Activities of Hydrogen Peroxide-Scavenging Enzymes in Germinating Wheat Seeds

During imbibition and germination of wheat (Triticum aestivum)in the dark over 72 h, activities of the enzymes of the ascorbate(AsA)-dependent H₂O₂-scavenging pathway, AsA peroxidase, monodehydroascorbate(MDAsA) reductase, dehydroascorbate (DHAsA) reductase and glutathione(GSSG) reductase as well as superoxide dismutase (SOD), catalaseand guaiacol peroxidase were determined both in whole grainsand in isolated embryos and endosperm. With the exception of DHAsA reductase, activities of the otherenzymes assayed increased in germinating seeds, especially duringradicle emergence (between 24–48 h of imbibition). Theseincreases, particularly for AsA peroxidase, were much higherin the embryo than in the endosperm. Within 72 h of imbibition,activities per seed increased 116-fold for AsA peroxidase, 19-foldfor guaiacol peroxidase, 5-fold for catalase and only 1·4-foldfor SOD. In contrast to the decreases in DHAsA reductase, theother AsA recycling enzyme, MDAsA reductase, increased 5-foldwithin 72 h. The results indicate that, in wheat seeds, imbibition and germinationis associated with enhanced cellular capacity to detoxify H₂O₂.For this detoxification the operation of AsA peroxidase togetherwith the AsA-regenerating enzymes appears to be of particularimportance. Key words: Ascorbate peroxidase, germination, hydrogen peroxide detoxification, inhibition, wheat     

Adenosine Phosphates in Germinating Radish (Raphanus sativus L.) Seeds

Changes in concentrations of adenosine phosphates (AMP, ADP, and ATP), oxygen utilization, and fresh weights were measured during the first 48 hours after imbibition of water by quiescent radish seeds (Raphanus sativus L.) at 22.5 C. The changes in ATP concentrations, oxygen utilization, and fresh weights followed a triphasic time course, characterized by a rapid initial increase, which extended from 0 to approximately 1.5 hours, a lag phase from 1.5 to 16 hours, and a sharp linear increase from 16 to 48 hours. In unimbibed seeds, the concentrations of ATP, ADP, and AMP were <0.1, 0.9, and 2.2 nmoles/seed, respectively. After imbibition of water by the quiescent seeds, for 1 hour, the ATP concentration had increased to 2.5, and ADP and AMP concentrations had decreased to 0.3 and 0.1 nmole/seed, respectively. These early changes occurred also in seeds maintained under anaerobic conditions (argon), or when treated with either 5 mm fluoroacetate, or 5 mm iodoacetate. The concentrations of ADP and AMP did not change significantly from 1 to 48 hours. The termination of the lag phase at 16 hours correlated with radicle emergence. Cell division in the radicles was initiated at approximately 28 hours. ATP concentrations in seeds maintained under argon or treated with fluoroacetate remained relatively constant from approximately 2 to 48 hours. In contrast, the ATP concentration of iodoacetate-treated seeds decreased curvilinearly from 4 to 48 hours. Oxidative phosphorylation was estimated to have contributed 15, 20, and 65% of the pool ATP at 1.5, 16, and 48 hours, respectively.     

Responses of Proteolytic Enzymes in Embryonic Axes of Germinating Bean Seeds under Copper Stress

The changes in protease activities in embryonic axes during the first days of bean (Phaseolus vulgaris L.) seed germination were investigated in response to copper stress. Synthetic substrates and specific protease inhibitors have been used to define qualitatively and quantitatively different catalytic classes, particularly endoproteases (EP), carboxypeptidases (CP) and aminopeptidases (AP), then identify which ones were affected in the presence of copper. In fact, a failure in storage proteins mobilization and a disorder of nitrogen supply at enzymatic level occurred in Cu. In fact, Cu inhibited azocaseinolytic activity (ACA) and cysteine-, aspartic-, serine-, and metallo-endopeptidases activities (Cys-EP, Asp-EP, Ser-Ep, and Met-EP, respectively). Besides, Cu affected leucine- and proline-aminopeptidases (LAP and PAP, respectively) and glycine-carboxypeptidases (Gly-CP). The proteolytic responses might also be associated with the decrease in defense capacity in the Cu-treated embryos.     

