Lipid biochemistry of echinochloa crus-galli during anaerobic germination

Seven day old seedlings of Echinochloa crus-galli var. oryzicola (Vasing) had a higher total lipid content when germinated under N₂ than in air, although ungerminated seeds contained more lipid than either seedling. The triacylglycerol pool was not depleted under anaerobiosis as it was in air and only air-grown seedlings showed a net increase in free fatty acids and polar lipids. Concentrations of most of the individual acids of the total fatty acid profile declined during germination in air and in the free acid and polar lipid fractions of these seedlings the relative proportion of polyunsaturated fatty acids increased. Compared to air-grown seedlings, ungerminated seeds and N₂-grown seedlings had a similar qualitative and quantitative lipid composition. Our results show that mobilization of storage lipids was apparently severely inhibited under anoxia. The importance of lipid metabolism to the germination and growth of Echinochloa during anoxia is discussed in terms of maintaining membrane integrity and serving (indirectly) to reoxidize pyridine nucleotides.     

Amylolytic activity and carbohydrate levels in relation to coleoptile anoxic elongation in Oryza sativa genotypes

Among starchy seeds, rice has the unique capacity to germinate successfully under complete anaerobiosis. In this conditions, starch degradation is supported by a complete set of starch-degrading enzymes that are absent or inactive in cereals except rice. A characterization of carbohydrate metabolism and starch-degrading enzyme activity across twenty-nine genotypes of Oryza sativa L. is presented here. The zymogram of amylolytic activities present in rice embryos and endosperms under anaerobic conditions seven days after sowing (DAS) revealed marked differences among cultivars. Coleoptile elongation was positively correlated with total amylolytic activities and α-amylase activity in embryos, and negatively correlated with α-amylase activity in endosperm. Moreover, carbohydrate content in embryos was found to be positively correlated with total amylolytic activities under anaerobic conditions, while a negative relationship was recorded in the endosperm. Carbohydrate status in rice seedlings has a primary importance in sustaining coleoptile elongation towards the surface. The relationship between carbohydrate level in embryo and anoxic germination, as well as with total amylolytic activities present in rice embryo under anaerobic condition 7 DAS, is consistent with the role of sugar metabolism to support rice germination under oxygen-deprived environment.     

Elevated CO2 decreases seed germination in Arabidopsis thaliana

The impact of elevated [CO₂] on seed germination was studied in different genotypes of Arabidopsis thaliana from natural populations. Two generations of seeds were studied: the maternal generation was produced in the greenhouse (present-day conditions), the offspring generation was produced in two chambers where the CO₂ concentration was either the present atmospheric concentration (about 350 ppm) or elevated (700 ppm). The seeds were tested for proportion of germinated seeds and mean germination time in both chambers to study the impact of elevated [CO₂] during seed production and germination. Elevated [CO₂] during maturation of seeds on the mother-plants decreased the proportion of germinated seeds, while elevated [CO₂] during germination had no effect on the proportion of germinated seeds. However, when seeds were both produced and germinated under elevated [CO₂] (situation expected by the end of next century), germination was slow and low. Moreover, the effect of the [CO₂] treatment differs among genotypes of Arabidopsis: there is a strong treatment × genotype interaction. This means that there is ample genetic variance for a selective response modiying the effects of high levels of [CO₂] in natural populations of Arabidopsis thaliana. The outcome at the community level will depend on what seeds are available, when they germinate and the resulting competition following germination.     

Distribution of alcohol dehydrogenase in roots and shoots of rice (Oryza sativa) and Echinochloa seedlings

Abstract Aerobically germinated seedlings of rice and Echinochloa were found to survive when placed in an anaerobic environment for 4 d, whereas pea and maize seedlings did not. Although root and shoot growth were inhibited in rice and Echinochloa under anaerobiosis, growth resumed when the seedlings were returned to aerobic conditions. Alcohol dehydrogenase (ADH) activity increased more, and protein synthesis was greater, in the shoots than in the roots under anaerobic conditions. These results suggest that, in anaerobiosis-tolerant species, ADH activity and protein synthesis in the shoots represents or results from metabolic adaptations to low oxygen. These results are discussed in terms of plant establishment and growth in a low-oxygen environment.     

