Ageing of isolated embryos and endosperms of durum wheat: An analysis of chromosome damage

Embryos (EM) and endosperms (END) were isolated from durum wheat seeds and aged separately for 3 years. Analysis of chromosome damage was carried out on root tip meristems of transplants in which aged (a) embryos and/or endosperms were used. The aim of the experiments was to investigate the ageing process when embryos and endosperms age separately and to evaluate the possible influence of the endosperm during storage of the seed.The results obtained indicate that (i) both aged embryo and endosperm produce mutagenic substances capable of inducing nuclear damage in the radicle meristem and (ii) age-induced damage in the embryo is not the consequence of endosperm ageing. The relationships between chromosome damage produced by aged embryos and endosperms in the transplants are discussed.     

Metabolic Changes in Partially Dormant Wheat Seeds during Storage

Changes in germination, seedling growth, respiration, response to applied gibberellic acid, and glucose-U-¹⁴C utilization were investigated in partially dormant wheat (Triticum aestivum L., Pa 151 × 107) seeds which were stored under various conditions for periods up to 1 year. Only seeds stored at −20 C and 12.4% moisture maintained partial dormancy, which was overcome by germinating in 10⁻³m gibberellic acid. Germination and seedling growth of seeds stored at 25 C and 15.1% moisture declined within 12 weeks and the percentage of seeds infected with storage fungi increased. Gibberellic acid produced faster growing seedlings, particularly from those seeds with partial dormancy, but did not overcome growth reduction which was caused by deterioration. Seeds kept under laboratory conditions (B), 25 C and 12.1% moisture (C), and 25 C and 15.1% moisture (D) for 12 weeks utilized 35, 55, and 80% less glucose, respectively, than those stored at −20 C and 12.4% moisture (A). Seeds stored under B and C consistently had higher germination, growth, and respiratory rates than seeds from A and D. The respiratory rate declined as deterioration advanced under D. Respiratory quotients ranged from 1.0 for seeds stored under A to 1.6 for seeds stored under D.     

施肥方式对紫色土土壤异养呼吸的影响

采用静态暗箱-气相色谱法于2010年12月至2011年10月对不同施肥方式下的紫色土土壤呼吸进行了研究,以揭示施肥方式对紫色土异养呼吸的影响。结果表明:施肥可对土壤异养呼吸产生激发效应。施肥后第5天出现峰值,猪厩肥处理的异养呼吸峰值为2356.8 mg CO2m-2h-1,显著高于秸秆配施氮磷钾(970.1 mgCO2m-2h-1)和常规氮磷钾处理(406.8 mgCO2m-2h-1)(P0.01);小麦季常规氮磷钾、猪厩肥和秸秆配施氮磷钾处理的平均土壤异养呼吸速率为212.9、285.8和305.8mgCO2m-2h-1,CO2排放量为255.1、342.3和369.5 gC/m2,玉米季为408.2、642.8和446.4 mgCO2m-2h-1,CO2排放量为344.7、542.8和376.9 gC/m2,玉米季土壤异养呼吸平均速率及CO2排放量均高于小麦季。全年平均土壤异养呼吸速率分别为310.6、446.3和377.4 mg CO2m-2h-1,CO2排放总量分别为599.8、885.1和746.4 gC/m2。猪厩肥对土壤异养呼吸速率和CO2排放量的影响最大,秸秆配施氮磷钾肥次之,氮磷钾肥最小,说明有机物料的投入是紫色土土壤异养呼吸速率的主要调控措施,低碳氮比的有机物料能促进土壤异养呼吸和CO2的排放。猪厩肥和秸秆配施氮磷钾肥处理相应地表和地下5 cm温度的Q10值分别为2.64、1.88和2.77、1.99,表明低碳氮比的有机物料还能增加土壤异养呼吸Q10值,使土壤异养呼吸速率对温度的敏感性加强。     

Growth and respiration patterns of snap bean fruits

The relationship of respiration and growth of seed, pericarp tissue and whole fruit of snap beans (Phaseolus vulgaris L.) was studied. The whole fruit exhibited an apparent climacteric type of respiration pattern. This pattern resulted from an increase in CO₂ production by the enlarging seed followed by a rapid decrease in CO₂ evolution by the pericarp tissue, and the pattern was not associated with any concomitant increase in ethylene production. Therefore, the apparent climacteric respiration pattern of a developing bean fruit is not comparable to the phenomenon that occurs in other ripening fruits.     

