Anaerobic Metabolism in Germinating Seeds of Echinochloa crus-galli (Barnyard Grass) : METABOLITE AND ENZYME STUDIES

Echinochloa crus-galli, a problem weed in rice fields, has the rare ability to germinate and to grow in a totally oxygen-free environment. After 7 days growth in the light or dark under N₂, E. crus-galli var. oryzicola produces a 2- to 3-centimeter nonpigmented shoot.     

Germination of Echinochloa crus-galli (Barnyard Grass) Seeds under Anaerobic Conditions : Respiration and Response to Metabolic Inhibitors

Echinochloa crus-galli L. Beauv., a rice-field weed, can germinate and grow for extended periods of time in an anaerobic environment. Compared to pea, which does not germinate under anaerobiosis, the evolution of CO₂ in Echinochloa and rice is lower and the peak rate of CO₂ evolution is delayed when germinated without oxygen. The plants studied also differ with respect to their respiration ratio ([CO₂] N₂/[CO₂] air) and metabolism used during the early stages of germination. Echinochloa does not increase its glycolytic rate under anaerobiosis, whereas pentose phosphate pathway activity appears to increase during the first 40 to 50 hours of germination.

Based on its response to metabolic inhibitors (NaF, dinitrophenol, and malonate), anaerobic metabolism in Echinochloa proceeds primarily through glycolysis, with partial operation of the tricarboxylic acid cycle and little or no oxidative phosphorylation. Also, Echinochloa is sensitive to CN during aerobic germination, whereas rice appears to be able to shift to CN-insensitive electron transport. Finally, the effectiveness of cyanide and azide on inhibiting germination of Echinochloa in N₂, but not CO, suggests that cytochrome oxidase is not used to reoxidize pyridine nucleotides in the absence of oxygen. The possible existence of an alternate electron acceptor is discussed.

     

Allelopathic Substance Exuded from a Serious Weed, Germinating Barnyard Grass (Echinochloa crus-galli L.), Roots

The allelopathy of a serious weed, barnyard grass (Echinochloa crus-galli L.), was investigated. Root exudates of young barnyard grass showed allelopathic effects and plant-selective activity and inhibited root elongation of all plants tested. With respect to shoot growth, the exudates did not show inhibition of barnyard grass only. The allelopathic substance was isolated and identified as p-hydroxymandelic acid by NMR. p-Hydroxymandelic acid strongly inhibited shoot growth and root elongation of all plants tested. The effects of three congeners of p-hydroxymandelic acid were tested on rice shoot growth. In the biological activity exhibited in rice, shoot growth was related to the hydroxyl groups. Received October 7, 1998; accepted March 29, 1999     

Distribution and Production of trans-Aconitic Acid in Barnyard Grass (Echinochloa crus-galli var. oryzicola) as Putative Antifeedant against Brown Planthoppers

A quantitative analysis of organic acid in leaves of barnyardgrass revealed that the contents of trans-aconitic acid werehigh, suggesting that this compound may act as an antifeedantagainst brown planthoppers. However, trans-aconitic acid couldnot be detected in the phloem sap which was considered to bethe main nutrient source for brown planthoppers. trans-Aconiticacid was formed in vitro from cis-aconitic acid through theaconitate isomerase activity which was detected only in theleaf sheath, but not in the leaf blade. (Received August 10, 1992; Accepted November 30, 1992)     

Development of Photosynthetic Activity following Anaerobic Germination in Rice-Mimic Grass (Echinochloa crus-galli var oryzicola)

Shoots of anaerobically germinated Echinochloa crus-galli var oryzicola are nonpigmented whether germinated in light or dark, and chlorophyll synthesis is minimal for the first 12 to 18 hours of greening after exposure to ambient conditions. When chlorophyll development is compared between greening anoxic and etiolated shoots, there is a 100-fold difference in chlorophyll levels at 8 hours, an 8-fold difference at 24 hours, but roughly equal amounts at 60 hours. The chlorophyll a/b ratio approaches 3 earlier in greening anoxic shoots than in greening etiolated shoots, relative to total chlorophyll. The long lag in chlorophyll synthesis can be shortened by giving dark-grown anoxic shoots a 24-hour midtreatment of air before light.

