Zeitschriftenartikel zum Thema „Barley Seeds Physiology“

AbstractModern hulless wheats, Triticum aestivum L., are more susceptible to the wheat midge, Sitodiplosis mosellana (Géhin), than the hulled, wild, ancestral species. Hulless cultivars of barley, Hordeum vulgare L., are becoming more widely grown in western Canada than in the past. Hulled and hulless cultivars of two-rowed and six-rowed barleys were tested for their susceptibility to wheat midge, to determine if this midge might become a serious pest of barley and to assess which plant traits might affect host suitability. In the field, larval populations on 10 barley cultivars were much lower than on wheat. In the laboratory, when the flag leaf sheath was peeled back to expose preflowering spikes, female midges readily oviposited on spikes of barley, although less so on younger spikes. Few larvae were able to develop on barley when eggs were laid after spikes had flowered. All barleys completed flowering, or nearly so, before spikes emerged from the flag leaf sheath, with two-rowed cultivars flowering earlier than six-rowed barleys. No differences in larval densities were found between hulless and hulled barleys, and therefore, factors other than the hulled trait must account for reduced susceptibility of barley. Because barley flowers within the flag leaf sheath, its period of susceptibility to infestation is much shorter than for wheat, as evidenced by reduced infestation of earlier-flowering two-rowed cultivars compared with later-flowering six-rowed cultivars. Also, the tight closure of the leaf-like glumes that form the florets of barley probably makes access to young seeds more difficult for newly hatched larvae than is the case for wheat. At comparable crop growth stages, larval densities on all the barleys were < 10% of those on spring wheat. The introduction of hulless barley for production in Canada is unlikely to increase wheat midge damage on barley to an economic level.

AbstractPeroxiredoxins are thiol–requiring antioxidants found in organisms ranging from bacteria to humans. They can be divided into two subgroups with either one or two conserved cysteine residues. In plants, 1–Cys peroxiredoxins have been identified in a number of grasses and cereals, and in the dicotyledonous speciesArabidopsis thaliana. In contrast to other antioxidants, the 1–Cys peroxiredoxin genes are expressed solely in seeds, and only in the parts of the seeds surviving desiccation, i.e. the embryo and the aleurone layer. The expression pattern is characteristic of late embryogenesis–abundant genes. The PER1 protein of barley is present in high concentrations in the nucleus at the onset of desiccation. 1–Cys genes are expressed in a dormancy–related manner in mature seeds, in that transcript levels are high in imbibed dormant seeds, but disappear upon germination of their non–dormant counterparts. 1–Cys transcript levels can be up–regulated by ABA and osmotic stresses and suppressed by gibberellic acid. Two hypotheses have been put forward on the function of 1–Cys peroxiredoxins in seed physiology. First, these proteins might protect macromolecules of embryo and aleurone cells against damaging reactive oxygen species during seed desiccation and early imbibition. And second, seed peroxiredoxins might play a role in the maintenance of dormancy. These hypotheses are discussed, taking into account present knowledge of the biochemistry and molecular biology of peroxiredoxins.

AbstractDusky wireworms, Agriotes obscurus (L.), aggregated in similar numbers at wheat [Triticum aestivum L. (Gramineae) ‘Max’], oat [Avena sativa L. (Gramineae) ‘Walderen’], barley [Hordeum vulgare L. (Gramineae) ‘Verdin’], and fall rye [Secale cereale L. (Gramineae) ‘Wheeler’ and ‘Prima’] cultivar bait stations containing 100 seeds planted 3 cm deep in 127-cm2 circular bait stations. Similar levels of aggregation also occurred at 11 varieties of wheat planted at 100 seeds/127 cm2. When wheat, oat, barley, and the fall rye cultivars were planted at increasing density (0–180 seeds per bait station), aggregation by A. obscurus increased initially, but reached a plateau at numbers and at seeding rates specific to each grain variety as determined using the asymptotic equation y = B0(1 – e–B1x). Except for barley, this equation predicted wireworm densities within 11% of the densities actually observed at bait stations with 100 seeds/127 cm2. It was concluded that any of the wheat, oat, barley, or fall rye varieties would be suitable for monitoring A. obscurus wireworm populations if planted in bait stations at 100 seeds/127 cm2, as well as for aggregating wireworms by means of a trap crop.

Background. In cereals, photosynthetically active parts of the ear significantly contribute to seed size and weight at the grain-filling stage. In barley, ear tissues may accumulate melanin pigments synthesized in chloroplast-derived melanoplasts. Effects of such pigments on yield parameters of seeds have not been evaluated to date.Materials and methods. Seed weight and size assessed by image analysis were compared between two near-isogenic barley lines differing in alleles of the Blp1 gene, which determines melanin accumulation in ear tissues. Data on grainrelated parameters were collected during 6 years and include data on seeds grown either in the field or under greenhouse conditions.Results and discussion. A negative effect of the Blp1 locus on the weight of 1000 seeds harvested in the field but not in the greenhouse was revealed. To determine whether this effect is related to grain size, a comparison of two-dimensional linear parameters of seeds between the lines was performed. It was shown that unlike the length and the area of seeds, the width of seeds was also negatively affected by the Blp1 locus. Although the same factors affected the weight of 1000 seeds and the width of seeds, a correlation between them was not found, implying a dependence of seed weight on other factors such as thickness and its related parameter, seed volume.Conclusion. Effects of barely ear pigmentation and of the gene controlling it on yield-related parameters of seeds were studied here for the first time. The observed negative impact of the Blp1 locus on seed weight and size may be mediated by an interfering chloroplast activity and/or accumulation of assimilates via melanogenesis. Additional studies are necessary to test this supposition and to investigate the interaction of melanin synthesis and photosynthetic activity of the tissues accumulating this pigment.

AbstractSoil salinity is one of the major abiotic stresses affecting seed germination, crop growth and productivity. In this study, seeds of three wheat (Triticum aestiveum L.) and three barley (Hordeum vulgare L.) cultivars were treated with different concentrations of NaCl to investigate the effect of salt on seed germination physiology and metabolism through the characterization of seed germination pattern, gamma-aminobutyric acid (GABA) shunt metabolite accumulation [GABA, glutamate (Glu) and alanine (Ala)] and glutamate decarboxylase (GAD) expression using RT-PCR. A trend of decreasing germination percentage with increasing NaCl concentrations was observed. Under all salt stress treatments, data showed significant increase with positive correlation (r = 0.50–0.99) between abundance of GABA shunt metabolites and salt concentration in all wheat and barley cultivars for 5 days. Increased GABA content was associated with a small but significant increase in Ala and Glu content in all cultivars. In all NaCl treatments, the transcription of GAD in terms of RNA abundance showed a significant increase in all cultivars with positive correlation (r = 0.50–0.98). Data showed significant association between GAD RNA transcription and the response of germinating seeds to salt stress in terms of GABA shunt metabolite accumulation. The elevated expression of GAD under salinity suggests the need for elevated activity of the GAD-mediated conversion of Glu to GABA during seed germination, which provides alternative metabolic routes to the respiratory machinery, balancing carbon and nitrogen metabolism and osmolyte synthesis in germinating seeds of wheat and barley under salt stress.

