David J. Longstreth

Changes in cell structure during the formation of root aerenchyma inSAGITTARIA LANCIFOLIA (Alismataceae).

In many wetland species, root aerenchyma is produced by the predictable collapse of root cortex cells, indicating a programmed cell death (PCD). The objective of this study was to characterize the cellular changes that accompany this PCD in the marsh species Sagittaria lancifolia. Structural changes in membranes and organelles were examined during development of root cortex cells to compare with previous examples of PCD. The organization of cortical microtubule (CMT) arrays in root cells from S. lancifolia was also evaluated as a possible predictor of cell lysis. Nuclear fragmentation and condensation were the earliest changes observed in cells undergoing lysis. Breakdown of the tonoplast and other organelles and disruption of the plasma membrane followed. After loss of cytoplasm, cells collapsed to form gas spaces. These results were compared to collapse of root cortical cells of Zea mays and Oryza sativa during aerenchyma development. Changes in the appearance of the cytoplasm of all three species were similar at later stages of aerenchyma development. The relative timing of disintegration of the tonoplast and middle lamella appeared to differ among the three species. Changes in the organization of CMT arrays did not appear to be a predictor of PCD in S. lancifolia. Aerenchyma production in plants involves a type of PCD that is morphologically distinct from PCD described from many animals.

Photorespiration and carbon concentrating mechanisms: two adaptations to high O2, low CO2 conditions

This review presents an overview of the two ways that cyanobacteria, algae, and plants have adapted to high O2 and low CO2 concentrations in the environment. First, the process of photorespiration enables photosynthetic organisms to recycle phosphoglycolate formed by the oxygenase reaction catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Second, there are a number of carbon concentrating mechanisms that increase the CO2 concentration around Rubisco which increases the carboxylase reaction enhancing CO2 fixation. This review also presents possibilities for the beneficial modification of these processes with the goal of improving future crop yields.

The carbon concentrating mechanism in Chlamydomonas reinhardtii : finding the missing pieces

The photosynthetic, unicellular green alga, Chlamydomonas reinhardtii, lives in environments that often contain low concentrations of CO2 and HCO3−, the utilizable forms of inorganic carbon (Ci). C. reinhardtii possesses a carbon concentrating mechanism (CCM) which can provide suitable amounts of Ci for growth and development. This CCM is induced when the CO2 concentration is at air levels or lower and is comprised of a set of proteins that allow the efficient uptake of Ci into the cell as well as its directed transport to the site where Rubisco fixes CO2 into biomolecules. While several components of the CCM have been identified in recent years, the picture is still far from complete. To further improve our knowledge of the CCM, we undertook a mutagenesis project where an antibiotic resistance cassette was randomly inserted into the C. reinhardtii genome resulting in the generation of 22,000 mutants. The mutant collection was screened using both a published PCR-based approach (Gonzalez-Ballester et al. 2011) and a phenotypic growth screen. The PCR-based screen did not rely on a colony having an altered growth phenotype and was used to identify colonies with disruptions in genes previously identified as being associated with the CCM-related gene. Eleven independent insertional mutations were identified in eight different genes showing the usefulness of this approach in generating mutations in CCM-related genes of interest as well as identifying new CCM components. Further improvements of this method are also discussed.

Photosynthesis and photorespiration in freshwater emergent and floating plants

Abstract Little is known about photosynthesis and photorespiration in freshwater emergent and floating species even though these plants can be very effective producers of biomass. Species for which carbon fixation has been examined in any detail are generally C3, although more studies are needed to draw firm conclusions. There appear to be no unique biochemical adaptations in the carbon fixation pathways of these species. There may be unusually high CO2 concentrations in the leaf intercellular spaces of at least some of these species. Gas movement from roots or rhizomes to leaves through spaces or aerenchyma produces these high concentrations. As a consequence, photosynthetic rates in situ may be higher than expected for C3 species because ambient CO2 concentrations are normally limiting to photosynthesis. Most emergent species have a relatively vertical leaf orientation and a high leaf-area index (leaf area per unit substrate surface area), characteristics that should optimize canopy carbon fixation. While hypoxia in standing water can severely affect plant physiology, species adapted to these conditions have a relatively unlimited water supply. Other environmental conditions such as temperature and photon flux density are generally favorable for carbon fixation in many of these wet habitats. Therefore, high productivity of at least some of these species may be caused by supplementation of atmospheric CO2 by substrate sources, optimal canopy architecture, and growth under favorable environmental conditions.

