11 research outputs found

    Mechanical Effects of Cellulose, Xyloglucan, and Pectins on Stomatal Guard Cells of Arabidopsis thaliana

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    Stomata function as osmotically tunable pores that facilitate gas exchange at the surface of plants. Stomatal opening and closure are regulated by turgor changes in guard cells that result in mechanically regulated deformations of guard cell walls. However, how the molecular, architectural, and mechanical heterogeneities that exist in guard cell walls affect stomatal dynamics is unclear. In this work, stomata of wild type Arabidopsis thaliana plants or of mutants lacking normal cellulose, hemicellulose, or pectins were experimentally induced to close or open. Three-dimensional images of these stomatal complexes were collected using confocal microscopy, images were landmarked, and three-dimensional finite element models (FEMs) were constructed for each complex. Stomatal opening was simulated with a 5 MPa turgor increase. By comparing experimentally measured and computationally modeled changes in stomatal geometry across genotypes, anisotropic mechanical properties of guard cell walls were determined and mapped to cell wall components. Deficiencies in cellulose or hemicellulose were both predicted to stiffen guard cell walls, but differentially affected stomatal pore area and the degree of stomatal opening. Additionally, reducing pectin molecular mass altered the anisotropy of calculated shear moduli in guard cell walls and enhanced stomatal opening. Based on the unique architecture of guard cell walls and our modeled changes in their mechanical properties in cell wall mutants, we discuss how each polysaccharide class contributes to wall architecture and mechanics in guard cells. This study provides new insights into how the walls of guard cells are constructed to meet the mechanical requirements of stomatal dynamics

    Balancing Strength and Flexibility: How the Synthesis, Organization, and Modification of Guard Cell Walls Govern Stomatal Development and Dynamics

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    Guard cells are pairs of epidermal cells that control gas diffusion by regulating the opening and closure of stomatal pores. Guard cells, like other types of plant cells, are surrounded by a three-dimensional, extracellular network of polysaccharide-based wall polymers. In contrast to the walls of diffusely growing cells, guard cell walls have been hypothesized to be uniquely strong and elastic to meet the functional requirements of withstanding high turgor and allowing for reversible stomatal movements. Although the walls of guard cells were long underexplored as compared to extensive studies of stomatal development and guard cell signaling, recent research has provided new genetic, cytological, and physiological data demonstrating that guard cell walls function centrally in stomatal development and dynamics. In this review, we highlight and discuss the latest evidence for how wall polysaccharides are synthesized, deposited, reorganized, modified, and degraded in guard cells, and how these processes influence stomatal form and function. We also raise open questions and provide a perspective on experimental approaches that could be used in the future to shed light on the composition and architecture of guard cell walls

    Universal Plant DNA Barcode Loci May Not Work in Complex Groups: A Case Study with Indian Berberis Species

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    BACKGROUND: The concept of DNA barcoding for species identification has gained considerable momentum in animals because of fairly successful species identification using cytochrome oxidase I (COI). In plants, matK and rbcL have been proposed as standard barcodes. However, barcoding in complex genera is a challenging task. METHODOLOGY AND PRINCIPAL FINDINGS: We investigated the species discriminatory power of four reportedly most promising plant DNA barcoding loci (one from nuclear genome--ITS, and three from plastid genome--trnH-psbA, rbcL and matK) in species of Indian Berberis L. (Berberidaceae) and two other genera, Ficus L. (Moraceae) and Gossypium L. (Malvaceae). Berberis species were delineated using morphological characters. These characters resulted in a well resolved species tree. Applying both nucleotide distance and nucleotide character-based approaches, we found that none of the loci, either singly or in combinations, could discriminate the species of Berberis. ITS resolved all the tested species of Ficus and Gossypium and trnH-psbA resolved 82% of the tested species in Ficus. The highly regarded matK and rbcL could not resolve all the species. Finally, we employed amplified fragment length polymorphism test in species of Berberis to determine their relationships. Using ten primer pair combinations in AFLP, the data demonstrated incomplete species resolution. Further, AFLP analysis showed that there was a tendency of the Berberis accessions to cluster according to their geographic origin rather than species affiliation. CONCLUSIONS/SIGNIFICANCE: We reconfirm the earlier reports that the concept of universal barcode in plants may not work in a number of genera. Our results also suggest that the matK and rbcL, recommended as universal barcode loci for plants, may not work in all the genera of land plants. Morphological, geographical and molecular data analyses of Indian species of Berberis suggest probable reticulate evolution and thus barcode markers may not work in this case

    Online Design Aid for Evaluating Manure Pit Ventilation Systems to Reduce Entry Risk

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    On-farm manure storage pits contain both toxic and asphyxiating gases such as hydrogen sulfide, carbon dioxide, methane and ammonia. Farmers and service personnel occasionally need to enter these pits to conduct repair and maintenance tasks. One intervention to reduce the toxic and asphyxiating gas exposure risk to farm workers when entering manure pits is manure pit ventilation. This article describes an online computational fluid dynamics based design aid for evaluating the effectiveness of manure pit ventilation systems to reduce the concentrations of toxic and asphyxiating gases in the manure pits. This design aid, developed by a team of agricultural engineering and agricultural safety specialists at Pennsylvania State University, represents the culmination of more than a decade of research and technology development effort. The article includes a summary of the research efforts leading to the online design aid development and describes protocols for using the online design aid, including procedures for data input and for accessing design aid results. Design aid results include gas concentration decay and oxygen replenishment curves inside the manure pit and inside the barns above the manure pits, as well as animated motion pictures of individual gas concentration decay and oxygen replenishment in selected horizontal and vertical cut plots in the manure pits and barns. These results allow the user to assess: (1) how long one needs to ventilate the pits to remove toxic and asphyxiating gases from the pit and barn, (2) from which portions of the barn and pit these gases are most and least readily evacuated, and (3) whether or not animals and personnel need to be removed from portions of the barn above the manure pit being ventilated
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