The Fine Structure of Young Tracheary Xylem Elements Arising by Redifferentiation of Parenchyma in Wounded Coleus Stem

The dense granulation characteristic of parenchyma cells redifferentiatinginto tracheary xylem elements was found to be due to a concentrationof organelles and associated vesicles in the cytoplasmic bandswithin which the secondary-wall thickenings form. Among theseorganelles are mitochondria, plastid-like bodies, elements ofthe endoplasmic reticulum, and Golgi-structures. Numerous smallvesicles occur in the cytoplasm adjacent to developing wallbands and occasionally within the bands themselves. As the secondary-wall reticulations develop, they stain moreintensely with permanganate, becoming increasingly more electrondense. The evidence suggests that this is due to their progressivelignification. The electron density appears also in the primarywall at the bases of the bands, and extends through it betweenopposite bands of adjacent cells.     

Cell Wall Thickening of the Xylem Tracheary Elements in Different Zones of the Adventitious Root of Allium cepa L.

Cell wall thickness of the xylem tracheary elements was measuredin the proto- and metaxylem of the Allium cepa L. adventitiousroot. Measurements were taken in root fragments of known age(1, 3, 5, 7 and 9 d) located in either the basal or medio-apicalzone. Tracheary elements in the protoxylem matured within ashorter period of time than those in the metaxylem. Final cellwall thickness was greater in metaxylem than in protoxylem components.The cell wall thickening in the tracheary elements in both proto-and metaxylem was more rapid in the basal zone of the root thanin the medio-apical zone. Additionally, cell walls of the maturetracheary elements were thicker in the basal zone than in areasfurther from the bulb. Allium cepa, onion, root, cell wall, xylem maturation     

