Determining the internal orientation, degree of ordering, and volume of elongated nanocavities by NMR: Application to studies of plant stem.

This study investigates the fibril nanostructure of fresh celery samples by modeling the anisotropic behavior of the transverse relaxation time (T2) in nuclear magnetic resonance (NMR). Experimental results are interpreted within the framework of a previously developed theory, which was successfully used to model the nanostructures of several biological tissues as a set of water filled nanocavities, hence explaining the anisotropy the T2 relaxation time in vivo. An important feature of this theory is to determine the degree of orientational ordering of the nanocavities, their characteristic volume, and their average direction with respect to the macroscopic sample. Results exhibit good agreement between theory and experimental data, which are, moreover, supported by optical microscopic resolution. The quantitative NMR approach presented herein can be potentially used to determine the internal ordering of biological tissues noninvasively.

Functional characterization of hydroxyproline-O-galactosyltransferases for Arabidopsis arabinogalactan-protein synthesis.

BACKGROUND: Arabinogalactan-proteins (AGPs) are structurally complex hydroxyproline-rich cell wall glycoproteins ubiquitous in the plant kingdom. AGPs biosynthesis involves a series of post-translational modifications including the addition of type II arabinogalactans to non-contiguous Hyp residues. To date, eight Hyp-galactosyltransferases (Hyp-GALTs; GALT2-GALT9) belonging to CAZy GT31, are known to catalyze the addition of the first galactose residues to AGP protein backbones and enable subsequent AGP glycosylation. The extent of genetic redundancy, however, remains to be elucidated for the Hyp-GALT gene family. RESULTS: To examine their gene redundancy and functions, we generated various multiple gene knock-outs, including a triple mutant (galt5 galt8 galt9), two quadruple mutants (galt2 galt5 galt7 galt8, galt2 galt5 galt7 galt9), and one quintuple mutant (galt2 galt5 galt7 galt8 galt9), and comprehensively examined their biochemical and physiological phenotypes. The key findings include: AGP precipitations with ß-Yariv reagent showed that GALT2, GALT5, GALT7, GALT8 and GALT9 act redundantly with respect to AGP glycosylation in cauline and rosette leaves, while the activity of GALT7, GALT8 and GALT9 dominate in the stem, silique and flowers. Monosaccharide composition analysis showed that galactose was decreased in the silique and root AGPs of the Hyp-GALT mutants. TEM analysis of 25789 quintuple mutant stems indicated cell wall defects coincident with the observed developmental and growth impairment in these Hyp-GALT mutants. Correlated with expression patterns, galt2, galt5, galt7, galt8, and galt9 display equal additive effects on insensitivity to ß-Yariv-induced growth inhibition, silique length, plant height, and pollen viability. Interestingly, galt7, galt8, and galt9 contributed more to primary root growth and root tip swelling under salt stress, whereas galt2 and galt5 played more important roles in seed morphology, germination defects and seed set. Pollen defects likely contributed to the reduced seed set in these mutants. CONCLUSION: Additive and pleiotropic effects of GALT2, GALT5, GALT7, GALT8 and GALT9 on vegetative and reproductive growth phenotypes were teased apart via generation of different combinations of Hyp-GALT knock-out mutants. Taken together, the generation of higher order Hyp-GALT mutants demonstrate the functional importance of AG polysaccharides decorating the AGPs with respect to various aspects of plant growth and development.

Sulfur Induces Resistance against Canker Caused by Pseudomonas syringae pv. actinidae via Phenolic Components Increase and Morphological Structure Modification in the Kiwifruit Stems.

Bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) has led to considerable losses in all major kiwifruit-growing areas. There are no commercial products in the market to effectively control this disease. Therefore, the defense resistance of host plants is a prospective option. In our previous study, sulfur could improve the resistance of kiwifruit to Psa infection. However, the mechanisms of inducing resistance remain largely unclear. In this study, disease severity and protection efficiency were tested after applying sulfur, with different concentrations in the field. The results indicated that sulfur could reduce the disease index by 30.26 and 31.6 and recorded high protection efficiency of 76.67% and 77.00% after one and two years, respectively, when the concentration of induction treatments was 2.0 kg/m3. Ultrastructural changes in kiwifruit stems after induction were demonstrated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and the activities of phenylalanine ammonia-lyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO), and the accumulation of lignin were determined by biochemical analyses. Our results showed that the morphological characteristics of trichomes and lenticels of kiwifruit stem were in the best defensive state respectively when the sulfur concentration was 3.0 kg/m3 and 1.5 kg/m3. Meanwhile, in the range of 0.5 to 2.0 kg/m3, the sulfur could promote the chloroplast and mitochondria of kiwifruit stems infected with Psa to gradually return to health status, increasing the thickness of the cell wall. In addition, sulfur increased the activities of PAL, POD and PPO, and promoted the accumulation of lignin in kiwifruit stems. Moreover, the sulfur protection efficiency was positively correlated with PPO activity (p < 0.05) and lignin content (p < 0.01), which revealed that the synergistic effect of protective enzyme activity and the phenolic metabolism pathway was the physiological effect of sulfur-induced kiwifruit resistance to Psa. This evidence highlights the importance of lignin content in kiwifruit stems as a defense mechanism in sulfur-induced resistance. These results suggest that sulfur enhances kiwifruit canker resistance via an increase in phenolic components and morphology structure modification in the kiwifruit stems. Therefore, this study could provide insights into sulfur to control kiwifruit canker caused by Psa.

A microscopic study on scattering in tissue section of Alternanthera philoxeroides under polarized light.

Like any other biological tissue, plant tissue also exhibits optical properties like refraction, transmission, absorption, coloration, scattering and so on. Several studies have been conducted using different parts of plants such as leaves, seedlings, roots, stems and so on, and their optical properties have been analyzed to study plant physiology, influence of environmental cues on plant metabolism, light propagation through plant parts and the like. Thus, it is essential to study in detail the optical properties of several plant parts to determine their structural relationship. In this backdrop, an experimental study was conducted to observe and analyze the optical properties of node and inter-nodal tissue cross-sections of the plant Alternanthera philoxeroides under a polarizing microscope constructed and standardized in the laboratory. The observed optical properties of the microscopic tissue sections have been then studied to determine a significant structural relationship between nodal and inter-nodal tissue arrangement patterns as a whole. Tissue sections that have undergone a sort of biological perturbation like loss of water (dried in air for 15 min) have also been studied to study the change in the pattern of tissue optical property when compared with that of normal plant-tissue cross-sections under a polarizing microscope. This type of biological perturbation was chosen for the study because water plays an important role in maintenance of the normal physiological processes in plants and most other forms of life.

Multiscale characterization and micromechanical modeling of crop stem materials.

An essential prerequisite for the efficient biomechanical tailoring of crops is to accurately relate mechanical behavior to compositional and morphological properties across different length scales. In this article, we develop a multiscale approach to predict macroscale stiffness and strength properties of crop stem materials from their hierarchical microstructure. We first discuss the experimental multiscale characterization based on microimaging (micro-CT, light microscopy, transmission electron microscopy) and chemical analysis, with a particular focus on oat stems. We then derive in detail a general micromechanics-based model of macroscale stiffness and strength. We specify our model for oats and validate it against a series of bending experiments that we conducted with oat stem samples. In the context of biomechanical tailoring, we demonstrate that our model can predict the effects of genetic modifications of microscale composition and morphology on macroscale mechanical properties of thale cress that is available in the literature.

Histone methyltransferase ATX1 dynamically regulates fiber secondary cell wall biosynthesis in Arabidopsis inflorescence stem.

Secondary wall thickening in the sclerenchyma cells is strictly controlled by a complex network of transcription factors in vascular plants. However, little is known about the epigenetic mechanism regulating secondary wall biosynthesis. In this study, we identified that ARABIDOPSIS HOMOLOG of TRITHORAX1 (ATX1), a H3K4-histone methyltransferase, mediates the regulation of fiber cell wall development in inflorescence stems of Arabidopsis thaliana. Genome-wide analysis revealed that the up-regulation of genes involved in secondary wall formation during stem development is largely coordinated by increasing level of H3K4 tri-methylation. Among all histone methyltransferases for H3K4me3 in Arabidopsis, ATX1 is markedly increased during the inflorescence stem development and loss-of-function mutant atx1 was impaired in secondary wall thickening in interfascicular fibers. Genetic analysis showed that ATX1 positively regulates secondary wall deposition through activating the expression of secondary wall NAC master switch genes, SECONDARY WALL-ASSOCIATED NAC DOMAIN PROTEIN1 (SND1) and NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1). We further identified that ATX1 directly binds the loci of SND1 and NST1, and activates their expression by increasing H3K4me3 levels at these loci. Taken together, our results reveal that ATX1 plays a key role in the regulation of secondary wall biosynthesis in interfascicular fibers during inflorescence stem development of Arabidopsis.