Development of photosynthetic activity in etiolated shoots, determined by CO₂ gas exchange, ¹⁴CO₂ uptake, and activity of carboxylating enzymes closely parallels development of chlorophylls. However, development of photosynthetic capability in greening anoxic shoots does not parallel chlorophyll development; ability to fix carbon lags behind chlorophyll synthesis. A reason for this lag is the very low activity of RuBP carboxylase during the first 36 hours of greening in anoxic shoots. The activity of phosphoenolpyruvate carboxylase is also delayed, but its kinetics more closely match those of chlorophyll development.

     

The Effects of Oxygen, Carbon Dioxide and Ethylene on Ethylene Biosynthesis in Relation to Shoot Extension in Seedlings of Rice (Oryza sativa) and Barnyard Grass (Echinochloa oryzoides)

Exposing dark-grown seedlings for 3 d to oxygen deficiency (0or 5 kPa) or to additions of carbon dioxide (10 kPa) or ethylene(0·1 Pa) slowed shoot extension in Echinochloa oryzoides,while in rice it was promoted by these treatments, except that5 kPa oxygen was without effect. In E. oryzoides this was dueto reduced growth of the mesocotyl, and in rice to enhancedgrowth of the coleoptile. These responses to carbon dioxideand oxygen deficiency were not consequences of increased ethyleneproduction, since this remained unchanged by carbon dioxideand depressed by oxygen shortage in both species. Furthermore,exogenous ethylene and the ethylene action inhibitor 2,5-norbornadieneeach failed to influence extension in anoxic seedlings, indicatingno regulatory role for ethylene in the absence of oxygen. However,concentrations of the ethylene precursor 1 -aminocyclopropane-1-carboxylic acid (ACC) were increased by carbon dioxide and0 kPa or 5 kPa oxygen, although after 72 h without oxygen totalACC production (i.e. changes in ethylene + ACC + MACC) was suppressedin both species. There was little effect on bound ACC [putativemalonyl-ACC (MACC)] formation. Transferring anaerobic (0 kPa)seedlings to oxygenated conditions (21 kPa) resulted in abnormallyfast rates of ethylene formation, possibly due to the accumulationof ACC under anoxia. This post-anoxic ethylene may have contributedto the faster extension by rice coleoptiles and slower extensionby mesocotyls of E. oryzoides compared with those of seedlingsmaintained continuously in air. Echinochloa oryzoides [Ard.] Fritsch, barnyard grass, Oryza sativa L, rice, oxygen shortage, carbon dioxide, ethylene biosynthesis, shoot extension, 1-aminocyclopropane-1-carboxylic acid (ACC), malonyl-ACC, GC-MS     

Sensitivity of Echinochloa muricata and Echinochloa crus-galli to HPPD- and ALS-inhibiting herbicides in corn

