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.

A Hyperspectral Imaging Approach for Classifying Geographical Origins of Rhizoma Atractylodis Macrocephalae Using the Fusion of Spectrum-Image in VNIR and SWIR Ranges (VNIR-SWIR-FuSI).

Hyperspectral data processing technique has gained increasing interests in the field of chemical and biomedical analysis. However, appropriate approaches to fusing features of hyperspectral data-cube are still lacking. In this paper, a new data fusion approach was proposed and applied to discriminate Rhizoma Atractylodis Macrocephalae (RAM) slices from different geographical origins using hyperspectral imaging. Spectral and image features were extracted from hyperspectral data in visible and near-infrared (VNIR, 435-1042 nm) and short-wave infrared (SWIR, 898-1751 nm) ranges, respectively. Effective wavelengths were extracted from pre-processed spectral data by successive projection algorithm (SPA). Meanwhile, gray-level co-occurrence matrix (GLCM) and gray-level run-length matrix (GLRLM) were employed to extract textural variables. The fusion of spectrum-image in VNIR and SWIR ranges (VNIR-SWIR-FuSI) was implemented to integrate those features on three fusion dimensions, i.e., VNIR and SWIR fusion, spectrum and image fusion, and all data fusion. Based on data fusion, partial least squares-discriminant analysis (PLS-DA) and support vector machine (SVM) were utilized to establish calibration models. The results demonstrated that VNIR-SWIR-FuSI could achieve the best accuracies on both full bands (97.3%) and SPA bands (93.2%). In particular, VNIR-SWIR-FuSI on SPA bands achieved a classification accuracy of 93.2% with only 23 bands, which was significantly better than those based on spectra (80.9%) or images (79.7%). Thus it is more rapid and possible for industry applications. The current study demonstrated that hyperspectral imaging technique with data fusion holds the potential for rapid and nondestructive sorting of traditional Chinese medicines (TCMs).

Analysis of Rhizome Development in Oryza longistaminata, a Wild Rice Species.

Vegetative reproduction is a form of asexual propagation in plants. A wide range of plants develop rhizomes, modified stems that grow underground horizontally, as a means of vegetative reproduction. In rhizomatous species, despite their distinct developmental patterns, both rhizomes and aerial shoots derive from axillary buds. Therefore, it is of interest to understand the basis of rhizome initiation and development. Oryza longistaminata, a wild rice species, develops rhizomes. We analyzed bud initiation and growth of O. longistaminata rhizomes using various methods of morphological observation. We show that, unlike aerial shoot buds that contain a few leaves only, rhizome buds initiate several leaves and bend to grow at right angles to the original rhizome. Rhizomes are maintained in the juvenile phase irrespective of the developmental phase of the aerial shoot. Stem elongation and reproductive transition are tightly linked in the aerial shoots, but are uncoupled in the rhizome. Our findings indicate that developmental programs operate independently in the rhizomes and aerial shoots. Temporal modification of the developmental pathways that are common to rhizomes and aerial shoots may be the source of developmental plasticity. Furthermore, the creation of new developmental systems appears to be necessary for rhizome development.

The localization of NADPH oxidase and reactive oxygen species in in vitro-cultured Mesembryanthemum crystallinum L. hypocotyls discloses their differing roles in rhizogenesis.