Organ specific analysis of the anaerobic primary metabolism in rice and wheat seedlings II: Light exposure reduces needs for fermentation and extends survival during anaerobiosis

Low oxygen stress in plants can occur during flooding and compromise the availability and utilization of carbohydrates in root and shoot tissues. Low-oxygen-tolerant rice and -sensitive wheat plants were analyzed under anaerobiosis in light to evaluate main factors of the primary metabolism that affect sensitivity against oxygen deprivation: activity of glycolysis and the rate of photosynthesis. Relatively stable ATP contents (93 and 58% of aerated control levels after 24 h anaerobiosis) in illuminated shoot tissues account for enhanced tolerance of rice and wheat seedlings to anaerobiosis upon light exposure in comparison to anoxia in darkness. Although the photosynthetic process was inhibited during low oxygen stress, which was partly due to CO₂ deficiency, more light-exposed than dark-incubated seedlings survived. Illuminated plants could tolerate a 70% lower anaerobic ethanol production in shoots in comparison to darkness, although still an 18-times higher ethanol production rate was determined in rice than in wheat leaves. In conclusion, light-exposed plants grown under anaerobiosis may recycle low amounts of generated oxygen between photosynthesis and dissimilation and generate additional energy not only from substrate phosphorylation during glycolysis but also from other sources like cyclic electron transport.     

Fermentative Metabolism of Hydrogen-evolving Chlamydomonas moewusii

The anaerobic metabolism of Chlamydomonas moewusii under both light (160 lux) and dark conditions has been examined using manometric and enzymic techniques. During anaerobiosis starch is broken down to glycerol, acetate, ethanol, CO₂, and H₂. The release of CO₂ and H₂ comes to an end when the starch pool is depleted.

There are only slight differences in the ratio of the end products of fermentation between light and dark metabolism. In the light, glycerol production is diminished and H₂ evolution is enhanced, whereas the production rate of all other end products generally does not change.

     

The Role of C2H4 in anaerobic induction of cocklebur seed germination

Non-dormant small cocklebur seeds (Xanthium pennsylvanicum Wallr.)are potentiated to germinate, if they are subjected to anaerobiccondition for certain time periods after being sufficientlypre-soaked under aerobic conditions. This is termed "anaerobicinduction" of seed germination. Such induction was slightlyinhibited by CO₂ applied during anaerobiosis, but markedly promotedby C²H⁴ Thus, C²H⁴ can exert its action even in anaerobiosis,but does not enhance the fermentative CO₂ evolution. No actualanaerobic induction occurred when over 1? O₂ was present, evenif C²H⁴ had been applied. Therefore, anaerobic induction seemsto be due to a concerted action of some anaerobically proceedingevents and the anaerobically produced C²H⁴. (Received May 31, 1976; )     

Imbibition and Germination: Influence of Hard Seed Coats on RNA Metabolism

RNA metabolism of Indian rice grass seeds was studied in relation to imbibition and germination. These seeds germinate only after scarification of the seed coats. The hard seed coats restrict germination but not water intake or changes in the quantity and quality of RNA formed during early hours of soaking. These changes differ markedly from those in scarified (H₂SO₄ treated) seeds which are able to germinate. Gibberellic acid hastens germination of scarified seeds and causes changes in the population of RNA transcribed.     

QTL mapping and confirmation for tolerance of anaerobic conditions during germination derived from the rice landrace Ma-Zhan Red

Wide adoption of direct-seeded rice practices has been hindered by poorly leveled fields, heavy rainfall and poor drainage, which cause accumulation of water in the fields shortly after sowing, leading to poor crop establishment. This is due to the inability of most rice varieties to germinate and reach the water surface under complete submergence. Hence, tolerance of anaerobic conditions during germination is an essential trait for direct-seeded rice cultivation in both rainfed and irrigated ecosystems. A QTL study was conducted to unravel the genetic basis of tolerance of anaerobic conditions during germination using a population derived from a cross between IR42, a susceptible variety, and Ma-Zhan Red, a tolerant landrace from China. Phenotypic data was collected based on the survival rates of the seedlings at 21 days after sowing of dry seeds under 10 cm of water. QTL analysis of the mapping population consisting of 175 F_2:3 families genotyped with 118 SSR markers identified six significant QTLs on chromosomes 2, 5, 6, and 7, and in all cases the tolerant alleles were contributed by Ma-Zhan Red. The largest QTL on chromosome 7, having a LOD score of 14.5 and an R ² of 31.7 %, was confirmed using a BC₂F₃ population. The QTLs detected in this study provide promising targets for further genetic characterization and for use in marker-assisted selection to rapidly develop varieties with improved tolerance to anaerobic condition during germination. Ultimately, this trait can be combined with other abiotic stress tolerance QTLs to provide resilient varieties for direct-seeded systems.     