Economy of water, carbon, and nitrogen in the developing cowpea fruit

The nutritional economy of the fruit of cowpea (Vigna unguiculata (L.) Walp cv Vita 3) was assessed quantitatively from intake and utilization of carbon, nitrogen, and water. Fruits failed to make net gains of CO₂ from the atmosphere during daytime, although pod photosynthesis did play a role in the fruit's carbon economy by refixing a proportion of the fruit's respired CO₂. Of every 100 units by weight of carbon entering the fruit, 70.4 were finally incorporated into seeds, 10.3 remained as nonmobilizable material in pod walls, and the remaining 19.3 were lost in fruit respiration. Phloem supplied 97% of the fruit's carbon and 72% of its nitrogen. The xylem contribution of nitrogen occurred mainly in early growth. Ninety-six% of the fruit's nitrogen was incorporated into seeds, approximately 10% of this mobilized from the senescing pod. The mean transpiration ratio of the fruit was very low—8 milliliters water transpired per gram dry matter accumulated. Models of carbon, nitrogen, and water flow were constructed for the two consecutive 11 day periods of fruit development, and indicated a considerably greater entry of water through xylem and phloem than could be accounted for in changes in fruit tissue water and transpiration loss. This discrepancy was greater in the second half of fruit growth and was interpreted as evidence that a significant fraction of the water entering the fruit through phloem cycled back to the parent plant via the xylem.     

Endosperms and embryos of mature dry castor bean seeds contain active ribosomes

Mature dry endosperms and embryos of castor bean contain ribosomes which are capable of catalysing protein synthesis in vitro. This observation contradicts previous reports that ribosomes are absent from mature dry endosperms. On hydration of the mature seeds the polyribosome content of both embryos and endosperms increases. These polyribosomes are lost on subsequent dehydration but, again, ribosomes are conserved. REFERENCES     

Brassinosteroids regulate grain filling in rice

Genes controlling hormone levels have been used to increase grain yields in wheat (Triticum aestivum) and rice (Oryza sativa). We created transgenic rice plants expressing maize (Zea mays), rice, or Arabidopsis thaliana genes encoding sterol C-22 hydroxylases that control brassinosteroid (BR) hormone levels using a promoter that is active in only the stems, leaves, and roots. The transgenic plants produced more tillers and more seed than wild-type plants. The seed were heavier as well, especially the seed at the bases of the spikes that fill the least. These phenotypic changes brought about 15 to 44% increases in grain yield per plant relative to wild-type plants in greenhouse and field trials. Expression of the Arabidopsis C-22 hydroxylase in the embryos or endosperms themselves had no apparent effect on seed weight. These results suggested that BRs stimulate the flow of assimilate from the source to the sink. Microarray and photosynthesis analysis of transgenic plants revealed evidence of enhanced CO₂ assimilation, enlarged glucose pools in the flag leaves, and increased assimilation of glucose to starch in the seed. These results further suggested that BRs stimulate the flow of assimilate. Plants have not been bred directly for seed filling traits, suggesting that genes that control seed filling could be used to further increase grain yield in crop plants.     

