Studies upon the copper fungicides: VIII. The penetration of copper into germinating peas

Three varieties of pea seed showed less damage when treated with solutions of simple cupric salts than with solutions of equivalent strength in which the copper could occur in complex form. On adding supplementary pea exudate to copper sulphate solution, thereby converting some cupric to complex copper, greater damage was caused. In spite of the greater phytotoxicity of complex forms of copper, the copper intake by the seeds was less. Sealing of the micropyle had little effect on the water and copper intake of peas.
Colorimetric tissue tests on treated peas indicated a high concentration of copper in the testa and the embryonic radical with only small amounts in the cotyledonary tissues. These observations were largely confirmed by chemical analyses-on the test as, cotyledons and embryos of copper treated peas.
The results suggest that when copper dissolves from dressings on pea seeds much becomes fixed on the testa, so providing fungicidal protection. The soluble complex forms of copper produced by pea exudate are markedly phytotoxic and injury to the embryonic radical is regarded as the main cause of damage to peas.     

Seed treatments with benomyl and other fungicides for the control of Ascochyta pisi on peas

In three experiments, using laboratory, glasshouse and field tests, seed dressings of benomyl-1½ (42.4 g) to 2 oz (56.6 g) Benlate 50% w.p. per 28 lb (12.7 kg) of seed-gave complete control of Ascochyta infection of pea seeds. Mixtures of benomyl and captan and benomyl and thiram were also effective. Carboxin, captan, and thiram used as seed dressings, either singly or in mixtures, did not eliminate the pathogen from peas. Germination, emergence and seedling vigour were not impaired by any dressing.     

Muscle and liver protein metabolism in rats fed raw or heat-treated pea seeds

Raw or extruded pea (Pisum sativum, cv. Ballet) diets with or without supplementary amino acids were fed for 15 days to young growing rats and the effects on tissue weights, liver and muscle protein metabolism and hormone levels monitored. Body weight gain, liver and gastrocnemius muscle weights and protein contents were reduced and some key hormones altered when rats were fed unsupplemented raw pea diets. This appeared to be a result of amino acid deficiencies in the diet, the action of antinutritional factors and the refractory nature of the reserve proteins and other seed components. However, this did not in itself improve the nutritional performance of the rats due to the overriding effects of the amino acid deficiencies in the pea diets. After supplementation, extruded peas supported much higher rates of growth and skeletal muscle deposition than did supplemented raw peas. Despite this, the weight gains remained less than achieved on a high quality control diet. Protein synthesis and degradation rates in skeletal muscles and total protein contents were similar to control values. The lower growth rate did not appear to be due to impaired deposition of skeletal muscle. Deposition of other body components, possibly lipids, may have been lowered by supplemented extruded pea diets. Liver protein levels were reduced in rats fed supplemented raw peas and blood corticosterone was elevated. In conclusion, extrusion treatment of peas in combination with amino acid supplementation appeared to abolish the negative effects of peas on skeletal muscle deposition.     

Biochemical changes associated with cadmium and copper stress in germinating pea seeds (Pisum sativum L.)

Seeds of pea (Pisum sativum L.) were germinated for four days over two sheets of filter paper moistened with H2O (control) and 5 mM Cd(NO3)2 or CuSO4 (treated). The relationship between heavy-metal stress and breakdown of storage compounds was studied. Germination rate and growth of radicle decreased, while the water content in stressed seeds remained around the control values. Cotyledons changed their biochemical constituents: disorders in the contents of micronutrients (Fe, Mn, Zn), free amino acids and soluble sugars were found. Decline of alpha-amylase activity as well as acid phosphatase were also observed, whereas beta-amylase and alkaline phosphatase ones were not modified by heavy-metal treatments. These results suggest that the inhibition of seed germinations after exposure to cadmium or copper is not the consequence of starvation in water uptake by seed tissues, but may be due to a failure in the reserve mobilization process from cotyledons.     

