Association among starch storage, metabolism, related genes and growth of Moso bamboo (Phyllostachys heterocycla) shoots.

BACKGROUND: Both underground rhizomes/buds and above-ground Moso bamboo (Phyllostachys heterocycla) shoots/culms/branches are connected together into a close inter-connecting system in which nutrients are transported and shared among each organ. However, the starch storage and utilization mechanisms during bamboo shoot growth remain unclear. This study aimed to reveal in which organs starch was stored, how carbohydrates were transformed among each organ, and how the expression of key genes was regulated during bamboo shoot growth and developmental stages which should lay a foundation for developing new theoretical techniques for bamboo cultivation. RESULTS: Based on changes of the NSC content, starch metabolism-related enzyme activity and gene expression from S0 to S3, we observed that starch grains were mainly elliptical in shape and proliferated through budding and constriction. Content of both soluble sugar and starch in bamboo shoot peaked at S0, in which the former decreased gradually, and the latter initially decreased and then increased as shoots grew. Starch synthesis-related enzymes (AGPase, GBSS and SBE) and starch hydrolase (α-amylase and ß-amylase) activities exhibited the same dynamic change patterns as those of the starch content. From S0 to S3, the activity of starch synthesis-related enzyme and starch amylase in bamboo rhizome was significantly higher than that in bamboo shoot, while the NSC content in rhizomes was obviously lower than that in bamboo shoots. It was revealed by the comparative transcriptome analysis that the expression of starch synthesis-related enzyme-encoding genes were increased at S0, but reduced thereafter, with almost the same dynamic change tendency as the starch content and metabolism-related enzymes, especially during S0 and S1. It was revealed by the gene interaction analysis that AGPase and SBE were core genes for the starch and sucrose metabolism pathway. CONCLUSIONS: Bamboo shoots were the main organ in which starch was stored, while bamboo rhizome should be mainly functioned as a carbohydrate transportation channel and the second carbohydrate sink. Starch metabolism-related genes were expressed at the transcriptional level during underground growth, but at the post-transcriptional level during above-ground growth. It may be possible to enhance edible bamboo shoot quality for an alternative starch source through genetic engineering.

Coleataenia prionitis, a C4-like species in the Poaceae.

C4-like plants represent the penultimate stage of evolution from C3 to C4 plants. Although Coleataenia prionitis (formerly Panicum prionitis) has been described as a C4 plant, its leaf anatomy and gas exchange traits suggest that it may be a C4-like plant. Here, we reexamined the leaf structure and biochemical and physiological traits of photosynthesis in this grass. The large vascular bundles were surrounded by two layers of bundle sheath (BS): a colorless outer BS and a chloroplast-rich inner BS. Small vascular bundles, which generally had a single BS layer with various vascular structures, also occurred throughout the mesophyll together with BS cells not associated with vascular tissue. The mesophyll cells did not show a radial arrangement typical of Kranz anatomy. These features suggest that the leaf anatomy of C. prionitis is on the evolutionary pathway to a complete C4 Kranz type. Phosphoenolpyruvate carboxylase (PEPC) and pyruvate, Pi dikinase occurred in the mesophyll and outer BS. Glycine decarboxylase was confined to the inner BS. Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) accumulated in the mesophyll and both BSs. C. prionitis had biochemical traits of NADP-malic enzyme type, whereas its gas exchange traits were close to those of C4-like intermediate plants rather than C4 plants. A gas exchange study with a PEPC inhibitor suggested that Rubisco in the mesophyll could fix atmospheric CO2. These data demonstrate that C. prionitis is not a true C4 plant but should be considered as a C4-like plant.

Sustainable Drying and Torrefaction Processes of Miscanthus for Use as a Pelletized Solid Biofuel and Biocarbon-Carrier for Fertilizers.