Starch-hydrolyzing Enzymes in Germinating Kidney Bean

Enzymatic production of D-Glu was investigated by the succesive reactions of a glutamate racemase (EC 5.1.1.3) and a glutamate decarboxylase (EC 4.1.1.15) on L-Glu.Lactobacillus brevis ATCC8287 was chosen as a source of glutamate racemase. This strain produced a glutamate decarboxylase simultaneously. The glutamate racemase activity in the cell free extracts was 0.035 units/mg protein. The enzyme kept its activity even at 500 Mm of L-Glu (74g/liter). The optimum pHs of the racemase and the decarboxylase were at around 8.5 and below 4.0, respectively. Both enzymes had no activity at the optimum pH for the other enzyme. L-Glu was racemized first by the glutamate racemase at pH 8.5, then the pH was shifted to 4.0 at which L-Glu was decarboxylated by the glutamate decarboxylase. Starting from 100 g/liter of L-Glu, 50 g/liter of D-Glu was produced and no L-Glu remained in the reaction mixture.     

Oxidative Phosphorylation in Germinating Lettuce Seeds

The effect of thiourea and coumarin, in vivo and in vitro, onthe phosphorylating activity of lettuce mitochondria was investigated,as well as the effect of coumarin on the P/O ratio. It was shownthat both substances in vitro inhibit phosphorylation; whilein vivo coumarin inhibits but thiourea under certain circumstancesstimulates. Coumarin was also shown to decrease the P/O ratioand therefore may be considered as an uncoupler. The difficulties in considering these effects of the substancesas a primary mechanism controlling germination are pointed outand discussed.     

萌发大麦种子水溶性蛋白组分提取

采用二次回归正交旋转组合试验方法,对大麦种子中水溶性蛋白组分的提取工艺进行了研究,得出了大麦种子水溶蛋白组分提取量与料液浓度,浸提温度和浸提时间的数学模型,确定了大麦籽粒水溶蛋白组分最佳提取条件为料液浓度9．1％,浸提温度20℃,浸提时间3h;通过连续检测萌发大麦种子水溶蛋白含量发现：随萌发时间增长,蛋白含量逐渐升高,麦芽中水溶蛋白含量约为大麦的4倍．     

Stronium Mobility in Germinating Seeds and Plants

The uptake of strontium in the bean plant (Phaseolus vulgaris) was linear for the first 34 hr during continuous exposure to radiostrontium. After 35 hr there was a sharp increase in the rate of uptake to 48 hr. Radioactivity could be detected in the plant as early as 1 hr after addition of radiostrontium to the growth medium.     

Ion Transport in the Testa of Germinating Seeds

Levengood, W. C. 1985, Ion transport in the testa of germinatingseeds.—J. exp. Bot. 36: 1053–1063. The current flow produced when an electrical potential is appliedto a partially hydrated seed is drastically altered by the applicationof a thermal pulse. Specific responses to thermally inducedchanges in electrical activity are related to the cell wallstructure of the seed coat and its state of hydration. It issuggested that expansion and contraction of the micropores inthe cell wall matrix provide a model based on a diffusion anddehydration during a thermal pulse and an ion-gating effectimmediately following the pulse. Mechanical flexing producedoscillatory behaviour in the electrical current flow throughseed coat tissues in a manner predicted by the thermal responses. Key words: Ion transport, testa, germinating seeds     

Soil Water Movement to Germinating Seeds

Water movement to germinating seeds of several range plantswas measured with a gammaray attenuation technique. Seeds wereplaced at a depth of 1.5 cm in soil at several water contentsand the soil water content of thin (3 mm) layers above and belowthe seeds was measured daily. It was found that the distancefrom which water was taken up by the seeds did not exceed 1cm and was not affected by soil water content. Germination wasfound to be dependent on seed water uptake, which was determinedby soil water content. The rate of seedling root growth wasnot affected by initial soil water content, but shoot growthwas strongly affected. Solutions of the water flow equation were applied to the caseof spheric geometry and infinite medium and to the case of linearflow and semi-infinite medium, in order to calculate water movementto a germinating seed or to a layer of seeds. Both the calculationsand the measurements showed that water uptake by a 2-mm-diameterseed from distances exceeding 1 cm was negligible.     

