vitality of trees may be associated with reduced habitat quality to wood-associated endophytes.
A panel of aluminum-tolerant (AlRes) mutants was isolated by protoplast-based T-DNA activation tagging in the tobacco cultivar SR1. The mutants fell into two phenotypic classes: a minority of the mutants were fertile and developed similarly to the wild type (type I), the majority was male-sterile and grew as semi-dwarfs (type II). These traits, along with the aluminum tolerance, were inherited in a monogenic dominant manner. Both types of mutants were characterized by excessive bundling of actin microfilaments and by a strongly increased abundance of actin, a phenotype that could be partially phenocopied in the wild type by treatment with aluminum chloride. The actin bundles could be dissociated into finer strands by addition of exogenous auxin in both types of mutants. However, actin microfilaments and leaf expansion were sensitive to blockers of actin assembly in the wild type and in the mutants of type I, whereas they were more tolerant in the mutants of type II. The mutants of type II displayed a hypertrophic development of vasculature, manifest in form of supernumerary leaf veins and extended xylem layers in stems and petioles. Whereas mutants of type I were characterized by a normal, but aluminum-tolerant polar auxin-transport, auxin-transport was strongly promoted in the mutants of type II. The phenotype of these mutants is discussed in terms of reduced endocytosis leading, concomitantly with aluminum tolerance, to changes in polar auxin transport.
Experiments with several Arabidopsis thaliana mutants have revealed a web of interactions between hormonal signaling. Here, we show that the Arabidopsis mutant radical-induced cell death1 (rcd1), although hypersensitive to apoplastic superoxide and ozone, is more resistant to chloroplastic superoxide formation, exhibits reduced sensitivity to abscisic acid, ethylene, and methyl jasmonate, and has altered expression of several hormonally regulated genes. Furthermore, rcd1 has higher stomatal conductance than the wild type. The rcd1-1 mutation was mapped to the gene At1g32230 where it disrupts an intron splice site resulting in a truncated protein. RCD1 belongs to the (ADP-ribosyl)transferase domain–containing subfamily of the WWE protein–protein interaction domain protein family. The results suggest that RCD1 could act as an integrative node in hormonal signaling and in the regulation of several stress-responsive genes.
Senescence is the last stage of development in plants. During foliar senescence, theleaves experience changes in metabolism, cell structures and gene expression. In thisprocess the limiting nutrients of leaves are relocated from senescing organs to thedeveloping leaves, fruits and buds. The genetic screening to identify senescence mutantsis of utmost importance both for scientific research and for future biotechnology becausethe stay green mutants could increase crop yields.In our experimental setup, the senescence was induced by darkening the individual leavesof Arabidopsis thaliana plants with "mittens" for six days. We have screened 1064Arabidopsis plants for the mutants with delayed senescence phenotypes (stay-green,light-green and slightly light green plants) by measuring chlorophyll content, proteincontent and change in protein levels of the rubisco LSU, COXII, GS1 and GS2 duringdark induced senescence. After senescence induction, the wild type plants had showntypical yellowing and drying of leaves coupled with chlorophyll and protein degradation,while the foliar senescence progresses more slowly in selected stay green mutants. Theselected stay green mutants (19/29, 19/1043 and 19/531) delayed the degradation ofchlorophyll and protein contents even after 6 days of dark treatment. Furthermore, theretention of chloroplast and mitochondrial protein (rubisco LSU and GS2, and COXIIrespectively) and their higher chlorophyll contents may suggest that 19/29, 19/531and19/1043 plants may maintain their photosynthetic and respiratory activities. Thesemutants delay the functional leaf senescence and could be candidates for functional staygreen mutants for upcoming experiments.
Recent efforts to sequence the genomes and transcriptomes of several gymnosperm species have revealed an increased complexity in certain gene families in gymnosperms as compared to angiosperms. One example of this is the gymnosperm sister Glade to angiosperm TM3-like MADS-box genes, which at least in the conifer lineage has expanded in number of genes. We have previously identified a member of this subclade, the conifer gene DEFICIENS AGAMOUS LIKE 19 (DAL19), as being specifically upregulated in cone-setting shoots. Here, we show through Sanger sequencing of mRNA-derived cDNA and mapping to assembled conifer genomic sequences that DAL19 produces six mature mRNA splice variants in Picea abies. These splice variants use alternate first and last exons, while their four central exons constitute a core region present in all six transcripts. Thus, they are likely to be transcript isoforms. Quantitative Real-Time PCR revealed that two mutually exclusive first DAL19 exons are differentially expressed across meristems that will form either male or female cones, or vegetative shoots. Furthermore, mRNA in situ hybridization revealed that two mutually exclusive last DAL19 exons were expressed in a cell-specific pattern within bud meristems. Based on these findings in DAL19, we developed a sensitive approach to transcript isoform assembly from short-read sequencing of mRNA. We applied this method to 42 putative MADS-box core regions in P abies, from which we assembled 1084 putative transcripts. We manually curated these transcripts to arrive at 933 assembled transcript isoforms of 38 putative MADS-box genes. 152 of these isoforms, which we assign to 28 putative MADS-box genes, were differentially expressed across eight female, male, and vegetative buds. We further provide evidence of the expression of 16 out of the 38 putative MADS-box genes by mapping PacBio Iso-Seq circular consensus reads derived from pooled sample sequencing to assembled transcripts. In summary, our analyses reveal the use of mutually exclusive exons of MADS-box gene isoforms during early bud development in P. abies, and we find that the large number of identified MADS-box transcripts in P. abies results not only from expansion of the gene family through gene duplication events but also from the generation of numerous splice variants.
Intact cells of Chlamydomonas reinhardtii as well as isolated thylakoid membranes and photosystem II complexes were used to examine a possible mechanism of anthracene (ANT) interaction with the photosynthetic apparatus. Since ANT concentrations above 1 mM were required to significantly inhibit the rate of oxygen evolution in PS II membrane fragments it may indicate that the toxicant did not directly interact with this photosystem. On the other hand, stimulation of oxygen uptake by ANT-treated thylakoids suggested that ANT could either act as an artificial electron acceptor in the photosynthetic electron transport chain or function as an uncoupler. Electron transfer from excited chlorophyll to ANT is impossible due to the very low reduction potential of ANT and therefore we propose that toxic concentrations of ANT increase the thylakoid membrane permeability and thereby function as an uncoupler, enhancing electron transport in vitro. Hence, its unspecific interference with photosynthetic membranes in vitro suggests that the inhibitory effect observed on intact cell photosynthesis is caused by uncoupling of phosphorylation.
Olive, representing one of the most important fruit crops of the Mediterranean area, is characterized by a general low fruit yield, due to numerous constraints, including alternate bearing, low flower viability, male-sterility, inter-incompatibility, and self-incompatibility (SI). Early efforts to clarify the genetic control of SI in olive gave conflicting results, and only recently, the genetic control of SI has been disclosed, revealing that olive possesses an unconventional homomorphic sporophytic diallelic system of SI, dissimilar from other described plants. This system, characterized by the presence of two SI groups, prevents self-fertilization and regulates inter-compatibility between cultivars, such that cultivars bearing the same incompatibility group are incompatible. Despite the presence of a functional SI, some varieties, in particular conditions, are able to set seeds following self-fertilization, a mechanism known as pseudo-self-compatibility (PSC), as widely reported in previous literature. Here, we summarize the results of previous works on SI in olive, particularly focusing on the occurrence of self-fertility, and offer a new perspective in view of the recent elucidation of the genetic architecture of the SI system in olive. Recent advances in research aimed at unraveling the molecular bases of SI and its breakdown in olive are also presented. The clarification of these mechanisms may have a huge impact on orchard management and will provide fundamental information for the future of olive breeding programs.
Abstract. 1. Many Salicaceae species naturally form hybrid swarms with parental and hybrid taxa that differ in secondary chemical profile and in resistance to herbivores. Theoretically, the differential mortality in the seedling stage can lead to changes in trait expression and alter subsequent interactions between plants and herbivores. This study examines whether herbivory by the generalist slug Arion subfuscus, which causes extensive mortality in young willow seedlings, causes shifts in (a) the foliar chemistry of F2 willow hybrids (Salix sericea and Salix eriocephala), and (b) the subsequent susceptibility to Japanese Beetles, Popillia japonica.
2. In 2001, two populations of F2 seedlings were generated: those that survived slug herbivory (80–90% of seedlings placed in the field were killed by the slugs) were designated as S-plants, whereas C-plants (controls) experienced no mortality.
3. Common garden experiments with cuttings from these populations, in 2001 and 2002, revealed extensive variation in the phenolic chemistry of F2 hybrids, but revealed no significant difference between S- and C-plants, although the levels of foliar nutrients, proteins and nitrogen tended to be higher in S-plants.
4. Concentrations of salicortin and 2'-cinnamoylsalicortin explained 55 and 38% of the the variation in leaf damage caused by Japanese beetles, and secondary chemistry was highly correlated within replicate clones (salicortin R2 = 0.85, 2-cinnamoylsalicortin R2 = 0.77, condensed tannins R2 = 0.68).