Until recently Echinochloa muricata var. microstachya Wiegand (rough barnyardgrass), an alien species native to North America, was completely overlooked in Belgium due to its close morphological resemblance to Echinochloa crus-galli (L.) P. Beauv. (barnyardgrass). E. muricata var. microstachya has gradually spread and is now locally naturalized and abundant in and along maize fields. One of the possible reasons for its expansion in maize fields, besides e.g. the lack of crop rotation, might be a lower sensitivity to postemergence herbicides acting against panicoid grasses, in particular 4-hydroxyphenyl pyruvate dioxygenase (HPPD)-inhibiting herbicides and acetolactate synthase (ALS) inhibiting herbicides. Dose-response pot experiments were conducted in the greenhouse to evaluate the effectiveness of four HPPD-inhibitor herbicides [topramezone (ARIETTA), mesotrione (CALLISTO), tembotrione (LAUDIS), sulcotrione (MIKADO) and the ALS-inhibitor herbicide nicosulfuron (KELVIN) for controlling local populations of E. crus-galli and E. muricata. Pots were planted with 25 seeds, thinned afterwards to 5 plants (one week after sowing) and irrigated by overhead sprinklers. Herbicides were applied at the 3-4 leaf stage (BBCH stage 13-14). Fresh biomass was harvested 28 d after treatment. In another dose-response pot experiment, the influence of leaf stage at time of herbicide application on efficacy of topramezone for (rough) barnyardgrass control was evaluated. Sensitivity to HPPD-inhibitor herbicides topramezone and sulcotrione was significantly lower for E. muricata populations than for E. crus-galli populations. However, nicosulfuron sensitivity of both species was similar. Compared to E. crus-galli, sensitivity of E. muricata to topramezone was more dependent on leaf stage. Due to the intragenus variability in sensitivity to HPPD-inhibitor herbicides, higher awareness is required for presence of E. muricata plants in maize fields in order to avoid insufficient "barnyardgrass" control.     

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]     

The Metabolism of Ethanol in Germinating Pea Seedlings

Considerable losses of ethanol occurred during the germinationof green pea seeds which could not be ascribed to losses dueto the volatility of the alcohol. Changes in the contents ofacetaldehyde, acetone, organic acids, and in the gas exchangessuggested that the alcohol was oxidatively metabolized. Feedingethanol to slices of pea cotyledon tissue also indicated ethanolconversion to acetaldehyde and the interconversion of acetaldehydeand acetone. Feeding ethanol 2:¹⁴C to the slices confirmed thatthe ethanol was metabolized, giving similar changes in the contentof carbonyl compounds and organic acids to those observed inthe intact germinating pea seedlings. Thus the endogenous ethanolwhich accumulated when pea seed imbibed water prior to germinationmay be metabolized subsequently by the germinating seedling.     

Bipolaris eleusines,a potential mycoherbicide candidate for control of barnyardgrass (Echinochloa crus-galli)

An isolate of Bipolaris eleusines was investigated as a potential candidate for biocontrol of barnyardgrass and additional weeds in paddy rice fields and for safety to selected crop species under greenhouse conditions. Barnyardgrass appeared more susceptible at the three-leaf stage than older plants, and disease severity increased as the fungal inoculum increased from 1 × 10⁵ to 1 × 10⁷ conidia/ml when sprayed till run-off. The high application rate caused 73% mortality of barnyardgrass, relative to the non-treated control, but increasing application rate to >1 × 10⁷ conidia/ml did not enhance efficacy (P >.05). This B. eleusines isolate showed no pathogenicity to rice (Oryza sativa spp. indica, O. sativa spp. japonica and an O. sativa hybrid), corn, wheat or any dicot crop species tested while causing slight infection on sorghum and barley. We conclude that B. eleusines, with high efficacy against barnyardgrass and demonstrated safety to rice, is a promising mycoherbicide candidate worthy of further evaluation and development for control of barnyardgrass in paddy rice fields.     

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.     

Interactions of Alcohols and Increased Air Pressure on Echinochloa crus-galli (L) Beauv. Seed Germination

In contrast to previous findings, where stimulation of E. crus-galliseed germination by n-propanol was reversed by simultaneoustreatment with 0.965 MPa air pressure, similar pressure wasunable to reverse stimulation by n-butanol. When pressure wasapplied up to 2 d prior to n-propanol, stimulation by n-propanolwas still prevented. Pressure appeared to exert its effect rapidly;only 15 min exposure gave the maximum effect. If temperatureduring treatment with pressure and n-propanol was 15 °C,rather than 35 °C, the inhibitory effect was not observed.The inhibitory action of pressure can persist but is not lethalsince drying the seeds after n-propanol and pressure treatmentrestored high levels of responsiveness to red irradiation. Theseseveral facts suggest that pressure may not simply prevent theaction of alcohols (anaesthetics) as previously thought, butmay have an action of its own, which may only be apparent uponinteraction with specific compounds, e.g. n-propanol. Phytochrome, anaesthesia, dormancy, membrane, Echinochloa crus-galli (L.) Beauv., barnyardgrass     