Miscanthus is resistant to dry, frosty winters in Poland and most European Union countries. Miscanthus gives higher yields compared to native species. Farmers can produce Miscanthus pellets after drying it for their own heating purposes. From the third year, the most efficient plant development begins, resulting in a yield of 25-30 tons of dry matter from an area of 1 hectare. Laboratory scale tests were carried out on the processes of drying, compacting, and torrefaction of this biomass type. The analysis of the drying process was conducted at three temperature levels of the drying agent (60, 100, and 140 °C). Compaction on a hydraulic press was carried out in the pressure range characteristic of a pressure agglomeration (130.8-457.8 MPa) at different moisture contents of the raw material (0.5% and 10%). The main interest in this part was to assess the influence of drying temperature, moisture content, and compaction pressure on the specific densities (DE) and the mechanical durability of the pellets (DU). In the next step, laboratory analyses of the torrefaction process were carried out, initially using the Thermogravimetric Analysis TGA and Differential Scaning Calorimeter DSC techniques (to assess activation energy (EA)), followed by a flow reactor operating at five temperature levels (225, 250, 275, 300, and 525 °C). A SEM analysis of Miscanthus after torrefaction processes at three different temperatures was performed. Both the parameters of biochar (proximate and ultimate analysis) and the quality of the torgas (volatile organic content (VOC)) were analyzed. The results show that both drying temperature and moisture level will affect the quality of the pellets. Analysis of the torrefaction process shows clearly that the optimum process temperature would be around 300-340 °C from a mass loss ratio and economical perspective.

Phragmites australis (Reed) as an Efficient, Eco-Friendly Adsorbent for Brackish Water Pre-Treatment in Reverse Osmosis: A Kinetic Study.

A cost-effective adsorbent was prepared by carbonization of pre-treated Phragmites australis reed at 500 °C. Phragmites australis was characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Brunauer-Emmett-Teller (BET) surface analyses. XRD of the as-prepared adsorbent exhibited a partially crystalline structure with a specific surface area of 211.6 m2/g and an average pore diameter of 4.2 nm. The biosorption potential of novel biosorbent Phragmites australis reed was investigated with a batch scale and continuous flow study. The study was conducted at different constraints to obtain optimum pH conditions, adsorbent dose, contact time, agitation speed, and initial TDS concentration. In order to analyze the properties of the procedure and determine the amount of sodium removal, Langmuir, Freundlich, and Dubinin-Radushkevich isotherms were tested. The optimal values of contact time, pH, and adsorbent dose were found to be 150 min, 4, and 10 g/L, respectively, with an agitation speed of 300 rpm at room temperature (27 °C). The three tested isotherms show that the adsorption of Na+ onto the prepared adsorbent is a hybrid process from physi- and chemisorption. For industrial application, the adsorbent was tested using the adsorbent column technique. Pseudo-first-order, pseudo-second-order, and diffusion models were connected, and it was discovered that the information fit best to the pseudo-second-arrange active model. According to the intraparticle diffusion model, the mechanism goes through four stages before reaching equilibrium. The periodicity test shows that the adsorption ability of Phragmites australis can be recovered by washing with 0.1 M HCl.

Sucrose phosphate synthase (SPS), sucrose synthase (SUS) and their products in the leaves of Miscanthus × giganteus and Zea mays at low temperature.

MAIN CONCLUSION: The changes in the expression of key sugar metabolism enzymes (SPS and SUS), sucrose content and arrangement of chloroplast starch may play a significant role in the cold response in M. giganteus and maize plants. To understand the mechanism of the chilling-response of two closely-related C4 plants, we investigated the changes in the expression of sucrose phosphate synthase (SPS) and sucrose synthase (SUS) as well as changes in their potential products: sucrose, cellulose and starch in the leaves of Miscanthus × giganteus and Zea mays. Low temperature (12-14 °C) increased SPS content in Miscanthus (MG) and chilling-sensitive maize line (Zm-S), but not in chilling-tolerant one (Zm-T). In Zm-S line, chilling also caused the higher intensity of labelling of SPS in the cytoplasm of mesophyll cells, as demonstrated by electron microscopy. SUS labelling was also increased by cold stress only in MG plants what was observed in the secondary wall between mesophyll and bundle sheath cells, as well as in the vacuoles of companion cells. Cold led to a marked increase in total starch grain area in the chloroplasts of Zm-S line. In turn, Fourier transform infrared spectroscopy (FTIR) showed a slight shift in the cellulose band position, which may indicate the formation of more compact cellulose arrangement in Zm-T maize line. In conclusion, this work presents new findings supporting diversified cold-response, not only between two C4 plant species but also within one species of maize.

