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PubMed1996

Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase from Brassica napus.

Seven distinct partial cDNAs, similar in sequence to previously described polygalacturonases (PGs), were amplified from cDNA derived from rape pod wall, dehiscence zone and leaves by the polymerase chain reaction. Northern analysis showed that one clone, PG35-8, was expressed at low levels in the dehiscence zone during the first five weeks after anthesis but was very abundantly expressed at week 6. In contrast, no PG35-8-related RNA was detected in the pod wall. Our data suggest that there are temporal and spatial correlations between the breakdown of the middle lamella, of the dehiscence zone cells and the pattern of synthesis of PG35-8 transcripts which may indicate a role for this particular PG in rape pod dehiscence. PG35-8 was used to isolate five cDNA clones from a rape dehiscence zone cDNA library. Restriction enzyme analysis and partial sequencing revealed that they were derived from four highly homologous transcripts which are probably allelic forms of a single gene. One full-length clone, RDPG1, was completely sequenced. The predicted protein of RDPG1 showed its highest identity with PG from apple fruit with an identity of 52%.

Amino Acid Sequence↗
PubMed1978

Electron microscopic studies in cultivated plants. I. Green pods of Phaseolus vulgaris var. nanus L.

Scanning (SEM) and transmission (TEM) electron microscopic studies in the legumen of Phaseolus vulgaris var. nanus L. (kidney beans) were carried out. In this work emphasis was laid on the analysis of the morphological structure of the parenchyma tissue, being the chief component of the pod at edible maturity. The leaf character of the pod becomes especially evident by the occurence of stomata, trichomes, and typical cuticle structures on the outside of the pod (SEM). The cells of the mesophyll-like parenchyma tissue are distinguished by a great range of variation of their plastids (chloroplasts--chloroamyloplasts--amyloplasts) and some special cytological features such as a strongly developed rough ER, big nuclei, and numerous plasmodesmata (TEM).

Cell Nucleus↗
PubMed1983

Chitosans and pectic polysaccharides both induce the accumulation of the antifungal phytoalexin pisatin in pea pods and antinutrient proteinase inhibitors in tomato leaves.

The Proteinase Inhibitor Inducing Factor, PIIF, a pectic polysaccharide that induces synthesis and accumulation of proteinase inhibitor proteins in tomato and potato leaves, is an effective elicitor of the phytoalexin pisatin in pea pod tissues. The levels of pisatin induced by PIIF, and the time course of elicitation, are similar to those induced by chitosans, beta-1,4 glucosamine polymers, which are potent elicitors of pisatin in pea pods. Similarly, the chitosans, found in both insect and fungal cell walls, are the most potent inducers yet found of proteinase inhibitor accumulation in excised tomato cotyledons. The similarity in the induction of synthesis of proteinase inhibitors in tomato cotyledons and of pisatin in pea pods by pectic polysaccharides and chitosans suggests that the two polysaccharide types may be triggering a similar fundamental system present in pea and tomato plants that regulates the expression of genes for natural protection systems.

Benzopyrans↗
PubMed1996

Polygalacturonase inhibitors in bean pods.

The amount of polygalacturonase-inhibiting protein (PGIP) was 14 times higher in bean pods than in etiolated hypocotyls. The PGIP was extracted from bean pods and partially purified by chromatography on columns of S-Sepharose. DEAE-Sephadex A-50, and Sephadex G-75. Further purification by ion-exchange chromatography on a Mono Q column separated two isoforms of the inhibitor. The two PGIPs were similar in most properties but differed slightly in pI values. They also differed in one residue of the N-terminal amino acid sequences. Both bean pod PGIPs differed in two and possibly three residues of the deduced N-terminal amino acid sequence for hypocotyl PGIP. Small alterations in the structure of PGIP may represent a strategy in bean plants for resistance to a variety of pathogens.

Amino Acid Sequence↗
PubMed2002

Antioxidant activity of polyphenols in carob pods.