5. Interestingly, Japanese beetle damage and condensed tannins were positively correlated within the S-plants, but not in the C-plants, suggesting that slugs had selected for plants with a positive relationship between tannins and P. japonica damage. This is unlikely to be a consequence of a preference for tannins, but is suggested to be related to the elevated nutrient levels in the S-plants, perhaps in combination with the complex-binding properties of tannins.
6. The damage was highly correlated within replicate clones and a model choice analysis suggested that Japanese beetle damage may be explained by four factors: concentrations of salicortin, condensed tannins, and nitrogen, as well as the specific leaf area (thick leaves were damaged less).
According to the geographic mosaic theory of coevolution (GMTC), clines of traits reflecting local co-adaptation (including resistance genes) should be common between a host and its parasite and should persist across time. To test the GMTC-assumption of persistent clinal patterns we compared the natural prevalence of two parasites on aspen Populus tremula trees: mining moths of the genus Phyllocnistis and leaf rust Melampsora spp. Damage data were collated from the Swedish National Forest Damage Inventory (2004–2006). In addition, occurrence of the parasites was scored in field conditions in two common gardens in the north and south of Sweden over five growing seasons (2004–2008), then related to biomass (stem height and diameter) and to concentrations of eleven leaf phenolics. Phyllocnistis mainly occurred in the northern garden, a distribution range which was confirmed by the countrywide inventory, although Phyllocnistis was more abundant on southern clones, providing evidence for possible local maladaptation. Melampsora occurred all over the country and in both gardens, but built up more quickly on northern clones, which suggests a centre of local clone maladaptation in the north. Stem growth also followed a clinal pattern as did the concentration of three phenolic compounds: benzoic acid, catechin and cinnamic acid. However, only benzoic acid was related to parasite presence: negatively to Phyllocnistis and positively to Melampsora and it could thus be a potential trait under selection.
In conclusion, clines of Phyllocnistis were stronger and more persistent compared to Melampsora, which showed contrasting clines of varying strength. Our data thus support the assumption of the GMTC model that clines exist in the border between hot and cold spots and that they may be less persistent for parasites with an elevated gene flow, and/or for parasites which cover relatively larger hot spots surrounded by fewer cold spots.
Although phenological matching between two and three trophic interactions has received some attention, it has largely been disregarded in explaining the lack of strong cascade dynamics in terrestrial systems. We studied the response of the specialist seed predator, Paroxyna plantaginis (Tephritidae) and associated generalist parasitoids (Chalcidoidea) to controlled fertilisation of individuals of naturally growing Tripolium vulgare (Asteraceae) on four island populations (Skeppsvik Archipelago, Sweden). We consistently found evidence of nutrient limitation: fertilised plants increased their biomass, produced more capitula (the oviposition units for tephritid flies), were more at risk of attack by the tephritids, and puparia were heavier in fertilised plants. During some parts of the season tephritids became more heavily parasitized, supporting the presence of cascade dynamics, however net parasitism over season decreased in response to nutrient addition. We found no evidence that capitulum size complicated parasitoid access to the tephritids, however the extended bud production prolonged the flowering season. Thus, tephritids utilized the surplus production of capitula throughout the entire season, while parasitoids did not expand their oviposition time window accordingly. Implications for top down regulation and cascade dynamics in the system are discussed.
Salicinoid phenolic glycosides are common defence substances in salicaceous trees and specialist leaf beetles use these compounds for their own defence against predators. Salicinoids vary qualitatively and qualitatively in aspen (Populus tremula) and this variation has a genetic basis. The foliar endophyte mycobiome is plentiful and we hypothesised that it is related to plant genotype, potentially mediated by salicinoid composition, and that interactions with the leaf beetle Chrysomela tremula may alter this relationship. We studied these three-way interactions in controlled greenhouse experiments. Endophytic fungi were isolated from sterilised leaf tissues with and without beetle damage, and from beetles. We confirmed that endophyte composition was influenced by host genotype. Beetle activity added generalist morphs to the mycobiome that overrode the initial host association. Yeast-like genera (Cryptococcus and Rhodotorula) were isolated only from beetle-damaged tissues and from beetles, whereas fast-growing filamentous fungi dominated beetle-free control plants. Competition experiments between filamentous fungi of plant origin and beetle-related yeasts suggested interaction of both stimulating and inhibiting modes of action amongst the fungi. As a result, we detected examples of amensalism, commensalism, parasitism and competition between the morphs tested, but we found no evidence of mutualism, and consequently no co-evolutionary relationship could be demonstrated, between yeasts carried by beetles, host genotype and associated filamentous morphs. Endophyte studies are method-dependent and high-throughput sequencing technology best define the fungal mycobiome, culturing however continues to be a cheap way to provide fundamental ecological insights and it is also required for experimental studies.
Vyacheslav Vasilevich (V.V.) Klimov (or Slava, as most of us called him) was born on January 12, 1945 and passed away on May 9, 2017. He began his scientific career at the Bach Institute of Biochemistry of the USSR Academy of Sciences (Akademy Nauk (AN) SSSR), Moscow, Russia, and then, he was associated with the Institute of Photosynthesis, Pushchino, Moscow Region, for about 50 years. He worked in the field of biochemistry and biophysics of photosynthesis. He is known for his studies on the molecular organization of photosystem II (PSII). He was an eminent scientist in the field of photobiology, a well-respected professor, and, above all, an outstanding researcher. Further, he was one of the founding members of the Institute of Photosynthesis in Pushchino, Russia. To most, Slava Klimov was a great human being. He was one of the pioneers of research on the understanding of the mechanism of light energy conversion and of water oxidation in photosynthesis. Slava had many collaborations all over the world, and he is (and will be) very much missed by the scientific community and friends in Russia as well as around the World. We present here a brief biography and some comments on his research in photosynthesis. We remember him as a friendly and enthusiastic person who had an unflagging curiosity and energy to conduct outstanding research in many aspects of photosynthesis, especially that related to PSII.
Photosynthetic organisms experience wide fluctuations in light intensity and regulate light harvesting accordingly to prevent damage from excess energy. The antenna quenching component qH is a sustained form of energy dissipation that protects the photosynthetic apparatus under stress conditions. This photoprotective mechanism requires the plastid lipocalin LCNP and is prevented by SUPPRESSOR OF QUENCHING1 (SOQ1) under non-stress conditions. However, the molecular mechanism of qH relaxation has yet to be resolved. Here, we isolated and characterized RELAXATION OF QH1 (ROQH1), an atypical short-chain dehydrogenase–reductase that functions as a qH-relaxation factor in Arabidopsis. The ROQH1 gene belongs to the GreenCut2 inventory specific to photosynthetic organisms, and the ROQH1 protein localizes to the chloroplast stroma lamellae membrane. After a cold and high-light treatment, qH does not relax in roqh1 mutants and qH does not occur in leaves overexpressing ROQH1. When the soq1 and roqh1 mutations are combined, qH can neither be prevented nor relaxed and soq1 roqh1 displays constitutive qH and light-limited growth. We propose that LCNP and ROQH1 perform dosage-dependent, antagonistic functions to protect the photosynthetic apparatus and maintain light-harvesting efficiency in plants.
The effect of nutrient enrichment on the distribution of polychlorinated biphenyl's (PCBs) in the microbial food web and the residence time of PCBs in seawater was studied in an experimental mesocosm system. Two 5 m high temperature and light controlled mesocosm tubes (empty set = 0.5 m) were filled with seawater from the northern Baltic Sea. Inorganic phosphorus and nitrogen were added daily to one mesocosm, while the other served as a control. Experiments were conducted at 5, 10 and 20 degrees C. Three C-14-labelled PCBs of different degree of chlorination were added to subsamples of the mesocosms: 4-chlorobiphenyl (MCB), IUPAC # 3, 2,2',5,5'-tetrachlorobiphenyl (TCB), IUPAC # 52 and 2,2',4,4',5,5'-hexachlorobiphenyl (HCB) IUPAC # 153. The biomasses and growth rates of the microorganisms as well as the sedimentation rate of particulate organic material increased with nutrient enrichment. The size distribution of the microorganisms changed with nutrient status, from dominance of picoplankton (< 2 mu m) in the control towards increased importance of micro (> 10 mu m) and nanoplankton (2-10 mu m) in nutrient enrichment. The specific growth rate of the bacterial community was found to be more temperature dependent than that of the phytoplankton community. The relative proportion of PCBs in the > 2 mu m fraction was observed to be in the order MCB < TCB < HCB, while the opposite distribution prevailed in the < 2 mu m fraction. We hypothesize that this is due to the combined effect of the different K-ow values of the PCBs and a different composition of the particulate organic carbon in the > 2 mu m and < 2 mu m fractions (e.g. different lipid composition). The residence time of the PCBs in the mesocosm generally decreased with nutrient enrichment, but was dependent on the degree of chlorination of the PCB. Our results indicate that the transport of organic pollutants up through the food web is more important in nutrient poor than in nutrient rich waters and that the importance of sedimentation is higher in eutrophic ecosystems.