Nonthermal plasma (NTP), or cold plasma, has shown many advantages in the agriculture sector as it enables removal of pesticides and contaminants from the seed surface, increases shelf life of crops, improves germination and resistance to abiotic stress. Recent studies show that plasma treatment indeed offers unique and environmentally friendly processing of different seeds, such as wheat, beans, corn, soybeans, barley, peanuts, rice and Arabidopsis thaliana, which could reduce the use of agricultural chemicals and has a high potential in ecological farming. This review covers the main concepts and underlying principles of plasma treatment techniques and their interaction with seeds. Different plasma generation methods and setups are presented and the influence of plasma treatment on DNA damage, gene expression, enzymatic activity, morphological and chemical changes, germination and resistance to stress, is explained. Important plasma treatment parameters and interactions of plasma species with the seed surface are presented and critically discussed in correlation with recent advances in this field. Although plasma agriculture is a relatively new field of research, and the complex mechanisms of interactions are not fully understood, it holds great promise for the future. This overview aims to present the advantages and limitations of different nonthermal plasma setups and discuss their possible future applications.

Abstract Barley possesses a branchless, spike-shaped inflorescence where determinate spikelets attach directly to the main axis, but the developmental mechanism of spikelet identity remains largely unknown. Here we report the functional analysis of the barley gene BRANCHED AND INDETERMINATE SPIKELET 1 (BDI1), which encodes a TCP transcription factor and plays a crucial role in determining barley inflorescence architecture and spikelet development. The bdi1 mutant exhibited indeterminate spikelet meristems that continued to grow and differentiate after producing a floret meristem; some spikelet meristems at the base of the spike formed two fully developed seeds or converted to branched spikelets, producing a branched inflorescence. Map-based cloning analysis showed that this mutant has a deletion of ~600 kb on chromosome 5H containing three putative genes. Expression analysis and virus-induced gene silencing confirmed that the causative gene, BDI1, encodes a CYC/TB1-type TCP transcription factor and is highly conserved in both wild and cultivated barley. Transcriptome and regulatory network analysis demonstrated that BDI1 may integrate regulation of gene transcription cell wall modification and known trehalose-6-phosphate homeostasis to control spikelet development. Together, our findings reveal that BDI1 represents a key regulator of inflorescence architecture and meristem determinacy in cereal crop plants.

Abstract Abscisic acid (ABA) is a phytohormone and a major determinant of seed dormancy in plants. Seed dormancy is gradually lost during dry storage, a process known as ‘after-ripening’, and this dormancy decay is related to a decline in ABA content and sensitivity in seeds after imbibition. In this study, we aimed at investigating the effect of after-ripening on ABA signaling in barley, our cereal model species. Phosphosignaling networks in barley grains were investigated by a large-scale analysis of phosphopeptides to examine potential changes in response pathways to after-ripening. We used freshly harvested (FH) and after-ripened (AR) barley grains which showed different ABA sensitivity. A total of 1,730 phosphopeptides were identified in barley embryos isolated from half-cut grains. A comparative analysis showed that 329 and 235 phosphopeptides were upregulated or downregulated, respectively after ABA treatment, and phosphopeptides profiles were quite different between FH and AR embryos. These results were supported by peptide motif analysis which suggested that different sets of protein kinases are active in FH and AR grains. Furthermore, in vitro phosphorylation assays confirmed that some phosphopeptides were phosphorylated by SnRK2s, which are major protein kinases involved in ABA signaling. Taken together, our results revealed very distinctive phosphosignaling networks in FH and AR embryos of barley, and suggested that the after-ripening of barley grains is associated with differential regulation of phosphosignaling pathways leading to a decay of ABA signaling.

Abstract Leaf senescence is an important process for plants to remobilize a variety of metabolites and nutrients to sink tissues, such as developing leaves, fruits and seeds. It has been suggested that reactive oxygen species (ROS) play an important role in the initiation of leaf senescence. Flag leaves of two different barley varieties, cv. Lomerit and cv. Carina, showed differences in the loss of photosystems and in the production of ROS at a late stage of senescence after significant loss of chlorophyll (Krieger-Liszkay et al. 2015). Here, we investigated photosynthetic electron transport and ROS production in primary leaves of these two varieties at earlier stages of senescence. Comparisons were made between plants grown outside in natural light and temperatures and plants grown in temperature-controlled growth chambers under low light intensity. Alterations in the content of photoactive P700, ferredoxin and plastocyanin (PC) photosynthetic electron transport were analyzed using in vivo near-infrared absorbance changes and chlorophyll fluorescence, while ROS were measured with spin-trapping electron paramagnetic resonance spectroscopy. Differences in ROS production between the two varieties were only observed in outdoor plants, whereas a loss of PC was common in both barley varieties regardless of growth conditions. We conclude that the loss of PC is the earliest detectable photosynthetic parameter of leaf senescence while differences in the production of individual ROS species occur later and depend on environmental factors.

ABSTRACT Quinclorac is a systemic herbicide absorbed by germinating seeds, roots and leaves of seedlings. It is a selective compound for crops such as rice, canola, barley, corn, sorghum, and pasture. Quinclorac can be used to control various monocots and dicotyledonous weed species. The biochemical function of this herbicide in the plant has intrigued scientists for nearly four decades. The objectives of this review are to present evidence of three hypotheses on the biochemical functioning of quinclorac and to propose an integrative mode of action. The first theory on the mode of action of quinclorac is supported by evidence of inhibition of incorporation of C14-glucose into cellulose and hemicellulose, thus, affecting the cell wall synthesis. The second hypothesis suggests that quinclorac acts as an auxin in broadleaved weed species. In grass species, however, this herbicide appears to stimulate the activity of the 1-aminocyclopropane-1-carboxylate synthase enzyme and, subsequently, to increase the ethylene production; also, it seems to increase the cyanide acid content to phytotoxic levels. A third hypothesis to explain the harmful effect in some plant species is the formation of reactive oxygen species (ROS). Apparently, these processes are not mutually exclusive; therefore, an integrative theory for the action of quinclorac is suggested. It is theorized that the aforementioned biochemical activities are interconnected and can be the phytotoxic backbone to explain the herbicidal effect depending on the plant species and the plant growth stage, among other factors.