We extracted polyphenols from carob (Ceratonia siliqua L.) pods, and evaluated the in vitro antioxidant activity of the crude polyphenol fraction (CPP). The total polyphenol content in CPP determined by the Folin-Ciocalteu method was 19.2%. The condensed tannin content determined by the vanillin and proanthocyanidin assay systems was 4.37% and 1.36%, respectively. beta-Carotene bleaching, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, inhibition of lipid peroxidation by the erythrocyte ghost, and microsomal assay systems were used to evaluate the antioxidant activity. CPP showed a stronger inhibitory effect against the discoloration of beta-carotene than other polyphenol compounds such as catechins and procyanidins. CPP had weaker antioxidant activity in the DPPH free radical scavenging, the erythrocyte ghost, and microsomal systems than authentic polyphenol compounds at the same concentrations. The activity adjusted by the polyphenol concentration was, however, comparable to that of authentic polyphenol compounds. Considering most carob pods are discarded and not effectively utilized at present, these results suggested that carob pods could be utilized as a functional food or food ingredient.

Antioxidants↗
PubMed2002

Cocoa-based media for culturing Phytophthora palmivora (Butl.) Butl., causal agent of black pod disease of cocoa.

Green cocoa pod husk agar (GCPA), ripe cocoa pod husk agar (RCPA), green cocoa bean agar (GCBA), ripe cocoa bean agar (RCBA), green cocoa mucilage agar (GCMA) and ripe cocoa mucilage agar (RCMA) were prepared and assessd for their clarity and for potential to support mycelial growth and sporulation of P. palmivora. Oatmeal agar (OMA), potato-dextrose agar (PDA), vegetable 8 juice agar (V8JA) and pineapple crown agar (PCA) were included for comparison. The highest radial growth rates of 8.3 and 7.2 mm/day were recorded, respectively, on OMA and GCPA but these were not significantly different (P < 0.05) from each other. The two media also supported good aerial mycelial growth but were not clear. Radial mycelial growth rates of 6.5, 7.0 and 6.6 mm/day were obtained on GCMA, RCPA and V8JA, respectively, and these rates were also not significantly different from each other. Of the three media, only the GCMA was clear and supported the best aerial mycelial growth. In comparison, the RCMA supported a significantly lower radial growth (4.6 mm/day) of P. palmivora than the three media. Growth rates were least on RCBA, PCA and PDA but sporulation was poorest on PDA, PCA and V8JA. GCMA was found to be the best medium based on all the growth parameters and media characteristics. GCMA has been used effectively to isolate/detect P. palmivora from infected cocoa pod tissues. Apart from differences in radial growth rate, both the GCMA and RCMA were similar in all other respects and are recommended for culturing P. palmivora.

Agar↗
PubMed1998

Nutritive value of hot water- or cocoa-pod ash solution-treated cocoa bean cake for broiler chicks.

1. The effectiveness of boiling (90 degrees C for 75 min) in water and alkali (cocoa-pod ash CPA solution) treatments for reducing the antinutritional factor (theobromine) in cocoa bean cake (CBC) and the influence of these treatments on the chemical composition, gross energy and metabolisable energy (ME) of CBC to the broiler chicken were examined. 2. Boiling did not affect crude protein content but resulted in lower ash, crude fibre, ether extract and gross energy values, and a 56% reduction in theobromine, while the use of increasing concentrations of cocoa-pod ash solution decreased crude protein, ether extract, theobromine and gross energy. 3. Mineral elements were significantly affected both by boiling and by cocoa-pod ash solution treatments. 4. There were no significant effects of hot water-treated or 5 g CPA/kg treated CBC on ME or MEn. However, efficiency of energy utilisation was improved. 5. Nutritional evaluation showed that there was a general decrease in performance at high dietary concentrations of CBC (treated or untreated). However, both forms of treatment (hot water-treated and 5 g CPA/kg treated CBC) significantly improved food utilisation and retention of nitrogen and dry matter.

Animal Nutritional Physiological Phenomena↗
PubMed1999

Human Daxx regulates Fas-induced apoptosis from nuclear PML oncogenic domains (PODs).

Daxx was first identified as a protein that binds the cytosolic domain of Fas and links this receptor to an apoptosis pathway involving activation of Jun N-terminal kinase (JNK). We show here that cells overexpressing the human homolog of Daxx (hDaxx) display enhanced sensitivity to apoptosis induced by Fas but not by several other cell death stimuli. hDaxx-mediated enhancement of Fas-induced apoptosis was correlated with accelerated activation of caspases but not with JNK induction. Although specifically enhancing Fas function, hDaxx does not bind Fas and instead is found in the nucleus where it localizes to PML oncogenic domains (PODs). Moreover, the hDaxx protein also exhibits the ability to repress transcription. Mutagenesis studies demonstrated a correlation between the localization of hDaxx to PODs and its ability to enhance Fas-induced cell death. Arsenic trioxide (As(2)O(3)), an agent that accentuates POD formation, collaborated synergistically with overexpression of hDaxx to increase cellular sensitivity to Fas-induced apoptosis. Taken together, these findings argue that hDaxx promotes sensitivity to Fas from a nuclear location, probably by modulating the transcription of genes involved in Fas-induced caspase activation and apoptosis.

