Inhibitory Effects of Oxygen on the Germination of Oryza sativa L. Seeds

Some workers have reported that the breaking of seed dormancyin rice (O. sativa L.) is usually enhanced by higher oxygentension, whereas others have shown that rice seed dormancy canbe broken by incubation under anaerobic conditions. This articleaims to clarify this paradox. The results show that high oxygentensions inhibit seed germination for a certain period afterharvest in Japonica rice, whereas Indica rice cultivars arenot inhibited by oxygen at any stage. Oxygen inhibition, seed germination, aquatic plants, Japonica rice, Indica rice     

Comparative genetic analysis of a wheat seed dormancy QTL with rice and <Emphasis Type="Italic">Brachypodium</Emphasis> identifies candidate genes for ABA perception and calcium signaling

Wheat preharvest sprouting (PHS) occurs when seed germinates on the plant before harvest resulting in reduced grain quality. In wheat, PHS susceptibility is correlated with low levels of seed dormancy. A previous mapping of quantitative trait loci (QTL) revealed a major PHS/seed dormancy QTL, QPhs.cnl-2B.1, located on wheat chromosome 2B. A comparative genetic study with the related grass species rice (Oryza sativa L.) and Brachypodium distachyon at the homologous region to the QPhs.cnl-2B.1 interval was used to identify the candidate genes for marker development and subsequent fine mapping. Expressed sequence tags and a comparative mapping were used to design 278 primer pairs, of which 22 produced polymorphic amplicons that mapped to the group 2 chromosomes. Fourteen mapped to chromosome 2B, and ten were located in the QTL interval. A comparative analysis revealed good macrocollinearity between the PHS interval and 3 million base pair (mb) region on rice chromosomes 7 and 3, and a 2.7-mb region on Brachypodium Bd1. The comparative intervals in rice were found to contain three previously identified rice seed dormancy QTL. Further analyses of the interval in rice identified genes that are known to play a role in seed dormancy, including a homologue for the putative Arabidopsis ABA receptor ABAR/GUN5. Additional candidate genes involved in calcium signaling were identified and were placed in a functional protein association network that includes additional proteins critical for ABA signaling and germination. This study provides promising candidate genes for seed dormancy in both wheat and rice as well as excellent molecular markers for further comparative and fine mapping.     

Effect of calcium peroxide on the growth and proliferation ofMicrocystis aerusinosa, a water-blooming cyanobacterium

The potential of calcium peroxide to act as an agent for waterblooming control was investigated by examining the growth inhibition ofMicrocystis aerusinosa. Due to the chemical nature of calcium peroxide, it can remove dissolved phosphate by forming an insoluble precipitate, generating radicals, coagulant, and oxygen as byproducts as it dissolves in water. The growth ofM. aerusinosa was severely inhibited and the chlorophyll-a concentration was drastically decreased in the presence of calcium peroxide. With 200 ppm of calcium peroxide dosage, a chlorophyll-a concentration of 1,700 mg/m³ was lowered to below 10% of its initial concentration after 4 days. One possible explanation for this growth inhibition is the removal of the available phosphate by calcium peroxide.     

Dormancy in Rice Seed: III. THE INFLUENCE OF TEMPERATURE, MOISTURE, AND GASEOUS ENVIRONMENT

The influence of storage conditions—temperature, moisturecontent, oxygen, nitrogen, and carbon dioxide—on dormancyin rice seed has been investigated. The effects of temperatureand oxygen when the seeds have been set to germinate have alsobeen studied. Storage in oxygen accelerates the breaking of dormancy; at lowertemperatures, the effect of oxygen is more noticeable. Carbondioxide and nitrogen have little or no effect except in so faras they exclude oxygen. It is shown that there is a negativelinear relationship between storage temperature and log. meandormancy period of intact seeds over the range 27°C. to47°C. In the variety tested, the Q₁₀ for the rate of breakingdormancy is 3.38. Variation of moisture content over the range12.0–14.5 per cent, has little effect at 27°C. andno detectable effect at higher temperatures. When dormant seeds are soaked in water, they attain a moisturecontent close to 30 per cent. Pre-soaking seeds to achieve moisturecontents in this region can stimulate the breaking of dormancywhen compared with dry storage. At laboratory temperature (meanabout 27°C.) the stimulation caused by pre-soaking intactseeds is usually small and sometimes non-existent, but at alow temperature (3°C.) the stimulation is increased; butdry storage at 3°C. markedly delays the breaking of dormancy.When seeds are dehusked, a large proportion lose their dormancy.Much of the residual dormancy of dehusked seeds can be brokenby soaking at laboratory temperature. But in the sample of dehuskedseeds used, low temperature did not increase the effect of thepre-soaking, but slightly decreased the stimulation caused bythe soaking treatment in this case. The effect of temperature on wet seed has also been investigatedwhen the seed has been set to germinate. Maximum germinationof a partially dormant population of seed is achieved at anoptimum temperature which is near or somewhat below 27°C.In the sample tested, no germination occurred at 42°C.,although in populations which have completely broken dormancyfull germination will occur at this temperature. Non-dormantpopulations of rice seed can germinate at very low oxygen tensionsor probably even in the absence of oxygen, but germination ofa partially dormant sample is reduced under these circumstances.When oxygen tension is very low there is less germination at27°C. than at 37°C. In some varieties, at least, there is evidence that the seedsgo through a stage when they will germinate in daylight butnot in the dark. A simple practical method for rapidly breakingthe dormancy of intact seeds is described. The significanceof these results is discussed in relation to the published workon seeds of other species.     