Lipids in seawater act as solvents and transporters of lipophilic organic pollutants. To investigate a possible transport route of lipophilic pollutants, the vertical flux of lipid was quantified during an annual cycle in the northern Baltic Sea. The lipid content in both sedimenting material and different size fractions of seawater was analyzed. During the year, 8 g lipid m-2 sedimented out from the photic zone to the benthic system. The sedimentation of lipid accounted for 300 to 400 % of the average standing stock of pelagic lipid and was concentrated in the spring bloom period (April-June) when 70 % of the total lipid sedimentation occurred. About 30 % of the produced pelagic lipid settled out from the system. In seawater the lipid maximum occurred at the end of the spring bloom, shortly after nutrient depletion, indicating a stress response in the algae. Since lipid sedimentation is concentrated in the spring bloom, removal of lipophilic organic pollutants may be important during this period.
Background We have developed genomic tools to allow the genus Populus (aspens and cottonwoods) to be exploited as a full-featured model for investigating fundamental aspects of tree biology. We have undertaken large-scale expressed sequence tag (EST) sequencing programs and created Populus microarrays with significant gene coverage. One of the important aspects of plant biology that cannot be studied in annual plants is the gene activity involved in the induction of autumn leaf senescence. Results On the basis of 36,354 Populus ESTs, obtained from seven cDNA libraries, we have created a DNA microarray consisting of 13,490 clones, spotted in duplicate. Of these clones, 12,376 (92%) were confirmed by resequencing and all sequences were annotated and functionally classified. Here we have used the microarray to study transcript abundance in leaves of a free-growing aspen tree (Populus tremula) in northern Sweden during natural autumn senescence. Of the 13,490 spotted clones, 3,792 represented genes with significant expression in all leaf samples from the seven studied dates. Conclusions We observed a major shift in gene expression, coinciding with massive chlorophyll degradation, that reflected a shift from photosynthetic competence to energy generation by mitochondrial respiration, oxidation of fatty acids and nutrient mobilization. Autumn senescence had much in common with senescence in annual plants; for example many proteases were induced. We also found evidence for increased transcriptional activity before the appearance of visible signs of senescence, presumably preparing the leaf for degradation of its components.
In photosynthesis, sunlight is converted into chemical energy that is stored mainly as carbohydrates and supplies basically all life on Earth with energy.
In order to efficiently absorb the light energy, plants have developed the outer light harvesting antenna, which is composed of ten different protein subunits (LHC) that bind chlorophyll a and b as well as different carotenoids. In addition to the light harvesting function, the antenna has the capacity to dissipate excess energy as heat (feedback de-excitation or qE), which is crucial to avoid oxidative damage under conditions of high excitation pressure. Another regulatory function in the antenna is the state transitions in which the distribution of the trimeric LHC II between photosystem I (PS I) and II is controlled. The same ten antenna proteins are conserved in all higher plants and based on evolutionary arguments this has led to the suggestion that each protein has a specific function.
I have investigated the functions of individual antenna proteins of PS II (Lhcb proteins) by antisense inhibition in the model plant Arabidopsis thaliana. Four antisense lines were obtained, in which the target proteins were reduced, in some cases beyond detection level, in other cases small amounts remained.
The results show that CP29 has a unique function as organising the antenna. CP26 can form trimers that substitute for Lhcb1 and Lhcb2 in the antenna structure, but the trimers that accumulate as a response to the lack of Lhcb1 and Lhcb2 cannot take over the LHC II function in state transitions. It has been argued that LHC II is essential for grana stacking, but antisense plants without Lhcb1 and Lhcb2 do form grana. Furthermore, LHC II is necessary to maintain growth rates in very low light.
Numerous biochemical evidences have suggested that CP29 and/or CP26 were crucial for feedback de-excitation. Analysis of two antisense lines each lacking one of these proteins clearly shows that there is no direct involvement of either CP29 or CP26 in this process. Investigation of the other antisense lines shows that no Lhcb protein is indispensable for qE. A model for feedback de-excitation is presented in which PsbS plays a major role.
The positions of the minor antenna proteins in the PS II supercomplex were established by comparisons of transmission electron micrographs of supercomplexes from the wild type and antisense plants.
A fitness experiment was conducted where the antisense plants were grown in the field and seed production was used to estimate the fitness of the different genotypes. Based on the results from this experiment it is concluded that each Lhcb protein is important, because all antisense lines show reduced fitness in the field.
The specific roles of the chlorophyll a/b binding proteins CP29 and CP26 in light harvesting and energy dissipation within the photosynthetic apparatus have been investigated. Arabidopsis was transformed with antisense constructs against the genes encoding the CP29 or CP26 apoprotein, which gave rise to several transgenic lines with remarkably low amounts of the antisense target proteins. The decrease in the level of CP24 protein in the CP29 antisense lines indicates a physical interaction between these complexes. Analysis of chlorophyll fluorescence showed that removal of the proteins affected photosystem II function, probably as a result of changes in the organization of the light-harvesting antenna. However, whole plant measurements showed that overall photosynthetic rates were similar to those in the wild type. Both antisense lines were capable of the qE type of nonphotochemical fluorescence quenching, although there were minor changes in the capacity for quenching and in its induction kinetics. High-light-induced violaxanthin deepoxidation to zeaxanthin was not affected, although the pool size of these pigments was decreased slightly. We conclude that CP29 and CP26 are unlikely to be sites for nonphotochemical quenching.
We have constructed Arabidopsis thaliana plants that are virtually devoid of the major light-harvesting complex, LHC II. This was accomplished by introducing the Lhcb2.1 coding region in the antisense orientation into the genome by Agrobacterium-mediated transformation. Lhcb1 and Lhcb2 were absent, while Lhcb3, a protein present in LHC II associated with photosystem (PS) II, was retained. Plants had a pale green appearance and showed reduced chlorophyll content and an elevated chlorophyll a/b ratio. The content of PS II reaction centres was unchanged on a leaf area basis, but there was evidence for increases in the relative levels of other light harvesting proteins, notably CP26, associated with PS II, and Lhca4, associated with PS I. Electron microscopy showed the presence of grana. Photosynthetic rates at saturating irradiance were the same in wild-type and antisense plants, but there was a 10-15% reduction in quantum yield that reflected the decrease in light absorption by the leaf. The antisense plants were not able to perform state transitions, and their capacity for non-photochemical quenching was reduced. There was no difference in growth between wild-type and antisense plants under controlled climate conditions, but the antisense plants performed worse compared to the wild type in the field, with decreases in seed production of up to 70%.
The effect of pH (from 4.8 to 9.8) on the production of pilosine and pilocarpine and on their partition between cell and medium was studied in two lineages (P and PP) of Pilocarpus microphyllus cell suspension cultures. Highest mass accumulation was observed at high pHs and both lineages produced pilocarpine while only lineage PP produced pilosine. Both alkaloids were released in the medium but higher accumulation occurred in the cells. The highest production of pilocarpine was at pH 8.8-9.8 in both cell lineages. Other imidazole alkaloids were also identified in both lineages. At all pHs tested, the pH in the media cultures tended to stabilize around 6 after 10-15 days of cultivation. NO(3) (-) and NH(4) (+) variation in the media might partially explain the pH stabilization.
In annual plants, flowering related genes were initially thought to have the only function to coordinate and execute reproductive development. In perennial species like poplar, however, it became clear that these genes seem to have acquired additional functions in the regulation of the yearly growth cycle. The two poplar orthologs of the Arabidopsis "florigen" FLOWERING LOCUS T (FT), PtFT1 and PtFT2, are associated with flowering as well as other aspects of phenology. After duplication they have diverged in expression pattern, and maybe also in function and their gene regulatory networks. In this study I have characterized the role of PtFT1 and PtFT2 during the yearly growth cyle, as well as the interaction between, FT-like genes, poplar homologs of the Arabidopsis flowering time repressor TFL1 and PHYTOCHROME A in the regulation of tree flowering. The preliminary data show new unexpected functional relations between these genes and new previously undescribed effects on plant growth and development.
The photosynthetic apparatus is composed of proteins encoded by genes from both the nucleus and the chloroplast. To ensure that the photosynthetic complexes are assembled stoichiometrically and to enable their rapid reorganization in response to a changing environment, the plastids emit signals that regulate nuclear gene expression to match the status of the plastids. One of the plastid signals, the chlorophyll intermediate Mg-ProtoporphyrinIX (Mg-ProtoIX) accumulates under stress conditions and acts as a negative regulator of photosynthetic gene expression. By taking advantage of the photoreactive property of tetrapyrroles, Mg-ProtoIX could be visualized in the cells using confocal laser scanning spectroscopy. Our results demonstrate that Mg-ProtoIX accumulated both in the chloroplast and in the cytosol during stress conditions. Thus, the signaling metabolite is exported from the chloroplast, transmitting the plastid signal to the cytosol. Our results from the Mg-ProtoIX over- and underaccumulating mutants copper response defect and genome uncoupled5, respectively, demonstrate that the expression of both nuclear- and plastid-encoded photosynthesis genes is regulated by the accumulation of Mg-ProtoIX. Thus, stress-induced accumulation of the signaling metabolite Mg-ProtoIX coordinates nuclear and plastidic photosynthetic gene expression.