Microstructural and histochemical analysis of aboveground organs of Centaurea cyanus used in herbal medicine.

Centaurea cyanus L. is a valuable source of many different bioactive substances. It is used in herbal medicine, but the structure of its organs used as raw material and secretory tissues has been insufficiently examined. The aim of this paper was to investigate the microstructure of C. cyanus flowers, bracts, stems and leaves with particular emphasis on secretory structures. Moreover, the main classes of secondary metabolites present in the secretion were identified and the taxonomic value of some micromorphological and anatomical features was analysed. Histochemical, micromorphological and ultrastructural analyses of aboveground organs of C. cyanus were carried out using light, fluorescence, scanning and transmission electron microscopy. The analyses revealed the presence of petal papillae and a characteristic cuticular pattern on the petals, stamens and stylar hairs. There were four types of non-glandular trichomes on the bracts, leaves and stem surfaces. The epidermal cells of the bracts contained prismatic calcium oxalate crystals. Two kinds of secretory structures, i.e. glandular trichomes and ducts, were observed in the C. cyanus organs. The glandular trichomes were located on the bract and stem surfaces, and the ducts were detected in the leaves and stems. Ultrastructural analyses of the epithelium of the ducts showed the presence of strongly osmiophilic insoluble phenolic material in vacuoles as well as moderately osmiophilic insoluble lipidic material in elaioplasts and vesicles. The results of histochemical assays showed a heterogeneous nature of the duct secretion, which contained essential oil, lipids, flavonoids, tannins and terpenes containing steroids.

Anatomical and ultrastructural responses of Hordeum sativum to the soil spiked by copper.

Effects of Cu toxicity from contaminated soil were analysed in spring barley (Hordeum sativum distichum), a widely cultivated species in South Russia. In this study, H. sativum was planted outdoors in one of the most fertile soils-Haplic Chernozem spiked with high concentration of Cu and examined between the boot and head emergence phase of growth. Copper toxicity was observed to cause slow ontogenetic development of plants, changing their morphometric parameters (shape, size, colour). To the best of our knowledge, the ultrastructural changes in roots, stems and leaves of H. sativum induced by excess Cu were fully characterized for the first time using transmission electron microscopy. The plant roots were the most effected, showing degradation of the epidermis, reduced number of parenchyma cells, as well as a significant decrease in the diameter of the stele and a disruption and modification to its cell structure. The comparative analysis of the ultrastructure of control plants and plants exposed to the toxic effects of Cu has made it possible to reveal significant disruption of the integrity of the cell wall and cytoplasmic membranes in the root with deposition of electron-dense material. The changes in the ultrastructure of the main cytoplasmic organelles-endoplasmic reticulum, mitochondria, chloroplasts and peroxisomes-in the stem and leaves were found. The cellular Cu deposition, anatomical and ultrastructural modifications could mainly account for the primary impact points of metal toxicity. Therefore, this work extends the available knowledge of the mechanisms of the Cu effect tolerance of barley.

Leaf and stem micromorphology of Byrsonima sericea DC. by light and scanning electron microscopy.

Micromorphological studies were carried out using multiple microscopic techniques on the leaves and stem bark of Byrsonima sericea DC. (Malpighiaceae), a species popularly known as "murici" and used medicinally, in order to identify both qualitative and quantitative features of leaf and stem anatomy and histochemistry as differential parameters to support both the quality control of its ethnodrugs and the taxonomy of the genus. The study was conducted using traditional techniques of plant anatomy, histochemical tests, and the stomatal index (SI). Byrsonima sericea has hypostomatic leaves, anomocytic stomata, and its epidermal walls are anticlinal and straight on the adaxial and curved on the abaxial faces. T-shaped trichomes were observed mainly on the abaxial surface. The leaf epidermis showed waxes syntopism on both surfaces, with the occurrence of different crystalloid forms on a single phylloplane. The mesophyll is dorsiventral, with 3-4 collateral vascular bundles. Phenolic compounds, starch, and proteins were identified in the petiole and stem. The SI was 14.5 ± 0.53% (p < .05), but did not showed significant variations. A set of characters were found to be distinctive for the studied species, however, constituting parameters that could be used to separate B. sericea from other species of the genus.