This work demonstrated how reactive oxygen species (ROS) are involved in the regulation of rhizogenesis from hypocotyls of Mesembryanthemum crystallinum L. cultured on a medium containing 1-naphthaleneacetic acid (NAA). The increase of NADPH oxidase activity was correlated with an increase of hydrogen peroxide (H2O2) content and induction of mitotic activity in vascular cylinder cells, leading to root formation from cultured hypocotyls. Diphenylene iodonium (DPI), an inhibitor of NADPH oxidase, inhibited H2O2 production and blocked rhizogenesis. Ultrastructural studies revealed differences in H2O2 localization between the vascular cylinder cells and cortex parenchyma cells of cultured explants. We suggest that NADPH oxidase is responsible for H2O2 level regulation in vascular cylinder cells, while peroxidase (POD) participates in H2O2 level regulation in cortex cells. Blue formazan (NBT) precipitates indicating superoxide radical (O2 (•-)) accumulation were localized within the vascular cylinder cells during the early stages of rhizogenesis and at the tip of root primordia, as well as in the distal and middle parts of newly formed organs. 3,3'-diaminobenzidine (DAB) staining of H2O2 was more intense in vascular bundle cells and in cortex cells. In newly formed roots, H2O2 was localized in vascular tissue. Adding DPI to the medium led to a decrease in the intensity of NBT and DAB staining in cultured explants. Accumulation of O2 (•-) was then limited to epidermis cells, while H2O2 was accumulated only in vascular tissue. These results indicate that O2 (•-) is engaged in processes of rhizogenesis induction involving division of competent cells, while H2O2 is engaged in developmental processes mainly involving cell growth.

[Anatomical structure and histochemical localization about rhizomes of five species of Polygonatum].

OBJECTIVE: The microscopic structure and histochemical localization of the Polygonatum sibiricum, P. cytonema, P. filipes, P. zanlanscianense and P. odoratum was studied. METHOD: Scanning electron microscope (SEM) and digital biological microscope (DBM) were applied in the research. RESULT: Venation bundle showed in three types included collateral, amphivasal and incomplete amphivasal bundles. Morphological differences of venation bundles could be seen under the fluorescence microscope. Calcium oxalate existed in the mucilage cells looked similar to columnar crystals under SEM. In rhizome of Polygonatum, polysaccharides presented in mucilage cells, saponins and volatile oil were found in ground tissue. CONCLUSION: Microstructure difference of rhizomes would be used for identification of the Polygonatum plants. Polysaccharides and saponins are distributed in different cells of Polygonatum plants.

Profiling of secondary metabolites in tissues from Rheum palmatum L. using laser microdissection and liquid chromatography mass spectrometry.

Evaluating the quality of herbal medicines by morphological features is a convenient, quick, and practical method compared with other methods that mostly depend on modern instruments. Here, laser microdissection and ultra-performance liquid chromatography are combined with mass spectrometry to map the distribution of secondary metabolites in cells or tissues of a herb itself for correlating its bioactive components and morphological features. The root and rhizome of Rheum palmatum L. were taken as research target, which is the Chinese medicine, Radix et Rhizoma Rhei. According to fluorescent microscopic characteristics, 12 herbal cells or tissues of Radix et Rhizoma Rhei were separated by laser microdissection. Thirty-eight compounds were identified or tentatively characterized in the microdissected tissues. (+)-Catechin, 1-O-galloyl-2-O-cinnamoyl-ß-D-glucose, and emodin were found to be the major components in most of the tissues. The brown ergastic substances found in rays of normal and anomalous vascular bundles as well as the parenchymatous cells of rhizome pith and the parenchymatous cells of root xylem contained higher than average amounts of these three components and more kinds of secondary metabolites. Overall, results suggest that Radix et Rhizoma Rhei of larger size and with conspicuous "brocaded patterns" and star spots are of higher quality as they tend to have greater contents of bioactive components. The study provides quantitative and specific criteria by which the quality of Radix et Rhizoma Rhei can be judged. This research also established a new, reliable, and practical method for direct profiling and imaging of secondary metabolites in any herbal tissue.

Ultrastructure and subcellular distribution of Cr in Iris pseudacorus L. using TEM and X-ray microanalysis.