Although the plant Phosphorylated Pathway of L-Ser Biosynthesis (PPSB) is essential for embryo and pollen development, and for root growth, its metabolic implications have not been fully investigated. A transcriptomics analysis of Arabidopsis (Arabidopsis thaliana) PPSB-deficient mutants at night, when PPSB activity is thought to be more important, suggested interaction with the sulfate assimilation process. Because sulfate assimilation occurs mainly in the light, we also investigated it in PPSB-deficient lines in the day. Key genes in the sulfate starvation response, such as the adenosine 5'phosphosulfate reductase genes, along with sulfate transporters, especially those involved in sulfate translocation in the plant, were induced in the PPSB-deficient lines. However, sulfate content was not reduced in these lines as compared with wild-type plants; besides the glutathione (GSH) steady-state levels in roots of PPSB-deficient lines were even higher than in wild type. This suggested that PPSB deficiency perturbs the sulfate assimilation process between tissues/organs. Alteration of thiol distribution in leaves from different developmental stages, and between aerial parts and roots in plants with reduced PPSB activity, provided evidence supporting this idea. Diminished PPSB activity caused an enhanced flux of S-35 into thiol biosynthesis, especially in roots. GSH turnover also accelerated in the PPSB-deficient lines, supporting the notion that not only biosynthesis, but also transport and allocation, of thiols were perturbed in the PPSB mutants. Our results suggest that PPSB is required for sulfide assimilation in specific heterotrophic tissues and that a lack of PPSB activity perturbs sulfur homeostasis between photosynthetic and nonphotosynthetic tissues.
The rate of meiotic recombination is one of the central factors determining genome-wide levels of linkage disequilibrium which has important consequences for the efficiency of natural selection and for the dissection of quantitative traits. Here we present a new, high-resolution linkage map for Populus tremula that we use to anchor approximately two thirds of the P. tremula draft genome assembly on to the expected 19 chromosomes, providing us with the first chromosome-scale assembly for P. tremula (Table 2). We then use this resource to estimate variation in recombination rates across the P. tremula genome and compare these results to recombination rates based on linkage disequilibrium in a large number of unrelated individuals. We also assess how variation in recombination rates is associated with a number of genomic features, such as gene density, repeat density and methylation levels. We find that recombination rates obtained from the two methods largely agree, although the LD-based method identifies a number of genomic regions with very high recombination rates that the map-based method fails to detect. Linkage map and LD-based estimates of recombination rates are positively correlated and show similar correlations with other genomic features, showing that both methods can accurately infer recombination rate variation across the genome. Recombination rates are positively correlated with gene density and negatively correlated with repeat density and methylation levels, suggesting that recombination is largely directed toward gene regions in P. tremula.
Chlamydomonas reinhardtii is a unicellular green alga that can survive at a wide range of inorganic carbon (Ci) concentrations by regulating the activity of a CO2-concentrating mechanism (CCM) as well as other cellular functions. Under CO2 limited conditions, C. reinhardtii cells display a wide range of adaptive responses including changes in photosynthetic electron transport, mitochondria localization in the cells, the structure of the pyrenoid starch sheath, and primary metabolism. In addition to these functional and structural changes, gene and protein expression are also affected. Several physiological aspects of the CO2 response mechanism have been studied in detail. However, the regulatory components (transcription factors and transcriptional regulators) involved in this process are not fully characterized. Here we report a comprehensive analysis of the C. reinhardtii nuclear proteome using liquid chromatography electrospray ionization spectrometry (LC-ESI-MS). The study aims to identify the proteins that govern adaptation to varying CO2 concentrations in Chlamydomonas. The nuclear proteome of C. reinhardtii cells grown in the air at high (5%) and low (0.04%) CO2 concentrations were analyzed. Using this approach, we identified 1378 proteins in total, including 90 putative transcription factors and 27 transcriptional regulators. Characterization of these new regulatory components could shed light on the molecular mechanisms underlying acclimation to CO2 stress.
Today, bioenergy is a practical and increasingly widely available option for heating that is being adopted by many industries and households that are looking to use more sustainable energy sources. Biomass‐derived fuels may be solids, liquids, or gases. They can be classified based on the processes used in their production, which include physical upgrading, microbiological processes, thermochemical processes, and chemical processes. The production of solid fuels from renewable resources has become more important due to the growing global demand for energy and environmental concerns. Various microbial processes can be exploited to utilize energy that has been stored in biomass by photosynthesis. Methanol, ethanol, and butanol are liquid biofuels that can be synthesized from biomass and used in both four‐stroke gasoline and diesel engines. These alcohols can be prepared from sugarcane, sugar beet, wheat, barley, corn, switch grass, agricultural residues, wood, and many other industrial wastes.
Background: Map based cloning in Arabidopsis thaliana can be a difficult and time-consuming process,specifically if the phenotype is subtle and scoring labour intensive. An alternative to map basedcloning would be to directly sequence the whole genome of a mutant to uncover the mutationresponsible for the phenotype.
Results: Here, we have re-sequenced the 120 Mb genome of a novel Arabidopsis clock mutant earlybird (ebi-1), using massively parallel sequencing by ligation. This process was further complicated by the fact that ebi-1 is in Wassilewskija (Ws-2), not the reference accession ofArabidopsis. The approach reveals evidence of DNA strand bias in the ethyl methanesulfonate(EMS) mutation process. We have demonstrated the utility of sequencing a backcrossed line andusing gene expression data to limit the number of SNP considered. Using new SNP informationwe have excluded a previously identified clock gene, PRR7. Finally, we have identified a SNPin the gene AtNFXL-2 as the likely cause of the ebi-1 phenotype and validated this bycharacterising a further allele.
Conclusion: In Arabidopsis, as in other organisms, the (EMS) mutation load can be high. Here wedescribe how sequencing a backcrossed line, using functional genomics and analysing new SNPinformation can be used to reduce the number EMS mutations for consideration. Moreover, theapproach we describe here does not require out-crossing and scoring F2 mapping populations, anapproach which can be compromised by background effects. The strategy has broad utility andwill be an extremely useful tool to identify causative SNP in other organisms.
The response of plant respiration (R) to temperature is an important component of the biosphere's response to climate change. At present, most global models assume that R increases exponentially with temperature and does not thermally acclimate. Although we now know that acclimation does occur, quantitative incorporation of acclimation into models has been lacking. Using a dataset for 19 species grown at four temperatures (7, 14, 21, and 28 °C), we have assessed whether sustained differences in growth temperature systematically alter the slope and/or intercepts of the generalized log–log plots of leaf R vs. leaf mass per unit leaf area (LMA) and vs. leaf nitrogen (N) concentration. The extent to which variations in growth temperature account for the scatter observed in log–log R–LMA–N scaling relationships was also assessed. We show that thermal history accounts for up to 20% of the scatter in scaling relationships used to predict R, with the impact of thermal history on R–LMA–N generalized scaling relationships being highly predictable. This finding enabled us to quantitatively incorporate acclimation of R into a coupled global climate–vegetation model. We show that accounting for acclimation of R has negligible impact on predicted annual rates of global R, net primary productivity (NPP) or future atmospheric CO2 concentrations. However, our analysis suggests that accounting for acclimation is important when considering carbon fluxes among thermally contrasting biomes (e.g. accounting for acclimation decreases predicted rates of R by up to 20% in high-temperature biomes). We conclude that acclimation of R needs to be accounted for when predicting potential responses of terrestrial carbon exchange to climatic change at a regional level.
When predicting the effects of climate change, global carbon circulation models that include a positive feedback effect of climate warming on the carbon cycle often assume that (1) plant respiration increases exponentially with temperature (with a constant Q10) and (2) that there is no acclimation of respiration to long-term changes in temperature. In this review, we show that these two assumptions are incorrect. While Q10 does not respond systematically to elevated atmospheric CO2 concentrations, other factors such as temperature, light, and water availability all have the potential to influence the temperature sensitivity of respiratory CO2 efflux. Roots and leaves can also differ in their Q10 values, as can upper and lower canopy leaves. The consequences of such variable Q10 values need to be fully explored in carbon modelling. Here, we consider the extent of variability in the degree of thermal acclimation of respiration, and discuss in detail the biochemical mechanisms underpinning this variability; the response of respiration to long-term changes in temperature is highly dependent on the effect of temperature on plant development, and on interactive effects of temperature and other abiotic factors (e.g. irradiance, drought and nutrient availability). Rather than acclimating to the daily mean temperature, recent studies suggest that other components of the daily temperature regime can be important (e.g. daily minimum and / or night temperature). In some cases, acclimation may simply reflect a passive response to changes in respiratory substrate availability, whereas in others acclimation may be critical in helping plants grow and survive at contrasting temperatures. We also consider the impact of acclimation on the balance between respiration and photosynthesis; although environmental factors such as water availability can alter the balance between these two processes, the available data suggests that temperature-mediated differences in dark leaf respiration are closely linked to concomitant differences in leaf photosynthesis. We conclude by highlighting the need for a greater process-based understanding of thermal acclimation of respiration if we are to successfully predict future ecosystem CO2 fluxes and potential feedbacks on atmospheric CO2 concentrations.