Phylogenetic relationships within Parianinae (Poaceae: Bambusoideae: Olyreae) with emphasis on Eremitis: Evidence from nuclear and plastid DNA sequences, macromorphology, and pollen ectexine patterns.

Eremitis, Pariana, and Parianella are herbaceous bamboos (tribe Olyreae) included in the subtribe Parianinae, which is characterized by the presence of fimbriae at the apex of the leaf sheaths and exclusively spiciform synflorescences. We analyzed 43 samples of herbaceous and woody bamboos in order to infer relationships within the Parianinae, based on combined data from the nuclear ribosomal internal transcribed spacer (ITS) and plastid DNA (rpl32-trnL and trnD-trnT spacers). Bayesian inference, maximum likelihood, and maximum parsimony methods were applied, and macro- and micromorphological aspects were also analyzed, including the ectexine patterns of pollen grains. Parianinae is represented by three well-supported lineages in our analyses: (1) Parianella, endemic to southern Bahia, Brazil; (2) Pariana sensu stricto with a broad distribution in southern Central America and northern South America, especially in the Amazon region; and (3) Eremitis, endemic to the Brazilian Atlantic Forest, from the states of Pernambuco to Rio de Janeiro, including one species previously described as a member of Pariana. Our molecular phylogeny showed that Pariana, as historically circumscribed, is not monophyletic, by recovering Pariana sensu stricto as strongly supported and sister to Eremitis + Pariana multiflora, with Parianella sister to the Pariana-Eremitis clade. Morphological features of their synflorescences and differences in ectexine patterns characterize each lineage. Based on all these characters and the phylogenetic results, Pariana multiflora, endemic to the state of Espírito Santo, Brazil, is transferred to Eremitis.

Decay of the water reed Phragmites communis caused by the white-rot fungus Phlebia tremellosa and the influence of some environmental factors.

The strain Phlebia tremellosa SBUG 1630 isolated from a thatched roof in Northern Germany is capable of colonizing and degrading effectively the water reed Phragmites communis. Within 96 h after inoculation, mycelia covered both the outer and the inner surface of reed shoot fragments as observed by scanning electron microscopy. Interestingly, top culm sections and culm edges were particularly susceptible towards fungal degradation. The weight loss of culms reached 20-73% depending on the environmental conditions applied during the incubation of 70 days. Reed degradation was stable at pH 4 to pH 8 and optimal between 25 and 30 °C. Short-term incubation at elevated temperatures (37 to 55 °C) affected the fungal reed degradation to only a minor extent, whereas > 18 h at 55 °C completely inhibited fungal growth and reed degradation. Supplementation with 43 mM NH4Cl enhanced the reed degradation up to 9%. In contrast, the addition of diammonium tartrate increased the weight loss of the samples considerably up to 16% at 344 mM. Furthermore, reed degradation by P. tremellosa was increased by supplementing the test medium with Mn (99 to 1584 µM), Cu (150 to 300 µM), and less significantly phosphate (4 mM), Zn (37 to 74 µM), and Ag (76 µM) after 70 days. In addition, activities of the ligninolytic enzymes laccase (max. 27.4 nmol ml-1 min-1) and lignin peroxidase (max. 22.8 nmol ml-1 min-1) were rather low in nitrogen-limited medium, whereas considerably higher levels of manganese peroxidase (max. 635.9 nmol ml-1 min-1) were observed.

Exploring key cellular processes and candidate genes regulating the primary thickening growth of Moso underground shoots.

The primary thickening growth of Moso (Phyllostachys edulis) underground shoots largely determines the culm circumference. However, its developmental mechanisms remain largely unknown. Using an integrated anatomy, mathematics and genomics approach, we systematically studied cellular and molecular mechanisms underlying the growth of Moso underground shoots. We discovered that the growth displayed a spiral pattern and pith played an important role in promoting the primary thickening process of Moso underground shoots and driving the evolution of culms with different sizes among different bamboo species. Different with model plants, the shoot apical meristem (SAM) of Moso is composed of six layers of cells. Comparative transcriptome analysis identified a large number of genes related to the vascular tissue formation that were significantly upregulated in a thick wall variant with narrow pith cavity, mildly spiral growth, and flat and enlarged SAM, including those related to plant hormones and those involved in cell wall development. These results provide a systematic perspective on the primary thickening growth of Moso underground shoots, and support a plausible mechanism resulting in the narrow pith cavity, weak spiral growth but increased vascular bundle of the thick wall Moso.