The relationship between cuticular lipids and associated gene expression in above ground organs of Thellungiella salsugineum (Pall.) Al-Shehbaz & Warwick.

The cuticle plays a critical role as barrier between plant and environment. Here, cuticular wax morphology, cuticular wax and cutin monomer composition, and expression of associated genes in five above ground organs were examined in model extremophyte Thellungiella salsugineum. Alkanes, ketones, and 2-alcohols were the predominant wax constitutes in rosette leaves, inflorescence stem leaves, stems, and siliques, whereas alkanes and acids were the predominant cuticular lipids in whole flowers. Unsubstituted acids were the most abundant cutin monomers in vegetative organs, especially C18:2 dioic acids, which reached the highest levels in stems. Hydroxy fatty acids were the predominant cutin monomers in flowers, especially 16-OH C16:0 and diOH C16:0. High-throughput RNA-Seq analysis using the Hiseq4000 platform was performed on these five above organs of T. salsugineum, and the differentially expressed lipid-associated genes and their associated metabolic pathways were identified. Expression of genes associated in previous reports to cuticle production, including those having roles in cuticle lipid biosynthesis, transport, and regulation were examined. The association of cuticle lipid composition and gene expression within different organs of T. salsugineum, and potential relationships between T. salsugineum's extreme cuticle and its adaptation to extreme environments is discussed.

The F-Box Protein SAGL1 and ECERIFERUM3 Regulate Cuticular Wax Biosynthesis in Response to Changes in Humidity in Arabidopsis.

Cuticular waxes, which cover the aboveground parts of land plants, are essential for plant survival in terrestrial environments. However, little is known about the regulatory mechanisms underlying cuticular wax biosynthesis in response to changes in ambient humidity. Here, we report that the Arabidopsis (Arabidopsis thaliana) Kelch repeat F-box protein SMALL AND GLOSSY LEAVES1 (SAGL1) mediates proteasome-dependent degradation of ECERIFERUM3 (CER3), a biosynthetic enzyme involved in the production of very long chain alkanes (the major components of wax), thereby negatively regulating cuticular wax biosynthesis. Disruption of SAGL1 led to severe growth retardation, enhanced drought tolerance, and increased wax accumulation in stems, leaves, and roots. Cytoplasmic SAGL1 physically interacts with CER3 and targets it for degradation. ß­glucuronidase (GUS) expression was observed in the roots of pSAGL1:GUS plants but was barely detected in aerial organs. High humidity-induced GUS activity and SAGL1 transcript levels were reduced in response to abscisic acid treatment and water deficit. SAGL1 levels increase under high humidity, and the stability of this protein is regulated by the 26S proteasome. These findings indicate that the SAGL1-CER3 module negatively regulates cuticular wax biosynthesis in Arabidopsis in response to changes to humidity, and they highlight the importance of permeable cuticle formation in terrestrial plants under high humidity conditions.

A model for extracellular freezing based on observations on Equisetum hyemale.

In frost hardy plants, the lethal intracellular formation of ice crystals has to be prevented during frost periods. Besides the ability for supercooling and pre-frost dehydration of tissues, extracellular ice formation is another strategy to control ice development in tissues. During extracellular ice formation, partially large ice bodies accumulate in intercellular spaces, often at preferred sites which can also be expandable. In this contribution, the physico-chemical processes underlying the water movements towards the sites of extracellular ice formation are studied theoretically, based on observations on the frost hardy horsetail species Equisetum hyemale, with the overall aim to obtain a better understanding of the physical processes involved in extracellular ice formation. In E. hyemale, ice accumulates in the extensive internal canal system. The study focuses on the processes which are triggered in the cellular osmotic-mechanic system by falling, and especially subzero temperatures. It can be shown that when the temperature falls, (1) water flow out of cells is actuated and (2) "stiff-walled" cells lose less water than "soft-walled" cells. Furthermore, (3) cell water loss increases with increasing (= less negative) turgor loss point. These processes are not related to any specific activities of the cell but are solely a consequence of the structure of the cellular osmotic system. On this basis, a directed water flow can be initiated triggered by subzero temperatures. The suggested mechanism may be quite common in frost hardy species with extracellular ice formation.