AbstractAssays were done in the laboratory and field to monitor the response of Pacific coast wireworm, Limonius canus LeConte, to food baits. A glass-plate assay was used in laboratory trials to study movement of wireworm larvae through soil in response to several food baits, including germinating seeds of grains, rolled oats [Avenu sauva L. (Poaceae)], carrot [Daucus carota L. (Umbelliferae)], and potato [Solanum tuberosum L. (Solanaceae)]. Studies were also done with these baits to determine effectiveness under field conditions. In both laboratory and field trials, germinating seed of wheat [Triticum aestivum L. (Poaceae)] and barley [Hordeum vulgare L. (Poaceae)] ranked higher in response by larvae than the remaining food baits. In the glass-plate assays, 65–70% of wireworms contacted the wheat or barley seed baits within a 2-h assay period. Corn [Zea mays L. (Poaceae)] seed and sliced carrot were also effective, with 60% of larvae contacting the baits. Rice [Oryza sativa L. (Poaceae)], rye [Secale cereale L. (Poaceae)], and potato were contacted by 30–45% of larvae. In the majority of assays, contact with the bait occurred within 30 min of the start of the assay. Trail lengths varied substantially among larvae (0–70 cm). The field studies showed that all food baits captured more wireworms than unbaited traps. A second assay conducted in the laboratory showed that moistened rolled oats were contacted with a higher probability if oats were aged 72 h following wetting (51% of larvae contacted the bait) than oats used immediately following wetting (28% of larvae). Field trials using baits composed of different volumes of rolled oats showed little evidence of a dose response in capture rates of L. canus, although all baits captured more wireworms than unbaited traps. These studies showed that L. canus is attracted to food baits in both laboratory and field trials, and that baits may prove useful to monitor populations of this pest in the field.

Barley (Hordeum vulgare L.) is considered one of the most important cereals, such as maize, rice and wheat, due to its multiplicity of usage, as much for human consumption as for feed. The Nitrogen is one of the main nutrients that influences the quality and productivity of barley seeds because it participates in numerous metabolic routes. It is a limiting factor of the production, the lack or the excess can bring consequence to the culture. In this manner, the present study aims to evaluate the influence of different nitrogen doses on the physiologic quality and biochemical characters of brewing barley seeds produced in two growing environments. The experiment was conducted in the 2017 crop season, in a randomized block design, organized in a factorial scheme, being 2 cultivation environment &times; 2 brewing barley cultivars &times; 4 nitrogen doses, disposed in four replicates. The physiological quality of the seeds was evaluated by germination, first counting of germination, accelerated aging, field emergence, shoot and root length, dry mass of shoot and root of seedlings, electrical conductivity (3, 6 and 24 hours) and isoenzymes. The growing environment promotes differences in the barley seeds physiologic quality. The nitrogen fertilizing, at dose of 120 kg ha-1, results in superiority in the physiologic quality of seeds. The cultivar BRS Cau&ecirc; in both environments presented higher vigor. The nitrogen fertilizing alters the isoenzymatic expression of barley seedlings, creating variation in the bands intensity, at different nitrogen doses.

Barley plants carrying a mutation in the Mlo (barley [Hordeum vulgare L.] cultivar Ingrid) locus conferring a durable resistance against powdery mildew are hypersusceptible to the rice blast fungus Magnaporthe grisea. It has been speculated that a functional Mlo gene is required for the expression of basic pathogen resistance and that the loss of Mlo function mediating powdery mildew resistance is an exception for this particular disease. Here, we report that the onset of acquired resistance (AR) after chemical as well as biological treatments is sufficient to overcome the hypersusceptible phenotype of backcross line BCIngridmlo5 (mlo) barley plants against M. grisea. Moreover, even barley plants bearing a functional Mlo gene and thus showing a moderate infection phenotype against rice blast exhibit a further enhanced resistance after induction of AR. Cytological investigations reveal that acquired resistance in mlo genotypes is manifested by the restoration of the ability to form an effective papilla at sites of attempted penetration, similarly to wild-type Mlo plants. In addition, the rate of effective papillae formation in Mlo plants was further enhanced after the onset of AR. These results demonstrate that treatments leading to the AR state in barley function independently of the Mlo/mlo phenotype and suggest that the Mlo protein is not a component of the AR signaling network. Moreover, it seems that only concomitant action of Mlo together with AR permits high level resistance in barley against blast. Higher steady state levels of PR1 and barley chemically induced mRNA correlate with higher disease severity rather than with the degree of resistance observed in this particular interaction.

In Arabidopsis, both pathogen invasion and benzothiadiazole (BTH) treatment activate the nonexpresser of pathogenesis-related genes 1 (NPR1)-mediated systemic acquired resistance, which provides broad-spectrum disease resistance to secondary pathogen infection. However, the BTH-induced resistance in Triticeae crops of wheat and barley seems to be accomplished through an NPR1-independent pathway. In the current investigation, we applied transcriptome analysis on barley transgenic lines overexpressing wheat wNPR1 (wNPR1-OE) and knocking down barley HvNPR1 (HvNPR1-Kd) to reveal the role of NPR1 during the BTH-induced resistance. Most of the previously designated barley chemical-induced (BCI) genes were upregulated in an NPR1-independent manner, whereas the expression levels of several pathogenesis-related (PR) genes were elevated upon BTH treatment only in wNPR1-OE. Two barley WRKY transcription factors, HvWRKY6 and HvWRKY70, were predicted and further validated as key regulators shared by the BTH-induced resistance and the NPR1-mediated acquired resistance. Wheat transgenic lines overexpressing HvWRKY6 and HvWRKY70 showed different degrees of enhanced resistance to Puccinia striiformis f. sp. tritici pathotype CYR32 and Blumeria graminis f. sp. tritici pathotype E20. In conclusion, the transcriptional changes of BTH-induced resistance in barley were initially profiled, and the identified key regulators would be valuable resources for the genetic improvement of broad-spectrum disease resistance in wheat. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Young barley seems to be a promising material for use as nutricosmetic due to the presence of many biologically active compounds. The aim of this study was to evaluate the effect of Hordeum vulgare L. extracts on human skin fibroblasts exposed to ultraviolet radiation B (UVB) radiation. Analysis of the chemical composition showed a predominance of 9,12,15-octadecatrienoic acid. The quality assessment showed that young barley preparations have high total polyphenolic content (TPC) and favourable total antioxidant status (TAS). They also contain antioxidant elements such as zinc, copper, and selenium. Furthermore, the analyzed products were found to be safe in terms of toxic elements (lead, cadmium and mercury) and lack of cytotoxic effect of young barley extracts on cells. In vitro bioactivity assays showed that young barley extract increased the survival rate and accelerated the migration of fibroblasts in research models with UVB radiation. The application of both extracts caused an increase in DNA biosynthesis, and in the number of cells arrested in S phase. Moreover, an inhibitory effect of the tested extracts on the expression of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) was observed. The results indicate that young barley extracts, due to protective as well as restorative effect, could potentially be used in the production of nutricosmetics and skin care products.