Adaptor Proteins, Signal Transducing↗
PubMed1994

Purification of the Major Soybean Leaf Acid Phosphatase That Is Increased by Seed-Pod Removal.

Fruit removal for 5 weeks after flowering increased acid phosphatase activity 10-fold in soybean (Glycine max L. Merr. Var Hobbit) leaves compared with normal seed-pod-bearing plants. The major acid phosphatase activity in leaves was purified over 2700-fold, yielding a single polypeptide of 51 kD with a specific activity of 1353 units/mg protein using p-nitrophenylphosphate as the substrate. Isoelectric focusing demonstrated that the purified protein co-migrated with a majority of the activity that increased in leaves following seed-pod removal. Immunoblot analysis demonstrated that at least part of the increased activity was due to an increased abundance of the phosphatase protein. In situ enzyme activity staining localized most of the total phosphatase activity to vascular tissues, the leaf paraveinal mesophyll cell layer, and the lower epidermis. This distribution and the response to seed-pod removal paralleled previous results for soybean vegetative storage protein (VSP) [alpha] and [beta]. However, in a native polyacrylamide gel the VSP detected by immunological staining of electrophoretically transferred protein did not migrate with the majority of the phosphatase activity. Fractionation of crude leaf protein on concanavalin A-Sepharose yielded a fraction containing 97% of the total VSP but only 0.1% of the total acid phosphatase activity.

Journal Article↗
PubMed1994

Comparative study of cellulases associated with adventitious root initiation, apical buds, and leaf, flower, and pod abscission zones in soybean.

Cellulase activity was measured in soybean (Glycine max) leaf abscission zones, flower abscission zones, pod abscission zones, apical buds, and adventitious rooting hypocotyls. Immunoprecipitation data showed that a cellulase immunologically similar to the bean abscission cellulase (isoelectric point 9.5) is present in soybean leaf, flower, and pod abscission zones, but is not present in soybean apical buds or rooting hypocotyls. cDNA and genomic clones for two different soybean genes were identified and show sequence similarity with the bean abscission cellulase clone pBAC10. The cDNA clone pSAC1, isolated from a soybean abscission cDNA library, hybridized to transcripts in soybean leaf, flower, and pod abscission zones. Although ethylene has been shown to play a role in the increase in cellulase activity associated with both abscission and adventitious root initiation, no signal was seen for hybridization of the soybean abscission cellulase clone, pSAC1, to RNA from soybean adventitious rooting hypocotyls. In addition, no soybean abscission cellulase transcripts were detected in apical buds. Transcripts for a second soybean cellulase gene (SC2) were not detected in any of the tissues surveyed.

Amino Acid Sequence↗
PubMed1999

Changes in cell wall polysaccharides of green bean pods during development.

The changes in cell wall polysaccharides and selected cell wall-modifying enzymes were studied during the development of green bean (Phaseolus vulgaris L.) pods. An overall increase of cell wall material on a dry-weight basis was observed during pod development. Major changes were detected in the pectic polymers. Young, exponentially growing cell walls contained large amounts of neutral, sugar-rich pectic polymers (rhamnogalacturonan), which were water insoluble and relatively tightly connected to the cell wall. During elongation, more galactose-rich pectic polymers were deposited into the cell wall. In addition, the level of branched rhamnogalacturonan remained constant, while the level of linear homogalacturonan steadily increased. During maturation of the pods, galactose-rich pectic polymers were degraded, while the accumulation of soluble homogalacturonan continued. During senescence there was an increase in the amount of ionically complexed pectins, mainly at the expense of freely soluble pectins. The most abundant of the enzymes tested for was pectin methylesterase. Peroxidase, beta-galactosidase, and alpha-arabinosidase were also detected in appreciable amounts. Polygalacturonase was detected only in very small amounts throughout development. The relationship between endogenous enzyme levels and the properties of cell wall polymers is discussed with respect to cell wall synthesis and degradation.