Stomatal cell wall composition: distinctive structural patterns associated with different phylogenetic groups.

Background and Aims: Stomatal morphology and function have remained largely conserved throughout ∼400 million years of plant evolution. However, plant cell wall composition has evolved and changed. Here stomatal cell wall composition was investigated in different vascular plant groups in attempt to understand their possible effect on stomatal function. Methods: A renewed look at stomatal cell walls was attempted utilizing digitalized polar microscopy, confocal microscopy, histology and a numerical finite-elements simulation. The six species of vascular plants chosen for this study cover a broad structural, ecophysiological and evolutionary spectrum: ferns ( Asplenium nidus and Platycerium bifurcatum ) and angiosperms ( Arabidopsis thaliana and Commelina erecta ) with kidney-shaped stomata, and grasses (angiosperms, family Poaceae) with dumbbell-shaped stomata ( Sorghum bicolor and Triticum aestivum ). Key Results: Three distinct patterns of cellulose crystallinity in stomatal cell walls were observed: Type I (kidney-shaped stomata, ferns), Type II (kidney-shaped stomata, angiosperms) and Type III (dumbbell-shaped stomata, grasses). The different stomatal cell wall attributes investigated (cellulose crystallinity, pectins, lignin, phenolics) exhibited taxon-specific patterns, with reciprocal substitution of structural elements in the end-walls of kidney-shaped stomata. According to a numerical bio-mechanical model, the end walls of kidney-shaped stomata develop the highest stresses during opening. Conclusions: The data presented demonstrate for the first time the existence of distinct spatial patterns of varying cellulose crystallinity in guard cell walls. It is also highly intriguing that in angiosperms crystalline cellulose appears to have replaced lignin that occurs in the stomatal end-walls of ferns serving a similar wall strengthening function. Such taxon-specific spatial patterns of cell wall components could imply different biomechanical functions, which in turn could be a consequence of differences in environmental selection along the course of plant evolution.

Museomics resolve the systematics of an endangered grass lineage endemic to north-western Madagascar.

BACKGROUND AND AIMS: Recent developments in DNA sequencing, so-called next-generation sequencing (NGS) methods, can help the study of rare lineages that are known from museum specimens. Here, the taxonomy and evolution of the Malagasy grass lineage Chasechloa was investigated with the aid of NGS. METHODS: Full chloroplast genome data and some nuclear sequences were produced by NGS from old herbarium specimens, while some selected markers were generated from recently collected Malagasy grasses. In addition, a scanning electron microscopy analysis of the upper floret and cross-sections of the rachilla appendages followed by staining with Sudan IV were performed on Chasechloa to examine the morphology of the upper floret and the presence of oils in the appendages. KEY RESULTS: Chasechloa was recovered within tribe Paniceae, sub-tribe Boivinellinae, contrary to its previous placement as a member of the New World genus Echinolaena (tribe Paspaleae). Chasechloa originated in Madagascar between the Upper Miocene and the Pliocene. It comprises two species, one of them collected only once in 1851. The genus is restricted to north-western seasonally dry deciduous forests. The appendages at the base of the upper floret of Chasechloa have been confirmed as elaiosomes, an evolutionary adaptation for myrmecochory. CONCLUSIONS: Chasechloa is reinstated at the generic level and a taxonomic treatment is presented, including conservation assessments of its species. Our study also highlights the power of NGS technology to analyse relictual or probably extinct groups.

Intracellular position of mitochondria in mesophyll cells differs between C3 and C4 grasses.