Mobilization of respiratory metabolism in potato tubers by carbon dioxide

Applying high concentrations of CO₂ to whole potato tubers stimulated a rapid and pronounced respiratory gas exchange, which persisted for a prolonged time. The upsurge in respiration was proportional to the applied CO₂ concentrations and was further augmented by high O₂ levels. Tests using whole potatoes, or potato tissue slices from tubers previously treated with CO₂, indicated that the rapid CO₂-induced respiration is sensitive to cyanide during the first 24 hours of CO₂ application. The respiratory rise cannot be attributed to the emergence of a cyanide-resistant alternative electron transport pathway, although prolonged applications of CO₂, up to 72 hours, led to a gradual development of the pathway. CO₂-stimulated respiration was accompanied by a pronounced decline in the content of starch and glucose 6-phosphate, suggesting an active utilization of respiratory substrates. The ATP content in the CO₂-treated potatoes increased markedly, resembling similar increases in tissues undergoing respiratory upsurge.     

Nutrition of a developing legume fruit: functional economy in terms of carbon, nitrogen, water

The economy of functioning of the developing fruit of white lupin (Lupinus albus L.) is assessed quantitatively in relation to intake and usage of carbon, nitrogen, and water. Of every 100 units of carbon imported from the parent plant, 52 are incorporated into seeds, 37 into nonmobilizable material of the pod, and the remaining 11 lost as CO₂ to the atmosphere. An illuminated fruit can make net gains of CO₂ from the atmosphere during the photoperiods of all but the last 2 weeks of its life, suggesting that it is active in assimilation of CO₂ respired from pods and seeds. This conservation activity is important to carbon economy.     

A debranching enzyme deficiency in endosperms of the sugary-1 mutants of maize

Many of the sugary-1 mutants of maize (Zea mays L.) have the highly branched water-soluble polysaccharide, phytoglycogen, in quantities equal to or greater than starch as an endosperm storage product in mature seeds. We find that all sugary mutants investigated are deficient in debranching enzyme [α-(1, 6)-glucosidase] activity in endosperm tissue 23 days postpollination and suggest that this deficiency is the primary biochemical lesion leading to phytoglycogen accumulation in sugary endosperms. This would indicate that the amylopectin component of starch depends on an equilibrium between the activities of branching enzymes introducing α-1,6 branch points into the linear α-1,4 glucans and debranching enzymes. The debranching enzyme activities from nonsugary endosperms can be separated into three peaks on a hydroxyapatite column. The sugary endosperm extracts lack one of these peaks of activity while the other two fractions have much reduced activity. The embryos of developing seeds (23 days after pollination) from both sugary and nonsugary genotypes have equivalent debranching activity. The debranching enzyme activity of developing endosperms is proportional to the number of copies (0 to 3) of the nonmutant (Su) allele present suggesting that the Su allele may be the structural gene for this debranching enzyme, although this is not definitive. This identification of debranching enzyme activity as being the biochemical lesion in sugary endosperms is consistent with several previous observations on the mutant.     

Root and nodule respiration in relation to acetylene reduction in intact nodulated peas

Inoculated pea plants (Pisum sativum L.) were grown with N-free nutrients in a controlled environment room and rates of respiratory CO₂ evolution and C₂H₂ reduction by the intact nodulated roots were determined. Experiments followed changes related to diurnal cycles, light and dark treatments, partial defoliation, aging of plants and NH₄NO₃ addition. In all experiments, changes in C₂H₂ reduction were associated with parallel changes in the respiration rate, although in all but the defoliation experiment there was a basal level of respiration which was independent of the rate of C₂H₂ reduction. In conditions which affected growth or plant size as well as C₂H₂ reduction, respiration changed by an average of 0.42 mg CO₂ (μmol C₂H₂ reduced)⁻¹. However, some treatments decreased C₂H₂ reduction without greatly changing the growth and in these conditions respiration was decreased by an average of 0.27 mg CO₂ (μmol C₂H₂ reduced)⁻¹. While this value may also include some respiration associated with other processes, it is proposed that it more closely estimates respiration directly associated with energy utilization for acetylene reduction; whereas the higher value includes respiration related to maintenance and growth processes as well.     