Chromium pollution of freshwater is hazardous for humans and other organisms, and places a limitation on the use of polluted water sources. Phytoremediation, the use of plants to remove pollutants from the environment, is a cost-effective, environmentally friendly approach for water decontamination. To improve the efficiency of the process, it is essential to increase the current knowledge about Cr accumulation in macrophytes. Plants of Iris pseudacorus L. were treated with Cr(III) at 0.75 mM for 5 weeks to investigate Cr localization by means of transmission electron microscopy and energy dispersive X-ray analysis. Chromium induced severe ultrastructural alterations in the rhizodermis (cell wall disorganisation, thickening, plasmolysis, and electron-dense inclusions) and rhizome parenchyma (reduced cell size, cell wall detachment, vacuolation, and opaque granules). The highest Cr contents were found in the cell walls of the cortex in the roots and in the cytoplasm and intercellular spaces of the rhizome. The Cr concentration in root tissues was in the order cortex>rhizodermis>stele, whereas in the rhizome, Cr was evenly distributed. It is proposed that root and rhizome have distinct functions in the response of I. pseudacorus to Cr. The rhizodermis limits Cr uptake by means of Si deposition and cell wall thickening. The rhizome cortex generates vacuoles and granules where Cr co-occurs with S, indicating Cr sequestration by metal-binding proteins.

Influence of confocal scanning laser microscopy specific acquisition parameters on the detection and matching of speeded-up robust features.

The robustness and distinctiveness of local features to various object or scene deformations and to modifications of the acquisition parameters play key roles in the design of many computer vision applications. In this paper we present the results of our experiments on the behavior of a recently developed technique for local feature detection and description, Speeded-Up Robust Features (SURF), regarding image modifications specific to Confocal Scanning Laser Microscopy (CSLM). We analyze the repeatability of detected SURF keypoints and the precision-recall of their matching under modifications of three important CSLM parameters: pinhole aperture, photomultiplier (PMT) gain and laser beam power. During any investigation by CSLM these three parameters have to be modified, individually or together, in order to optimize the contrast and the Signal Noise Ratio (SNR), being also inherently modified when changing the microscope objective. Our experiments show that an important amount of SURF features can be detected at the same physical locations in images collected at different values of the pinhole aperture, PMT gain and laser beam power, and further on can be successfully matched based on their descriptors. In the final part, we exemplify the potential of SURF in CSLM imaging by presenting a SURF-based computer vision application that deals with the mosaicing of images collected by this technique.

[Comparative identification on Dai medicine "Pokou" and its processing product].

OBJECTIVE: To research identification methods of the Dai Medicine "Pokou" (the rhizome of Homalomena gigantea) and its processing product, and provide basis for identification of the drug in further research and application. METHODS: Macroscopic, microscopic observation and TLC and FTIR techniques were used to authenticate this raw medicine and its processing product. RESULTS: There were certain differences in the macroscopic features. The TLC result and infrared spectra of the samples had also obvious differences. The methods for identification of this raw medicine and its processing product were established, The detailed tissue and powder of this medicine were drawn. CONCLUSION: The results provided the basis for identification of the medicine and establishment of its quality standard.

Rhizotoxic effects of silver in cowpea seedlings.

Silver (Ag) is highly toxic to aquatic organisms, including algae, invertebrate animals, and fish, but little information exists on Ag rhizotoxicity in higher plants. In two solution culture experiments with approximately 1,000 microM Ca(NO3)2 and 5 microM H3BO3 (pH 5.4), 20 to 80% of added Ag (< or =2 microM) was lost from solution within approximately 30 min, with a further decrease after 48 h root growth. Using measured Ag concentrations at the start of the experiments, the median effective concentration (EC50) for root elongation rate of cowpea (Vigna unguiculata [L.] Walp. cv. Caloona) was 0.010 microM Ag in the first 4 h of exposure (0.021 microM in the first 8 h). This demonstrates that Ag (as Ag+) is rapidly rhizotoxic to cowpea seedlings at concentrations similar to those that are toxic to freshwater biota. Rupturing of rhizodermal and outer cortical layers was evident after 48 h with 0.13 to 0.57 microM Ag initially in solution, being most severe at 0.13 or 0.25 microM Ag. An additional experiment showed that ruptures were first evident after 20 h exposure to 0.17 microM Ag, with increased severity of rupturing over time. The rhizotoxic effects of Ag are similar to those of some other trace metals (e.g., Cu, Al, La) that bind strongly to hard ligands and weakly to soft ligands. The similarity of rupturing effects, despite the difference in strong binding to soft ligands by Ag and to hard ligands by the other metals, suggests a distinctive metabolic effect of Ag that binds only weakly to hard ligands.