* The arbuscular mycorrhizal (AM) symbiosis is ubiquitous, and the fungus represents a major pathway for carbon movement in the soil-plant system. Here, we investigated the impacts of AM colonization of Plantago lanceolata and temperature on the regulation of root respiration (R). * Warm-grown AM plants exhibited higher rates of R than did nonAM plants, irrespective of root mass. AM plants exhibited higher maximal rates of R (R(max)-R measured in the presence of an uncoupler and exogenous substrate) and greater proportional use of R(max) as a result of increased energy demand and/or substrate supply. The higher R values exhibited by AM plants were not associated with higher maximal rates of cytochrome c oxidase (COX) or protein abundance of either the COX or the alternative oxidase. * Arbuscular mycorrhizal colonization had no effect on the short-term temperature dependence (Q(10)) of R. Cold-acclimated nonAM plants exhibited higher rates of R than their warm-grown nonAM counterparts. By contrast, chilling had a negligible effect on R of AM-plants. Thus, AM plants exhibited less cold acclimation than their nonAM counterparts. * Overall, these results highlight the way in which AM colonization alters the underlying components of respiratory metabolism and the response of root R to sustained changes in growth temperature.
1. Scaling relationships linking photosynthesis (A) to leaf traits are important for predicting vegetation patterns and plant-atmosphere carbon fluxes. Here, we investigated the impact of growth temperature on such scaling relationships.
2. We assessed whether changes in growth temperature systematically altered the slope and/or intercepts of log-log plots of A vs leaf mass per unit leaf area (LMA), nitrogen and phosphorus concentrations for 19 contrasting plant species grown hydroponically at four temperatures (7, 14, 21 and 28 degrees C) in controlled environment cabinets. Responses of 21 degrees C-grown pre-existing (PE) leaves experiencing a 10 day growth temperature (7, 14, 21 and 28 degrees C) treatment, and newly-developed (ND) leaves formed at each of the four new growth temperatures, were quantified. Irrespective of the growth temperature treatment, rates of light-saturated photosynthesis (A) were measured at 21 degrees C.
3. Changes in growth temperature altered the scaling between A and leaf traits in pre-existing (PE) leaves, with thermal history accounting for up to 17% and 31% of the variation on a mass and area basis, respectively. However, growth temperature played almost no role in accounting for scatter when comparisons were made of newly-developed (ND) leaves that form at each growth temperature.
4. Photosynthetic nitrogen and phosphorus use efficiency (PNUE and PPUE, respectively) decreased with increasing LMA. No systematic differences in temperature-mediated reductions in PNUE or PPUE of PE leaves were found among species.
5. Overall, these results highlight the importance of leaf development in determining the effects of sustained changes in growth temperature on scaling relationships linking photosynthesis to other leaf traits.
Climate is changing and it is certain that this change is due to human activities. Atmospheric greenhouse gases have been rising in an unprecedented way during the last two centuries, although the land biosphere has dampened their increase by absorbing CO2 emitted by anthropogenic activities. However, it is unclear if this will continue in the future. This uncertainty makes it difficult to predict future climate changes and to determine how much greenhouse gas emissions must be reduced to protect climate.
To understand the future role of plants in limiting the atmospheric CO2 level, the effect of increasing CO2 on plant photosynthesis and productivity has been studied. However, studies on trees showed contradictory results, which depended on the duration of the experiment. This revealed that an initial strong CO2 fertilization may be a transient response that disappears after a few years. Because climate changes over centuries, we must explore the response of vegetation to increasing CO2 on this time scale. Studying tree rings is a good alternative to impractical decade-long experiments, because trees have experienced the CO2 increase during the last 200 years and may already have responded to it.
This thesis shows that the intramolecular distribution of the stable hydrogen isotope deuterium (deuterium isotopomer distribution, DID) of tree rings is a reliable tool to study long-term plant-climate adaptations. The premise for this is that the deuterium abundance in tree rings depends on environmental as well as physiological factors. Using newly developed methodology for DID measurements, the influences of both factors can be separated. Applied to tree rings, separating both factors opens a strategy for simultaneous reconstruction of climate and of physiological responses.
The results presented show that DIDs are influenced by kinetic isotope effects of enzymes, allowing studies of metabolic regulation. We show that the abundances of specific D isotopomers in tree-ring cellulose indeed allow identifying environmental and physiological factors. For example, the D2 isotopomer is mostly influenced by environment, its abundance should allow better reconstruction of past temperature. On the other hand, the abundance ratio of two isotopomers (D6R and D6S) depends on atmospheric CO2, and might serve as a measure of the efficiency of photosynthesis (ratio of photorespiration to assimilation). The presence of this dependence in all species tested and in tree-ring cellulose allows studying adaptations of plants to increasing CO2 on long time scales, using tree-ring series or other remnant plant material.
he deuterium (D) abundance of tree-ring cellulose archives past climatic conditions, but previous attempts to access this archive have led to conflicting results. Based on an overview of D fractionation mechanisms in plants, we explain why past measurements of D abundance yield unreliable paleo signals. Our data show that variation in D abundance among the C–H groups (isotopomer variation) of tree-ring cellulose is generally greater than variation in D abundance due to climatic influences. A comparison of the D isotopomer abundances of soluble sugars of annual plants and of trees, and of tree-ring cellulose shows that an "isotopomer pattern" of the C3 photosynthetic pathway is transmitted from soluble sugars to tree-ring cellulose. Differences in this pattern between oaks and conifers appear to be related to differences in metabolism. Furthermore, the patterns are modified by hydrogen isotope exchange between C–H groups and source water during cellulose synthesis. Based on these results, we propose a strategy to simultaneously reconstruct climate signals and signals related to tree physiology from D isotopomers of tree rings. Combination of climate signals and physiological signals may allow the detection of century-time-scale adaptations of trees to past environmental change, and help to forecast future adaptations.
• The abundance of the hydrogen isotope deuterium (D) in tree rings is an attractive record of climate; however, use of this record has proved difficult so far, presumably because climatic and physiological influences on D abundance are difficult to distinguish.
• Using D labelling, we created a D gradient in trees. Leaf soluble sugars of relatively low D abundance entered cellulose synthesis in stems containing strongly D-labelled water. We used nuclear magnetic resonance (NMR) spectroscopy to quantify D in the C-H groups of leaf glucose and of tree-ring cellulose.
• Ratios of D abundances of individual C-H groups of leaf glucose depended only weakly on leaf D labelling, indicating that the D abundance pattern was determined by physiological influences. The D abundance pattern of tree-ring cellulose revealed C-H groups that exchanged strongly (C(2)-H) or weakly (C(6)-H2) with water during cellulose synthesis.
• We propose that strongly exchanging C-H groups of tree-ring cellulose adopt a climate signal stemming from the D abundance of source water. C-H groups that exchange weakly retain their D abundance established in leaf glucose, which reflects physiological influences. Combining both types of groups may allow simultaneous reconstruction of climate and physiology from tree rings.
DNA methylation plays important roles in many biological processes, such as silencing of transposable elements, imprinting, and regulating gene expression. Many studies of DNA methylation have shown its essential roles in angiosperms (flowering plants). However, few studies have examined the roles and patterns of DNA methylation in gymnosperms. Here, we present genome-wide high coverage single-base resolution methylation maps of Norway spruce (Picea abies) from both needles and somatic embryogenesis culture cells via whole genome bisulfite sequencing. On average, DNA methylation levels of CG and CHG of Norway spruce were higher than most other plants studied. CHH methylation was found at a relatively low level; however, at least one copy of most of the RNA-directed DNA methylation pathway genes was found in Norway spruce, and CHH methylation was correlated with levels of siRNAs. In comparison with needles, somatic embryogenesis culture cells that are used for clonally propagating spruce trees showed lower levels of CG and CHG methylation but higher level of CHH methylation, suggesting that like in other species, these culture cells show abnormal methylation patterns.
The molecular basis of short-day-induced growth cessation and dormancy in the meristems of perennial plants (e.g., forest trees growing in temperate and high-latitude regions) is poorly understood. Using global transcript profiling, we show distinct stage-specific alterations in auxin responsiveness of the transcriptome in the stem tissues during short-day-induced growth cessation and both the transition to and establishment of dormancy in the cambial meristem of hybrid aspen trees. This stage-specific modulation of auxin signaling appears to be controlled via distinct mechanisms. Whereas the induction of growth cessation in the cambium could involve induction of repressor auxin response factors (ARFs) and down-regulation of activator ARFs, dormancy is associated with perturbation of the activity of the SKP-Cullin-F-box(TIR) (SCF(TIR)) complex, leading to potential stabilization of repressor auxin (AUX)/indole-3-acetic acid (IAA) proteins. Although the role of hormones, such as abscisic acid (ABA) and gibberellic acid (GA), in growth cessation and dormancy is well established, our data now implicate auxin in this process. Importantly, in contrast to most developmental processes in which regulation by auxin involves changes in cellular auxin contents, day-length-regulated induction of cambial growth cessation and dormancy involves changes in auxin responses rather than auxin content.