钙盐及模拟干旱互作对大花套唇苣苔种子萌发和幼苗生长的影响

苦苣苔科原大花旋蒴苣苔(former Boea clarkeana Hemsl.)为我国特有的复苏植物,在最近的修订中被并入套唇苣苔属(Damrongia Kerr),更名为大花套唇苣苔[D. clarkeana(Hemsl.)C. Puglisi]。为了探究该植物对喀斯特特殊生境的适应机制,该文以大花套唇苣苔成熟种子为材料,通过观察钙盐和模拟干旱互作试验,研究了干旱和盐胁迫共同作用对其种子萌发和幼苗生长的影响。结果表明:钙盐及模拟干旱互作时,低浓度钙盐可减轻干旱对种子萌发和幼苗生长的抑制作用,且低程度的干旱与低浓度的钙盐相互作用时反而会促进种子萌发和幼苗生长; 低钙盐低干旱复合条件对大花套唇苣苔生长的促进作用体现了该物种对喀斯特地区保水性差的石灰岩土壤基质的专适性,避免了和其他植物直接生态位的竞争。     

Effects of temperature and water potential on the germination of muskmelon cultivars

The rate and final germination of four muskmelon cultivars (Cucumis melo) were examined in response to incubation temperatures of 20, 26 and 32°C. Germination was also characterised at 26°C pr 32°C over a range of water potentials from 0 to - 1000 kPa achieved with solutions of polyethylene glycol. The germination of one cultivar, TAM-Uvalde, was consistently slower at 20°C than at 26°C or 32°C. The other three cultivars, Perlita, TAM-Dew and Greenflesh, were inhibited by incubation at 32°C. However, the germination responses of cvs Perlita, TAM-Dew and Greenflesh at 26°C or 32°C improved as water potentials were reduced from 0 to – 200 or – 400 kPa. Cv. TAM-Uvalde was extremely sensitive to water stress and failed to germinate at water potentials below – 600 kPa when incubated at 26°C. The inhibition of germination at low water potentials was partially reversed in all cultivars by increasing the incubation temperature from 26°C to 32°C. It is suggested that the inhibition of germination at 0 kPa (distilled water) was due to a seed coat-mediated barrier to oxygen that could be reversed by removal of the seed coat or exposure to an oxygen-enriched atmosphere.     

The effect of excess moisture on the germination of Spinacia oleracea L.

Summary The reversible inhibition of the germination of spinach (Spinacia oleracea L.) seeds in conditions which are even slightly wetter than optimal has been traced to the production, in a wet environment, of a layer of mucilage around and within the fruit coat which surrounds the true seed. Such wet seeds may however germinate readily when the temperature is lowered, or the oxygen pressure of the environment is raised, or the intact seeds are placed for a short time in hydrogen peroxide before being transferred to what normally would be an excess of water. Even in the absence of an increased oxygen supply the seeds will germinate under water provided the fruit coat, or even a small part of it where it covers the radicle, is crefully removed. No evidence has been found of a water soluble inhibitor and the findings are consistent with the hypothesis that germination is dependent on a sufficiently high rate of supply of oxygen to the sites embryonic respiration. The mucilage which is formed under wet conditions forms a barrier which prevents the transfer of oxygen to the embryo by gaseous diffusion or aqueous convection currents and restricts it to the process of aqueous diffusion, and under these conditions the rate of oxygen supply may not reach the threshold level required for germination.     