Integration of Transcriptome, Proteome, and Metabolome Provides Insights into How Calcium Enhances the Mechanical Strength of Herbaceous Peony Inflorescence Stems.

Weak stem mechanical strength severely restrains cut flowers quality and stem weakness can be alleviated by calcium (Ca) treatment, but the mechanisms underlying Ca-mediated enhancement of stem mechanical strength remain largely unknown. In this study, we performed a comparative transcriptomic, proteomic, and metabolomic analysis of herbaceous peony (Paeonia lactiflora Pall.) inflorescence stems treated with nanometer Ca carbonate (Nano-CaCO3). In total, 2643 differentially expressed genes (DEGs) and 892 differentially expressed proteins (DEPs) were detected between the Control and nano-CaCO3 treatment. Among the 892 DEPs, 152 were coregulated at both the proteomic and transcriptomic levels, and 24 DEPs related to the secondary cell wall were involved in signal transduction, energy metabolism, carbohydrate metabolism and lignin biosynthesis, most of which were upregulated after nano-CaCO3 treatment during the development of inflorescence stems. Among these four pathways, numerous differentially expressed metabolites (DEMs) related to lignin biosynthesis were identified. Furthermore, structural observations revealed the thickening of the sclerenchyma cell walls, and the main wall constitutive component lignin accumulated significantly in response to nano-CaCO3 treatment, thereby indicating that Ca can enhance the mechanical strength of the inflorescence stems by increasing the lignin accumulation. These results provided insights into how Ca treatment enhances the mechanical strength of inflorescence stems in P. lactiflora.

Nanoscale Structural Organization of Plant Epicuticular Wax Probed by Atomic Force Microscope Infrared Spectroscopy.

The cuticle covers external surfaces of plants, protecting them from biotic and abiotic stress factors. Epicuticular wax on the outer surface of the cuticle modifies reflectance and water loss from plant surfaces and has direct and indirect effects on photosynthesis. Variation in epicuticular wax accumulation, composition, and nanoscale structural organization impacts its biological function. Atomic force microscope infrared spectroscopy (AFM-IR) was utilized to investigate the internal and external surfaces of the cuticle of Sorghum bicolor, an important drought-tolerant cereal, forage, and high-biomass crop. AFM-IR revealed striking heterogeneity in chemical composition within and between the surfaces of the cuticle. The wax aggregate crystallinity and distribution of chemical functional groups across the surfaces was also probed and compared. These results, along with the noninvasive nondestructive nature of the method, suggest that AFM-IR can be used to investigate mechanisms of wax deposition and transport of charged molecules through the plant cuticle.

How the activity of natural enemies changes the structure and metabolism of the nutritive tissue in galls? Evidence from the Palaeomystella oligophaga (Lepidoptera) -Macairea radula (Metastomataceae) system.

Insect-induced galls usually develop nutritional cells, which they induce and consume directly, and any metabolic modification of those cells may reflect changes of the insect's own metabolism. The system Palaeomystella oligophaga (Lepidoptera)-Macairea radula (Melastomataceae) presents a series of natural enemies, including parasitoids and cecidophages that can function as a natural experiment, respectively removing the specific galling feeding stimulus and providing a nonspecific one. Considering that the process of induction and maintenance of gall tissues strictly depends on the constant specific stimulus of galling, question I:What kind of metabolic changes these different groups of natural enemies can promote in chemical and structural composition of these galls? II: How the specialized tissues are metabolically dependent on the constant specific stimulus of galling in latter stages of gall development? Galls without natural enemies, with parasitoids or cecidophages in larvae or pupae stages were analyzed through histochemistry and cytological profiles and all compared to galls in natural senescence state. The analysis revealed the accumulation of proteins and lipids in typical nutritive tissue and starch in the storage tissue, as well a high integrity of cellular organelles and membrane systems on galls with gallings in the larval stage. Both parasitoids and cecidophages stop galling feeding activities, which resulted in the paralysis of the stimulus that maintain the metabolism of gall tissues, leading to generalized collapse. We demonstrate that the development and metabolic maintenance of a typical nutritive tissue in these galls are completely dependent on constant larval stimulus.