Structure of Cellulose Microfibrils in Primary Cell Walls from Collenchyma

In the primary walls of growing plant cells, the glucose polymer cellulose is assembled into long microfibrils a few nanometers in diameter. The rigidity and orientation of these microfibrils control cell expansion; therefore, cellulose synthesis is a key factor in the growth and morphogenesis of plants. Celery (Apium graveolens) collenchyma is a useful model system for the study of primary wall microfibril structure because its microfibrils are oriented with unusual uniformity, facilitating spectroscopic and diffraction experiments. Using a combination of x-ray and neutron scattering methods with vibrational and nuclear magnetic resonance spectroscopy, we show that celery collenchyma microfibrils were 2.9 to 3.0 nm in mean diameter, with a most probable structure containing 24 chains in cross section, arranged in eight hydrogen-bonded sheets of three chains, with extensive disorder in lateral packing, conformation, and hydrogen bonding. A similar 18-chain structure, and 24-chain structures of different shape, fitted the data less well. Conformational disorder was largely restricted to the surface chains, but disorder in chain packing was not. That is, in position and orientation, the surface chains conformed to the disordered lattice constituting the core of each microfibril. There was evidence that adjacent microfibrils were noncovalently aggregated together over part of their length, suggesting that the need to disrupt these aggregates might be a constraining factor in growth and in the hydrolysis of cellulose for biofuel production.Growth and form in plants are controlled by the precisely oriented expansion of the walls of individual cells. The driving force for cell expansion is osmotic, but the rate and direction of expansion are controlled by the mechanical properties of the cell wall (Szymanski and Cosgrove, 2009). Expanding, primary cell walls are nanocomposite materials in which long microfibrils of cellulose, a few nanometers in diameter, run through a hydrated matrix of xyloglucans, pectins, and other polymers (Knox, 2008; Mohnen, 2008; Szymanski and Cosgrove, 2009; Scheller and Ulvskov, 2010). Native cellulose microfibrils are partially crystalline (Nishiyama, 2009; Fernandes et al., 2011). Formerly, primary wall cellulose was thought to have a unique crystal structure called cellulose IV₁ (Dinand et al., 1996), but NMR evidence suggests the presence of forms similar to the better characterized cellulose Iα and Iβ crystalline forms together with large quantities of less ordered cellulose (Wickholm et al., 1998; Sturcová et al., 2004; Wada et al., 2004). Nevertheless, cellulose is much more ordered than any other component of the primary cell wall (Bootten et al., 2004), in keeping with its key role of providing strength and controlling growth.The stiffness of the cell wall is greatest in the direction of the cellulose microfibrils, where growth is directional and the predominant microfibril orientation is usually transverse to the growth direction (Green, 1999; MacKinnon et al., 2006; Szymanski and Cosgrove, 2009). Expansion of the cell wall then requires either widening of the spacing between microfibrils (Marga et al., 2005) or slippage between them (Cosgrove, 2005), or both, and the microfibrils reorient toward the direction of growth (Anderson et al., 2010). Polymer cross bridges between microfibrils (McCann et al., 1990) are thought to resist these deformations of the cell wall nanostructure and, thus, to control the rate of growth. Until recently, most attention was focused on bridging xyloglucans, hydrogen bonded to microfibril surfaces (Scheller and Ulvskov, 2010). However, there is evidence that not all xyloglucans are appropriately positioned (Fujino et al., 2000; Park and Cosgrove, 2012a) and that other bridging polymers may be involved (Zykwinska et al., 2007). It has also been suggested that bundles of aggregated microfibrils, not single microfibrils, might be the key structural units in primary cell walls (Anderson et al., 2010), as in wood (Fahlén and Salmén, 2005; Fernandes et al., 2011). If so, single microfibrils could bridge between microfibril bundles. In summary, the growth of plant cells is not well understood, and we need more information on how cellulose orientation is controlled and on the nature of the bridging polymers, the cellulose surfaces to which these polymers bind, and the cohesion between microfibril surfaces that might mediate aggregation.Cellulose microfibrils are synthesized at the cell surface by large enzyme complexes having hexagonal symmetry, sometimes called “rosettes” (Somerville, 2006). Each complex contains multiple cellulose synthases that differ between primary cell walls and wood, although the appearance of the complexes is similar (Somerville, 2006; Atanassov et al., 2009). The simultaneous synthesis, from the same end, of all the chains in a native cellulose microfibril is why they are parallel (Nishiyama et al., 2002, 2003), in contrast to the entropically favored antiparallel structure found in man-made celluloses like rayon (Langan et al., 2001). The number of chains in a microfibril and the number of cellulose synthases in the synthetic complex are evidently related. It is commonly assumed that the number of chains is divisible by six, matching the hexagonal rosette symmetry, and 36-chain models (Himmel et al., 2007) bounded by the hydrophilic [110] and [1-10] crystal faces, as in algal celluloses (Bergenstråhle et al., 2008), have been widely adopted. The assembly and orientation of cellulose are connected, as several cellulose synthase mutants have phenotypes defective in cellulose orientation and plant form as well as depleted in cellulose content (Paredez et al., 2008). In certain other mutant lines, the crystallinity of the microfibrils appears to be affected (Fujita et al., 2011; Harris et al., 2012; Sánchez-Rodríguez et al., 2012).Therefore, a detailed understanding of the structure of primary wall cellulose microfibrils would help us to understand cellulose synthesis as well as the growth and structural mechanics of living plants (Burgert and Fratzl, 2009). Primary cell walls and their cellulose skeletons also affect food quality characteristics like the crispness of salad vegetables and apples (Malus domestica; Jarvis, 2011). When biofuels are produced from lignocellulosic biomass, lignification leads to recalcitrance (Himmel et al., 2007), but some of the cell types in Miscanthus spp., switchgrass (Panicum virgatum), and arable crop residues have only primary walls with no lignin, and recalcitrance then depends on the nature of the cellulose microfibrils (Beckham et al., 2011).A relatively detailed structure has recently been proposed for the microfibrils of spruce (Picea spp.) wood (Fernandes et al., 2011), which are 3.0 nm in diameter, allowing space for only about 24 cellulose chains. Evidence from x-ray diffraction supported a “rectangular” shape (Matthews et al., 2006) bounded by the [010] and [200] faces. There was considerable disorder increasing toward the surface, and the microfibrils were aggregated into bundles about 15 to 20 nm across, with some, but not all, of the lateral interfaces being resistant to water (Fernandes et al., 2011). Disordered domains are a feature of other strong biological materials such as spider silk (van Beek et al., 2002).Therefore, it is of interest whether any of these features of wood cellulose might also be found in the cellulose microfibrils of primary (growing) cell walls. It would be particularly useful to characterize the disorder known to be present in primary wall microfibrils, that is, to define how cellulose that is not measured as “crystalline” differs from crystalline cellulose. Many of the experiments leading toward a structure for wood cellulose were dependent on exceptionally uniform orientation of the cellulose microfibrils (Sturcová et al., 2004; Fernandes et al., 2011). However, in growing cell walls, the microfibrils are not uniformly oriented. When microfibrils are first laid down at the inner face of the primary cell wall, their orientation is normally transverse to the direction of growth, but as the cell wall expands, the microfibrils reorient so that the orientation distribution, integrated across the thickness of the expanded cell wall, becomes progressively closer to random (Cosgrove, 2005; MacKinnon et al., 2006).This technical problem does not apply to the cell walls of celery (Apium graveolens) collenchyma, which are similar in composition to other primary cell walls but have their microfibrils oriented relatively uniformly along the cell axis (Sturcová et al., 2004; Kennedy et al., 2007a, 2007b). Some structural information on celery collenchyma cellulose has already been derived from spectroscopic and scattering experiments (Sturcová et al., 2004; Kennedy et al., 2007a, 2007b), confirming the disorder expected in a primary wall cellulose. Some of these experiments were analogous to what has been done on spruce cellulose (Fernandes et al., 2011), but insufficient data are available to specify the number of chains in each primary wall microfibril, the nature and location of the disorder, and the presence or absence of direct contact between microfibrils. Here, we report x-ray and neutron scattering and spectroscopic experiments addressing these questions and leading to a proposed structure for primary wall cellulose microfibrils. Characterizing a structure containing so much disorder presented unusual challenges, but disorder appears to be central to the enigmatic capacity of primary wall cellulose to provide high strength and yet to permit and control growth.     