Carboxylic Ester Hydrolases↗
PubMed1974

Mode of Pisatin Induction: Increased Template Activity and Dye-binding Capacity of Chromatin Isolated from Polypeptide-treated Pea Pods.

Increases in phenylalanine ammonia lyase activity and pisatin synthesis were induced in excised pea pods (a) by basic polypeptides such as protamine, histone, lysozyme, cytochrome c, and ribonuclease; (b) by the polyamines spermine, spermidine, cadaverine, and putrescine, and (c) by the synthetic oligopeptides poly-l-lysine, poly-dl-ornithine, and poly-l-arginine.Poly-l-lysine (1 milligram per milliliter, molecular weight 7,200) was utilized as a model inducer of pisatin and phenylalanine ammonia lyase. The poly-l-lysine-induced responses could be inhibited by adding the RNA synthesis inhibitors cordycepin or alpha-amanitin to the pods prior to or at the time of inducer application. Cordycepin added 1.5 hours after inducer no longer completely inhibited induction. The application of poly-l-lysine was shown to characteristically change the rate of RNA synthesis within 30 minutes. Ultrastructural changes in pea nuclei were detected within 3 hours, and gross changes in nuclear morphology were apparent at 14 hours after inducer application. The physical appearance of uranyl acetate-stained chromatin isolated from poly-l-lysine 2 hours after inducer application differed from that of water-treated tissues. The template properties of chromatin extracted from pods 3 hours after inducer application were consistently superior to control chromatin when assayed with Escherichia coli RNA polymerase (without sigma factor). Chromatin from poly-l-lysine-induced tissue also bound 49% more actinomycin D-(3)H.The DNA-complexing properties of inducer compounds and the induced changes in the template and dye-binding properties of pea chromatin formed the basis for a proposed mode of action for phytoalexin induction.

Journal Article↗
PubMed1975

Growth and Development of Soybean (Glycine max [L.] Merr.) Pods: CO(2) Exchange and Enzyme Studies.

The rates of CO(2) exchange and (14)CO(2) incorporation in the light and dark and the activities of several photosynthetic, photorespiratory, and respiratory enzymes of soybean (Glycine max [L.] Merr. cv. Wye) reproductive structures were determined at weekly intervals from anthesis to pod maturity. At all stages of pod development soybean reproductive structures were found to be incapable of net photosynthesis under the experimental conditions employed, but capable of gross photosynthesis and light-induced (14)CO(2) uptake. Consistent with the lack of net photosynthesis throughout the development of the reproductive structure, the maximum in vitro activity of ribulose 1,5-bisphosphate carboxylase (EC 4.1.1.39) in pod tissue was only 3% of that in leaf extracts when expressed on a fresh weight basis. We concluded that the major role of the reproductive structure of the soybean with respect to photosynthetic carbon metabolism is the reassimilation of its respiratory CO(2).

Journal Article↗
PubMed1980

Stomatal closure and photosynthetic inhibition in soybean leaves induced by petiole girdling and pod removal.

The presence of strong sinks of photoassimilates is thought to stimulate photosynthesis by minimizing photosynthetic end product accumulation in leaves. This hypothesis was examined in soybeans (Glycine max [L] Merr.) with treatments designed to alter the phloem translocation of photoassimilates out of source leaves. Pod removal and petiole girdling resulted in 70 and 90% reductions, respectively, in leaf CO(2) exchange rate. Reductions of similar magnitude also were observed in stomatal diffusive conductivity.Time course data showed that CO(2) exchange rates as well as stomatal conductivities were significantly reduced 0.5 hour after girdling and 5 hours after pod removal, thus suggesting that stomatal closure was the cause of the reduced rates of photosynthesis.Mesophyll conductivities to CO(2), calculated from gas exchange data, were not affected by the treatments. Radiocarbon assimilation rates by leaf slices floated in KH(14)CO(3) solutions were not reduced by pod removal and were reduced only 10% by girdling. It was concluded that treatments which block or slow translocation from source leaves reduce their photosynthetic rate by inducing stomatal closure.

Journal Article↗
PubMed1990

Foliar boron applications increase the final number of branches and pods on branches of field-grown soybeans.