Changes in Hexokinase Activity in Echinochloa phyllopogon and Echinochloa crus-pavonis in Response to Abiotic Stress

Hexokinase (HXK; EC 2.7.1.1) regulates carbohydrate entry into glycolysis and is known to be a sensor for sugar-responsive gene expression. The effect of abiotic stresses on HXK activity was determined in seedlings of the flood-tolerant plant Echinochloa phyllopogon (Stev.) Koss and the flood-intolerant plant Echinochloa crus-pavonis (H.B.K.) Schult grown aerobically for 5 d before being subjected to anaerobic, chilling, heat, or salt stress. HXK activity was stimulated in shoots of E. phyllopogon only by anaerobic stress. HXK activity was only transiently elevated in E. crus-pavonis shoots during anaerobiosis. In roots of both species, anoxia and chilling stimulated HXK activity. Thus, HXK is not a general stress protein but is specifically induced by anoxia and chilling in E. phyllopogon and E. crus-pavonis. In both species HXK exhibited an optimum pH between 8.5 and 9.0, but the range was extended to pH 7.0 in air-grown E. phyllopogon to 6.5 in N₂-grown E. phyllopogon. At physiologically relevant pHs (6.8 and 7.3, N₂ and O₂ conditions, respectively), N₂-grown seedlings retained greater HXK activity at the lower pH. The pH response suggests that in N₂-grown seedlings HXK can function in a more acidic environment and that a specific isozyme may be important for regulating glycolytic activity during anaerobic metabolism in E. phyllopogon.     

Carbon dioxide effects on ethanol production, pyruvate decarboxylase, and alcohol dehydrogenase activities in anaerobic sweet potato roots

The effect of varied anaerobic atmospheres on the metabolism of sweet potato (Ipomoea batatas [L.] Lam.) roots was studied. The internal gas atmospheres of storage roots changed rapidly when the roots were submerged under water. O₂ and N₂ gases disappeared quickly and were replaced by CO₂. There were no appreciable differences in gas composition among the four cultivars that were studied. Under different anaerobic conditions, ethanol concentration in the roots was highest in a CO₂ environment, followed by submergence and a N₂ environment in all the cultivars except one. A positive relationship was found between ethanol production and pyruvate decarboxylase activity from both 100% CO₂-treated and 100% N₂-treated roots. CO₂ atmospheres also resulted in higher pyruvate decarboxylase activity than did N₂ atmospheres. Concentrations of CO₂ were higher within anaerobic roots than those in the ambient anaerobic atmosphere. The level of pyruvate decarboxylase and ethanol in anaerobic roots was proportional to the ambient CO₂ concentration. The measurable activity of pyruvate decarboxylase that was present in the roots was about 100 times less than that of alcohol dehydrogenase. Considering these observations, it is suggested that the rate-limiting enzyme for ethanol biosynthesis in sweet potato storage roots under anoxia is likely to be pyruvate decarboxylase rather than alcohol dehydrogenase.     

Effects of anaerobiosis on carbohydrate oxidation by roots of Pisum sativum

The aim of this work was to discover the effects of anaerobiosis on the breakdown of sugars by the apical 6 mm of the roots of 5-day-old seedlings of Pisum sativum. Estimates of the maximum catalytic activities of alcohol dehydrogenase, lactate dehydrogenase, phosphoenolpyruvate carboxylase and NADP-specific malic enzyme showed them to be comparable to that of phosphofructokinase. Metabolism of sucrose-[U-¹⁴C] by excised apices was restricted by anoxia mainly to conversion to ethanol, CO₂ alanine and glycolytic intermediates. Measurements of metabolites over a period of 240 min after transfer of excised apices to nitrogen showed a marked and continual accumulation of ethanol, a smaller continual accumulation of alanine, a small initial rise in lactate and no detectable accumulation of malate or pyruvate. The rates of CO₂ production, of accumulation of ethanol and alanine, and of the labelling of these compounds by sucrose-[¹⁴C] declined markedly during the first 240 min of anaerobiosis. The conclusion is that under anaerobic conditions carbohydrate metabolism in the pea root apex is largely restricted to alcoholic fermentation, and, to a lesser degree, to alanine production.     