Carbohydrate metabolism in germinating caryopses of <Emphasis Type="Italic">Oryza sativa</Emphasis> L. exposed to prolonged anoxia

Anoxia tolerance can be evaluated not only in terms of growth or survival of plant organs during oxygen deprivation, but also in relation to carbohydrate utilization in the context of a well-modulated fermentative metabolism. Rice (Oryza spp.) is unique among cereals, in that it has the distinctive ability to germinate under complete anaerobiosis by using the starchy reserves in its seeds to fuel the anaerobic metabolism. The aim of the present study was to evaluate the ability of germinating rice seedlings to survive a long-term oxygen deficiency [40 days after sowing (DAS)] and the effects on sugar metabolism, focusing on starch degradation as well as soluble sugars transport and storage under anoxia. No significant decline in vitality occurred until 30 DAS though no recovery was detected following longer anoxic treatments. Growth arrest was observed following anoxic treatments longer that 20 DAS, in concomitance with considerably lower ethanol production. Amylolytic activity in embryos and endosperms had similar responses to anoxia, reaching maximum content 30 days after the onset of stress, following which the levels declined for the remainder of the experiment. Under anoxia, average amylolytic activity was twofold higher in embryos than endosperms. Efficient starch degradation was observed in rice under anoxia at the onset of the treatment but it decreased over time and did not lead to a complete depletion. Our analysis of α-amylase activity did not support the hypothesis that starch degradation plays a critical role in explaining differences in vitality and coleoptile growth under prolonged oxygen deprivation.     

Gibberellin-induced hydrolysis of endosperm cell walls in gibberellin-deficient tomato seeds prior to radicle protrusion

The weakening of the mechanical restraint of the endosperm layer in tomato (Lycopersicon esculentum Mill.) seeds, a prerequisite for germination, has been studied with the use of seeds of the gibberellin (GA)-deficientgib-1 mutant. Incubation ofgib-1 endosperms, including part of the testa, in 10 M GA₄₊₇, resulted within 12 h in the release of fructose, glucose, galactose and mannose into the incubation medium. Only small amounts of sugars diffused out of thegib-1 endosperms during incubation in water. Chemical hydrolysis of endosperm cell walls ofgib-1 seeds showed that they are mainly composed of mannose, and smaller quantities of glucose and galactose. Treatment with GA₄₊₇ induced in the endosperms the production of endo--mannanase activity that was not detectable during incubation in water, and also increased the activities of mannohydrolase and -galactosidase as compared with the water controls. No cellulase activity was found. It is concluded that in tomato seeds the weakening of endosperms prior to radicle protrusion is mediated by a GA-induced enzymatic degradation of the mannan-rich cell walls.Abbreviation GA(s) gibberellin(s)     

Control of Seed Respiration and Growth in Vicia faba by Oxygen and Temperature: No Evidence for an Oxygen Diffusion Barrier

The rate of dry matter accumulation by seeds of Vicia faba L. cv. Minica increases with temperature in the range of 16 to 26°C. The duration of dry matter accumulation decreases with temperature, resulting in a decrease of final seed dry weight. In this study we test the hypothesis that a diffusion barrier for O₂, located in the seed coat, inhibits seed respiration and growth. The rate of O₂ uptake of intact seeds and of excised embryos and seed coats (separated seeds) was measured in air and buffer at 16, 20, and/or 26°C at various O₂ concentrations and developmental stages. Oxygen uptake rates of intact seeds in buffer were only 9 to 15% of those in air. In buffer, the respiration rate of intact seeds decreased at a pO₂ below air saturation (21 kilopascals), whereas separated seeds showed a decline of O₂ uptake only below 80% of air saturation. In air, embryo excision had no effect on the sensitivity of seed respiration to pO₂, at both 20 and 26°C. In air at 20°C, separated and intact seeds showed similar rates of O₂ uptake. Oxygen uptake by intact seeds, both halfway and beyond the linear growth phase, showed a temperature coefficient Q₁₀ of 2.3 and was insensitive to pO₂ in the range of 80 to 100% of ambient. These results indicate that V. faba seed respiration in air is not limited by the diffusion of O₂ into the seed.     