With advances in plant genomics, plant phenotyping has become a new bottleneck in plant breeding and the need for reliable high-throughput plant phenotyping techniques has emerged. In the face of future climatic challenges, it does not seem appropriate to continue to solely select for grain yield and a few agronomically important traits. Therefore, new sensor-based high-throughput phenotyping has been increasingly used in plant breeding research, with the potential to provide non-destructive, objective and continuous plant characterization that reveals the formation of the final grain yield and provides insights into the physiology of the plant during the growth phase. In this context, we present the comparison of two sensor systems, Red-Green-Blue (RGB) and multispectral cameras, attached to unmanned aerial vehicles (UAV), and investigate their suitability for yield prediction using different modelling approaches in a segregating barley introgression population at three environments with weekly data collection during the entire vegetation period. In addition to vegetation indices, morphological traits such as canopy height, vegetation cover and growth dynamics traits were used for yield prediction. Repeatability analyses and genotype association studies of sensor-based traits were compared with reference values from ground-based phenotyping to test the use of conventional and new traits for barley breeding. The relative height estimation of the canopy by UAV achieved high precision (up to r = 0.93) and repeatability (up to R2 = 0.98). In addition, we found a great overlap of detected significant genotypes between the reference heights and sensor-based heights. The yield prediction accuracy of both sensor systems was at the same level and reached a maximum prediction accuracy of r2 = 0.82 with a continuous increase in precision throughout the entire vegetation period. Due to the lower costs and the consumer-friendly handling of image acquisition and processing, the RGB imagery seems to be more suitable for yield prediction in this study.

Abstract Ten doubled haploid (DH) lines of winter barley with an increased range of freezing/drought tolerance were used to identify phytohormones involved in plant stress acclimation. Cold hardening and drought stress were applied at the most critical stages of plant development on young seedlings and heading plants, respectively. The level of the phytohormones was significantly higher at heading, more than 5-fold in respect of salicylic acid (SA) and total brassinosteroids (BRs) and 1.7-fold in the case of abscisic acid (ABA). Moreover, the spectrum of detectable BRs increased from one—homocastasterone (HCS)—found in seedlings to four BRs identified in heading plants [HCS, castasterone (CS), teasterone and dolicholide], with the last one detected for the first time in cereal species. To some extent freezing tolerance seems to be determined by native hormonal status as control seedlings of tolerant DH lines contained 1.4- and 2.3-fold lower amount of ABA and HCS and 2.3-fold higher amount of SA in comparison to freezing-sensitive ones. Such dependency was not observed in heading plants as significant variation in CS content was the only detected difference. Under stress treatments, tolerant DH lines accumulated significantly lower (75–81%) amount of ABA, which probably reflected lower stress intensity resulting from another defence strategy. In contrast, stress-induced significant almost 2-fold increase in HCS/CS and 2–3-fold decrease in SA content specific for tolerant DH lines of barley suggest the involvement of these molecules in freezing/drought defence. Detected correlations suggest their interaction with nonspecific peroxidase and low molecular weight antioxidants.

Abstract In C3 cereals such as wheat and barley, grain filling was traditionally explained as being sustained by assimilates from concurrent leaf photosynthesis and remobilization from the stem. In recent decades, a role for ear photosynthesis as a contributor to grain filling has emerged. This review analyzes several aspects of this topic: (i) methodological approaches for estimation of ear photosynthetic contribution to grain filling; (ii) the existence of genetic variability in the contribution of the ear, and evidence of genetic gains in the past; (iii) the controversy of the existence of C4 metabolism in the ear; (iv) the response of ear photosynthesis to water deficit; and (v) morphological and physiological traits possibly related to ear temperature and thermal balance of the ear. The main conclusions are: (i) there are a number of methodologies to quantify ear photosynthetic activity (e.g. gas exchange and chlorophyll fluorescence) and the contribution of the ear to grain filling (individual ear shading, ear emergence in shaded canopies, and isotope composition); (ii) the contribution of ear photosynthesis seems to have increased in modern wheat germplasm; (iii) the contribution of the ear to grain filling increases under resource-limitation (water deficit, defoliation, or pathogen infection); (iv) there is genetic variability in the contribution of the ear in wheat, opening up the possibility to use this trait to ameliorate grain yield; (v) current evidence supports the existence of C3 metabolism rather than C4 metabolism; (vi) the ear is a ‘dehydration avoider organ’ under drought; and (vii) thermal balance in the ear is a relevant issue to explore, and more research is needed to clarify the underlying morphological and physiological traits.

Challenge by a nonadapted powdery mildew fungal pathogen leads to the formation of a local cell-wall apposition (papilla) beneath the point of attempted penetration. Several plasma membrane (PM) proteins with opposing roles in powdery mildew infection, including Arabidopsis thaliana PENETRATION1 (PEN1) and barley (Hordeum vulgare) MILDEW RESISTANCE LOCUS O (MLO), are localized to the site of powdery mildew attack. PEN1 contributes to penetration resistance to nonadapted powdery mildews, whereas MLO is a susceptibility factor required by adapted powdery mildew pathogens for host cell entry. Our previous studies have demonstrated that the vesicle and endosomal trafficking inhibitors, brefeldin A and wortmannin, have opposite effects on the penetration rates of adapted and nonadapted powdery mildews on grapevine. These findings prompted us to study the pathogen-induced intracellular trafficking of grapevine variants of MLO and PEN1. We first identified grapevine (Vitis vinifera) VvPEN1 and VvMLO orthologs that rescue Arabidopsis Atpen1 and Atmlo2 mlo6 mlo12 null mutants, respectively. By using endomembrane trafficking inhibitors in combination with fluorescence microscopy, we demonstrate that VvMLO3/VvMLO4 and VvPEN1 are co-trafficked together from the PM to the site of powdery mildew challenge. This focal accumulation of VvMLO3/VvMLO4 and VvPEN1 to the site of attack seems to be required for their opposing functions during powdery mildew attack, because their subcellular localization is correlated with the outcome of attempted powdery mildew penetration.

In a cell-free system from neutrophil cytosol GTP(γ)S can induce an increase in the number of free filament barbed ends and massive actin polymerisation and cross-linking. GTP(γ)S stimulation was susceptible to an excess of GDP, but not Bordetella pertussis toxin and could not be mimicked by aluminium fluoride, myristoylated GTPgammaS. Gialpha2 or Gbeta1gamma2 subunits of trimeric G proteins. In contrast, RhoGDI and Clostridium difficile toxin B (inactivating Rho family proteins) completely abrogated the effect of GTPgammaS. When recombinant, constitutively activated and GTPgammaS-loaded Rac1, RhoA, or Cdc42 proteins alone or in combination were probed at concentrations &gt;100 times the endogenous, however, they were ineffective. Purified Cdc42/Rac-interactive binding (CRIB) domain of WASP or C3 transferase did not prevent actin polymerisation by GTPgammaS. The action of GTPgammaS was blocked by mM [Mg2+], unless a heat- and trypsin-sensitive component present in neutrophil plasma membrane was added. Liberation of barbed ends seems therefore to be mediated by a toxin B-sensitive cytosolic Rho-family protein, requiring a membrane-associated guanine nucleotide exchange factor (GEF) for its activation by GTPgammaS under physiologic conditions. The inefficiency of various protein kinase and phosphatase inhibitors (staurosporine, genistein, wortmannin, okadaic acid and vanadate) and removal of ATP by apyrase, suggests that phosphate transfer reactions are not required for the downstream propagation of the GTPgammaS signal. Moreover, exogenously added phosphoinositides failed to induce actin polymerisation and a PtdIns(4,5)P2-binding peptide did not interfere with the response to GTPgammaS. The speed and simplicity of the presented assay applicable to protein purification techniques will facilitate the further elucidation of the molecular partners involved in actin polymerisation.