The influence of the phosphorus and nitrogen nutrition of pea plants on the growth of their progeny

Summary In glasshouse experiments with peas grown in nutrient cultures, seed from phosphorus-deficient plants contained much lower phosphorus concentrations than seed from plants not deficient in this element, but differences in the supply of mineral nitrogen to the plants (which were nodulated) had little effect on the chemical composition of the seed.Lower haulm weights and pea yields were obtained from sowings of low-phosphorus as compared with high-phosphorus seed when the plants were grown in cultures deficient in phosphorus. In cultures not deficient in phosphorus, there were no such differences between the performance of the seed.In field experiments carried out on a fertile soil, low-phosphorus seeds gave 20 to 25 per cent lower yields of both haulms and peas than were obtained from sowings of high-phosphorus seed.It appeared that the differences in the performance of the seeds weer related to differences in the concentration of phosphorus they contained rather than to any other variation in their composition or size.     

Influence of selenium on uptake and toxicity of copper and cadmium in pea (Pisum sativum) and wheat (Triticum aestivum)

The detoxifying effect of selenium on animals toxicated with heavy metals is well known. In this study we examine if there is a similar effect in plants. Wheat ( Triticum aestivum L. cv. Sunny) and pea ( Pisum sativum L. cv. Fenomen) were grown for 21 days on a nutrient solution based on the nutrient proportions in healthy plants. Nutrients along with cadmium, copper, selenite, selenate or selenite and selenate in combinations with copper or cadmium were supplied in small amounts with a daily incremental increase of 0.12 (wheat) and 0.20 (pea). The metal and selenium uptake and distribution in the plants as well as the effects on growth were investigated.
The results show that selenium does not reduce the toxicity of heavy metals to plants. Instead, selenium enhances metal uptake and toxicity, especially in peas grown in the presence of metal and selenate. Selenite increased cadmium concentrations of pea roots up to 300% and selenate that of wheat shoots up to 50%.     

玉兰肉质外种皮挥发性成分与功能的初步研究

以玉兰(Magnolia denudata Desr.)发育成熟的种子为材料,采用水蒸气蒸馏法和GC-MS技术对其肉质外种皮化学成分的种类和含量进行了分析研究,并就玉兰肉质外种皮对其种子吸水和失水的影响以及化感作用进行比较研究,以探讨其相应的生态学和生物学功能。结果表明:(1)玉兰肉质外种皮占整个完整种子重量的48.25%,有无肉质外种皮对其种子自身吸水没有明显影响,但在失水过程中有肉质外种皮的种子失水较慢,具有一定的保水作用。(2)玉兰种子肉质外种皮中萜烯类和醇类的种类较多,分别有12种和11种;但酮类和酯类的相对含量较高,分别为32.84%和18.03%,其中含量较多的成分依次是甲基庚烯酮(31.95%)、乙酸丁酯(17.69%)、对伞花烃(13.64%)和芳樟醇(6.61%)。(3)玉兰肉质外种皮浸提液对豌豆、白菜种子萌发具有明显的抑制作用,且随着浸提液浓度增加,豌豆种子发芽率由86.7%下降为6.0%,白菜种子发芽率由28.0%下降为0。研究推测,玉兰种子的肉质外种皮在功能上可能是一种类似肉质果果肉的结构,具有丰富的化学成分,不仅可以吸引鸟类传播其种子,也具有一定的化感作用。     

The solubilization and degradation of pumpkin seed globulin during germination

Pumpkin seed globulin decreased 88% during the first 4 daysof germination. This decrease was concomitant with a 2.5 foldincrease in water soluble protein which arose directly fromthe water insoluble globulin. The sequence of solubilizationand breakdown of the globulin was followed through 12 days ofgermination. Pumpkin seed globulin was determined to have subunitsof 56,000 daltons, while the new water soluble protein consistedof two proteins, as determined by sodium dodecyl sulfate polyacrylamideelectrophoresis; one having a molecular weight of 42,000 daltonsand the other 28,000 daltons. Trypsin mimicked the first stepof the breakdown by solubilizing pumpkin seed globulin to yieldidentical digestion products as were obtained in vitro. Maximumproteolytic acitvity, as measured by the release of ninhydrinpositive, materials occurred at 6 days of germination, at whichtime both the concentration of free amino acids and the incorporationof ¹⁴C into amino acids increased rapidly. A second proteolyticenzyme system which solubilized the pumpkin seed globulin butdid not act on hemoglobin, casein, or bovine serum albumin reachedits maximum activity at two days of germination. (Received April 17, 1978; )     