Growth and maintenance respiration in stems of Quercus alba after four years of CO2 enrichment

Atmospheric CO₂ enrichment is increasingly being reported to inhibit leaf and whole-plant respiration. It is not known, however, whether this response is unique to foliage or whether woody-tissue respiration might be affected as well. This was examined for mid-canopy stem segments of white oak (Quercus alba L.) trees that had been grown in open-top field chambers and exposed to either ambient or ambient + 300 µmol mol^?1 CO₂ over a 4-year period. Stem respiration measurements were made throughout 1992 by using an infrared gas analyzer and a specially designed in situ cuvette. Rates of woody-tissue respiration were similar between CO₂ treatments prior to leaf initiation and after leaf senescence, but were several fold greater for saplings grown at elevated concentrations of CO₂ during much of the growing season. These effects were most evident on 7 July when stem respiration rates for trees exposed to elevated CO₂ concentrations were 7.25 compared to 3.44 µmol CO₂ m^?2 s^?1 for ambient-grown saplings. While other explanations must be explored, greater rates of stem respiration for saplings grown at elevated CO₂ concentrations were consistent with greater rates of stem growth and more stem-wood volume present at the time of measurement. When rates of stem growth were at their maximum (7 July to 3 August), growth respiration accounted for about 80 to 85% of the total respiratory costs of stems at both CO₂ treatments, while 15 to 20% supported the costs of stem-wood maintenance. Integrating growth and maintenance respiration throughout the season, taking into account treatment differences in stem growth and volume, indicated that there were no significant effects of elevated CO₂ concentration on either respiratory process. Quantitative estimates that could be used in modeling the costs of woody-tissue growth and maintenance respiration are provided.     

Osservazioni Sull'effetto Inibente di Elevate Pressioni Osmotiche su Diversi Tipi di Crescita

Abstract

On the inhibition of seeds germination and of growth by cell enlargement by the osmotic pressure of the medium. — The mechanism of inhibition by osmotic pressure (O.P.) of the medium on growth and respiration of germinating wheat, castor bean and lettuce sèeds and of etiolated pea internode segments was investigated.

The following results were obtained:

1 - External osmotic pressure (up to 0.3 M) of various substances such as mannitol, urea, glucose, NaCl, was shown to inhibit the germination and growth of lettuce, wheat and castor bean seeds.

2 - a) A remarkable decrease of the development of respiration during the first 48 h of germination was demonstrated in embryos of wheat seeds germinated and maintained in mannitol solutions at concentration from 0,2 to 0,3 M.

b) A slight but reproducible inhibition of óxygen uptake by O.P. was also observed in embryos isolated from wheat seeds germinated in water for 24 and 34 h and transported respectively in water or into 0,2 M mannitol solutions.

This is interpreted as indicating that high external O.P. inhibits both the respiratory metabolism and the development with time of enzyme systems supporting respiration.

3 - Mannitol solutions (0,2–0,3 M) inhibited completely growth by cell enlargement in pea internode sections, while they did not at all affect oxygen uptake and protein synthesis ( ¹⁴ C - leucine incorporation). The stimulatory effect of auxin on pea elongation was almost completely suppressed by mannitol, whereas the hormone stimulation of respiration remained unchanged.

These data are interpreted as indicating that in tissues, presenting an advanced differentiation, high external O.P. inhibits growth by a direct physico-chemical mechanism; while the inhibitory effect in embrional tissues seems to comprehend, besides this direct effect, a complicated metabolic component, apparently influencing protein synthesis.     