Activity of the Pentose Phosphate and Glycolytic Pathways During Anaerobic Germination of Echinochloa crus-galli (barnyard grass) Seeds

Changes in the activity of key enzymes in glycolysis and theoxidative pentose phosphate pathway were studied in Echinochloacrus-galli (L.) Beauv. var. oryzicola seeds during germinationin air or nitrogen. In addition, the metabolism of specificallylabelled [^I4C]glucose was followed to evaluate the activityof both pathways during anaerobic germination. During the 7 d time period studied there was no difference betweenair and nitrogen in phosphofructokinase activity. Under anaerobicconditions, fructose-1, 6-bisphosphate aldolase increased morethan two-fold in 7 d; whereas in air, it decreased. The activityof the pentose phosphate pathway enzyme, glucose-6-phosphatedehydrogenase, increased under N₂ until day three, when it levelledoff, whilst it continued to increase up to day seven in air. Incubation of Echinochloa seedlings with specifically labelledglucose also resulted in differences between anaerobic- andair-grown seedlings. Labelling of phosphorylated sugars andlipids predominated under N₂; whereas in air, malate and fumaratewere the most heavily labelled compounds. In both air and N₂,there was a greater percentage of label in CO2 from [l-¹⁴C]glucose,while [6-14C] resulted in a greater percentage label in ethanol.These differences were more pronounced under N2, especiallyduring the first 24 h of imbibition, suggesting increased activityof the pentose phosphate pathway. Key words: Echinochloa, Anaerobic metabolism, Oxidative pentose phosphate pathway     

Comparison of Growth Responses of Barnyard Grass (Echinochloa oryzoides) and Rice (Oryza sativa) to Submergence, Ethylene, Carbon Dioxide and Oxygen Shortage

Unlike germination of wheat (Triticum aestivum), millet (Eleucinecoracana), and sorghum (Sorghum caudatum), that of Echinochloaoryzoides (barnyard grass) and Oryza satwa (rice) was not inhibitedby poorly aerated solutions with 11 k Pa oxygen (equilibriumpartial pressure) or less In the dark, seedling shoots of riceincluded a coleoptile, and in Echinochloa, a mesocotyl alsoGrowth in fresh and dry weight of shoots was strongly depressedby poorly aerated solutions in both rice and Echinochloa butthe effects on extension differed in the two species in rice,coleoptile extension was promoted by solutions partly depletedof oxygen, and also by the absence of oxygen The stimulationin partly de-oxygenated solutions resulted from the combinedpromoting effects of small oxygen partial pressures, carbondioxide, ethylene and buoyant tension in contrast, these treatmentsneither promoted nor inhibited elongation by the Echinochloacoleoptile while severely inhibiting extension of the mesocotyl Overall, poorly aerated solutions lengthened the shoot of riceand shortened it in Echinochloa when compared with those submergedin well-aerated solutions These opposite effects were broughtabout by the same gaseous changes, i e oxygen shortage, elevatedethylene and carbon dioxide The effect on Echinochloa was almostentirely restricted to the mesocotyl, coleoptile extension beingremarkably insensitive to large increases in ethylene and carbondioxide, or to extreme oxygen shortage Seedlings of the twospecies thus have contrasting strategies for survival Stress, submergence, ethylene, oxygen shortage, carbon dioxide, adaptation, anaerobiosis, rice (Oryza sativa L), barnyard grass [Echinochloa oryzoides (Ard) Fritseh]     

On Autotropism in Botrytis: Measurement Technique and Control by CO(2)

The mutual orientation of the germination of nearby pairs of Botrytis spores growing in a simple, dilute medium was studied. If the medium is equilibrated with room air, they show a very strong tendency to germinate both toward each other and in a cis arrangement, i.e., toward the same side of the line joining their centers. If the medium is equilibrated with air enriched with 0.3% or 3% CO₂, (i.e., 10 or 100 times the normal CO₂ concentration) then the cells show an equally strong tendency to germinate away from each other. The interaction shows little dependence upon pH.     

