Day-night changes of energy-rich compounds in crassulacean acid metabolism (CAM) species utilizing hexose and starch. (1/45)

BACKGROUND AND AIMS: Plants with crassulacean acid metabolism (CAM) can be divided into two groups according to the major carbohydrates used for malic acid synthesis, either polysaccharide (starch) or monosaccharide (hexose). This is related to the mechanism and affects energy metabolism in the two groups. In Kalanchoe pinnata and K. daigremontiana, which utilize starch, ATP-dependent phosphofructokinase (tonoplast inorganic pyrophosphatase) activity is greater than inorganic pyrophosphate-dependent phosphofructokinase (tonoplast adenosine triphosphatase) activity, but the reverse is the case in pineapple (Ananas comosus) utilizing hexose. To test the hypothesis that the energy metabolism of the two groups differs, day-night changes in the contents of ATP, ADP, AMP, inorganic phosphate (Pi), phosphoenolpyruvate (PEP) and inorganic pyrophosphate (PPi) in K. pinnata and K. daigremontiana leaves and in pineapple chlorenchyma were analysed. METHODS: The contents of energy-rich compounds were measured spectrophotometrically in extracts of tissue sampled in the light and dark, using potted plants, kept for 15 d before the experiments in a growth chamber. KEY RESULTS: In the three species, ATP content and adenylate energy charge (AEC) increased in the dark and decreased in the light, in contrast to ADP and AMP. Changes in ATP and AEC were greater in Kalanchoe leaves than in pineapple chlorenchyma. PPi content in the three species increased in the dark, but on illumination it decreased rapidly and substantially, remaining little changed through the rest of the light period. Pi content of Kalanchoe leaves did not change between dark and light, whereas Pi in pineapple chlorenchyma increased in the dark and decreased in the light, and the changes were far greater than in Kalanchoe leaves. Light-dark changes in PEP content in the three species were similar. CONCLUSIONS: These results corroborate our hypothesis that day-night changes in the contents of energy-rich compounds differ between CAM species and are related to the carbohydrate used for malic acid synthesis.  (+info)

Molecular cloning, expression, and functional characterization of a cystatin from pineapple stem. (2/45)

A cDNA fragment encoding the cysteine protease inhibitor, cystatin, was cloned from pineapple (Ananas comosus) stem. This clone was constructed in a fusion vector and was easily over-expressed in Escherichia coli; satisfactory over-expression of non-fusion cystatin was achieved after an additional start codon was inserted prior to its coding sequence. Both recombinant cystatins were predominately found in the soluble fraction of the cell extract, and were demonstrated to be functionally active in a reverse zymographic assay. The fusion and non-fusion cystatins were separately purified to homogeneity via a His-tag or papain-coupling affinity column. Effective inhibitory activity against papain was detected with both the fusion and non-fusion cystatins with comparable K(i) values of 1.18 x 10(-10) M and 9.53 x 10(-11) M, respectively. The recombinant cystatins were found to be thermally stable up to 60 degrees C. Inhibition of the endogenous protease activity in minced fish muscle revealed that the recombinant pineapple cystatins might be an adequate stabilizer to prevent protein degradation during industrial food processing.  (+info)

Respiratory properties and malate metabolism in Percoll-purified mitochondria isolated from pineapple, Ananas comosus (L.) Merr. cv. smooth cayenne. (3/45)

An investigation was made of the respiratory properties and the role of the mitochondria isolated from one phosphoenolpyruvate carboxykinase (PCK)-CAM plant Ananas comosus (pineapple) in malate metabolism during CAM phase III. Pineapple mitochondria showed very high malate dehydrogenase (MDH), and low malic enzyme (ME) and glutamate-oxaloacetate transaminase (GOT) activities. The mitochondria readily oxidized succinate and NADH with high rates and coupling, while they only oxidized NADPH in the presence of Ca(2+). Pineapple mitochondria oxidized malate with low rates under most assay conditions, despite increasing malate concentrations, optimizing pH, providing cofactors such as coenzyme A, thiamine pyrophosphate, and NAD(+), and supplying individually external glutamate or GOT. However, providing glutamate and GOT simultaneously strongly increased the rates of malate oxidation. The OAA easily permeated the mitochondrial membranes to import into or export out of pineapple mitochondria during malate oxidation, but the mitochondria did not consume external Asp or alpha-KG. These results suggest that OAA played a significant role in the mitochondrial malate metabolism of pineapple, in which malate was mainly oxidized by active mMDH to produce OAA which could be exported outside the mitochondria via a malate-OAA shuttle. Cytosolic GOT then consumed OAA by transamination in the presence of glutamate, leading to a large increase in respiration rates. The malate-OAA shuttle might operate as a supporting system for decarboxylation in phase III of PCK-CAM pineapple. This shuttle system may be important in pineapple to provide a source of energy and substrate OAA for cytosolic PCK activity during the day when cytosolic OAA and ATP was limited for the overall decarboxylation process.  (+info)

Developing pineapple fruit has a small transcriptome dominated by metallothionein. (4/45)