AbstractWestern boxelder bugs, Boisea rubrolineata (Barber), form large aggregations on pistillate boxelder, Acer negundo L. (Aceraceae), host trees with maturing seeds, and cluster on warm, sunlit surfaces prior to overwintering. We have recently shown that B. rubrolineata is attracted to the host-tree semiochemicals phenylacetonitrile and 2-phenethyl acetate. We report results of chemical analyses and laboratory bioassays suggesting that aggregation and sexual communication in B. rubrolineata are mediated by 2-phenylethanol. This compound serves as an aggregation pheromone for females, males, and 5th-instar nymphs in midsummer, and in males it appears to serve as a sex-attractant pheromone in early spring. As an aggregation pheromone, 2-phenylethanol originates from the feces of seed-feeding females and males and (or) the ventral abdominal gland of males. As a sex-attractant pheromone, it originates from the ventral abdominal gland of males that emerge from overwintering diapause. Aggregations of B. rubrolineata in the fall and winter are mediated by other as yet unknown pheromones.

Apoptotic hepatocytes have been demonstrated to represent an important signal for transmigration of leukocytes sequestered in sinusoids during endotoxemia in vivo. Beside leukocytes, platelets and their adhesion to endothelial cells and leukocytes have been implicated in inflammatory liver injury. Using in vivo multifluorescence microscopy, we examined the possibility that hepatocellular apoptosis causes both leukocytes and platelets to colocalize within the sinusoidal microvasculature of endotoxemic livers. We further addressed the issue whether cellular colocalization with apoptotic hepatocytes is cause or consequence of apoptosis. Intraperitoneal exposure of rats with LPS (5 mg/kg) induced liver injury after 6 and 16 h, as given by nutritive perfusion failure (20 ± 2 and 21 ± 2%), intrahepatic leukocyte (60 ± 10 and 121 ± 48 cells/mm2), and platelet (12 ± 4 and 34 ± 4 cells/mm2) accumulation as well as parenchymal cell apoptosis (4 ± 1 and 11 ± 2 cells/mm2) and caspase cleavage (4.7 ± 2.4- and 7.0 ± 3.0-fold increase; P < 0.05 vs. saline-exposed controls). Higher doses of LPS (10 mg/kg ip) further increased intrahepatic leukocyte and platelet accumulation but not the extent of parenchymal apoptosis. Detailed spatial analysis revealed colocalization of leukocytes (range 12–24%) but barely of platelets (<6%) with apoptotic hepatocytes in all endotoxemic groups studied. It is of interest, however, that platelets were found at increasing rates in colocalization with leukocytes at 6 and 16 h after LPS exposure (5 mg/kg LPS: 7 ± 3 and 25 ± 6%; 10 mg/kg LPS: 11 ± 4 and 14 ± 1%). Platelet-leukocyte events significantly correlated with the extent of caspase cleavage as an indicator of tissue apoptosis ( P < 0.05; r = 0.82). Blockade of apoptosis by a pan-caspase inhibitor caused a significant reduction of leukocyte adherence and platelet-leukocyte colocalization on LPS exposure. On the other hand, leukocytopenic animals revealed reduced hepatocyte apoptosis, although values still exceeded those of controls, and in leuko- and thrombocytopenic animals, hepatocyte apoptosis was found reduced to control values. Taken together, LPS-associated hepatocyte apoptosis seems to be initiated by circulating blood cells that become adherent within the liver but might also contribute to further sustain the inflammatory cell-cell response.

Abstract Seeds are complex biological systems comprising three genetically distinct tissues: embryo, endosperm, and maternal tissues (including seed coats and pericarp) nested inside one another. Cereal grains represent a special type of seeds, with the largest part formed by the endosperm, a specialized triploid tissue ensuring embryo protection and nourishment. We investigated dynamic changes in DNA content in three of the major seed tissues from the time of pollination up to the dry seed. We show that the cell cycle is under strict developmental control in different seed compartments. After an initial wave of active cell division, cells switch to endocycle and most endoreduplication events are observed in the endosperm and seed maternal tissues. Using different barley cultivars, we show that there is natural variation in the kinetics of this process. During the terminal stages of seed development, specific and selective loss of endoreduplicated nuclei occurs in the endosperm. This is accompanied by reduced stability of the nuclear genome, progressive loss of cell viability, and finally programmed cell death. In summary, our study shows that endopolyploidization and cell death are linked phenomena that frame barley grain development.