Factors affecting the growth of Eucalyptus delegatensis seedlings in inhibitory forest and grassland soils

In many highland forests of Eucalyptus delegatensis in Tasmania the establishment and healthy growth of eucalypts is promoted and maintained by fire. In the absence of fire, secondary succession from eucalypt forest to rainforest occurs, during which the eucalypts decline and die prematurely. On sites that are prone to radiation frost severe reduction or removal of a tree canopy allows a sward of tussock grasses to develop, in competition with which seedlings of eucalypts decline in growth and a high proportion dies.Factors of the soil that could contribute to these phenomena were investigated by means of pot experiments that used soils from:o     

蹄盖蕨科5种植物管状分子的研究

采用扫描电镜观察了国产蹄盖蕨科3属5种植物的管状分子,发现安蕨属、拟鳞毛蕨属和蹄盖蕨属管状分子结构类似,具体可分为3种类型:(1)梯状穿孔板,无穿孔板二型性现象;(2)梯状穿孔板,有穿孔板二型性现象;(3)梯状-网状混合穿孔板.除长江蹄盖蕨不具有梯状穿孔板,有穿孔板二型性现象外,其余4种均具有上述3种类型.该结果支持安蕨属、拟鳞毛蕨属和蹄盖蕨属三者之间有亲密关系的观点,并在前人基础上提出了新的管胞定义:管胞是指维管植物木质部中存在的一类狭长中空,端部圆凸或尖削,不具有明显端壁,侧壁具有多个侧壁穿孔板,纹孔膜从缺失到不同程度存在的死细胞.     

植物管状细胞栓塞后的重新充注研究进展

在植物吸收水分以后 ,水分运输对于植物正常的生长发育是非常重要的。在干旱和冬季反复冻融循环以后 ,植物体内的管状细胞容易充满水蒸气和空气 ,形成腔隙和栓塞。腔隙和栓塞的形成对水分在植物体内的运输造成了很大的障碍 ,从而影响了植物的生长与发育。当植物重新获得水分时 ,已形成腔隙和栓塞的管状细胞的重新充注能使一部分管状细胞的输水功能得到恢复 ,从而保证了一些器官的生理功能的正常进行。近些年来 ,人们对植物管状细胞的重新充注涉及到的许多植物组织和生理过程进行深入的研究 ,并提出了各种机理。鉴于植物管状细胞形成栓塞后重新充注对植物水分运输的重要生理作用 ,本文对重新充注的许多机理进行了综合评述     