An Arabidopsis mutant that exhibited reduced root length was isolated from a population of activation-tagged T-DNA insertion lines in a screen for aberrant root growth. This mutant also exhibited reduced hypocotyl length as well as a delay in greening and altered leaf shape. Molecular genetic analysis of the mutant indicated a single T-DNA insertion in the gene RpoT;2 encoding a homolog of the phage-type RNA polymerase (RNAP), that is targeted to both mitochondria and plastids. A second T-DNA-tagged allele also showed a similar phenotype. The mutation in RpoT;2 affected the light-induced accumulation of several plastid mRNAs and proteins and resulted in a lower photosynthetic efficiency. In contrast to the alterations in the plastid gene expression, no major effect of the rpoT;2 mutation on the accumulation of examined mitochondrial gene transcripts and proteins was observed. The rpoT;2 mutant exhibited tissue-specific alterations in the transcript levels of two other organelle-directed nuclear-encoded RNAPs, RpoT;1 and RpoT;3. This suggests the existence of cross-talk between the regulatory pathways of the three RNAPs through organelle to nucleus communication. These data provide an important information on a role of RpoT;2 in plastid gene expression and early plant development.


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Important! This job ad is no longer active
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Instructors & Trainers, Flight Instructor
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BAA Training is looking for a Visual Flight Instructor (Spain)
We are BAA Training, one of the TOP 3 largest independent aviation training centers in Europe. Our people are united by passion for aviation and a desire to ensure efficient training solutions for both pilots and airlines. A comprehensive list of EASA approved training programs and extensive experience on the market allow us to meet the training needs of the global aviation.
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Graduation exercises, May 17, 2013, 7 p.m., Gordo High School Football Stadium
Ken Holder
Baccalaureate ceremonies
May 12, 2013, 6 p.m.
Gordo Elementary Auditorium
Dr. Mark Searby
Graduation exercises
May 17, 2013, 7 p.m.
Gordo High School Football Stadium
Valedictorian and salutatorian
Victoria Mika Eatman, daughter of Christie Eatman and Stan Eatman. She has a 4.037 GPA.
Kongzong Elijah Yang, son of Mr. and Mrs. Long Yang. He has a 3.926 GPA.
Class officers
Caroline Turner, president
Macy Hoggle, vice president
Jessalyn Wallace, secretary
School color
Green and gold
Class motto
"Our lives are before us… Our pasts are behind us… But our memories are forever with us."
Candidates for graduation include Anna Leigh Ashmore, Tyesha Marie Bonner, Tyree Marcus Bonner, Jonah Elisha Bowers, Andrew Taylor Brown, William Blake Burkhalter, Ashley Nicole Burton, Irlanda Vanessa Cano Rojo, Montanna Faye Cargile, Lance Keith Carpenter, Kenny Wayne Cobb, Aaron Devontae Cousette, Jarron Deontae Cousette, Shaquille Jamar Cousette, Steven West Cowart, Derrian Tyrie Davidson, La'Tatayana Marcel Doughty, Victoria Mika Eatman, Bassam Abdullha Elnaham, Timothy Ryan Fulgham, Chancellor Shervonte Glass, Jonna McCall Gray, James Dylan Gregg, Taylor Lynn Gregory, Christopher Dewayne Guyton, Natazhia Lashe' Handy, Rachel Cheyenne Hodge, Kaylee Nicole Hodgkins, Macy Jewel Hoggle, Ahujia Shantea' Howard, Erick Lee Hubbard, Marcus Deshun Jones, Demario Devonte Kennedy, Peyton Elizabeth Kent, Lance Phillip Koon, Iesha Shambre Latham, Sequoia Lashea LeGrone, Sandi Diane Lockhart, Christopher Purnell Logan, Jamarcus Keon Martin, J. Elias Martinez, Zachary Ryan Matyjasik, Sarah Diane McDaniel, Cody Lee Wayne McElroy, James Tyler Miner, Brandon Leon Mullenix, Cami Destin Nelson, Aliia Sherie Nicholson, Anna Gayle Norris, Alyssia RaShaud Nunn, Deanthony Rashon Pruitt, Kantreal Je'Nae Rice, Alexisua La'Paris Richardson, Tiffany Marie Scott, Lauren Celeste Shirley, Matthew Kyle Shirley, Robert Lewis Hunter Simpson, Lateeia Shambrell Spain, JaVaris WilShondria Spencer, T'Quann D'Miles-Ameer Stewart, Malcolm Demarest Storey, Samantha Alaine Sutton, Colton Renay Tant, Devin Wade Tilley, Caroline Lee Turner, Jessalyn Shay Wallace, Da'Michael Cortez White, De'Angelio Montreal White, Grace Nicole Wilder, Nayla Renee Windham, Kentrell De'Vonte Wright, Kongzong Elijah Yang.


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Consciousness of Christ: the Why of Everything
(Note: This article is wrapping up pretty much everything there is to say from my perspective. Yes, it's long, the first part maybe a bit technical, but the unsearchable riches of Christ, I believe, are here expressed! It was a feast for me to write it and may it be a sumptous feast for the hungry reader. More than one has told me it has been that for them.)
By Fred Pruitt
Someone recently wrote and asked me two questions: 1) By whose power did Adam and Eve decide to listen to Satan in the garden? and 2) Also in your book "Hearts of Flesh" you compare Abraham and Saul in that they were both doing God's work regardless of what it looked like – both were men of God. In other words, some Christians can look like a Saul and keep going down a road towards destruction and still be a Christian. Did I understand you correctly?
Here is the reply:
(This is a little different from what I usually write, and may not appeal to all readers. When I have spoken of these things like this in the past some have commented that I am too complicated. I apologize if some of this comes across that way. I believe the reader who perseveres all the way through will be satisfied — at least I hope so.)
You have an interesting way of phrasing your first question — "by whose power did Adam and Eve decide to listen to Satan in the garden? I don't think I've ever heard anyone phrase it that way before — "by whose power."
So that's an easy answer, since there is no power except the power which is God. Satan is a misuse of the One Power, using in a sense the substance/being/existence of God Himself to create for himself in singularity, or dense concentration of self, or what I might call a vortex of sucking self, like a black hole — a dark fire that has almost infinite heat but which emits no light or warmth, that exists only to feed itself by the consumption of everything that comes within its reach — to use it all to feed and build up himself by the death and destruction of all.
God is also Consuming Fire, but only consuming in the sense that everything which He attracts into Himself for consumption is re-emitted as Light, Heat, and Life (Luke 20:18). This is because God has eternally determined to be Love, exhibited eternally by the Son in the form of a Lamb Slain forever in the center of the Eternal Godhead and Throne. Satan seeks to bring only death and is anti-life, whereas God's principle includes death, first of all in a sense in Himself when in His out-going to express Himself in the Son, He eternally dies to or says "NO" to that sucking self-for-self which seeks to live by feeding on the death of others so that it is supreme and without competition. Then by that death (Cross) or "NO" to self-seeking, He rises as the Son, whose eternal characteristic is to give His Life as a ransom and means of Life to others. Satan's principle is death only; God in Trinity's Principle is death AND resurrection, both death to self-seeking (by the Cross), and resurrection in the Deity out of "NO" to "YES," by which Giving-Life is out-splayed with infinite energy and infinite power to inseminate the Universe with Life unto Life. The motion of the resurrection out of death (from the lowest depths to the highest heights) is what fuels the dynamo that empowers everything that is. It is like an atomic explosion without destruction, but instead an infinitely energetic perpetual effluence of light and life in every direction.
So I would think the question, then, isn't really "whose" power it is, but where does their freedom come from that allows them to choose or follow the wrong path, or to disobey?
And the answer there is again the same. The Bible mentions liberty and freedom often, especially in the New Testament. God is Free and IS Freedom, so therefore freedom, like all things, originates in God and is first of all His principle. Freedom is having the option to choose between alternatives. We may have a limited set of alternatives, but freedom is the divine principle of personhood — that which makes a person a person. A tree cannot choose anything for itself. Even a dog cannot do this. Because the type of choosing we are speaking of is the choice of what kind of person I'm going to be, and who I will be hooked up with.
What I say "Yes" to determines what I say "No" to, and visa versa. That "choice," as I explained above regarding the Deity, originated in God the Father in the eternal, so that eternally He has chosen or determined to be nothing but Love — Life which does not seek its own good primarily (a "NO" to self-seeking), but preeminently the good of another (a "YES" to other-love). Satan made the eternal choice to be for himself, which is also an eternal trap because he became a caricature of True Self, by rejecting union with the Light and death to self-seeking, to live in what is really an unreality, a false fantasy, a self-delusion of himself as just himself, all by himself alone, an island of self, a closed-up fortress, having his own power, his own beauty, his own goodness, out of his own fire. Since like all sentient beings God created, he only exists to be a re-expression in limited or particular form of the Unlimited Universal Deity, but out of the same freedom by which God chose to be love, Lucifer/Satan chose to be only himself and reject Another to be his life, and thereby "fell."
So that is the same freedom, and the same power, and the same set of alternatives that confronted Adam and Eve and us all in our infancy and lack of consciousness. With this fundamental and important difference. The temptation of Adam and Eve was in the temporal, given them (with God of course knowing the outcome and MEANING IT) for their ultimate redemption and subsequent consciousness of their identity in God.