IntroductionAccording to Friedrich Nietzsche, the death of Man follows the death of God. Man as a concept must be overcome. Yet Nietzsche extends humanism’s jargon of creativity that privileges Man over animal. To truly overcome the notion of Man, one must undercome Man, in other words go below Man. Once undercome, creativity devolves into a type of building and unbuilding, affording art the ability to conceive of the subject emptied of divine creation. This article will examine how Man is unbuilt in three works by three different artists: Francis Bacon’s “Study of a Baboon” (1953), Jan Švankmajer’s Darkness, Light, Darkness (1989), and Patricia Piccinini’s “The Young Family” (2002). All three artists evoke the animalistic in their depiction of what could be called the sub-subject, a diminished agent. Unbuilding the subject becomes the basis for building the sub-subject in these depictions of the human remainder. Man, from this vantage, will be examined as a cultural construct. Man largely means human, yet the Renaissance concept favoured a certain type of powerful male. Instead of rescuing Man, Bacon, Švankmajer and Piccinini, present the remnants of the human amidst the animal rather than the human subject detached from the animal. Such works challenge humanism, expressed in Giorgio Vasari’s analysis of art and creativity as indicative of Man’s closeness to the divine, which in a strange way, is extended in Nietzsche’s writings. These artists dismantle and build a subhuman form of subjectivity and thereby provide a challenge to traditional conceptions of creativity that historically favour Man as the creator beneath only God Himself. In the course of this article, I explore the violence of Bacon’s painted devolution, the deflationary animation of Švankmajer and Piccinini’s subhuman tenderness. I do not argue that we must abandon humanism altogether as there are a multiplicity of humanisms, or attempt to invalidate all the various posthumanisms, transhumanisms and antihumanisms. Rather, I attempt to show that Nietzsche’s posthumanism is a suprahumanism and that one possible way to frame the death of Man is through undercoming Man. Art, held in high esteem by Renaissance humanism, becomes a vehicle to imagine and engage with subhuman subjectivity.What Is Humanism? Humanism has numerous connotations from designating atheism to celebrating culture to privileging humans above other animals. The type of humanism I am interested in is not secular humanism, but rather humanism that celebrates and conceptualises Man’s place in the universe and does so through accentuating his (and I mean his given humanism’s often sexist, masculinist history) creativity and intellectual power. This celebration of creativity depends in part on a type of religious view, where Man is at the centre of God’s design. Such a view holds that Man’s power to shape nature’s materials resembles God. This type of humanism remains today but usually in a more humbled form, enfeebled by the scientific realisations that characterised the Enlightenment, namely the realisation that Man was not the centre of God’s universe. The Enlightenment is sometimes characterised as the birth of modern humanism, where the human subject undergoes estrangement from his surroundings through the conceptualisation of the subject–object division, and gains control over nature. A common narrative is that the subject’s autonomy and power came to extend to art itself, which in turn, became valued as possessing its own aesthetic legitimacy and yet also becoming an alienated commodity. Yet Cary Wolfe, in What Is Posthumanism?, echoes Michel Foucault’s claim that the Enlightenment could be viewed in tension to humanism (“Introduction” n.p.). Indeed, the Enlightenment’s creation of modern science would come to seriously challenge any view of humanity’s privileged status in this world. In contrast, Renaissance humanism conceived of Man as the centrepiece of God’s design and gifted with artistic creation and the ability to uncover truth. Renaissance HumanismRenaissance humanism is encapsulated by Vasari’s preface to The Lives of the Artists. In his preface, Vasari contends that God was the first artist, being both a painter and sculptor: God on High, having created the great body of the world and having decorated the heavens with its brightest lights, descended with His intellect further down into the clarity of the atmosphere and the solidity of the earth, and, shaping man, discovered in the pleasing invention of things the first form of sculpture and painting. (3)Interestingly, God discovers creation, which is a type of decoration, where the skies are decorated with bright lights—the stars. Giving colour, light and shade to the world and heavens, qualifies God as a painter. The human body, according to Vasari, is sculpted by God, which in turn inspires artists to depict the human form. Art and design—God’s design—is thereby ‘at the origin of all things’ and not merely painting and sculpture, though the reality we know is still the product of God’s painting and sculpture. According to Vasari, God privileges Man not for his intellect per se, but by bestowing him with the ability of creation and design. Indeed, creativity and design are for Vasari a part of all intellectual discovery. Intellect is the mode of discovering design, which for Vasari, is also creation. Vasari claims “that divine light infused in us by a special act of grace which has not only made us superior to other animals but even similar, if it is permitted to say so, to God Himself” (4). God is more than just a maker, he is a creator with an aesthetic sense. All intellectual human endeavours, claims Vasari, are aesthetic and creative, in their comprehension of God’s design of the world. Vasari’s emphasis on design became outmoded as Renaissance humanism was challenged by the Enlightenment’s interest in humans and other animals as machines. However, evolution challenges even some mechanistic understandings of the human subject, which sometimes presupposed that the human-machine had a maker, as with William Paley’s watchmaker theory. As Richard Dawkins put it in The Blind Watchmaker, nature “has no vision, no foresight, no sight at all. If [evolution] can be said to play the role of the watchmaker in nature, it is the blind watchmaker” (“Chapter One: Explaining the Very Improbable” n.p.). No longer was God’s universe designed for Man’s comprehension and appreciation, foretelling humanity’s own potential extinction.Man and God’s DeathThe idea that humanity was created by blind processes raises the question of what sort of depiction of the human subject is possible after the death of God and the Enlightenment’s tendency toward disenchantment? An art and self-understanding founded on atheism would be in sharp distinction to Vasari’s characterisation of the nature as an artwork coloured by the divine painter and sculptor in the heavens. Man’s creativity and design are, for the Renaissance humanist, part of discovery, the embodied realisations and iterations of the Platonic realm of divine forms. But such designs, wondrous for Vasari, can be viewed as shadows without origin in a post-God world. In Vasari, Platonism is still present where the artist’s creation becomes a way of discerning the origin of all forms, God himself. Yet, without divine origin, these forms are no longer discoveries and the possibility emerges that they are not even creations, emptied of the divine meaning that gave Man’s creative and scientific work value. Nietzsche understood that the loss of God called for the revaluation of all values. This is why Nietzsche claims that God’s death signifies the death of Man. For Nietzsche, the last Man was such an iteration, a shadow of what man had been (Thus Spoke Zarathustra 9-10). The Post-Man, the Übermensch, is one who extends the human power of creation and evaluation. In Vasari, Man is a model created by God. Nietzsche extends this logic: Man is his own creation as is God Man’s model. Man is capable of self-construction and overcoming without the hindrance of the divine. This freedom unlocked by auto-creation renders Man capable of making himself God. As such, art remains a source of sacred power for Nietzsche since it is a process of creative evaluation. The sacred is affirmed against secular profanity. For Nietzsche, God must be envisaged as Dionysus, a God that Nietzsche claims takes on a human form in Greek festivals dedicated to creation and fecundity. Mankind, in order to continue to have value after God’s death, “must become gods”, must take the place of God (The Gay Science 120). Nietzsche, All-Too HumanistNietzsche begins a project of rethinking Man as a category. Yet there is much in common with Renaissance humanism generated by Nietzsche’s Dionysian belief in a merger between God and Man. Man is overcome by a stronger and more creative figure, that of the Übermensch. By comparing Nietzsche with Vasari we can understand just how humanist Nietzsche remained. Indeed, Nietzsche fervently admired the Renaissance as a rebirth of paganism. Such an assessment of the rebirth of pagan art and values can almost be found in Vasari himself. Vasari claimed that pagan art, far from being blasphemous, brought Man closer to the divine in a tribute