Knockout of the S-acyltransferase Gene, PbPAT14, Confers the Dwarf Yellowing Phenotype in First Generation Pear by ABA Accumulation.

The development of dwarf fruit trees with smaller and compact characteristics leads to significantly increased fruit production, which is a major objective of pear (Pyrus bretschneideri) breeding. We identified the S-acylation activity of PbPAT14, an S-acyltransferase gene related to plant development, using a yeast (Saccharomyces cerevisiae) complementation assay, and also PbPAT14 could rescue the growth defect of the Arabidopsis mutant atpat14. We further studied the function of PbPAT14 by designing three guide RNAs for PbPAT14 to use in the CRISPR/Cas9 system. We obtained 22 positive transgenic pear lines via Agrobacterium-mediated transformation using cotyledons from seeds of Pyrus betulifolia ('Duli'). Six of these lines exhibited the dwarf yellowing phenotype and were homozygous mutations according to sequencing analysis. Ultrastructure analysis suggested that this dwarfism was manifested by shorter, thinner stems due to a reduction in cell number. A higher level of endogenous abscisic acid (ABA) and a higher transcript level of the ABA pathway genes in the mutant lines revealed that the PbPAT14 function was related to the ABA pathway. Overall, our experimental results increase the understanding of how PATs function in plants and help elucidate the mechanism of plant dwarfism.

Can climate and soil conditions change the morpho-anatomy among individuals from different localities? A case study in Aldama grandiflora (Asteraceae) / Condições climáticas e de solo podem alterar a morfo-anatomia entre indivíduos de diferentes localidades? Um estudo de caso em Aldama grandiflora (Asteraceae)

Abstract Vegetative aerial organs are considerably more exposed to environmental conditions and can reflect the specific adaptations of plants to their local environment. Aldama grandiflora species are known to be widely distributed in Brazil; therefore, individuals from different populations of this species are thought to be exposed to different abiotic and biotic conditions. Several anatomical studies conducted on Brazilian Aldama species have mainly focused on the qualitative anatomical characters or traits of these species, but not on their quantitative traits. In this study, we evaluated whether climate and soil conditions can change the morphometry among individuals of A. grandiflora collected from six sites in the Goiás State, Brazil, by assessing their anatomical characters. Further, soil sampling was performed, and climate data were collected from all the six sites. The analysis indicated few statistical differences among the populations evaluated, showing that A. grandiflora presented consistent leaf and stem anatomical characteristics. The small morpho-anatomical differences found among individuals of the different populations evaluated, reflected the soil conditions in which these populations were grown. Therefore, environmental factors have a significant influence on the morpho-anatomy of Aldama grandiflora.

Resumo Os órgãos vegetativos aéreos estão consideravelmente mais expostos às condições ambientais e podem refletir as adaptações específicas das plantas ao seu habitat. A espécie Aldama grandiflora é amplamente distribuída no Brasil e, dessa forma, indivíduos de diferentes populações podem estar expostos a diferentes condições ambientais. Vários estudos anatômicos realizados com espécies brasileiras do gênero Aldama têm abordado, principalmente, as características anatômicas qualitativas dessas espécies, mas não em suas características quantitativas. Neste estudo avaliamos se as condições climáticas e do solo podem alterar a morfometria entre os indivíduos de A. grandiflora coletados em seis populações do Estado de Goiás. Foram avaliados os caracteres anatômicos foliares e caulinares, além da amostragem do solo e coleta de dados climáticos, para os seis locais. A análise indicou algumas diferenças estatísticas entre as populações avaliadas, mostrando que A. grandiflora apresentou características anatômicas foliares e caulinares bastante consistentes. As pequenas diferenças morfo-anatômicas encontradas entre indivíduos das diferentes populações avaliadas, refletiram as condições do solo nos quais essas populações se desenvolveram. Assim sendo, fatores ambientais relacionados ao clima e condições do solo têm uma influência significativa sobre a morfo-anatomia de Aldama grandiflora.

Water availability dynamics have long-term effects on mature stem structure in Vitis vinifera.