Influence of waterlogging on the emergence and growth ofLolium perenne L. shoots from seed coated with calcium peroxide

Summary The emergence and seedling growth of cv. S. 24 perennial ryegrass plants was significantly reduced by two waterlogging treatments in a glasshouse environment. These adverse effects of temporary waterlogging were alleviated by coating the seed with calcium peroxide. This oxygenating agent did not enhance or reduce seedling establishment when the soil medium was not waterlogged. Calcium peroxide did not affect the germination of coated seeds when placed on wet filter paper.The establishment of plants in contrasting soils, which experienced water tables close to ground level at two upland field sites, was also improved by coating perennial ryegrass seed with calcium peroxide. Alleviation of the detrimental effects of excessively high water tables was not observed in plants which were established from seed coated with calcium carbonate.     

Expression of a fungal glucoamylase in transgenic rice seeds

Glucoamylase, which catalyses the hydrolysis of the α-1,4 glycosidic bonds of starch, is an important industrial enzyme used in starch enzymatic saccharification. In this study, a glucoamylase gene from Aspergillus awamori, under the control of the promoter of seed storage protein Gt1, was introduced into rice by Agrobacterium-mediated transformation. Significant glucoamylase activity was detected specifically in the seeds but not other tissues of the transgenic rice lines. The highest enzymatic activity was found in the transgenic line Bg17-2, which was estimated to have about 500 units per gram of seeds (one unit is defined as the amount of enzyme that produces 1 μmol of reducing sugar in 1 min at 60 °C using soluble starch as substrate). The optimum pH for the activity of the rice produced enzyme is 5.0–5.5, and the optimum temperature is around 60 °C. One part of this transgenic glucoamylase rice seed flour fully converted 25 parts of corn starch pre-liquefied by an α-amylase also produced by a transgenic rice into glucose in 16 h incubation. This study suggests that this hydrolysis enzyme may substitute commercial fermentation enzymes for industrial starch conversion.     

Hydrogen peroxide as an oxygen source for immobilized Gluconobacter oxydans converting glycerol to dihydroxyacetone

Summary A flow injection analysis (FIA) system with amperometric detection was developed for measuring hydrogen peroxide which was used as an oxygen source for immobilized cells. A constant concentration of peroxide in the reactor was maintained by processing the analytical signal in a computer programmed as a PI-regulator. The concentration of dissolved oxygen was followed using a commercial Clark-electrode. The simultaneous measurements of hydrogen peroxide and dissolved oxygen are discussed with respect to process control.Conversion of glycerol to dihydroxyacetone by Gluconobacter oxydans immobilized in calcium alginate was used as a model system.Initial specific productivity increased with increasing hydrogen peroxide concentration. However, decreases in viable counts, enzymatic activities and overall productivities were noted. Various techniques for improving operational stability are discussed.     

Glycollate metabolism of wheat and rice leaves during senescence and under the influence of growth regulators

The glycollate metabolism of wheat (Triticum vulgare Vill. cv. Sonalika) and rice (Oryza sativa L. ev. Jaya) leaves was studied during senescence by estimating the endogenous levels of glycollate and hydrogen peroxide (H₂O₂) and the activities of glycollate oxidase and catalase. In comparison with light incubation the incubation of excised leaves in the dark caused a decline in the glycollate content and in the activities of glycollate oxidase and catalase, and an increase in the H₂O₂ content, more marked in the leaves of rice than in the leaves of wheat. Glycollate oxidase activity gradually decreased with incubation time, and glycollate metabolism decreased during senescence. The glycollate oxidase in particular and glycollate metabolism of rice were more sensitive to incubation time than those of wheat. Kinetin increased the glycollate oxidase activity and glycollate metabolism during senescence, while ethrel (2-chloroethylpho-sphonic acid) and ABA (abscisic acid) reduced these activities in both plant species.     