to the creativity of God. Vasari’s criticism of Christianity is careful but present. Indeed, Vasari—in a way that anticipates Nietzsche’s view that secular sacrilege was merely an extension of Christian sacrilege—attacks Christian iconoclasm, noting that barbarians and Christians worked together to destroy sacred forms of art: not only did [early Christianity] ruin or cast to the ground all the marvellous statues, sculptures, paintings, mosaics, and ornaments of the false pagan gods, but it also did away with the memorials and testimonials to an infinite number of illustrious people, in whose honour statues and other memorials had been constructed in public places by the genius of antiquity. (5) In this respect, Vasari embodies the values Nietzsche so praised in the Italian Renaissance. Vasari emphasises the artistic creations that enshrine distinctions of value and social hierarchy. While Vasari continues Platonic notions that ideals exist before human creation, he nevertheless holds human creation as a realisation and embodiment of the ideal, which is not dissimilar to Nietzsche’s notion of divine embodiment. For Nietzsche and Vasari, Man is exulted when he can rise, like a god, above other men. Another possibility would be to lower Man to just another animal. One way to envision such a lowering would be to subvert the mode by which Man is deemed God-like. Art that engages with the death of Man helps conceptualise subhumanism and the way that the subject ceases to be raised above the animal. What follows are studies of artworks that unbuild the subject. Francis Bacon’s “Study of a Baboon”Francis Bacon’s work challenges the human subject by depicting nonhuman subjects, where the flesh is torn open and Man’s animal flesh is exposed. Sometimes Bacon does not merely disfigure the human form but violently abandons it to focus on animals that reveal animal qualities latent in the subject. Bacon’s “Study of a Baboon”, expresses a sense of human devolution: Man devolved to monkey. In the work, we see a baboon within an enclosure, sitting above a tree that simultaneously resembles a gothic shadow, a cross, and even a smear. The dark, cross-like tree may suggest the conquering of God by a baboon, a type of monkey, recalling the old slander of Darwin’s theory, namely that Darwinism entailed that humanity descended from monkeys (which Darwin’s theory does not claim). But far from victorious, the monkey is in a state of suffering. While the baboon is not crucified on or by the tree, suffering pervades the frame. Its head resembles some sort of skull. The body is faintly painted in a melancholy blue with smudges of purple and is translucent and ghostly—at once a lump of matter and a spectral absence. We do not see the baboon through the cage. Instead we see through the baboon at the cage. Indeed, its very physiology involves the encountering of trauma as the head of the baboon does not simply connect to the body but stabs through the body as a sharp bone, perhaps opaquely evoking the violence of evolution. Similarly, the baboon’s tail seems to stab through the tree. Its eye is an enlarged void and a pupil is indicated by a bluish white triangle splitting through the void. The tree has something of the menacing and looming quality of a shadow and there is a sense of wilderness confronted by death and entrapment, evoked through the background. The yellowy ground is suggestive of dead grass. While potentially gesturing to the psychical confusion and intensity of Vincent Van Gogh or Edvard Munch, the yellowed grass more likely evokes the empty, barren and hostile planes of the desert and contrasts with the darkened colours. The baboon sitting on the cross/tree may seem to have reached some sort of pinnacle but such a status is mocked by the tree that manages to continue outside the fence: the branches nightmarishly protrude through the fence to conquer the frame, which in turn furthers the sense of inescapable entrapment and threat. The baboon is thereby precluded from reaching a higher point on the tree, unable to climb the branches, and underscores the baboon’s confines. The painting is labelled a study, which may suggest it is unfinished. However, Bacon’s completed works preserve an unfinished quality. This unfinished quality conveys a sense in which Man and evolution are unfinished and that being finished in the sense of being completed is no longer possible. The idea that there can finished work of art, a work of art that preserves an eternal meaning, has been repeatedly subject to serious doubt, including by artists themselves. Indeed, Bacon’s work erases the potential for perfection and completion, and breaks down, through devolution, what has been achieved by Man and the forces that shaped him. The subject is lowered from that of human to that of a baboon and is therefore, by Vasari’s Renaissance reasoning, not a subject at all. Bacon’s sketch and study exist to evoke a sense of incompletion, involving pain without resolution. The animal state of pain is therefore married with existential entrapment and isolation as art ceases to express the Platonic ideal and aims to show the truth of the shadow—namely that humanity is without a God, a God that previously shed light on humanity’s condition and anchored the human subject. If there is a trace or echo of human nobility left, such a trace functions through the wild and violent quality of animal indignation. A scream of painful indignity is the last act approaching (or descending from) any dignity that is afforded. Jan Švankmajer’s Darkness, Light, DarknessAn even more extreme case of the subject no longer being the subject, of being broken and muted—so much so that animal protest is annulled—can be witnessed in Jan Švankmajer’s animated short Darkness, Light, Darkness. In the animation, green clay hands mould and form a human body in order to be part of it. But when complete, the human body is trapped, grotesquely out of proportion with its environment. The film begins in a darkened house. There is a knocking of the door, and then the first green hand opens the door and turns on the light. The hand falls to the floor, blindly making its way to another door on the opposite side of the house. The hand opens the door only for eyeballs to roll out. The eyes look around. The hand pushes its clay fingers against the eyeballs, and the eyeballs become attached to the fingers. Suddenly with sight, the hand is able to lift itself up. The hand discovers that another hand is knocking at the door. The first hand helps the second hand, and then goes to the window where a pair of ears are stuck together flapping like a moth. The hands work together and break the ears apart. The first hand, the one with eyes, attaches the ears to the second hand. Then a head with a snout, but missing eyes and ears, enters through the door. The hands pull the snout until it becomes a nose, suppressing and remoulding the animal until it becomes human. As with Bacon, the violence of evolution, of auto-construction is conveyed indirectly: in Bacon’s case, through painted devolution and, in the case of the claymation, through a violent construction based on mutilation and smashing body parts together.Although I have described only three minutes of the seven-minute film, it already presents an image of human construction devoid of art or divine design. Man, or rather the hands, become the blind watchman of evolution. The hands work contingently, with what they are provided. They shape themselves based on need. The body, after all, exists as parts, and the human body is made up of other life forms, both sustaining and being sustained by them. The hands work together, and sacrifice sight and hearing for the head. They tear off the ears and remove the eyes and give them to the head. Transcendence is exchanged for subsistence. The absurdity of this contingency becomes most apparent when the hands attempt to merge with the head, to be the head’s feet. Then the feet actually arrive and are attached to the head’s neck. The human subject in such a state is thereby deformed and incomplete. It is a frightened form, cowering when it hears banging at the door. It turns out that the banging is being produced by an angry erect penis pounding at the door. However, even this symbol of masculine potency is subdued, rendered harmless by the hands that splash a bucket of cold water on it. The introduction of the penis signifies the masculinist notions implicit in the term Man, but we only ever see the penis when it is flaccid. The human subject is able to be concluded when clay pours from both doors and the window. The hands sculpt the clay and make the body, which, when complete is oversized and barely fits within the house. The male subject is then trapped, cramped in a foetal position. With its head against the ceiling next to the light, breathing heavily, all it can do is turn out the light. The head opens its mouth either in horror or a state of exertion and gasps. The eyes bulge before one of the body’s hands turns switch, perhaps suggesting terror before death or simply the effort involved in turning off the light. Once completed and built, the human subject remains in the dark. Despite the evident quirky, playful humour, Švankmajer’s film reflects an exhaustion