Aerenchyma Carbon Dioxide Can Be Assimilated in Typha Iatifolia L. Leaves

Leaf structural characteristics and gas-exchange measurements were used to determine whether photosynthetic tissue of Typha Iatifolia L. (cattail) utilized CO2 from the aerenchyma gas spaces, part of an internal pathway for gas transport in this wetland species. The partial pressure of CO2 (pCO2) in these aerenchyma gas spaces can be more than 10 times atmospheric pCO2. The photosynthetic tissue occurred in structurally similar adaxial and abaxial palisades, which were distinctly separated from each other by the aerenchyma gas spaces. In each palisade there were three to four layers of tightly packed, nonchlorophyllous cells separating the photosynthetic tissue from the aerenchyma gas space. Different lines of evidence indicated that CO2 conductance in the light was significantly greater across the epidermal surface than across the internal surface of both palisades. However, at an epidermal pCO2 of 350 [mu]bars and an internal pCO2 of 820 [mu]bars, the net rates of CO2 uptake (PN) across the epidermal and internal surfaces were about equal. PN across the internal surface was greater than across the epidermal surface at higher internal pCO2. Gas space pCO2 can be greater than 820 [mu]bars in the field, and therefore, PN across the internal surface could be a significant proportion of epidermal surface PN.

The Cytoplasmic Carbonic Anhydrases βCA2 and βCA4 Are Required for Optimal Plant Growth at Low CO2

The removal of two cytoplasmic carbonic anhydrases, βCA2 and βCA4, from Arabidopsis causes reduced growth in plants grown in a low-CO2 environment. Carbonic anhydrases (CAs) are zinc metalloenzymes that interconvert CO2 and HCO3−. In plants, both α- and β-type CAs are present. We hypothesize that cytoplasmic βCAs are required to modulate inorganic carbon forms needed in leaf cells for carbon-requiring reactions such as photosynthesis and amino acid biosynthesis. In this report, we present evidence that βCA2 and βCA4 are the two most abundant cytoplasmic CAs in Arabidopsis (Arabidopsis thaliana) leaves. Previously, βCA4 was reported to be localized to the plasma membrane, but here, we show that two forms of βCA4 are expressed in a tissue-specific manner and that the two proteins encoded by βCA4 localize to two different regions of the cell. Comparing transfer DNA knockout lines with wild-type plants, there was no reduction in the growth rates of the single mutants, βca2 and βca4. However, the growth rate of the double mutant, βca2βca4, was reduced significantly when grown at 200 μL L−1 CO2. The reduction in growth of the double mutant was not linked to a reduction in photosynthetic rate. The amino acid content of leaves from the double mutant showed marked reduction in aspartate when compared with the wild type and the single mutants. This suggests the cytoplasmic CAs play an important but not previously appreciated role in amino acid biosynthesis.

Solutes Involved in Osmotic Adjustment to Increasing Salinity in Suspension Cells of Alternanthera philoxeroides Griseb

Cell recovery from osmotic stress was studied in suspension cell cultures from Alternanthera philoxeroides [Mart.] Griseb. Changes in different classes of cellular solutes were measured after cells were transferred from 0 to 200 mM NaCl (high salt) to obtain an integrated picture of the solute pools involved in osmotic adjustment. By 2 h, cellular [Na+] and [Cl−] had increased several-fold, potentially accounting for the osmotic adjustment that produced a rapid recovery of cell turgor. There was a four-fold increase in the concentration of quaternary ammonium compounds (QAC) by 12 h and a slower increase for several days afterward. Betaine aldehyde dehydrogenase (BADH) is required for synthesis of glycine betaine, a QAC produced by a range of organisms in response to osmotic stress. Western-blot analysis for BADH suggested that glycine betaine was a significant component of the QAC solutes. The amount of BADH was generally similar at different sampling times for control and high salt cells, unlike previous reports of stimulation by osmotic stress in intact plants of some species. Between 3 and 7 days after cell transfer to high salt, other organic solutes increased in concentration and [Na+] and [Cl−] decreased. In A. philoxeroides, high [Na+] and [Cl−] produce rapid osmotic adjustment but organic solutes apparently replace these potentially harmful inorganic ions after the recovery of turgor.