Effect of extrusion under controlled temperature and moisture conditions on ileal apparent amino acid and starch digestibility in peas determined with young broilers

The objective of the experimental programme was to examine the influence of extrusion of peas under controlled temperature and moisture conditions on coefficient of ileal apparent digestibility (CIAD) of amino acids and to examine the changes to starch structure (crystallisation) and subsequent effects on CIAD of starch in young broilers. Experiment 1 used one pea cultivar (2.94 mg trypsin inhibitor units (TIU)/g DM) processed under two extrusion temperatures (exit temperatures of 70 and 140 °C) and two moisture levels (0 and 1.96l water added/h) giving a 2 × 2 factorial design (with a 5th treatment being the raw pea sample). All pea samples were added at a rate of 500 g/kg into a purified diet where peas were the only source of protein/amino acids. Processing generally led to a reduction in CIAD of amino acids but CIAD of starch for Raw, Treatments 1 and 3 (added water) was lower than that for Treatments 2 and 4 (no water) with data of 0.730, 0.808 and 0.759 vs. 0.852 and 0.834, respectively. Experiment 2 evaluated two near-isogenic lines of peas (HA5 and HB5) processed under the same conditions as for Experiment 1. Extrusion at 70 °C/addition of water had no significant effect on trypsin inhibitor activity but the inhibitor was eliminated at 140 °C with or without addition of water (P<0.001). Pea line HA5 generally had higher CIAD than HB5 (P<0.05 for cystine, lysine, leucine, histidine, alanine; P<0.01 for threonine). There were significant temperature × water addition interactions for CIAD (P<0.001 for all amino acids except methionine; P=0.006). For example, data for lysine were 0.760, 0.828, 0.860 and 0.759, respectively for 140 °C/no water, 140 °C/1.96 l/h, 70 °C/no water and 70 °C/1.96 l/h. CIAD for starch improved following processing, attributed to the loss of crsytallinity. The data indicate that peas are sensitive to high processing temperatures which are associated with a reduction in nutritive value, in terms of amino acid digestibility, for young poultry. In contrast, the results reinforce the adverse effect of crystallinity on starch digestibility and show that extrusion in the absence of water improves the digestibility of raw starches due to granular disorganisation (as evidenced by X-ray diffraction patterns) and changes in crystallinity.     

Response to water deficit and high temperature of transgenic peas (Pisum sativum L.) containing a seed-specific alpha-amylase inhibitor and the subsequent effects on pea weevil (Bruchus pisorum L.) survival

The effects of water deficit and high temperature on the production of alpha-amylase inhibitor 1 (alpha-AI-1) were studied in transgenic peas (Pisum sativum L.) that were developed to control the seed-feeding pea weevil (Bruchus pisorum L., Coleoptera: Bruchidae). Transgenic and non-transgenic plants were subjected to water-deficit and high-temperature treatments under controlled conditions in the glasshouse and growth cabinet, beginning 1 week after the first pods were formed. In the water-deficit treatments, the peas were either adequately watered (control) or water was withheld after first pod formation. The high-temperature experiments were performed in two growth cabinets, one maintained at 27/22 degrees C (control) and one at 32/27 degrees C day/night temperatures, with the vapour pressure deficit maintained at 1.3 kPa. The plants exposure to high temperatures and water deficit produced 27% and 79% fewer seeds, respectively, than the controls. In the transgenic peas the level of alpha-AI-1 as a percentage of total protein was not influenced by water stress, but was reduced on average by 36.3% (the range in two experiments was 11-50%) in the high-temperature treatment. Transgenic and non-transgenic pods of plants grown at 27/22 degrees C and 32/27 degrees C were inoculated with pea weevil eggs to evaluate whether the reduction in level of alpha-AI-1 in the transgenic pea seeds affected pea weevil development and survival. At the higher temperatures, 39% of adult pea weevil emerged, compared to 1.2% in the transgenic peas grown at the lower temperatures, indicating that high temperature reduced the protective capacity of the transgenic peas.     