In C3 plants, part of the CO2 fixed during photosynthesis in chloroplasts is released from mitochondria during photorespiration by decarboxylation of glycine via glycine decarboxylase (GDC), thereby reducing photosynthetic efficiency. The apparent positioning of most mitochondria in the interior (vacuole side of chloroplasts) of mesophyll cells in C3 grasses would increase the efficiency of refixation of CO2 released from mitochondria by ribulose 1,5-bisphosphate carboxylase/​oxygenase (Rubisco) in chloroplasts. Therefore, in mesophyll cells of C4 grasses, which lack both GDC and Rubisco, the mitochondria ought not to be positioned the same way as in C3 mesophyll cells. To test this hypothesis, we investigated the intracellular position of mitochondria in mesophyll cells of 14 C4 grasses of different C4 subtypes and subfamilies (Chloridoideae, Micrairoideae, and Panicoideae) and a C3-C4 intermediate grass, Steinchisma hians, under an electron microscope. In C4 mesophyll cells, most mitochondria were positioned adjacent to the cell wall, which clearly differs from the positioning in C3 mesophyll cells. In S. hians mesophyll cells, the positioning was similar to that in C3 cells. These results suggest that the mitochondrial positioning in C4 mesophyll cells reflects the absence of both GDC and Rubisco in the mesophyll cells and the high activity of phosphoenolpyruvate carboxylase. In contrast, the relationship between the mitochondrial positioning and enzyme distribution in S. hians is complex, but the positioning may be related to the capture of respiratory CO2 by Rubisco. Our study provides new possible insight into the physiological role of mitochondrial positioning in photosynthetic cells.

Evolutionary divergence of ß-expansin structure and function in grasses parallels emergence of distinctive primary cell wall traits.

Expansins are wall-loosening proteins that promote the extension of primary cell walls without the hydrolysis of major structural components. Previously, proteins from the EXPA (α-expansin) family were found to loosen eudicot cell walls but to be less effective on grass cell walls, whereas the reverse pattern was found for EXPB (ß-expansin) proteins obtained from grass pollen. To understand the evolutionary and structural bases for the selectivity of EXPB action, we assessed the extension (creep) response of cell walls from diverse monocot families to EXPA and EXPB treatments. Cell walls from Cyperaceae and Juncaceae (families closely related to grasses) displayed a typical grass response ('ß-response'). Walls from more distant monocots, including some species that share with grasses high levels of arabinoxylan, responded preferentially to α-expansins ('α-response'), behaving in this regard like eudicots. An expansin with selective activity for grass cell walls was detected in Cyperaceae pollen, coinciding with the expression of genes from the divergent EXPB-I branch that includes grass pollen ß-expansins. The evolutionary origin of this branch was located within Poales on the basis of phylogenetic analyses and its association with the 'sigma' whole-genome duplication. Accelerated evolution in this branch has remodeled the protein surface in contact with the substrate, potentially for binding highly substituted arabinoxylan. We propose that the evolution of the divergent EXPB-I group made a fundamental change in the target and mechanism of wall loosening in the grass lineage possible, involving a new structural role for xylans and the expansins that target them.

Physiological significance of isoprenoids and phenylpropanoids in drought response of Arundinoideae species with contrasting habitats and metabolism.

Physiological, biochemical and morpho-anatomical traits that determine the phenotypic plasticity of plants under drought were tested in two Arundinoideae with contrasting habitats, growth traits and metabolism: the fast-growing Arundo donax, which also is a strong isoprene emitter, and the slow-growing Hakonechloa macra that does not invest on isoprene biosynthesis. In control conditions, A. donax displayed not only higher photosynthesis but also higher concentration of carotenoids and lower phenylpropanoid content than H. macra. In drought-stressed plants, photosynthesis was similarly inhibited in both species, but substantially recovered only in A. donax after rewatering. Decline of photochemical and biochemical parameters, increased concentration of CO2 inside leaves, and impairment of chloroplast ultrastructure were only observed in H. macra indicating damage of photosynthetic machinery under drought. It is suggested that volatile and non-volatile isoprenoids produced by A. donax efficiently preserve the chloroplasts from transient drought damage, while H. macra invests on phenylpropanoids that are less efficient in preserving photosynthesis but likely offer better antioxidant protection under prolonged stress.

Structural Insight into Cell Wall Architecture of Micanthus sinensis cv. using Correlative Microscopy Approaches.