with art itself. Human life becomes clay comically finding its own form. For Vasari, the ideal of the human form is realised first by God and then by Man through marble; for Švankmajer it is green clay. He demotes man back to the substance for a God to mould but, as there is no God to breathe life into it and give form, there is just the body to imperfectly mould itself. The film challenges both Vasari’s humanism and the suprahumanism of Nietzschean spectacle. Instead of the self-generating power and radical interdependence and agency of Übermensch, Švankmajer’s sub-subject is Man undercome—man beneath as opposed to over man, man mocked by its ambition, and with no space to stand high. Švankmajer thereby realises the anti-Nietzschean potential inherent within cinema’s anti-spectacular nature. Antonin Artaud, who extends the aesthetics advanced by Nietzsche in The Birth of Tragedy, contrasts the theatre’s sense of animal life with cinema. Artaud observes that movies “murder us with second-hand reproductions […] filtered through machines” (84). Thus, films murder creative and animal power as film flattens life to a dead realm of reproduction. Continuing Jacques Derrida’s hauntological framing of the screen, the animation theorist Alan Cholodenko has argued that the screen implies death. Motion is dead and replaced by illusion, a recording relayed back to us. What renders cinema haunting also renders it hauntological. For Cholodenko, cinema’s animation challenges ontology and metaphysics by eschewing stable ontologies through a process that entails both presence and absence. As Cholodenko points out, all film is a type of animation and reanimation, of making images move that are not in fact moving. Thus, one can argue that the animated-animation (such as Švankmajer’s claymation) becomes a refinement of death, a Frankesteinian reanimation of dead material. Indeed, Darkness, Light, Darkness accentuates the presence of death with the green clay almost resembling putrefaction. The fingerprints on the clay accentuate a lack of life, for the autonomous and dead matter that constructs and shapes a dead body from seemingly severed body parts. Even the title of the film, Darkness, Light, Darkness reflects an experience of cinema as deflation rather than joyous spectacle. One goes to a darkened space, watches light flicker on a screen and then the light goes out again. The cartoonish motions of the hands and body parts in the film look only half alive and therefore seem half-dead. Made in the decaying Communist state of Czechoslovakia, Švankmajer’s film aptly acknowledges the deflation of cinema, reflecting that illumination—the light of God, is put out, or more specifically, switched off. With the light of God switched off, creation becomes construction and construction becomes reconstruction, filtered through cinema’s machine processes as framed through Cholodenko. Still, Švankmajer’s animation is not unsympathetic to the plight of the hands. We do see the body parts work together. When a vulgar, meaty, non-claymation tongue comes out through the door, it goes straight to the other door to let the teeth in. The teeth and tongue are aided by the hands to complete the face. Indeed, what they produce is a human being, which has some sense of coherence and success—a success enmeshed with failure and entrapment. Piccinini’s “The Young Family”Patricia Piccinini’s sculptural works offer a more tender approach to the subject, especially when her works focus on the nonhuman animal with human characteristics. Piccinini is interested in the combinations of the animal and the machine, so her ideas can be seen almost as transhuman, where the human is extended beyond humanism. Her work is based on connection and connectedness, but does not emphasise the humanist values of innovation and self-creation often inherent to transhumanism. Indeed, the emphasis on connection is distinct from the entrapment of Bacon’s baboon and Švankmajer’s clay human, which half lament freedom’s negation.The way that Piccinini preserves aspects of humanism within a framework of subhumanism is evident in her work “The Young Family”. The hypperrealistic sculpture depicts a humanoid pig form, flopped, presumably exhausted, as piglet-babies suckle on her nipples. The work was inspired by a scientific proposal for pigs to be genetically modified to provide organs for humans (“Educational Resource” 5). Such a transhuman setting frames a subhuman aesthetic. Care is taken to render the scene with sentiment but without a sense of the ideal, without perfection. One baby-piglet tenderly grasps its foot with both hands and stares with love at its mother. We see two piglets enthusiastically sucking their mother’s teat, while a third baby/piglet’s bottom is visible, indicating that there is a third piglet scrambling for milk. The mother gazes at us, with her naked mammalian body visible. We see her wrinkles and veins. There is some fur on her head and some hair on her eyebrows humanising her. Indeed, her eyes are distinctly human and convey affection. Affection seems to be a motif that carries through to the materials (carefully crafted by Piccinini’s studio). The affection displayed in the artwork is trans-special, emphasising that human tenderness is in fact mammalian tenderness. Such tenderness conflates the human, the nonhuman animal and the material out of which the humanoid creature and its young are constructed. The sub-agency brings together the young and the old by displaying the closeness of the family. Something of this sub-subjectivity is theorised in Malcolm Bull’s Anti-Nietzsche, where he contrasts Nietzsche’s idea of the Übermensch with the idea of the subhuman. Bull writes that subhumanism involves giving up on “becoming more than a man and think[ing] only of becoming something less” (n.p.; Chapter 2, sec. “The Subhuman”). Piccinini depicts vulnerability and tenderness with life forms that are properly speaking subhuman, and reject the displays of strength of Nietzsche’s suprahumanism or Vasari’s emphasis on art commemorating great men. But Piccinini’s subhumanism preserves enough humanism to understand art’s ability to encourage an ethics of nurturing. In this respect, her works offer an alternative to Bull’s subhumanism that aims, so Bull argues, to devalue art altogether. Instead, Piccinini affirms imagination, but through its ability to conjure new ways to perceive animal affection. The sub-subject thereby functions to reveal states of emotion common to mammals (including humans) and other animals. ConclusionThese three artists therefore convey distinct, if related and intersecting, ways of visualising the sub-subject: Bacon through animal suffering, Švankmajer through adaptation that ultimately leads to the agent’s entrapment, and Piccinini who, instead of marrying anti-humanism with the subhumanism (the procedure of Švankmajer, and Bacon), integrates aspects of transhumanism and Renaissance humanism into her subhuman vision. As such, these works present a realisation of how we might think of the going under of the human subject after Darwin, Nietzsche and the deaths of God, Man and the diminishment of creativity. Such works remain not only antithetical to Vasari’s humanism but also to Nietzsche’s suprahumanism. These artists use art’s power to humble—not through overpowering awe but through the visible breakdown of the human agent, speaking for and to the sub-subject. Such art, by unbuilding and dismantling the subject, draws on prehuman trajectories of evolution, and in the case of Piccinini, transhuman trajectories. Art ceases to be about the grandiose evocations of power. Rather, more modestly, these works build a connection between the human with other mammals. Acknowledgements I wish to acknowledge Daniel Canaris for his valuable insights into Christianity and the Italian Renaissance, Alan Cholodenko for providing copies of his works that were central to my interpretation of Švankmajer, and Rachel Franks and Simon Dwyer for their invaluable assistance and finding very helpful reviewers. References Artaud, Antonin. The Theatre and Its Double. New York: Grove P, 1958.Art Gallery of South Australia. “Educational Resource Patricia Piccinini.” Adelaide: Art Gallery of South Australia. 11 Dec. 2016 <https://www.artgallery.sa.gov.au/agsa/home/Learning/docs/Online_Resources/Piccinini_online_resource.pdf>.Bacon, Francis. “Head I.” 1948. Oil on Canvas. 100.3 x 74.9cm. ———. “Study of a Baboon.” 1953. Oil on Canvas. 198.3 x 137.3cm. Bull, Malcolm. Anti-Nietzsche. New York: Verso, 2011. Cholodenko, Alan. “First Principles of Animation.” Animating Film Theory. Ed. Karen Beckman. Duke UP, 2014. 98-110.———. “The Crypt, the Haunted House, of Cinema.” Cultural Studies Review 10.2 (2004): 99-113. Darkness, Light, Darkness. Jan Švankmajer, 1990. 35mm. Nietzsche, Friedrich. The Birth of Tragedy and Other Writings. Cambridge: Cambridge UP, 2007. ———. The Gay Science. Cambridge: Cambridge UP, 2007. ———. Thus Spoke Zarathustra. Cambridge: Cambridge UP, 2006.Piccinini, Patricia. “The Young Family.” 2002. Silicone, Polyurethane, Leather, Plywood, Human Hair, 80 x 150 x 110cm. Vasari, Giorgio. The Lives of Artists. Oxford: Oxford UP, 1998.Wolfe, Cary. What Is Posthumanism? Minneapolis: U of Minnesota P, 2010.