The production of copper,zinc and molybdenum deficiencies in crop plants grown in sand culture with special reference to some effects of water supply and seed reserves

Summary (1) A method is described for the production of acute deficiencies of copper, zinc or molybdenum in large scale sand cultures in which an adequate and satisfactory supply of water was obtained by treating rain water with ion-exchange resins. Results compared favourably with the use of glass distilled water.(2) Visual symptoms of copper, zinc and molybdenum deficiencies in large seeded leguminous crops, and in other species are summarised.(3) Seed reserves played an important part in the incidence of copper and molybdenum deficiencies and to a lesser extent of zinc deficiency in pea, runner bean, dwarf bean and broad bean.(4) The deficient seed was obtained by harvesting seed from plants grown in the respective deficiency sand cultures, and was compared with seed obtained similarly from complete nutrient sand cultures or commercially from field-grown plants.(5) Seed reserves of molybdenum were usually sufficient for a complete generation but reserves of copper and zinc were not adequate for the whole copper or zinc requirements of the plants.(6) Analytical data showed a marked decrease in the copper or molybdenum content of deficient seed, and a less striking decrease in zinc.(7) Foliage of deficient plants showed decreased content of copper and molybdenum and often of zinc compared with plants given the complete nutrient treatment, but use of deficient seed only decreased yields without further appreciably decreasing the mineral content of the foliage.     

Role of various parameters in synthesis of insoluble agrobased xanthates for removal of copper from wastewater

The research identifies and studies the parameters involved in the synthesis of two types of insoluble agrobased xanthates, named insoluble baggase xanthate (IBX) and insoluble wood xanthate (IWX) and their ability to remove heavy metals from wastewater. Copper was selected as the model metal and the metal removal capacity of IBX and IWX were expressed by copper loading (mg of copper removed/g of IBX or IWX). It was observed that contact times of 3 h and alkali strengths of 4 mol/L were necessary during alkalization. The maximum base material to alkali ratio (w/w) was 1.125 without sacrificing proper mixing of the base material alkali mixture. Optimum contact time during xanthation was 4h with a carbon-disulfide to base material ratio (w/w) of 1.25. Only strong bases like NaOH, KOH contributed high copper loadings and the starting liquor was reused twice without sacrificing the copper removal capacity of IBX and IWX. Maximum copper loadings achieved with IBX and IWX were 26.9+/-0.15 and 27.8+/-0.39 mg/g, respectively.     

Determination of the taste threshold of copper in water

Copper effects on human health represent a relevant issue in modern nutrition. One of the difficulties in assessing the early, acute effects of copper ingested via drinking water is that the taste of copper may influence the response and the capacity to taste copper in different waters is unknown. The purpose of the study was to determine the taste threshold of copper in different types of water, using soluble and insoluble salts (copper sulfate and copper chloride). Copper-containing solutions (range 1.0-8.0 mg/l Cu) were prepared in tap water, distilled deionized water and uncarbonated mineral water. Sixty-one healthy volunteers (17-50 years of age), with no previous training for sensory evaluation, participated in the study. A modified triangle test was used to define the taste threshold value. The threshold was defined as the lowest copper concentration detected by 50% of the subjects assessed. To evaluate the olfactory input in the threshold value obtained, 15 of 61 subjects underwent a second set of triangle tests with the nose open and clamped, using distilled water with copper sulfate at a concentration corresponding to the individual's threshold. The taste threshold in tap water was 2.6 mg/l Cu for both copper sulfate and copper chloride. The corresponding values for distilled deionized water were 2.4 and 2.5 mg/l Cu for copper sulfate and copper chloride, respectively. In uncarbonated mineral water the threshold values were slightly higher, 3.5 and 3.8 mg/l Cu for copper sulfate and for copper chloride, respectively, which are significantly higher than those observed in tap and distilled waters (P < 0.01, Kruskal-Wallis test). The taste threshold did not change significantly when the nose was clamped. In conclusion, the median values for copper taste threshold were low, ranging between 2.4 and 3.8 mg/l Cu, depending on the type of water.     