Some Properties and Characterization of Rice Seed Hemagglutinin

The phytohemagglutinin of rice seed has been purified by a sequence of steps involving fractionation with ammonium sulfate and successive chromatography on DEAE-and eMcellulose and finally gel filtration on Bio-Gel P-100. The purified rice seed hemagglutinin was shown to be homogeneous by electrophoresis on polyacrylamide gel and its molecular weight was 10,000, calculated from both the Ve/Vo value of gel filtration on Bio-Gel P-100 and the sum of the individual constituents (amino acids, sugars and metals). In addition to amino acid, the rice seed hemagglutinin contained 26.8% covalently bound carbohydrate which was identified and quantitated by gas chromatography of the acetylated alditols. Glucose was the predominant sugar with lesser amounts of glucosamine, xylose, and mannose also being present. And the rice seed hemagglutinin contained 1 g atom of calcium per molecule. The molecular weight of the rice seed hemagglutinin is smallest compared with some of phytohemagglutinins isolated from leguminous seeds and other plant sources. The rice seed hemagglutinin has the blastogenetic activity for human peripheral lymphocytes as well as Phaseolus vulgaris phytohemagglutinins or concanavalin A, jack bean hemagglutinin.     

Mitochondrial hydrogen peroxide production alters oxygen consumption in an oxygen-concentration-dependent manner

Metabolic responses of mammalian cells toward declining oxygen concentration are generally thought to occur when oxygen limits mitochondrial ATP production. However, at oxygen concentrations markedly above those limiting to mitochondria, several mammalian cell types display reduced rates of oxygen consumption without energy stress or compensatory increases in glycolytic ATP production. We used mammalian Jurkat T cells as a model system to identify mechanisms responsible for these changes in metabolic rate. Oxygen consumption was 31% greater at high oxygen (150–200 μM) compared to low oxygen (5–10 μM). Hydrogen peroxide was implicated in the response as catalase prevented the increase in oxygen consumption normally associated with high oxygen. Cell-derived hydrogen peroxide, predominately from the mitochondria, was elevated with high oxygen. Oxygen consumption related to intracellular calcium turnover was shown, through EDTA chelation and dantrolene antagonism of the ryanodine receptor, to account for 70% of the response. Oligomycin inhibition of oxygen consumption indicated that mitochondrial proton leak was also sensitive to changes in oxygen concentration. Our results point toward a mechanism in which changes in oxygen concentration influence the rate of hydrogen peroxide production by mitochondria, which, in turn, alters cellular ATP use associated with intracellular calcium turnover and energy wastage through mitochondrial proton leak.     

Transgenic rice as bioreactor for production of the Candida antarctica lipase B

Candida antarctica lipase B (CALB) is a versatile biocatalyst used for a wide range of biotransformation. Methods for low cost production of this enzyme are highly desirable. Here, we report a mass production method of CALB using transgenic rice seeds as the bioreactor. The transgenic rice transformed with the CALB gene under the control of the promoter of the rice seed storage protein GT1 was found to have accumulated a large quantity of CALB in seeds. The transgenic line with the highest lipolytic activity reached to 85 units per gram of dry seeds. One unit is defined as the amount of lipase necessary to liberate 1 μmol p‐nitrophenol from p‐nitrophenyl butyrate in 1 min. The rice recombinant lipase (rOsCALB) from this line represents 40% of the total soluble proteins in the crude seed extracts. The enzyme purified from the rice seeds had an optimal temperature of 40 °C, and optimal pH of 8.5, similar to that of the fermentation products. Test of its conversion ability as a biocatalyst for biodiesel production suggested that rOsCALB is functionally identical to the fermentation products in its industrial application.     

Assay of nitrogen supplying capacity of tropical rice soils

Summary The nitrogen supplying capacity of 39 wetland rice soils evaluated by two anaerobic incubation methods and six chemical methods was compared with N uptake of IR 26 rice grown on these soils under flooded conditions in a greenhouse pot study. The uptake of N by rice correlated highly with the N supplying capacity determined by anaerobic incubation methods involving incubation of soils at 30°C for 2 weeks (r=0.84^**) or at 40°C for 1 week (r=0.82^**) as well as with the organic carbon (r=0.82^**) and total N (r=0.84^**) contents of soils. Among the chemical indexes, available N determined by the oxidative release of soil N by alkaline permanganate, acid permanganate, acid dichromate and hydrogen peroxide also provided good index of soil N availability to rice. According to these results soil organic carbon and total N contents seem to be good indexes of available nitrogen in tropical wetland rice soils.     

