(2004b). Rubisco activity limits photosynthesis at low temperatures. Rev. The bundle sheath in a leaf is a layer of compactly arranged parenchyma surrounding the vasculature (Esau, 1965) and is a conduit between the vasculature and the mesophyll cells. In addition, representative 2-DE gels from BS and M chloroplast stroma with associated information can be interrogated. Where needed, functional assignments (Bins) were modified based on information from the published literature. Thus, the size of bundle sheath compartments and the total volume of chloroplasts within them limit the space available for Rubisco. Five independent M and BS chloroplast preparations were resolved on five pairs of 2-DE gels. The Calvin cycle produces ________, which are in turn used in the light reactions. RuBP carboxylase (rubisco) is able to bind with both ________________. The yield of chloroplast preparations was inversely proportional to their purity. The two-step process by which 4-carbon compound oxaloacetate is produced in the mesophyll cells and transported into bundle sheath cells of chloroplast in Crassulacean acid metabolism (CAM) plants is called the C4 pathway in mesophyll cells. Which of the following is a disadvantage of C4 photosynthesis relative to C3 photosynthesis? Another three molecules of ATP are then used in the cycle. Although bundle sheath chloroplasts display normal internal ultrastructure and are photosynthetically active, they are smaller, fewer in number, and are positioned along the cell wall distal to the vasculature. C3 plants counter this limitation by increasing leaf Rubisco content, yet few C4 species do the same. The MS or MS/MS spectra were searched against the maize EST assembly from TIGR (http://www.tigr.org) (ZmGI, v1.4) and maize genome assemblies (AZM 4.0) as well as by homology-based searches against the annotated rice (Oryza sativa) genome (TIGR Rice Genome version 2), using an in-house installation of Mascot (www.matrixscience.com). Energy to drive the formation of ATP in photosynthesis is derived from ____________. Ambiguous matches were resolved by protein identification from MS. After background removal and normalization of each spot volume to the total gel spot volume, virtual average M and BS gels were created, where each spot volume represented the average of volumes of matched spots in M or BS gels. When the stomata close during heat stress, RuBP carboxylase is not exposed to the increase in oxygen concentration experienced in the leaf. Although a large collection of reticulate leaf mutants is Mesophyll and bundle sheath cells have structurally and functionally different chloroplasts. On average, preparations with <15% cross-contamination yielded ∼300 μg of protein for each M and BS fraction. The cell-specific C 4 chloroplast arrangement is established during cell … Carotenoids are found in high concentrations in yellow and orange leaves or vegetables. Plant Physiol. open stomata only at night, limiting water loss because of heat and low humidity. Subsequently, chloroplasts were further isolated, and the stromal proteome was released by osmotic shock, followed by removal of thylakoid and envelope membranes by ultracentrifugation, similarly as described by Peltier et al. How does ATP synthase obtain the energy to produce ATP? On average, we included gels only those matched spots that were present at least three times on M or BS gels. This plant very likely carries out C4 photosynthesis. CAM photosynthesis limits CO2 fixation to nighttime hours in order to _______________. In maize ( Zea mays ), Bundle Sheath Defective2 (BSD2) plays an essential role in Rubisco biogenesis and is required for correct bundle sheath (BS) cell differentiation. It summarizes the currently accepted views on metabolic compartmentation of biochemical reactions between the chloroplasts of bundle sheath cells and mesophyll cells of a C 4 plant. D) They do not have any intercellular spaces. Which stages of the Calvin cycle reactions require the use of energy from ATP? This pathway is also called Hatch and Slack pathway. In the first run, mass spectrometric parameters were setup favoring more time for the survey scans, and high-quality tandem MS spectra for peptides with higher signal intensity could also be obtained in this run. Sage et al. When needed, peak detection was manually adjusted to remove the influence of obvious satellite peak(s). In a redox reaction, reduction means ________, and oxidation means ________. Supplemental Table 3. What is the purpose of the three additional ATP molecules? Salvucci for polyclonal serum against PEPC. The … A plant that uses CAM photosynthesis is most likely to be successful in what type of environment? ↵ Peltier, and J. Ytterberg for numerous discussions and advice. These distinct lineages exhibit unique aspects that highlight their independent evolution from C3 ancestors. reflect and transmit yellow and orange wavelengths of light. Corresponding interactive gels with protein spot identities that were quantified are available in the PPDB. NADP-malic enzyme (NADP-ME) occurs in the chloroplasts of bundle-sheath cells and, like the other decarboxylases of C 4 photosynthesis, it releases CO 2, which can be fixed directly by Rubisco. The protein quantification was done by comparing the peak areas of corresponding peptides from BS or M. Masslynx 4.0 software (Waters) was employed to extract the peak areas of identified peptides of the corresponding single ion chromatograms. Functional Differentiation of Bundle Sheath and Mesophyll Maize Chloroplasts Determined by Comparative Proteomics. Yet, BSD2 RNA and protein levels are similar in mesophyll (M) and BS chloroplasts, although Rubisco accumulates only in BS chloroplasts. Which of the following is not derived from G3P? B Biol. The majority of spots (65% for M and 54% for BS gels) were represented at least three times either in M and/or in BS proteomes (45% for M and 39% for BS spots were represented at least four times). Bundle sheath cells constitute ∼15% of chloroplast-containing cells in an Arabidopsis leaf (Kinsman and Pyke, 1998), and they conduct fluxes of compounds both into the leaf, particularly during leaf development, and out of the leaf, during export of photosynthates and during senescence. Denatured apparent molecular masses were determined from molecular mass markers loaded onto the second dimension gels. The removal of electrons and/or hydrogen atoms from a substrate is called what? Before being converted into sucrose, starch, or cellulose, G3P must first be converted into which compound? It takes place prior to the calvin cycle. Therefore, only ZmGI was used for detailed protein quantification. Replacement electrons for the reaction center of photosystem II are derived from _____________. How many times must the Calvin cycle turn to yield a single molecule of glyceraldehyde 3-phosphate (G3P)? A new species of plant is discovered and you are asked to help classify it. The _______ from the light reactions is used to reduce CO2 to a carbohydrate using energy derived from ______. Briefly, purified BS and M stromal proteomes were reduced with DTT and alkylated using iodoacetamide, followed by in-solution digestion by trypsin. The flow-through containing the nonlabeled peptides was also collected since they were valuable for protein identification. During photosynthesis, ________ is oxidized, while ________ is reduced. The biotin-tagged and cystein-containing peptides (useful for quantification) were then purified on avidin affinity columns, and the majority of the tag (containing the bulky biotin tag but not the isotope labels) was removed by acid hydrolysis, according to the manufacturer's instructions. It is concluded that the structure of the specialized chloroplasts in bundle sheath cells of sugar cane is a result of reduction, and that the development of chloroplast dimorphism is related in some way to leaf cell differentiation. Hydrogen ions flow down a concentration gradient from the thylakoid space to the stroma through ATP synthase, releasing energy that can be used to produce ATP from ADP + Pi. 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