[Anatomy and adaptation to environment study of endangered alpine medical plant Neopicrorhiza scrophulariiflora].

OBJECTIVE: To study the anatomical structure of endangered alpine medical plant Neopicrorhiza scrophulariiflora and the high altitude adaptability. METHODS: The leaf epidermis character as well as section structure of leaf, aerial stem and rhizome were observed by light microscopical technique. RESULTS: The leaf surface of Neopicrorhiza scrophulariiflora was covered with two kinds of glandular hair, and the stommata was anomocytic type. Moreover, the leaf was isolateral and differed from most of alpine plant. The aerial stem had well-developed mechanical tissue. The rhizome was distributed by well-developed cork layers and collenchyma. Large numbers of aerenchymas distributed widely in leaf, aerial stem and rhizome. CONCLUSION: There existed characteristic traits in Neopicrorhiza scrophulariiflora that adapted the alpine environment, however, there still had some particular character different from other alpine plant. Thus, the adaptive style of alpine plant to high altitude environment was diversity.

[Distribution of legume arabinogalactanprotein-extensin (AGPE) glycoproteins in symbiotically defective pea mutants with abnormal infection threads].

The interface between the host cell and the microsymbiont is an important zone for development and differentiation during successive stages of the rhizobium-legume symbiosis. Legume root nodule extensins, otherwise known as arabinogalactanprotein-extensin (AGPE) are abundant components of the infection thread matrix. We have characterized the origin and distribution of these glycoproteins at the symbiotic interface of root nodules of symbiotically defective mutants of pea (Pisum sativum L.) using immunogold localization with MAC265 an anti-AGPE monoclonal antibody. For mutants with defective growth of infection threads, the AGPE epitope was abundant in the extracellular matrix surrounding infected host cells in the central infected tissue of the nodule as well as being present in the lumen of Rhizobium-induced infection threads. This suggests a mis-targetting of AGPE as a consequence of abnormal growth of the infection threads. Furthermore, mutants in gene sym33 showed reduced labelling with MAC265, and in infection threads and droplets label was completely absent, a phenomenon not observed in wild-type nodules. This suggests an alteration in the composition of the infection thread matrix for sym33 mutants which may be correlated with the absence of endocytosis of rhizobia into the host cytoplasm.

[Colonization of cucumber rhizosphere by phosphate-mobilizing bacteria of Bacillus genus].

An ability of phosphate-mobilizing bacteria, related to four species of Bacillus genus, to colonize the cucumber root zone has been investigated. It has been found that all the studied strains can adapt to the root zone of those plants colonizing its parts to different extent. Essential nonuniformity of the root surface populating with bacteria is noted. Microcolonies of these organisms are formed in some root zones. This peculiarity is probably caused both by the difference in the number and composition of exudates of some zones of the plant roots, and by peculiarities of trophic requirements of the strains of studied bacteria The paper is presented in Ukrainian.

Phenolic compound localisation in Polypodium vulgare L. rhizomes after mannitol-induced dehydration and controlled desiccation.