In a first step toward understanding the molecular basis of pineapple fruit development, a sequencing project was initiated to survey a range of expressed sequences from green unripe and yellow ripe fruit tissue. A highly abundant metallothionein transcript was identified during library construction, and was estimated to account for up to 50% of all EST library clones. Library clones with metallothionein subtracted were sequenced, and 408 unripe green and 1140 ripe yellow edited EST clone sequences were retrieved. Clone redundancy was high, with the combined 1548 clone sequences clustering into just 634 contigs comprising 191 consensus sequences and 443 singletons. Half of the EST clone sequences clustered within 13.5% and 9.3% of contigs from green unripe and yellow ripe libraries, respectively, indicating that a small subset of genes dominate the majority of the transcriptome. Furthermore, sequence cluster analysis, northern analysis, and functional classification revealed major differences between genes expressed in the unripe green and ripe yellow fruit tissues. Abundant genes identified from the green fruit include a fruit bromelain and a bromelain inhibitor. Abundant genes identified in the yellow fruit library include a MADS box gene, and several genes normally associated with protein synthesis, including homologues of ribosomal L10 and the translation factors SUI1 and eIF5A. Both the green unripe and yellow ripe libraries contained high proportions of clones associated with oxidative stress responses and the detoxification of free radicals.  (+info)

Towards sustainable use of potassium in pineapple waste. (5/45)

Due to the 1997/98 haze problem in South-East Asia and the increasing need for sustainable food production and development, the usual management of crop residues (including pineapple wastes) through burning is prohibited. As a result, the need for alternative uses of pineapple wastes in pineapple production has been emphasized. This study investigated an environmentally friendly means of recycling pineapple leaves for agricultural use. Pineapple leaves were shredded and composted in a composting drum for 30 days. Part of the shredded leaves was ashed in a muffle furnace for 4 h. Humic acid (HA), K-fulvate, and K in HA and compost were analyzed using standard procedures. An ash to water ratio of 1:7 was used to extract 0.1 molar (M) KOH from the shredded leaves. The 0.1 M KOH contained 50% K and was able to extract 20% HA from the composted pineapple leaves. Percent K in the fulvate using 0.1 M KOH was 43. Besides serving as a foliar spray (supplement soil application K fertilizers), source of K for freshwater fish (e.g., tilapia), the HA produced can be used as a soil conditioner. Studies show that between 0.05-0.01 g of HA per kg soil retards runoff by 36% in sandy and sandy loam soils. The K-fulvate can be used as a fluid fertilizer. In addition, the pH of 2 of the K-fulvate suggests it could be used to dissolve phosphate rocks, particularly those in the arid regions where high soil pH does not facilitate the dissolution of these important rocks that serve as one of the sources of phosphorus fertilizer in agriculture.  (+info)

Best evidence topic report. Proteolytic enzymes for oesophageal meat impaction. (6/45)

A short cut review was carried out to establish whether proteolytic enzymes are effective at resolving oesophageal meat impaction. Altogether 98 papers were found using the reported search, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated.  (+info)

Purification, characterization, and antifungal activity of chitinases from pineapple (Ananas comosus) leaf. (7/45)

Three chitinases, designated pineapple leaf chitinase (PL Chi)-A, -B, and -C were purified from the leaves of pineapple (Ananas comosus) using chitin affinity column chromatography followed by several column chromatographies. PL Chi-A is a class III chitinase having a molecular mass of 25 kDa and an isoelectric point of 4.4. PL Chi-B and -C are class I chitinases having molecular masses of 33 kDa and 39 kDa and isoelectric points of 7.9 and 4.6 respectively. PL Chi-C is a glycoprotein and the others are simple proteins. The optimum pHs of PL Chi-A, -B, and -C toward glycolchitin are pH 3, 4, and 9 respectively. The chitin-binding ability of PL Chi-C is higher than that of PL Chi-B, and PL Chi-A has lower chitin-binding ability than the others. At low ionic strength, PL Chi-B exhibits strong antifungal activity toward Trichoderma viride but the others do not. At high ionic strength, PL Chi-B and -C exhibit strong and weak antifungal activity respectively. PL Chi-A does not have antifungal activity.  (+info)

Applied K fertilizer use efficiency in pineapples grown on a tropical peat soil under residues removal. (8/45)

In Malaysia, pineapples are grown on peat soils, but most K fertilizer recommendations do not take into account K loss through leaching. The objective of this study was to determine applied K use efficiency under a conventionally recommended fertilization regime in pineapple cultivation with residues removal. Results showed that K recovery from applied K fertilizer in pineapple cultivation on tropical peat soil was low, estimated at 28%. At a depth of 0-10 cm, there was a sharp decrease of soil total K, exchangeable K, and soil solution K days after planting (DAP) for plots with K fertilizer. This decline continued until the end of the study. Soil total, exchangeable, and solution K at the end of the study were generally lower than prior values before the study. There was no significant accumulation of K at depths of 10-25 and 25-45 cm. However, K concentrations throughout the study period were generally lower or equal to their initial status in the soil indicating leaching of the applied K and partly explained the low K recovery. Potassium losses through leaching in pineapple cultivation on tropical peat soils need to be considered in fertilizer recommendations for efficient recovery of applied K.  (+info)

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