Our previous work demonstrated that boron (B) supplied to soybeans (Glycine max [L.] Merrill cv ;Williams 82') by a stem infusion technique increased the number of pods on branches and led to a significant yield increase. Therefore, research was continued to determine whether soil or foliar applications of B could be used to achieve the same results. Field experiments were completed with both soil and foliar applications of B. Only the foliar applications of B resulted in a significant increase in the number of pods/branch. When split foliar treatments were applied twice during flowering, the total application of 0.56 kilograms of B per hectare was the optimal treatment for increasing pods/branch. In a second field experiment in 1987, soybeans were treated weekly from flowering through podfill with six split foliar applications of aqueous H(3)BO(3) solutions so that total applications were either 0, 1.1, or 2.24 kilograms of B per hectare. Foliar applications increased the number of branches/plant at the end of the season and significantly stimulated the formation of pods on branches, with 1.12 kilograms of B per hectare being the optimal treatment for these variables. This rate also tended to increase the number of seeds/plant and seed yield/plant. A duplicate experiment with minor modifications was conducted during the summer of 1988, and again the 1.12 kilograms of B per hectare application rate resulted in significant increases in number of branches at harvest as well as number of pods on branches. The 2.24 kilograms of B per hectare application rate also significantly increased these parameters. Foliar B applications induced increases in leaf B concentration far above the 60 micrograms per gram level that was previously accepted as the upper level of tolerance for soybeans. Since optimal branching and per plant yield parameters were achieved by plants with B leaf concentrations greater than 160 micrograms per gram, the accepted range of soybean tolerance for B must be reconsidered when B is foliarly applied.

Journal Article↗
PubMed2006

Pod shatter-resistant Brassica fruit produced by ectopic expression of the FRUITFULL gene.

Arabidopsis has proven to be extremely useful as a reference organism for studies in plant biology, and huge efforts have been employed to unravel various mechanisms of Arabidopsis growth. A major challenge now is to demonstrate that this wealth of knowledge can be used for global agricultural and environmental improvement. Brassica species are closely related to Arabidopsis and represent ideal candidates for model-to-crop approaches as they include important crop plants, such as canola. Brassica plants normally disperse their seeds by a pod-shattering mechanism. Although this mechanism is an advantage in nature, unsynchronized pod shatter constitutes one of the biggest problems for canola farmers. Here, we show that ectopic expression of the Arabidopsis FRUITFULL gene in Brassica juncea is sufficient to produce pod shatter-resistant Brassica fruit and that the genetic pathway leading to valve margin specification is conserved between Arabidopsis and Brassica. These studies demonstrate a genetic strategy for the control of seed dispersal that should be generally applicable to diverse Brassica crop species to reduce seed loss.

Arabidopsis↗
PubMed2003

Intracellular bacterial biofilm-like pods in urinary tract infections.

Escherichia coli entry into the bladder is met with potent innate defenses, including neutrophil influx and epithelial exfoliation. Bacterial subversion of innate responses involves invasion into bladder superficial cells. We discovered that the intracellular bacteria matured into biofilms, creating pod-like bulges on the bladder surface. Pods contained bacteria encased in a polysaccharide-rich matrix surrounded by a protective shell of uroplakin. Within the biofilm, bacterial structures interacted extensively with the surrounding matrix, and biofilm associated factors had regional variation in expression. The discovery of intracellular biofilm-like pods explains how bladder infections can persist in the face of robust host defenses.

Adhesins, Bacterial↗
PubMed1965

EFFECT OF CAROB POD EXTRACT ON CELLULOLYSIS, PROTEOLYSIS, DEAMINATION, AND PROTEIN BIOSYNTHESIS IN AN ARTIFICIAL RUMEN.

Carob pod extract and its tannin and sugar fractions were compared with gallotannic acid and sucrose for their effect on the cellulolytic, proteolytic, protein biosynthetic, and deaminative activities of rumen microorganisms. The inhibitory effects of carob pod extract upon the cellulolysis and deamination were correlated mainly with its sugar, rather than its tannin components. On the other hand, proteolytic activity and protein biosynthesis were more significantly affected by the tannin fraction. In contrast to the tannin fraction of carob pod extract, gallotannic acid inhibited cellulolytic activity. The harmful effect of a low concentration of tannins on protein biosynthesis could be prevented by the addition of carbohydrates to the reaction mixture. At high tannin concentration (40 mug/ml), however, the addition of carbohydrates did not prevent the inhibition.

Ammonia↗