Spectroscopic studies on the copper(II)—Inosine system

Interactions of inosine derivatives with copper(II) were studied in the pH range 1.4–13 in 50% H₂O-50% DMSO solution. The distinct pH dependence of the optical spectra observed in copper(II)-inosine complexes are correlated to their respective EPR changes as a function of pH. It was concluded that a simple 1:1 complex of copper(II)-inosine is formed in the pH range 1.4–5.0 and bis complexes are present in the pH 5.0–6.2 region solutions of inosine and Cu(II). From pH 6.2 to 7.8 a diamagnetic, hydroxybridged complex dominates. At pH 7.8–9.2 an insoluble, oxybridged species is formed in addition to the soluble paramagnetic Cu(NI)₄ complex. Starting from pH 9.1 the N-polymeric complex is formed which is stable up to pH 12.5, and above pH 12.5 the only species is the Cu(ribose)₂ complex.     

Seed Production in Leafless and Conventional Phenotypes of Pisum sativum L. in Relation to Water Availability Within a Controlled Environment

Water requirements in relation to seed production was studiedin near-isogenic lines of leafless (afafstst) and conventional(++++) pea plants (Pisum sativum). The plants were grown toseed maturity in pots in a controlled environment under conditionsof high, medium and low irrigation levels. When each genotypewas irrigated independently and on demand and the soil moisturecontent maintained at 65–80 per cent of full capacitythere was no significant phenotypic difference in water useefficiency (WUE), defined as g d. wt seed per kg H₂O utilized.There existed genotypically-controlled upper and lower limitsto yield between which the total dry weight of seed per plantcan be determined by water availability. There was no significantdifferential effect of genotype or of irrigation treatment onthe number of pods, number of seed, unit seed dry weight andbiological yield per plant. There was significant interactionon stem length, and leaf area at specified nodes. When the wateractually required in relation to the water available was takeninto account, the leafless phenotype consistently utilized 33–38per cent less water and produced a correspondingly lower totaldry weight of seed than the conventional counterpart. Independentlyof regime the total dry weight seed per phenotype remained anear constant proportion of the above-ground biomass. Pisum sativum L., garden pea, leafless peas, seed production, water availability     

Oxidative Processes in Soybean and Pea Seeds: Effect of Light, Temperature, and Water Content

Oxidative processes are probable determinants of longevity of seeds in storage. Measurements of actual oxygen uptake rates were made for soybean and pea seeds as a comparison of short and long lived seeds when light, temperature, and moisture contents were varied. In both peas and soybeans, the oxygen uptake was depressed at low temperatures (<16°C) and low water contents (<0.25 gram H₂O per gram dry weight). Apparent activation energies under these conditions are very high, while apparent activation energies of seeds at higher water contents and at temperatures greater than 22°C are much less. Light enhances the level of oxygen uptake in pea, but reduces the level of oxygen uptake in soybean. The complexities of the interactions of oxygen uptake with environmental conditions in soybean compared to pea suggest that oxidative processes occur in soybean at low water contents, but are essentially absent in pea. It is suggested that the additional oxidative processes in soybean with moisture contents between 0.10 and 0.24 gram per gram may contribute to the poorer longevity of soybean seed compared to pea seed.     