Now I haven't mentioned consciousness at this point, but God's desire to be the Son is not limited to the Eternal Trinity which is an infinite circumincession within Themselves of love loving by going out of itself to extend and propagate love. Love in not seeking itself, must instead propagate itself, expand itself, go out of itself. In other words, for love to love, there must be something for love to love and expand into. Therefore the Son must not be just the One Eternal Son, but that the Son might be reproduced and expanded into a family, the whole family being individual unique expressions of that One Son, Who is both the Eternal Object of the Father's Love, and IS the Eternal Father's Love.
Now this is what has created the tremendous tension in the universe, which Paul alludes to in Romans 8:18-22, i.e. the whole universe groaning and travailing, awaiting the final manifestation of the Sons of God. And this is about consciousness — because for the Sons to be finally and fully manifest, they must know themselves as the Son in the sons, after having freely desired, sought and received the Sonship, raised by the Father/Son/Spirit into the full understanding and comprehension of the heights, depths, and fullness of Christ — in them! Unto a perfect MAN, as Ephesians says, raised into the Head, which is Christ. MAN raised fully into CHRIST!
And that is WHY there was a fall and WHY there was the giving of the Law and WHY Israel, as God's lamb for the whole world, had to rise and fall, rise and fall, and WHY there had to be a Saul followed by a David, and WHY, ultimately, the Son of God had to come to be the Son of Man, and WHY there had to be a Calvary in space and time as there is an Eternal Lamb Slain in the heart of eternity, and WHY there had to be the time of the Gentiles and WHY, even though the promises have been given and fulfilled there HAD to be a time when it continues to look like He has forsaken His People and has delayed His Coming, and WHY even though we are even now recipients of the Promises we still, for this time, wander in sheepskins and goatskins and find in this world no continuing city. It is all part of this ultimate tension which will explode into that final manifestation of God as His Fully Formed and Conscious Sons to the glory and unbounded benefit of the entire universal creation!
There is coming a time, and it now is, when God is beginning to peel back the layers, to give understanding, to reveal the mysteries of the kingdom. Jesus said He could not speak plainly the mysteries of the kingdom of heaven, but must speak them in dark sayings and parables, in order to hide them from the "wise and prudent" and only reveal them unto "babes." All the stories of the Old Testament have this quality to them. While infantile Christians are arguing with "the world" and trying to convince those whose natural minds cannot see the things of God, that these things are literal histories or descriptions of the creation scientifically, in their zeal they are straining at a gnat and swallowing a camel. They miss the tremendous riches of Christ, redemption, and the New Birth revealed in the stories — which are intended not just for information's sake, knowledge of the sake of knowledge, but for the purpose of CONSCIOUSNESS OF CHRIST, because as we see Him we know Him and these stories are all about HIS LIFE IN US.
Rather than focusing on Saul's behavior and whether he still attained heaven because of the way he turned out, as we all have at one time or another, the far greater riches are found in seeing Saul and David as counterparts of essentially the same man. And it is ALL ABOUT coming to consciousness. Because we are all like Jesus — we learn obedience through the things we suffer (Heb 5:8), and we come to maturity (raised consciousness) by the exercise of the senses discerning good and evil (Heb 5:14). Not good and evil in the temporal "be good to be accepted" sense that the world knows, but good and evil in the ultimate sense, i.e., that which is God and that which is anti-God, and that the "knowing" of that is really coming to oneness with the truth — as Adam "knew" Eve, became one flesh with her, so we "know" Christ in truth, and are One Spirit with Him. I've spoken much about what being "one" means elsewhere, so won't repeat that here.
There is a recurring theme in the Old Testament, which Jesus just summed up in, "The first shall be last, the last shall be first." You can see it beginning to be successively plainer in the Genesis stories; in Cain and Abel, Ishmael and Isaac, in Jacob and Esau, in Joseph and his brethren, in the generation of Israel born in Egypt which perished in the wilderness, and the generation born in the wilderness which went across the Jordan into the Promise, and so on. It is picked up again in Saul and David.
I can only briefly speak of it, because it is deep mystery but the Lord is unfolding it more and more day by day. This has been percolating in me for several years and I've tried to write and speak about it, but it is still fresh and unformed. So be patient with me.
First a bit about types and shadows. If we realize that the Scriptures were put together by the Holy Spirit for the revelation of Christ, and not thematically by men who were following an outline or plan, then we can also realize that they are not systematic in unfolding these mysteries. The mysteries of God are revealed by the revelation of the Spirit, not a systematic theology which always holds that a certain type remains the same type throughout. For instance, Esau in one sense by the Spirit represents fallen Adam whom God the Father (Old Isaac) would love to bless but cannot because he has spurned his birthright by selling it for a mess of pottage, as Adam traded his divine birthright (rights of the firstborn who gets the lion's share of the father's heritage) for an animal skin, and in other cases Esau represents the flesh which lusts against the things of the Spirit and therefore must be spurned. So the types change even within the life of a the same person. Sometimes Pharaoh represents God the Father, as in the story of Joseph, and sometimes he represents the devil who has stolen man and enslaved him, as the Pharaoh of Moses' time. Only the Spirit can reveal these things.
So then we come to Saul. You begin to see what is going on in the story when the people of Israel tell Samuel that they want a king, just like all the other nations around them have a king. In a way it is like Adam and Eve all over again, because they were under, at the time, the direct kingship of God, through the prophet and judge, Samuel. The prior years they had been the same — no king, but a series of judges and rescuers in their ongoing battles with the Philistines and their on-again/off-again relationship with the Baals and idolatries. That might not seem like such a hunky-dory situation, but when they seek a king "like the other nations," God's reaction is pretty negative. He says they are rejecting him as King by wanting a king they can see, feel, touch, etc.
Remember, keep our eyes on the prize — the goal here, for God and Israel (which God knows but Israel does not), is consciousness. Sons who know who they are by the exercise of their senses. In other words, they must find out by experience; they must know the wrong way in order to know the right way. And even though they were not very steadfast in their dealings with God under the judges — they were always falling away and worshipping other gods, etc. — still in essence they were under the kingship of God, Who they could not see — they had to believe and follow Him in His invisibility, and in their dim and infantile consciousness, it was necessary that they reject this invisibility by which God kept them in their unconsciousness, and find a visible palpable king that would protect them from their enemies and make them great. All of this to the benefit of us all. Their lack of consciousness leads to our consciousness when we see.
So what's the first thing God does? He tells them what's what, but He honors their request. He gives them the kind of king that they, in their immaturity, think they want. He gives them a great-looking guy, who stands a head taller than everybody else. A man impressive to the flesh, and intimidating to their enemies. Someone they can put their hopes and dreams into, who will take care of all their needs, protect the land, and give them peaceful lives in security and prosperity. (Something which mankind is still doing, apparently!)
When we consider this spiritually in our own lives, when we come to God and do not yet know who we are, thinking it's about being good and obeying rules, etc., this "Saul" is who we want to reign over us. It is ourselves, our selves that we have always known ourselves to be, helped and improved by God and made into a fit and proper king. Impressive to our friends for our spirituality, and hopefully intimidating to our enemy, the devil. "Make me, Lord, be this and that. Here is this person that I am, Lord. O make me better, give me more love, more patience, more kindness." We want to stand up and be the one who is a head taller than everyone else. In our unconsciousness and immaturity, still having only a consciousness of flesh or independent self, we cannot help but think and pray this way. We do not know that we are praying for the kingship of a false hope, a wisp of self that is a persona the devil has created. Oh, our real self is in there somewhere, and there is the anointing of God, but God is a God who grants our desires, even those which are wrong sometimes for us, in order to bring us to the fullness of Himself by our failure. And that self, like Saul, must fail.
Because that self thinks that it is where the holiness resides. It is a presumptuous self, a self that takes upon itself the right to sacrifice, thinking, wrongly, that itself is the life of God, in control, the decider.
Now let's step back a moment and consider Saul. He is first of all, as above, "goodlier" than any other in Israel. He looks the part, and at first is humbled by the fact he has been chosen of God. And God puts His Spirit upon him, and anoints him king. Now, why is this so, if God already knew Saul was not going to work out, which of course He did know?
This is where this purpose of God to bring us into consciousness of our sonship begins to be revealed. Saul, in ourselves, as I said above, is the false image of ourselves that we are born with, that we know as ourselves all our lives even into our new birth into Christ, and God honors it, anoints it, blesses it, leads it, thus granting our desires in order to finally bring that self to its own self-destruction, as Saul at Gilboa fell on his sword before he was overrun by the Philistines. The first generation out of Egypt into the wilderness is the same. They are given the law and they all agree to keep the law, which of course they cannot do. They are presumptuous, rebelling against Moses who they think is no better than they. They complain. Everytime they are challenged they revert to their former selves they have always been, but are rescued by God anyway. Even after they refuse to go into the Promised Land the first time, afraid of the giants and great walled cities thereby receiving God's curse on their generation, by which they must all perish in the wilderness and never see the Promise, STILL God leads them by His Spirit in the form of a cloudy pillar by day and a fiery pillar by night. God honors them, cares for them, leads them by His Spirit, feeds them bread from heaven, gives them Christ in the form of water out of a rock, yet they cannot come to the Promise. They were born in Egypt and are perpetually in consciousness only flesh, so that they cannot touch the mountain of God.