Structural organization of the plant cell wall is a key parameter for understanding anisotropic plant growth and mechanical behavior. Four imaging platforms were used to investigate the cell wall architecture of Miscanthus sinensis cv. internode tissue. Using transmission electron microscopy with potassium permanganate, we found a great degree of inhomogeneity in the layering structure (4-9 layers) of the sclerenchymatic fiber (Sf). However, the xylem vessel showed a single layer. Atomic force microscopy images revealed that the cellulose microfibrils (Mfs) deposited in the primary wall of the protoxylem vessel (Pxv) were disordered, while the secondary wall was composed of Mfs oriented in parallel in the cross and longitudinal section. Furthermore, Raman spectroscopy images indicated no variation in the Mf orientation of Pxv and the Mfs in Pxv were oriented more perpendicular to the cell axis than that of Sfs. Based on the integrated results, we have proposed an architectural model of Pxv composed of two layers: an outermost primary wall composed of a meshwork of Mfs and inner secondary wall containing parallel Mfs. This proposed model will support future ultrastructural analysis of plant cell walls in heterogeneous tissues, an area of increasing scientific interest particularly for liquid biofuel processing.

[Valve-like structures in motor cells of the leaf of Distichlis spicata "Yesen 4A" (Poaceae)].

The salt gland of the leaf of Distichlis directly contacts the cells of interfascicular aquiferous parenchyma (motor cells). The cytoplasmic strand of motor cells produce deep invaginations, which form with the participation of mitochondria. The constriction of the cytoplasmic strand at the site of the localization of mitochondria leads to the fusion of the tonoplast and plasmalemma with mitochondrial membranes and the formation of a thin one-layer plate, a valve. At this locus, vacuolar and apoplast spaces are separated only by a valve. The cytoplasm of motor cells is filled with electron dense granules, which are considered as contractile elements. It is assumed that the cytoplasmic strand is involved in the reduction of the volume, which results in the generation of pressure on the valve. This leads to the direct throw-in of water into the apoplast space adjacent to the salt gland.

Visualisation by high resolution synchrotron X-ray phase contrast micro-tomography of gas films on submerged superhydrophobic leaves.

Floods can completely submerge terrestrial plants but some wetland species can sustain O2 and CO2 exchange with the environment via gas films forming on superhydrophobic leaf surfaces. We used high resolution synchrotron X-ray phase contrast micro-tomography in a novel approach to visualise gas films on submerged leaves of common cordgrass (Spartina anglica). 3D tomograms enabled a hitherto unmatched level of detail regarding the micro-topography of leaf gas films. Gas films formed only on the superhydrophobic adaxial leaf side (water droplet contact angle, Φ=162°) but not on the abaxial side (Φ=135°). The adaxial side of the leaves of common cordgrass is plicate with a longitudinal system of parallel grooves and ridges and the vast majority of the gas film volume was found in large ∼180µm deep elongated triangular volumes in the grooves and these volumes were connected to each neighbouring groove via a fine network of gas tubules (∼1.7µm diameter) across the ridges. In addition to the gas film retained on the leaf exterior, the X-ray phase contrast micro-tomography also successfully distinguished gas spaces internally in the leaf tissues, and the tissue porosity (gas volume per unit tissue volume) ranged from 6.3% to 20.3% in tip and base leaf segments, respectively. We conclude that X-ray phase contrast micro-tomography is a powerful tool to obtain quantitative data of exterior gas features on biological samples because of the significant difference in electron density between air, biological tissues and water.

Bundle sheath suberization in grass leaves: multiple barriers to characterization.

High-yielding, stress-tolerant grass crops are essential to meet future food and energy demands. Efforts are underway to engineer improved varieties of the C3 cereal crop rice by introducing NADP-malic enzyme C4 photosynthesis using maize as a model system. However, several modifications to the rice leaf vasculature are potentially necessary, including the introduction of suberin lamellae into the bundle sheath cell walls. Suberized cell walls are ubiquitous in the root endodermis of all grasses, and developmental similarities are apparent between endodermis and bundle sheath cell walls. Nonetheless, there is considerable heterogeneity in sheath cell development and suberin composition both within and between grass taxa. The effect of this variation on physiological function remains ambiguous over forty years after suberin lamellae were initially proposed to regulate solute and photoassimilate fluxes and C4 gas exchange. Interspecies variation has confounded efforts to ascribe physiological differences specifically to the presence or absence of suberin lamellae. Thus, specific perturbation of suberization within a uniform genetic background is needed, but, until recently, the genetic resources to manipulate suberin composition in the grasses were largely unavailable. The recent dissection of the suberin biosynthesis pathway in model dicots and the identification of several promising candidate genes in model grasses will facilitate the characterization of the first suberin biosynthesis genes in a monocot. Much remains to be learned about the role of bundle sheath suberization in leaf physiology, but the stage is set for significant advances in the near future.