Proteomic Analysis of Microtubule Interacting Proteins over the Course of Xylem Tracheary Element Formation in Arabidopsis

Plant vascular cells, or tracheary elements (TEs), rely on circumferential secondary cell wall thickenings to maintain sap flow. The patterns in which TE thickenings are organized vary according to the underlying microtubule bundles that guide wall deposition. To identify microtubule interacting proteins present at defined stages of TE differentiation, we exploited the synchronous differentiation of TEs in Arabidopsis thaliana suspension cultures. Quantitative proteomic analysis of microtubule pull-downs, using ratiometric ¹⁴N/¹⁵N labeling, revealed 605 proteins exhibiting differential accumulation during TE differentiation. Microtubule interacting proteins associated with membrane trafficking, protein synthesis, DNA/RNA binding, and signal transduction peaked during secondary cell wall formation, while proteins associated with stress peaked when approaching TE cell death. In particular, CELLULOSE SYNTHASE-INTERACTING PROTEIN1, already associated with primary wall synthesis, was enriched during secondary cell wall formation. RNAi knockdown of genes encoding several of the identified proteins showed that secondary wall formation depends on the coordinated presence of microtubule interacting proteins with nonoverlapping functions: cell wall thickness, cell wall homogeneity, and the pattern and cortical location of the wall are dependent on different proteins. Altogether, proteins linking microtubules to a range of metabolic compartments vary specifically during TE differentiation and regulate different aspects of wall patterning.     

国产对囊蕨亚科（蹄盖蕨科）植物的管状分子

利用扫描电镜观察了国产蹄盖蕨科（Athyriaceae）对囊蕨亚科（Deparioideae）10种植物及双盖蕨属（Diplazium Sw．）3种植物根状茎的管状分子。结果显示,这些管状分子端壁和侧壁的形态及结构分别相同且侧壁具有穿孔板（多穿孔板）。根据穿孔板的形态特征,将该亚科的管状分子分为5种类型：（1）梯状穿孔板,无穿孔的二型性现象：（2）梯状穿孔板,有穿孔的二型性现象：（3）网状穿孔板：（4）梯状-网状混合的穿孔板：（5）大孔状穿孔板。按照纹孔膜残留的程度又可分为3种：部分区域有完整的纹孔膜、残留呈网状或线状以及很少或无纹孔膜残留。结合前人的研究资料,发现蕨类植物的管状分子与被子植物的导管分子在形态和输导机理上存在明显差异,管胞和导管分子不能仅仅根据纹孔膜的存在与否来确定,而应根据穿孔板存在于端壁还是侧壁进行判断,即穿孔板仅存在于端壁的管状分子为导管分子：端壁和侧壁形态及结构分别相同,有或无穿孔板的管状分子为管胞。由此可以推测蕨类植物和裸子植物中输导水分和矿物质的管状分子主要为管胞。单叶双盖蕨属（Triblemma（J．Sm．）Ching）与双盖蕨属管状分子的特征并不相似,显示了将单叶双盖蕨属从双盖蕨属独立出来归人对囊蕨亚科的合理性。根据管状分子的特征,推测假蹄盖蕨属（Athyriopsis Ching）和蛾眉蕨属（Lunathyrium Koidz．）可能是比较进化的属,而介蕨属（Dryoathyrium Ching）相对比较原始,单叶双盖蕨属的系统位置应介于假蹄盖蕨属与介蕨属之间。     

Development and Structure of Unusual Xylem Elements During Graft Union Formation in Picea sitchensis L.