The Biosynthesis of Cytokinins in Germinating Lupin Seeds

Imbibed intact seeds, and excised embryos and cotyledons ofyellow lupin (Lupinus luteus L. cv. Weiko III) have been incubatedwith [¹⁴C]-adenine to investigate cytokinin biosynthesis duringthe early stages of germination. Following incubation the tissueswere extracted and purified by solvent partition and chromatographyon cellulose phosphate, diethylaminoethyl cellulose and SephadexLH-20 columns. Using a variety of thin layer chromatographic,high performance liquid chromato-graphic and chemical procedures,incorporation of ¹⁴C into dihydrozeatin riboside and its nucleotidewas demonstrated in extracts of intact embryos, intact cotyledonsand excised embryos. However, radioactivity was not found associatedwith cytokinins in fractions derived from the isolated cotyledons.This is the first direct demonstration of cytokinin biosynthesisin germinating seeds and the results indicate that the capacityfor cytokinin biosynthesis is probably confined to the embryonicaxes. If this is so, the levels of [¹⁴CJ-dihydrozeatin ribosideassociated with intact embryo and intact cotyledon fractionsindicate that the synthesized cytokinin is transported to andaccumulates in the cotyledons. Key words: Lupinus luteus, cytokinin biosynthesis, seed germination     

The Movement of Calcium in Germinating Pea Seeds

Pea seeds contain less calcium than phosphorus, potassium ormagnesium; more than half of this calcium is located in thetesta. Peas at either end of a pod have more calcium than thosein the middle. When pea seeds are allowed to germinate in water,less than 30 per cent of the cotyledonary calcium moved to thegrowing axis during the first 15 days of germination, whereas70–90 per cent of magnesium, potassium and phosphate wasexported. Various attempts to increase the amount of calciumexported were not successful. When radioactive calcium was appliedto the cotyledons, essentially no movement to the axis was observedunder conditions where extensive movement of radioactive phosphateoccurred.     

Apparent Free Space of Germinating Pea Seeds

The apparent free space (AFS) of pea seeds was determined by measurement of the exodiffusion of solutes from the seeds previously equilibrated with radioactive solutions. AFS (per cent of volume of water taken up by dry seeds) varied with the kinds of solutes used, that is, values of 27–30% were obtained for mannitol or leucine and 12% for malonate, at pH 6. When the seeds were incubated in a radioactive solution at lower pH (3.5), higher AFS values were obtained, especially for malonate. The AFS of imbibing seeds showed a tendency to increase during the course of imbibition.     

Development of Pyrimidine-metabolizing Enzymes in Cotyledons of Germinating Peas

Mechanisms controlling conversion of orotic acid-6-¹⁴C to uridine-5′-phosphate in cotyledons of germinating Alaska peas (Pisum sativum L.) were investigated. The content of 5-phosphoribosyl-1-pyrophosphate was very low in dry seeds, increased to a maximum after about 12 hours of imbibition, and then rapidly declined. Orotidine-5′-phosphate pyrophosphorylase and orotidine-5′-phosphate decarboxylase activities more than doubled during the first 24 hours of germination and then also decreased. These results do not account for the continuous increases of orotate anabolism in such cotyledons as we observed previously. The initial increases in activities of these two enzymes were unaffected by cycloheximide, while the subsequent decreases were less rapid in the presence of this inhibitor. Activities of cotyledonary cytidine deaminase and uridine hydrolase also increased during imbibition, but the activity of only the latter showed a decrease after imbibition was completed. Cycloheximide inhibited the initial rapid increase in uridine hydrolase activity but had little effect on its subsequent decline. Cycloheximide had only slight inhibitory effects on the development of cytidine deaminase activity during the first 62 hours. The evidence suggests that uridine hydrolase might be synthesized de novo during the first few days of germination, but that the other three enzymes might not be.