PREMISE OF THE STUDY: The stem of Vitis vinifera, a climbing vine of global economic importance, is characterized by both wide and narrow vessels and high specific hydraulic conductivity. While the effect of drought stress has been studied in 1- and 2-yr-old stems, there are few data documenting effects of drought stress on the anatomical structure of the mature, woody stem near the base of the vine. Here we describe mature wood anatomical responses to two irrigation regimes on wood anatomy and specific hydraulic conductivity in Vitis vinifera Merlot vines. METHODS: For 4 years, irrigation was applied constantly at low, medium, or high levels, or at alternating levels at two different periods during the growing season, either early spring or late summer, resulting in late season or early spring deficits, respectively. The following variables were measured: trunk diameter, annual ring width and area, vessel diameter, specific hydraulic conductivity and stem water potential. KEY RESULTS: High water availability early in the season (late deficit) resulted in vigorous vegetative growth (greater trunk diameter, ring width and area), wider vessels and increased specific hydraulic conductivity. High water availability early in the season caused a shift of the vessel population towards the wider frequency classes. These late deficit vines showed more negative water potential values late in the season than vines that received low but relatively constant irrigation. CONCLUSIONS: We concluded that high water availability during vegetative growth period of Vitis increases vessels diameter and hydraulic conductivity and causes the vines to be more vulnerable to drought stress late in the season.

Transcriptomic and functional analysis of cucumber (Cucumis sativus L.) fruit phloem during early development.

The phloem of the Cucurbitaceae has long been a subject of interest due to its complex nature and the economic importance of the family. As in a limited number of other families, cucurbit phloem is bicollateral, i.e. with sieve tubes on both sides of the xylem. To date little is known about the specialized functions of the internal phloem (IP) and external phloem (EP). Here, a combination of microscopy, fluorescent dye transport analysis, micro-computed tomography, laser capture microdissection and RNA-sequencing (RNA-Seq) were used to study the functions of IP and EP in the vascular bundles (VBs) of cucumber fruit. There is one type of VB in the peduncle, but four in the fruit: peripheral (PeVB), main (MVB), carpel (CVB) and placental (PlVB). The VBs are bicollateral, except for the CVB and PlVB. Phloem mobile tracers and 14 C applied to leaves are transported primarily in the EP, and to a lesser extent in the IP. RNA-Seq data indicate preferential gene transcription in the IP related to differentiation/development, hormone transport, RNA or protein modification/processing/transport, and nitrogen compound metabolism and transport. The EP preferentially expresses genes for stimulus/stress, defense, ion transport and secondary metabolite biosynthesis. The MVB phloem is preferentially involved in photoassimilate transport, unloading and long-distance signaling, while the PeVB plays a more substantial role in morphogenesis and/or development and defense response. CVB and PlVB transcripts are biased toward development of reproductive organs. These findings provide an integrated view of the differentiated structure and function of the vascular tissue in cucumber fruit.

Chemical, molecular and structural studies of Boswellia species: ß-Boswellic Aldehyde and 3-epi-11ß-Dihydroxy BA as precursors in biosynthesis of boswellic acids.

The distribution and biosynthesis of boswellic acids (BAs) is scarce in current literature. Present study aims to elucidate the BAs biosynthetic and its diversity in the resins of Boswellia sacra and Boswellia papyrifera. Results revealed the isolation of new (3ß, 11ß-dihydroxy BA) and recently known (as new source, ß-boswellic aldehyde) precursors from B. sacra resin along with α-amyrin. Following this, a detailed nomenclature of BAs was elucidated. The quantification and distribution of amyrins (3-epi-α-amyrin, ß-amyrin and α-amyrin) and BAs in different Boswellia resins showed highest amyrin and BAs in B. sacra as compared with B. serrata and B. papyrifera. Distribution of BAs significantly varied in the resin of B. sacra collected from dry mountains than coastal trees. In B. sacra, high content of α-amyrin was found in the roots but it lacked ß-amyrin and BAs. The leaf part showed traces of ß-ABA and AKBA but was deficient in amyrins. This was further confirmed by lack of transcript accumulation of amyrin-related biosynthesis gene in leaf part. In contrast, the stem showed presence of all six BAs which are attributed to existence of resin-secretory canals. In conclusion, the boswellic acids are genus-specific chemical constituents for Boswellia species albeit the variation of the amounts among different Boswellia species and grades.