Kinetics and implications of seedling growth responses to 2-chloroethylphosphonic acid

The effects of soaking seed in 2-chloroethylphosphonic acid (CEPA) for 24 or for 48 h on the cumulative 5-day seedling growth ofCucumis sativus L. (cucumber) andPisum sativum L. (peas) were studied. Each cucumber seed absorbed an average of 0.015 ml of CEPA solution, while pea seed absorbed 0.365 ml, over a 24 h period. In cucumber, 240 mg l^?1 CEPA concentration decreased radicle length by 23%, regardless of soaking duration. The same concentration increased radicle weight in a 24 h soaking duration, but decreased radicle weight when soaking was for 48 h. At 48 h, CEPA concentrations of 0.24 and 2.4 mg l^?1 increased plumule growth by 26%. In peas, the 240 mg l^?1 decreased the length and the weight of both the radicle and the plumule in a 48 h soaking duration, but had no significant effect at a 24 h soaking. At the low concentration of 0.24 mg l^?1, seedling growth was stimulated by over 30%. Cucumber was 3 times more efficient than peas in the utilization of CEPA for seedling growth, in terms of total fresh weight of seedling per microgram of CEPA absorbed: 1 127 and 274 mg μg^?1 CEPA in cucumber and peas respectively. Extrapolative calculation, using cucumber responses as standard, suggests from this seedling study that about 12 mg l^?1 CEPA is likely to stimulate growth and/or yield in sprayed pea plants.     

Double-headed protease inhibitors from black-eyed peas. I. Purification of two new protease inhibitors and the endogenous protease by affinity chromatography.

Two new double-headed protease inhibitors have been isolated from black-eyed peas. The isoinhibitors can be purified to homogeneity with greater than 90% recovery in a four-step procedure by means of sequential affinity chromatography on trypsin-Sepharose and chymotrypsin-Sepharose affinity columns. The isoinhibitors both have molecular weights near 8,000 and both have the same NH1-terminal residue serine. Black-eyed pea chymotrypsin and trypsin inhibitor (BEPCI) has an isoelectric point of 5.1 and inhibits trypsin and chymotrypsin simultaneously. Black-eyed pea trypsin inhibitor (BEPTI) has an isoelectric point of 6.5 and inhibits 2 molecules of trypsin simultaneously. BEPTI binds to chymotrypsin-Sepharose above pH 6 but does not inhibit chymotrypsin in the standard inhibitor assay with 10-3 M substrate. These new inhibitors are distinct from the Ventura inhibitor isolated from Serido black-eyed peas. An endogenous seed protease has been isolated from black-eyed peas by affinity chromatography on soybean inhibitor-carboxymethylcellulose affinity columns. A protease-BEPCI complex has been isolated by ion exchange chromatography. A dual physiological function of inhibition and protection of the seed protease is suggested as a plausible role of seed protease inhibitors.     

Screening of the occurrence of copper amine oxidases in Fabaceae plants

Aim of this work was to find the best source for obtaining high amount of copper amine oxidase (EC 1.4.3.6) that can be further used for analytical or industrial applications. The study focused on plant enzymes, because they occur in much higher content in the starting material than the enzymes from other sources, have higher specific activity and are also more thermostable. Presence of the amine oxidase was tested in extracts from 4 to 7-d-old seedlings of thirty-four various Fabaceae plants. Amine oxidases from nine selected plants were purified by general method involving ammonium sulfate fractionation, controlled heat denaturation, and three chromatographic steps. Kinetic properties of the amine oxidases purified were tested with a wide range of substrates and inhibitors and were found to be very similar. Best purification yield, and total and specific activities were obtained for the enzyme from grass pea (Lathyrus sativus) throughout all purification steps. Hence, the grass pea extract was chosen as a suitable candidate for massive production of the amine oxidase. This revised version was published online in July 2006 with corrections to the Cover Date.     

The solubilization and partial characterization of pea RNA polymerases

A technique has been developed for solubilizing RNA polymerase from pea seedling, which employs the sonication of frozen cells and the addition of insoluble polyvinylpyrrolidone(Polyclar AT) to prevent inactivation of the enzyme. RNA polymerases from peas have properties similar to those of other eucaryotes.