Polypodium vulgare L. is a desiccation-tolerant fern that can withstand successive dry periods in its life cycle. To better understand this mechanism, the current study was undertaken to assess the role of phenolic compounds in rhizome dehydration and determine their localisation in the rhizome cells after enforced dehydration in mannitol solution or controlled desiccation with or without abscisic acid (ABA) pretreatment. Phenolic distribution at the subcellular level was studied using gold particle-complexed laccase. Cells from different tissues: cortical parenchyma, endodermis and stelar elements--pericycle, sieve cells and vascular parenchyma were observed under a transmission electron microscope (TEM). The content of phenolic compounds was greater in ABA-untreated rhizomes after enforced dehydration in mannitol solution and subsequent rehydration. After controlled desiccation the phenolic content significantly increased in ABA-untreated rhizomes. A large number of phenolic compound deposits were present in all types of rhizomatous cells. Phenolics were widely distributed in the vacuoles of all cells, and in the secondary cell walls of sieve cells, although scattered labelling was hardly ever observed in the primary cell walls. In dehydrated and plasmolysed cells from the cortex and endodermis, phenolic compounds were present in the apoplastic compartments between the plasma membranes and the cell walls. There is evidence that abscisic acid plays a role as a crucial antioxidant resulting in no damage and a lower level of phenolic increase as compared to ABA-untreated rhizomes. Moreover, the location of phenolics suggests a protective chemical barrier against environmental stresses.

Composition, speciation and distribution of iron minerals in Imperata cylindrica.

A comparative study of the roots, rhizomes and leaves of an iron hyperaccumulator plant, Imperata cylindrica, isolated from the banks of an extreme acidic environment, using complementary techniques: Mösbauer spectroscopy (MS), X-ray diffraction (XRD), scanning electron microscopy (SEM) coupled to energy-dispersive X-ray microanalysis (EDAX) and transmission electron microscopy (TEM), has shown that two main biominerals, jarosite and ferrihydrate-ferritin, accumulate in the different tissues. Jarosite accumulates mainly in roots and rhizomes, while ferritin has been detected in all the structures. A model of iron management in I. cylindrica is presented.

[Microscopic identification of medicinal species of Acorus].

A study on the identification of commercial drugs of medicinal species of Acorus collected from 10 different areas had been carried out. The result showed that the commercial drugs were mainly derived from three species, i. e., Acorus tatarinowii, A. calamus L. and A. macrospadicesu (Yamamoto) F. N. Wei et Y. K. Li. With the digital imaging technique, the paper visually reflected the microscopic structures of medicinal plants of Acorus, and can be used for identifying the market drugs.

Endophytic nitrogen fixation in dune grasses (Ammophila arenaria and Elymus mollis) from Oregon.

Several tropical grasses harbor symbiotic nitrogen-fixing bacteria within their stem and rhizome tissue that may contribute to the nitrogen nutrition of the host plant. We present evidence here that sand dune grasses (Ammophila arenaria and Elymus mollis) from Oregon also contain nitrogen-fixing bacteria. Surface-sterilized stem and rhizome tissue from these species possess acetylene reduction (nitrogen fixation) activity and large populations (10(5) to 10(6) cfu/g fresh weight) of bacteria. These bacteria were cultured on N-free media and identified by sequencing of 16S rRNA genes or by GC-FAME. Random sequencing of numerous colonies from the initial isolation plates of mixed isolates showed that pseudomonads (Stenotrophomonas and Pseudomonas) were by far the most common microorganism. One isolate -Burkholderia sp. strain Aa1 - reduced acetylene in culture with maximum activity at an O(2) concentration of 2% (v/v) in liquid media or 10% on solid media. PCR screening of all the isolates with nifH and nifD primers was positive only for this species. Immunolocalization studies with antibodies to nitrogenase resulted in labeling within plant cell walls of stems and rhizomes. Evidence for a similar nitrogen-fixing association was also detected in Uniola paniculata (sea oats) and Ammophila brevigulata (American beachgrass). We conclude that these grasses, and probably other dune grasses from temperate climates, contain endophytic, diazotrophic bacteria that may contribute to the phenomenal success of these grasses on nutrient-poor sand.