Effects of anaerobiosis on ethylene production, respiration and flowering in iris bulbs

Ethylene production of iris bulbs (Iris hollandica cv. Ideal) was very low. When stored at 30°C, production was 12–20 pmol C₂H₄ (kg fresh weight)^?1 h^?1. Higher temperatures (35°C, 40°C) enhanced the ethylene production; a treatment with 40°C for ca 7 days caused a 3 times higher ethylene production than at 30°. During anaerobic storage (in 100% N₂) ethylene production was equal to that of control bulbs. When after a 7 day period of anaerobiosis the N₂ was replaced by air, a burstlike ethylene production was observed. Twenty-four h after the replacement, ethylene production was equal to control values again. The effects of this production of ethylene on mitochondrial respiration and flowering were investigated. When mitochondria were isolated immediately after the anaerobic treatment (before the enhanced ethylene production) alternative pathway capacity was not detectable, a situation also occurring in control bulbs. When mitochondria were isolated 24 h after the end of the anaerobiosis (after the ethylene burst) uninhibited respiration did not change significantly, but a capacity of the alternative pathway was observed. The increase in alternative pathway capacity after anaerobiosis was partly inhibited by 2,5-norbornadiene (NBD), an ethylene antagonist. Fermentation occurred during anaerobiosis: ethanol concentrations increased during the treatment and decreased when air was supplied. When bulbs were exposed to ethanol vapour the alternative pathway was induced but only when very high ethanol levels in the bulbs were reached. The amount of ethanol accumulated in the bulbs during a 7 day anaerobic treatment was far too low to explain the observed induction of alternative pathway capacity. Flowering percentages were enhanced after a 24 h treatment with ethylene and after a 7 day anaerobic treatment. NBD significantly inhibited the effect of exogenous ethylene and of anaerobiosis on flowering. Ethanol was not able to induce flowering. The burst-like production of ethylene after anaerobiosis probably is responsible for the effects on respiration and flowering.     

Studies on the endogenous metabolism of Escherichia coli

1. The endogenous metabolism of Escherichia coli has been studied by examining changes in cellular composition and of the suspending fluid during starvation of washed suspensions of the organism, in water or in phosphate buffer, at 37° under aerobic and anaerobic conditions. 2. When E. coli is grown in glucose–ammonium salts media the cells contain glycogen, which is utilized rapidly during subsequent starvation of the cells. 3. Ammonia is released by starved cells only after a lag period, which corresponds to the time taken for the cellular glycogen to be almost completely utilized. 4. If cells are grown under conditions that permit incorporation of ¹⁴C into protein but not into glycogen and are then starved, release of ¹⁴CO₂ commences immediately and continues at a linear rate throughout the period of glycogen utilization; it is concluded that the presence of glycogen in the cell prevents the net degradation of nitrogenous materials but does not suppress protein turnover. 5. RNA is degraded by the cells immediately they are starved, ribose is oxidized and ultraviolet-absorbing materials are released to the suspending medium. 6. There is no significant utilization of lipid during the starvation of glucose-grown E. coli. 7. There is no loss of viability during the initial 12hr. period of starvation under either aerobic or anaerobic conditions, but thereafter the cells die more rapidly under conditions of anaerobiosis. 8. These results are discussed in relation to the known patterns of endogenous metabolism and survival of other bacteria.     

Indole-3-acetic acid and rice coleoptile elongation under anoxia

To investigate the presence of a possible synergistic effect of IAA and anaerobiosis on rice coleoptile elongation, excised coleoptiles grown in aerobic and anaerobic conditions were tested and compared with intact seedling aerial parts for response to exogenous IAA and for levels of endogenous IAA. Excised coleoptiles were fed with³H-IAA to study aerobic and anaerobic IAA metabolism. Our results can be summarized as follows. (1) IAA and anaerobiosis have no synergistic effect on rice coleoptile elongation. (2) This behavior is due not to an inhibition of IAA uptake but probably to a reduced and different IAA metabolism in coleoptile grown in the absence of oxygen. (3) In anaerobic rice coleoptiles, the conversion to inactive conjugate (IAA-Asp) could be adopted as means of detoxification in the case of abnormally high and unutilized IAA levels. (4) The increase in IAA level found in coleoptiles of intact seedlings during anaerobic treatment could be due, as in the roots, to a translocation from the endosperm, in which the hormone is contained in a great quantity.