Electron microscope studies of Sitka spruce have been undertakento observe the sequence of cellular development following grafting.Observations at the graft interface reveal a recovery and regenerationsequence associated with union formation. Xylem elements differentiateddirectly from the vascular cambia of the rootstock and scionare different from elements arising from parenchymatous callusderived from ray parenchyma, which may be produced in an attemptto establish a connecting water conducting system as rapidlyas possible. Failure to form such a system may be a primarycause of graft failure. Grafting, Picea sitchensis, xylen elements     

Decomposition of litter in sub-alpine forests of Eucalyptus delegatensis,E. pauciflora and E. dives

The rate of decomposition of summer leaf-fall (abscised leaves), winter leaf-fall (containing some green leaves) and mature green (picked) leaves was assessed in sub-alpine forests of E. delegatensis (R. T. Baker), E. pauciflora (Sieb. ex Spreng) and E. dives (Schau.) in the Brindabella Range, Australian Capital Territory, using litter bag and tethered leaf techniques. The relative contribution of leaching, microbial respiration and grazing by invertebrate macrofauna to loss of leaf weight was determined. The effect of leaching and microbial respiration was assessed in terms of weight loss per unit area of leaf (specific leaf weight), while losses due to macro-faunal grazing were assessed by measuring reductions in leaf area. Litter decomposition constants for litter components (leaf, bark, wood) and total litter were determined from long-term records of litterfall and accumulated litter. Weight losses of abscised leaves during the initial 12 months ranged from 25% for E. pauciflora to 39% for E. delegatensis and were almost entirely due to reduction in specific leaf weight. Losses in the weight of leaves falling in winter ranged from 38 to 49%, while green leaves lost 45 - 59%. Approximately 50% of the total weight loss of green leaves was due to a loss in leaf area caused by skeletonization by litter macrofauna. Thus abscised leaves rather than green leaves must be used for measuring litter decomposition rates since abscised leaves constitute most of the litterfall in eucalypt forests. Leaves placed in the field in autumn decomposed slowly during the first summer, while the rate increased during the second winter and summer. Low litter moisture content appears to limit decomposition in the initial summer period in all communities, after which litterfall provides a mulch which reduces the rate of desiccation of lower litter layers. A simple linear regression model relating decomposition rate to the number of days (D) when litter moisture content exceeded 60% ODW accounted for 63-83% of the variation in decomposition of leaves in the field. Inclusion of mean monthly air temperature (T) and the product of D and T (day degrees when litter was wet) in a multiple linear regression increased the variation in decomposition accounted for to 80 – 90%. The rate of weight loss showed a positive linear relationship with the initial concentration of nitrogen (N) or phosphorus (P) in the leaf. These concentrations are an index of the decomposability of leaf substrates (e.g. degree of sclerophylly or lignification). The rate of loss of specific weight was similar for tethered leaves and for leaves enclosed in mesh bags. Measured loss in specific leaf weight after 70 – 90 weeks was less than that predicted using decomposition constants (k).     

Temporal heterogeneity of outcrossing rates in alpine ash (Eucalyptus delegatensis R.T. Bak.)

Summary Three seed crops of a Eucalyptus delegatensis population were assayed for their allozyme genotype at three loci to determine estimates of mating system parameters. In the pollen the allelic frequencies at each of the three loci were similar to those in the parents and the progeny. Overall there was a significant amount of inbreeding (23%) in the population. The levels of outcrossing in each crop were significantly different from each other indicating apparent temporal variation in outcrossing rates. The outcrossing rate was greatest in the oldest crop (85%) and lowest in the most recent crop (66%). Mean heterozygosity in the progeny of all three crops was less than the heterozygosity in the parents indicating that selection favours heterozygotes during the life cycle. The implications of a balanced mixed mating system for a eucalypt breeding program are discussed.     

Nocturnal water loss in mature subalpine Eucalyptus delegatensis tall open forests and adjacent E. pauciflora woodlands

We measured sap flux (S) and environmental variables in four monospecific stands of alpine ash (Eucalyptus delegatensis R. Baker, AA) and snowgum (E. pauciflora Sieb. ex Spreng., SG) in Australia's Victorian Alps. Nocturnal S was 11.8 ± 0.8% of diel totals. We separated transpiration (E) and refilling components of S using a novel modeling approach based on refilling time constants. The nocturnal fraction of diel water loss (f(n)) averaged 8.6 ± 0.6% for AA and 9.8 ± 1.7% for SG; f(n) differed among sites but not species. Evaporative demand (D) was the strongest driver of nocturnal E (E(n)). The ratio E(n)/D (G(n)) was positively correlated to soil moisture in most cases, whereas correlations between wind speed and G(n) varied widely in sign and strength. Our results suggest (1) the large, mature trees at our subalpine sites have greater f(n) than the few Australian native tree species that have been studied at lower elevations, (2) AA and SG exhibit similar f(n) despite very different size and life history, and (3) f(n) may differ substantially among sites, so future work should be replicated across differing sites. Our novel approach to quantifying f(n) can be applied to S measurements obtained by any method.     

