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Histological analysis and 3D reconstruction of winter cereal crowns recovering from freezing: a unique response in oat (Avena sativa L.).

ABSTRACT: The crown is the below ground portion of the stem of a grass which contains meristematic cells that give rise to new shoots and roots following winter. To better understand mechanisms of survival from freezing, a histological analysis was performed on rye, wheat, barley and oat plants that had been frozen, thawed and allowed to resume growth under controlled conditions. Extensive tissue disruption and abnormal cell structure was noticed in the center of the crown of all 4 species with relatively normal cells on the outside edge of the crown. A unique visual response was found in oat in the shape of a ring of cells that stained red with Safranin. A tetrazolium analysis indicated that tissues immediately inside this ring were dead and those outside were alive. Fluorescence microscopy revealed that the barrier fluoresced with excitation between 405 and 445 nm. Three dimensional reconstruction of a cross sectional series of images indicated that the red staining cells took on a somewhat spherical shape with regions of no staining where roots entered the crown. Characterizing changes in plants recovering from freezing will help determine the genetic basis for mechanisms involved in this important aspect of winter hardiness.

SUBMITTER: Livingston DP

PROVIDER: S-EPMC3544926 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Histological analysis and 3D reconstruction of winter cereal crowns recovering from freezing: a unique response in oat (Avena sativa L.).

Livingston David P DP Henson Cynthia A CA Tuong Tan D TD Wise Mitchell L ML Tallury Shyamalrau P SP Duke Stanley H SH

PloS one 20130114 1

The crown is the below ground portion of the stem of a grass which contains meristematic cells that give rise to new shoots and roots following winter. To better understand mechanisms of survival from freezing, a histological analysis was performed on rye, wheat, barley and oat plants that had been frozen, thawed and allowed to resume growth under controlled conditions. Extensive tissue disruption and abnormal cell structure was noticed in the center of the crown of all 4 species with relatively ...[more]

PMID: 23341944

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Project description:BackgroundDeficiency of available forms of phosphorus is common in most soils and causes reduction of crop plants growth and yield. Recently, model plants responses to phosphate (Pi) deficiency have been intensively studied. However, acclimation mechanisms of cereals like oat (Avena sativa L.), to low Pi stress remains not fully understood. Oat plants have been usually cultured on poor soils, with a low nutrient content, but their responses to such conditions are not well known, therefore the main goal of the study was to investigate the mechanisms that enable oat plants to grow under low Pi conditions.MethodsFour oat cultivars (A. sativa, cv. Arab, Krezus, Rajtar and Szakal) were grown for three weeks in a nutrient media with various P sources: inorganic-KH2PO4 (control), organic-phytate (PA) and with no phosphate (-P). The effects of Pi deficiency on the level of P, oat growth parameters, intensity of photosynthesis, plant productivity, root exudation ability, localization, activity and isoforms of acid phosphatases, enzymes involved in Pi mobilization, were estimated. In addition, the effect of mycorrhization on plant growth was also observed.ResultsAll studied oat cultivars grown on Pi-deficient media had significantly decreased Pi content in the tissues. Pi deficiency caused inhibition of shoot growth, but generally it did not affect root elongation; root diameter was decreased, root/shoot ratios increased, whereas PA plants showed a similar growth to control. Photosynthesis rate and productivity parameters decreased under low Pi nutrition, however, sugar content generally increased. Studied oat cultivars did not respond to low Pi via increased exudation of carboxylates from the roots, as pH changes in the growth media were not observed. Pi starvation significantly increased the activity of extracellular and intracellular acid phosphatases (APases) in comparison to the control plants. Three major APase isoforms were detected in oat tissues and the isoform pattern was similar in all studied conditions, usually with a higher level of one of the isoforms under Pi starvation. Generally no significant effects of mycorrhizal colonization on growth of oat cultivars were observed.DiscussionWe postulated that acid phosphatases played the most important role in oat cultivars acclimation to Pi deficiency, especially extracellular enzymes involved in Pi acquisition from soil organic P esters. These APases are mainly located in the epidermis of young roots, and may be released to the rhizosphere. On the other hand, intracellular APases could be involved in fast Pi remobilization from internal sources. Our study showed that oat, in contrast to other plants, can use phytates as the sole source of P. The studied oat cultivars demonstrated similar acclimation mechanisms to Pi deficiency, however, depending on stress level, they can use different pools of acid phosphatases.

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Project description:One of the most important characteristics of successful plant defence is the ability to rapidly identify potential threats in the surrounding environment. Plants rely on the perception of microbe-derived molecular pattern chemicals for this recognition, which initiates a number of induced defence reactions that ultimately increase plant resistance. The metabolome acts as a metabolic fingerprint of the biochemical activities of a biological system under particular conditions, and therefore provides a functional readout of the cellular mechanisms involved. Untargeted metabolomics was applied to decipher the biochemical processes related to defence responses of oat plants inoculated with pathovars of Pseudomonas syringae (pathogenic and non-pathogenic on oat) and thereby identify signatory markers that are involved in host or nonhost defence responses. The strains were P. syringae pv. coronafaciens (Ps-c), P. syringae pv. tabaci, P. syringae pv. tomato DC3000 and the hrcC mutant of DC3000. At the seedling growth stage, metabolic alterations in the Dunnart oat cultivar (tolerant to Ps-c) in response to inoculation with the respective P. syringae pathovars were examined following perception and response assays. Following inoculation, plants were monitored for symptom development and harvested at 2-, 4- and 6 d.p.i. Methanolic leaf extracts were analysed by ultra-high-performance liquid chromatography (UHPLC) connected to high-definition mass spectrometry. Chemometric modelling and multivariate statistical analysis indicated time-related metabolic reconfigurations that point to host and nonhost interactions in response to bacterial inoculation/infection. Metabolic profiles derived from further multivariate data analyses revealed a range of metabolite classes involved in the respective defence responses, including fatty acids, amino acids, phenolic acids and phenolic amides, flavonoids, saponins, and alkaloids. The findings in this study allowed the elucidation of metabolic changes involved in oat defence responses to a range of pathovars of P. syringae and ultimately contribute to a more comprehensive view of the oat plant metabolism under biotic stress during host vs nonhost interactions.

Dataset Information

Histological analysis and 3D reconstruction of winter cereal crowns recovering from freezing: a unique response in oat (Avena sativa L.).

Publications

Histological analysis and 3D reconstruction of winter cereal crowns recovering from freezing: a unique response in oat (Avena sativa L.).

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