And Saul is this same first generation, ourselves in false independence but which we in our wrong consciousness think is what and who God blesses and uses. And for a time God goes along with that. He blesses this false self, loves it, anoints it, even while it presumes upon itself its own deity. Not consciously in the sense that it believes it is God, but unconsciously in a de facto sense, because its lifelong independence is something it cannot part with, it MUST remain the chooser, the doer, the decider, thereby making itself as God. The false self created by reaching out and taking the fruit cannot change. It can only die.
Then David enters the story. Now David is the type of the new man, Christ in us, who first sprouts as a very small seed within us. At first Saul is delighted with David. David will serve him, will do his bidding, will kill his enemies. He is glad to have such a champion. But things change as Saul begins to see the handwriting on the wall. He senses David is going to replace him as king, and he cannot accept this. It is his own posterity he protects and lifts up, and, forgetting that he is a servant of the Lord God, he begins persecuting David who clearly is God's man. His favor toward David turns to anger as his visible presumption of taking the things of God to himself causes him to lose the favor of God, which is transferred to David. Even though David comforts him with his songs when he is overtaken by his melancholia, which happens more and more often (this is the beginning of Romans 7 struggle), still he begins to hate David more and more. And so the struggle begins, with Saul some days loving David and some days trying to kill him. When he is confronted with his ill treatment of David and with David's continuing loyalty to him, even after David has been anointed to be the next king, Saul repents over and over, but he cannot change. He "rededicates," but, he still wants to be king. He still wants the people to follow him, to love him, to recognize that HE is their leader and is greater than David. This is a perfect picture of the wrong self that we have been deceived into believing all our lives IS our self. It is gearing up to know and finally accept its own death, but this struggle between Saul and David, between the mind of the flesh and the mind of the Spirit, is what finally brings him to that final self-destruction on Mt. Gilboa. Because finally it cannot do what it is supposed to do — defeat the enemies of the Lord. They are finally too much for Saul, and overrun him, so that he becomes the "O wretched man," and falls on his own sword. He takes the death that must be, "I am crucified with Christ," so that "it is no longer I, but Christ" can then take the throne.
Looking at it from David's perspective, this true man, this man who we really are in Christ, which is Christ in us as one with us, as I said above, starts in humble beginnings. He is the man of the Spirit, who we hardly know at first. He is ourselves, but our unified self in Christ, and because we are still carnal in our minds he is as if he is separate from us. At first we delight in him, but like Saul, we become suspicious of him. He is too free. He is dangerous. We are afraid to let him live.
David, unlike Saul, takes nothing to himself or for himself. Though he is continually tempted to reach out and take the kingdom from Saul, he knows he cannot touch Saul, for he is still God's beloved, still the "anointed of the Lord," and he cannot reach out and take for himself. Even though he knows that the kingdom is his, still he must receive it as a gift, and not as something he presumptuously takes. He becomes Saul's secret helper, in that he protects the borders of Israel from marauders, from the Philistines. Little by little the people begin to come to him, though he finds himself hiding in caves and in the desert. Many of his followers urge him to kill Saul and take the kingdom, and though he easily could do this, David continually refuses, knowing that if the kingdom is to be his, and he does know it, it will be given as a promise and a gift of grace, and that he won't gain it by his own hand.
That is akin to Abraham's story with Ishmael and Isaac. God gave the promise which was to be fulfilled in Sarah, but Abraham and Sarah decided to hurry things up a bit and bring a son by Hagar. Like Saul, God does bless Ishmael and has a plan for him, but he is not the son of Promise, but of the presumption of the flesh. All this is for our understanding and consciousness of this same point: the first birth, Adam, Cain, Ishmael, Esau, Reuben, the generation born in Egypt, Saul, etc., all point at the man of the flesh who is deceived into thinking he is the ruler and should reign on earth, while the second birth, Christ (2nd Adam), Abel, Isaac, Jacob, Joseph, the generation born in the wilderness, David, etc., all point at the new man, Christ in us as one with us, who as the Stone Cut out without Hands, smashes the image of iron and clay — our false selves in independence, so that that Stone fills the whole earth (Dan 2:34,35), and we finally recognize Christ in us as All in all, and therefore, as us.
This is the mystery of Christ revealed in simple stories by the Holy Spirit, that we might know that He does the same in us, to accomplish the same, to move us in consciousness from Cain to Abel (Christ Who died) and Seth (Christ Who rose), from Ishmael to Isaac, from Esau to Jacob, from Saul to David, that we might come in our understanding into the fullness of Him who fills all in all, in and through ourselves. All this is the fulfillment of Galatians 2:20, told over and over and over throughout the scriptures, because it is really all there is.
Once Christ is formed in us in consciousness, then we stand up as David, as Moses, as Paul, as the fathers John spoke of, living out of the eternal water of life that we now recognize flows out of us, not by anything we have done, could do, should do, but by the grace of the One Who favors us, WHo is pleased with us, Who sees us as "the beloved Son, in whom I am well pleased … hear ye Him!"
It is the inner mystery of the whole Scripture. It's all about Christ in us and how he comes to fullness in us. It is not something we instigate, make happen, help along, but solely and purely grace in action by His Spirit Who has come to dwell in us, his human temples, making us fruitful branches on the Vine which is Christ.


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Now that Julia is back to school and on the bus very early, I've had a chance to go outside and experience the glorious autumn early morning light. I love this time of year when the sun starts getting at a lower angle in the sky. The beautiful natural colors start taking on that gorgeous autumn glow.
Here is one of the little sheds that I have surrounded with hostas. It is filled with some odds and ends. It was Julia's playhouse when she was little but now is a catchall. Every morning the light is just spectacular around it and the hosta blooms dance in the air. The photo doesn't do it justice.
This year, I planted morning glories on both sides of the door. They are growing haphazardly around the door along strings. Every few days I have to rein them into the string set up. I think I will do this every year. I love how the dark blue of the Grandpa Ott morning glory bounces off the yellow color.
Here is a jumble of color in the little garden outside the studio.
Today, I am announcing the big Fall Craftsy sale. All classes are on sale for up to 50% off. Have you been thinking about learning Crewel Embroidery this winter? Maybe you used to do it and want to start stitching again? Here's your chance. CLICK HERE TO RECEIVE THE DISCOUNT. My class is called Stitch It with Wool: Crewel Embroidery
Craftsy is a great organization for me to be affiliated with. Not only are their classes stellar but they give me the opportunity to earn some extra money by helping them promote their classes. I am very thankful for that opportunity because I have learned so much from the different classes I have watched. So know that by clicking my link, you are also helping me to support my family with a bit of extra revenue. And you get to learn something too.
Thanks everyone again for your kind words and encouragement on the blog.


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Tag Archives: Decorate
Christmas Trees
Written by Gene Glotzer on December 16, 2018 — Leave a Comment
Charles Dickens wrote a piece called "A Christmas Tree" as a magazine article. In it, he describes a gathering of people admiring a Christmas tree. Then he goes on to describe all the memories that get triggered by the sight. He remembers childhood toys, food, and ghost stories among other things. Until this year, the […]


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The McGarvey Service Center is the main office of the French Creek Council. It also serves as the council's scout store for uniform, camping, and advancement needs. At the McGarvey Service Center you will find the council executive, district executives, financing staff, and other council administrators helping to make the Boy Scouts of America a successful experience for all of the French Creek Council.
The McGarvey Service Center is located at:
1815 Robison Road W
Erie, PA 16509
October 4, 2012
French Creek Council
- Council News
- Office & Scout shop closed to public
- Council Dinner 2020
- Scout Shop Closed
- Charter Renewal
- Events
- Join Scouting Today
- Recruitment Season
- Camp Leader Update
- On-Line Youth Applications
- Like to ride Motorcycles?
- Pennsylvania State Background checks for volunteers: PA Act 153 of 2014:
- Pinewood Derby Track Rental
- Silver Beaver Nomination Information
- BSA and United Way
- Scouts are more successful
- Council Needs and Wants List
- Historic Trails
- Events
- About French Creek Council
- McGarvey Service Center
- Media Center
- Online Charter Renewal


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On January 12th, 2010, a massive earthquake devastated Haiti, one of the poorest countries in the world, causing severe damages and casualties.
Based on its efficient experience with collecting funds for Asian tsunami survivors in 2004, Fondation de France immediately mobilized its aid campaign and set up at the same time a specific experts committee dedicated to "Haiti Solidarity".
All donations to Fondation de France through "Haiti Solidarity" are consequently redistributed to organizations helping the quake victims.
Fondation de France supports long-term rebuilding projects of French organizations or NGO working in partnership with Haitian organizations. These projects which may last for up to 48 months aim at rebuilding disaster areas and at helping the Haitian population to get back to normal life. These programs indeed enable the rebuilding of houses, schools, medical centers, sports centers and other community buildings, but they also aim at boosting local economic life. Last, but not least, some projects also provide with psychological help the most vulnerable people, until they can get back to normal life.
Each and every supported project will be evaluated on the ground and financially audited. According to its standard procedures, Fondation de France will lastly account for every expense granted over Haiti.