Graniferous Tracheary Elements in the Haustorium of Osyris arborea Wall. (Santalaceae)

In the haustorium of Osyris arborea (a non-host specific roothemi-parasite) a distinct interrupted zone is present abovethe vascular core. The majority of the xylem elements in thevascular core are perforated. Graniferous tracheary elementsin this species are recorded for the first time. Cytochemicaltests showed the granules to be proteinaceous. The suggestedfunction of graniferous tracheary elements in the regulationof pressure and flow of sap is discussed. Osyris arborea, root hemi-parasite, Santalaceae, haustorium, graniferous tracheary elements, protein granules     

Observations on the Fine Structure of Plant Cell Walls III. The Sieve Tube Wall in Cucurbita

The sieve tube wall in Cucurbita was examined in ultra-thinsections of petioles treated in different ways for the removalof non-cellulosic wall components. The sections were stainedwith permanganate. The microfibrillar components of the wallare arranged in concentric lamellae. The earliest (outermost)part of the wall is similar to that of ordinary parenchyma inhaving its lamellae composed of thinly-distributed microfibrilsreadily separated from one another by certain treatments suchas pectinase extraction. In the characteristically-thickenedinner (nacreous) layer the microfibrils are very densely packedand the lamellae do not separate readily. The microfibrils inthis layer of the wall are very close to transverse and the‘crossed fibrillar’ orientation is not easily discernible.     

洋蒲桃次生木质部中导管分子的解剖学

运用细胞图像分析系统和显微照相的方法对洋蒲桃（Syzygium samarangense）次生木质部导管分子进行了观察研究。次生木质部导管分子类型有：两端具尾导管、一端具尾导管和无尾导管。导管分子穿孔板存在着4种类型：两端均为具2个单穿孔的复穿孔板;一端为1个单穿孔板,另1端为具2个单穿孔的复穿孔板;两端均为单穿孔板：两单穿孔板位于同一端壁两侧相互对应以及一些过渡类型穿孔板。根据观察结果,分析了各类型穿孔板之间的演化关系。     

Breakdown of End Walls in Differentiating Vessels of Secondary Xylem in Quercus rubra L.

Formation and breakdown of end walls were studied in the vesselsof the secondary xylem of Quercus rubra L. The end walls inradially expanding vessel members were very thin but showeda three-layered structure— two peripheral layers and adarker central layer. When longitudinal walls of vessel membersformed secon dary walls, the end walls had thickened considerablyand acquired secondary walls on their periphery. The disintegrationof end walls occurred at about the same time as the disintegrationof the vessel proto plasm. Frequent observations of intermediatestages in the disintegrating end walls indicate that breakdownis a gradual process brought about by the activity of vesselmembers' protoplasm.     

In Situ Detection of nDNA Fragmentation during the Differentiation of Tracheary Elements in Higher Plants

Programmed cell death (pcd) is thought to occur during the autolysis of xylem vessels. Although several ultrastructural aspects of this differentiation process have been characterized, certain key aspects of this process remain unsolved. Here we demonstrate in pea (Pisum sativum) that nuclei of vessel elements undergoing pcd contain fragmented nDNA. This finding may provide evidence for the activation of a DNA degradation mechanism prior to the final disruption of the nucleus that occurs during the autolysis stage of this differentiation process. In situ detection of DNA fragmentation in nuclei of vessel elements undergoing pcd may therefore suggest that this death process involves the activation of a mechanism for DNA degradation, similar to that activated during apoptosis in animal cells. In addition, this differentiation process may serve as a useful positive control for the in situ detection of pcd in other developmental pathways and during the hypersensitive response of plants to avirulent pathogens.