Effects of Seedbed Management on Blast Development in Susceptible and Partially Resistant Rice Cultivars

Four rice cultivars, susceptible or partially resistant to Pyricularia grisea were evaluated for their apparent infection rates (r) and for terminal severity values in seedlings grown in seedbeds. Susceptibility of partially resistant cultivars decreased when seeded in wet seedbed compared to those grown in raised or upland seedbed. The degree of blast reduction due to flooding varied with rice genotype, seeding rate, and environmental conditions. Flooding was effective in managing seedling blast of partially resistant cultivars but not of susceptible cultivar. Use of a lower seed rate in comparison to a higher seed rate also led to reductions in r-values and terminal disease severities. However, in case of the susceptible cultivar IR50, effect of seed rate was more pronounced, when low amount and short duration of rainfall occurred. Flooding of wet beds with low seed rate further reduced the blast incidence. Results of this study suggest that with appropriate use of water, seed rate, and partial host resistance there are prospects for effective control of leaf blast in the tropics.     

Germination and seedling growth under anaerobic conditions in Echinochloa crus-galli (barnyard grass)*

Abstract Although rice has long been recognized to be uniquely adapted for growth in low oxygen environments of flooded rice fields, rice weeds of the Echinochloa crus-galli complex appear to be at least as well specialized for germination and growth under such unusual biological conditions. Seeds of two varieties of E. crus-galli germinate and grow for prolonged periods in a totally oxygen-free environment. E. crus-galli germinates as well as rice (Oryza sativa) under a total nitrogen atmosphere and produces as large a seedling in spite of its much smaller seed size. Like rice, the seedlings of E. crus-galli are unpigmented, the primary leaves do not emerge from the coleoptile and no root growth occurs without oxygen. Of particular interest is the ultrastructure of mitochondria from anaerobically-grown seedlings. Mitochondrial profiles from the primary leaf of seedlings grown continuously in nitrogen are very similar to those grown aerobically. The size and shape of the mitochondria are similar and the cristae are numerous and normal in appearance. This is in sharp contrast to previous studies of other species which have reported that mitochondria were vesiculate and tended to lose their normal fine-structure after similar periods without oxygen. Finally, based on ultrastructure and ¹⁴C labeling studies, anaerobically-grown seedlings are highly active metabolically, which may explain, at least for E. crus-galli var. oryzicola, its ability to germinate and emerge from flooded rice fields.     

Rice LTG1 is involved in adaptive growth and fitness under low ambient temperature

Low temperature (LT) is one of the most prevalent factors limiting the productivity and geographical distribution of rice (Oryza sativa L.). Although significant progress has been made in elucidating the effect of LT on seed germination and reproductive development in rice, the genetic component affecting vegetative growth under LT remains poorly understood. Here, we report that rice cultivars harboring the dominant LTG1 (Low Temperature Growth 1) allele are more tolerant to LT (15–25°C, a temperature range prevalent in high‐altitude, temperate zones and high‐latitude areas), than those with the ltg1 allele. Using a map‐based cloning strategy, we show that LTG1 encodes a casein kinase I. A functional nucleotide polymorphism was identified in the coding region of LTG1, causing a single amino acid substitution (I357K) that is associated with the growth rate, heading date and yield of rice plants grown at LT. We present evidence that LTG1 affects rice growth at LT via an auxin‐dependent process(es). Furthermore, phylogenetic analysis of this locus suggests that the ltg1 haplotype arose before the domestication of rice in tropical climates. Together, our data demonstrate that LTG1 plays an important role in the adaptive growth and fitness of rice cultivars under conditions of low ambient temperature.     

The effect of cytochrome P-450 and reduced glutathione on the ATP-dependent calcium pump of hepatic microsomes from male rats

In the presence of ATP hepatic microsomes sequester calcium. This sequestration is thought to be important in the modulation of free cytosolic calcium concentration. We find that on the addition of NADPH the uptake of calcium by the hepatic microsomes is inhibited 27-85%. This inhibition is reversed by the addition of 1 mM reduced glutathione (85-91% of control), incubation under a nitrogen atmosphere (112% of control), or incubation in a 80% carbon monoxide/20% oxygen atmosphere (75% of control). Superoxide dismutase had no effect on the inhibition, while catalase reversed the inhibition by 35%. The addition of 1 mM reduced glutathione at 2 and 5 min after the addition of NADPH led to uptakes of calcium which paralleled the uptake seen when the reduced glutathione was added at the beginning of the incubation. The effect of reduced glutathione showed saturation kinetics with a Km of 10 microM. Together these data suggest that cytochrome P-450 reduces the activity of the microsomal ATP-dependent calcium pump both by the production of hydrogen peroxide and by the direct oxidation of the protein thiols. The reversal of this effect by reduced glutathione appears to be enzymatically catalyzed.