Effects of temperature and CO2 enrichment on kinetic properties of NADP+-malate dehydrogenase in two ecotypes of Barnyard grass (Echinochloa crus-galli (L.) Beauv.) from contrasting climates

Summary The apparent energy of activation (E_a), Michaelis-Menten constant (K_mfor oxaloacetate), V_max/K_mratios and specific activities of NADP⁺-malate dehydrogenase (NADP⁺-MDH; EC 1.1.1.82) were analyzed in plants of Barnyard grass from Québec (QUE) and Mississippi (MISS) acclimated to two thermoperiods 28/22°C, 21/15°C, and grown under two CO₂ concentrations, 350 l l^-1 and 675 l l^-1. E_avalues of NADP⁺-MDH extracted from QUE plants were significantly lower than those of MISS plants. K_mvalues and V_max/K_mratios of the enzyme from both ecotypes were similar over the range of 10–30°C but reduced V_max/K_mratios were found for the enzyme of QUE plants at 30 and 40°C assays. MISS plants had higher enzyme activities when measured on a chlorophyll basis but this trend was reversed when activities were expressed per fresh weight leaf or per leaf surface area. Activities were significantly higher in plants of both populations acclimated to 22/28°C. CO₂ enrichment did not modify appreciably the catalytic properties of NADP⁺-MDH and did not have a compensatory effect upon catalysis or enzyme activity under cool acclimatory conditions. NADP⁺-MDH activities were always in excess of the amount required to support observed rates of CO₂ assimilation and these two parameters were significantly correlated. The enhanced photosynthetic performance of QUE plants under cold temperature conditions, as compared to that of MISS plants, cannot be attributed to kinetic differences of NADP⁺-malate dehydrogenase among these ecotypes.     

Anaesthetic Enhancement of Echinochloa crus-galli (L.) Beauv. Seed Germination: Possible Membrane Involvement

Taylorson, R. B. 1988. Anaesthetic enhancement of Echinochloacrus-galli (L.) Beauv. seed germination: possible membrane involvement.—J.exp. Bot 39: 50–58. Dormant E. crus-galli seeds can be stimulated to germinate bycontact with solutions of several anaesthetic-like substances.The order of activity was n-pentanol > benzyl alcohol >n-butanol > n-propanol > ethanol. 2-propanol is nearlyinactive. The relative activity of the substances as germinationstimulants is closely related to their membrane-buffer partitioncoefficients in a manner similar to their effectiveness as anaestheticsin animals. Thus, n-pentanol is a more active stimulant of germinationthan ethanol and its increased effectiveness is correlated witha higher membrane-buffer partition coefficient Similarly, ananaesthetic that stimulated germination (n-propanol) causedgreater leakage of electrolytes, U.V. absorbing substances andamino acids from E. crus-galli seeds than a substance that wasnot active as a germination stimulant (2-propanol). In animals,action of anaesthetics can be prevented by application of externalpressure, an argument for membrane action. Action of n-propanolin stimulating germination can be similarly prevented by pressurein E. crus-galli seeds. The several lines of evidence supporta membrane action for anaesthetic-like substances in seeds and,accordingly, a membrane site as the locus of seed dormancy. Key words: Anaesthetics, germination, membranes     

Effects of Temperature and CO(2) Enrichment on Carbon Translocation of Plants of the C(4) Grass Species Echinochloa crus-galli (L.) Beauv. from Cool and Warm Environments