Effect of mild alkali pretreatment on structural changes of reed (Phragmites communis Trinius) straw.

The effect of dilute sodium hydroxide (NaOH) on reed straw structural change at 105 degreeC temperature was evaluated in this study. Various concentrations of NaOH (1% to 2.5%) were used for pretreatment of reed straw at 105 degreeC for 10min. Scanning electron microscopy, atomic force microscopy and Fourier transform infrared spectroscopy studies showed that 2% and 2.5% NaOH pretreated sample exposed more cellulose fibers compared with other treatments. The cellulose crystalline index was increased by the 1% to 2.0% NaOH treatments and slightly lowered by the 2.5% NaOH treatment due to destructing cellulose fibres. Two per cent NaOH pretreatment caused 69.9% lignin removal, whereas 2.5% NaOH pretreatment removed 72.4% lignin. Besides, reed straw, when pretreated at 2% and 2.5% NaOH, resulted 56.4% and 60.5% hemicellulose removal, respectively. However, the difference in removal of lignin and hemicellulose between 2% and 2.5% NaOH treated reed straw was very marginal. In addition, very negligible increase of cellulose level was estimated, amounting 78.8% and 76.6% in 2.5% and 2% NaOH-treated sample, respectively. Moreover, after 72 h, reducing sugar yield was 81.2% and 83.3% using enzyme loading of 15 FPU (g dry biomass)-' and 30 IU (g dry biomass)- and xylanase 4 FXU (g dry biomass)-1 from 2% and 2.5% NaOH pretreated reed straw, respectively. Reducing sugar yield was increased very marginally when NaOH concentration increased from 2% to 2.5% for reed straw pretreatment. Therefore, 2% NaOH is supposed to be effective for reed straw pretreatment at this mentioned condition.

Effects of a steam explosion pretreatment on sugar production by enzymatic hydrolysis and structural properties of reed straw.

Reed lignocellulose was subjected to a steam explosion pretreatment to obtain a high conversion rate of sugar after subsequent enzymatic hydrolysis using a commercial cellulase mixture. Under conditions of differing temperature (200 °C, 220 °C and 240 °C) and residence time (2, 5, and 8 min), the effect of the pretreatment on the sugar yield from enzymatic hydrolysis was studied. The highest respective reducing sugar and glucose yields were 36.14% and 15.35% after 60-h enzymatic hydrolysis of reed straw that had been pretreated with a steam explosion at 220 °C for 5 min. Fourier transform infrared spectrophotometry (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used in this study to comprehensively investigate the steam explosion-induced changes in the organizational structure and morphological properties of reed straw to analyze the reason for the increased sugar yield from enzymatic hydrolysis after the steam explosion.

Autoimmune response and repression of mitotic cell division occur in inter-specific crosses between tetraploid wheat and Aegilops tauschii Coss. that show low temperature-induced hybrid necrosis.

Common wheat is an allohexaploid species originating from a naturally occurring inter-specific cross between tetraploid wheat and the diploid wild wheat Aegilops tauschii Coss. Artificial allopolyploidization can produce synthetic hexaploid wheat. However, synthetic triploid hybrids show four types of hybrid growth abnormalities: type II and III hybrid necrosis, hybrid chlorosis, and severe growth abortion. Of these hybrid abnormalities, type II necrosis is induced by low temperature. Under low temperature, elongation of stems and expansion of new leaves is repressed in type II necrosis lines, which later exhibit necrotic symptoms. Here, we characterize type II necrosis in detail. Comparative transcriptome analysis showed that a number of defense-related genes were highly up-regulated in seedling leaves that showed type II necrosis. Transmission electron microscopy revealed extensive cell death in the leaves under low-temperature conditions, accompanied by abundant generation of reactive oxygen species. In addition, down-regulation of cell cycle-related genes was observed in shoot apices of type II necrosis lines under low-temperature conditions. Quantitative RT-PCR and in situ hybridization showed repression of accumulation of histone H4 transcripts in the shoot apical meristem of type II necrosis lines. These results strongly suggest that an autoimmune response-like reaction and repression of cell division in the shoot apical meristem are associated with the abnormal growth phenotype in type II necrosis lines.