Plants of Echinochloa crus-galli from Québec and Mississippi were grown under two thermoperiods (28°C/22°C, 21°C/15°C) and two atmospheric CO₂ concentrations (350 and 675 microliters per liter) to examine possible differential responses of northern and southern populations of this C₄ grass species. Translocation was monitored using radioactive tracing with short-lived ¹¹C. CO₂ enrichment induced a decrease in the size of the export pool in plants of both populations. Other parameters did not strongly respond to elevated CO₂. Low temperature reduced translocation drastically for plants from Mississippi in normal CO₂ concentration, but this reduction was ameliorated at high CO₂. Overall, plants from Québec had a higher ¹¹C activity in leaf phloem and a higher percentage of ¹¹C exported, whereas these northern plants had lower turnover time and smaller pool size than plants from the southern population.     

Efficacy of Exserohilum longirostratum on barnyard grass (Echinochloa crus-galli spp. crusgalli) under field conditions

An isolate of Exserohilum longirostratum obtained from Rottboelia cochinchinensis in Malaysia was highly pathogenic to barnyardgrass (Echinocloa crus-galli,) a serious weed in rice fields in Malaysia. In glasshouse trials, high levels of barnyardgrass control were achieved when E. longirostratum was applied as a conidial concentration at 10⁵ conidia/mL. This conidial concentration may not be sufficient for field use; therefore, a mini plot trial was carried out to assess the bioherbicidal potential of the fungus and the efficacy of different inoculum types under field conditions. Excellent barnyard grass disease as indicated by Area Under Disease Progress Curve (AUDPC) was achieved with mycelium (AUDPC = 583.8 unit²); mycelium + pretichlaclor (AUDPC = 610.4 unit²) and conidia alone (AUDPC = 468.3 unit²) compared to conidia + pretichlaclor (AUDPC = 395.8 unit²). Although conidia caused lower disease severity on barnyard grass compared to mycelium, this inoculum equally reduced the competition from barnyard grass as indicated by lower mean dry weight and fewer barnyard grass tiller numbers. Positive correlations between AUDPC of rice and its tiller number and AUDPC of rice and its dry weight were recorded while negative correlations were observed for the AUDPC of barnyard grass and its tiller number and AUDPC of barnyard grass and its dry weight. These findings confirmed that E. longirostratum applied either in the form of conidia or mycelium was effective against barnyard grass under field conditions.     

盐胁迫下大麦根系多胺代谢与其耐盐性的关系

研究了0～300mmol/LNaCl对大麦(Hordeum-vulgare-L.)幼苗生长速率、根系游离和结合态多胺含量以及多胺生物合成关键酶活性的影响。结果表明,在0～200mmol/L NaCl处理下精氨酸脱羧酶（ADC）、多胺氧化酶(PAO)以及转谷酰胺酶(Tgase)活性明显提高,而在300 mmol/L NaCl处理下活性下降。与之对应,游离腐胺（Put）含量随处理盐浓度的提高一直呈上升趋势,亚精胺(Spd)和在根系内检测到的未知多胺(Pax)在低浓度盐处理时含量上升,随盐浓度的提高含量下降。盐处理前后精胺(Spm)含量变化不明显。低浓度盐处理时游离态（Spd+Pax）/Put上升,随盐浓度的提高比值明显下降。结合态Put、Spd和Pax含量以及结合态多胺总量均在低浓度盐处理时上升,随盐浓度的提高含量明显下降。统计分析显示,大麦相对生长速率与游离态（Spd+Pax）/Put和结合态多胺含量间均呈极显著正相关关系,与游离态多胺和结合态多胺的比值间均呈显著负相关关系,上述结果说明盐胁迫下大麦体内游离态Spd、Pax与Put以及结合态形式之间的平衡与大麦耐盐性关系密切,游离态Put向Spd 、Pax以及结合态形式转化均有利于大麦耐盐性的提高.     

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