Influence of Etherel and Gibberellic Acid on Carbon Metabolism, Growth, and Essential Oil Accumulation in Spearmint (Mentha Spicata)

Changes in growth parameters and ¹⁴CO₂ and [U-¹⁴C]-sucrose incorporation into the primary metabolic pools and essential oil were investigated in leaves and stems of M. spicata treated with etherel and gibberellic acid (GA). Compared to the control, GA and etherel treatments induced significant phenotypic changes and a decrease in chlorophyll content, CO₂ exchange rate, and stomatal conductance. Treatment with etherel led to increased total incorporation of ¹⁴CO₂ into the leaves wheras total incorporation from ¹⁴C sucrose was decreased. When ¹⁴CO₂ was fed, the incorporation into the ethanol soluble fraction, sugars, organic acids, and essential oil was significantly higher in etherel treated leaves than in the control. However, [U-¹⁴C]-sucrose feeding led to decreased label incorporation in the ethanol-soluble fraction, sugars, organic acids, and essential oils compared to the control. When ¹⁴CO₂ was fed to GA treated leaves, label incorporation in ethanol-insoluble fraction, sugars, and oils was significantly higher than in the control. In contrast, when [U-¹⁴C]-sucrose was fed the incorporation in the ethanol soluble fraction, sugars, organic acids, and oil was significantly lower than in the control. Hence the hormone treatment induces a differential utilization of precursors for oil biosynthesis and accumulation and differences in partitioning of label between leaf and stem. Etherel and GA influence the partitioning of primary photosynthetic metabolites and thus modify plant growth and essential oil accumulation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Root environment acidity as a regulatory factor in ammonium assimilation by the bean plant

Previous experiments have revealed a much greater efficiency of ammonium utilization by bean plants (Phaseolus vulgaris L.) when the acidity of the ambient medium was maintained at near-neutral conditions with carbonates or hydroxides. The present investigation, in which ¹⁵N-labeled ammonium was used, permitted an assessment of the origin of nitrogen in tissue nitrogen pools with and without acidity control (CaCO₃ treated and untreated, respectively) in the root environment. Control of acidity resulted in greater ammonium uptake and greater incorporation into the amino fraction, amide, and ethanol-insoluble nitrogen by the root tissue. These differences were clearly evident by the fifth day after ammonium nitrogen had been applied.     

13C isotope fractionation during rhizosphere respiration of C3 and C4 plants

Stable carbon isotopes are used extensively to partition total soil CO₂ efflux into root-derived rhizosphere respiration or autotrophic respiration and soil-derived heterotrophic respiration. However, it remains unclear whether CO₂ from rhizosphere respiration has the same δ¹³C value as root biomass. Here we investigated the magnitude of ¹³C isotope fractionation during rhizosphere respiration relative to root biomass in six plant species. Plants were grown in a carbon-free sand-perlite medium inoculated with microorganisms from a farm soil for 62 days inside a greenhouse. We measured the δ¹³C value of rhizosphere respiration using a closed-circulation 48-hour CO₂ trapping method during 40~42 and 60~62 days after sowing. We found a consistent depletion in ¹³C (0.9~1.7‰) of CO₂ from rhizosphere respiration relative to root biomass in three C₃ species (Glycine max L. Merr., Helianthus annuus L. and Triticum aestivum L.), but a relatively large depletion in ¹³C (3.7~7.0‰) in three C₄ species (Amaranthus tricolor L., Sorghum bicolor (L.) Moench and Zea mays L. ssp. mays). Overall, our results indicate that CO₂ from rhizosphere respiration is more ¹³C-depleted than root biomass. Therefore, accounting for this ¹³C fractionation is required for accurately partitioning total soil CO₂ efflux into root-derived and soil-derived components using natural abundance stable carbon isotope methods.     

Effects of flooding under hydrostatic pressure on the respiratory metabolism of germinated wheat seeds

Germinated wheat seeds ( Triticum aestivum L. cv. Barqai) that had been subjected to short hydrostatic pressure treatments (0.3–1.2 MPa) changed their normal metabolism into one which is characterized by a high ethanol production, a low O₂ consumption and a low CO₂ evolution. Alcoholic fermentation could account for ca half of the CO₂ evolved from the pressurized seeds. The level of acetaldehyde was low, though significantly higher in the pressurized seeds than in the controls. Subjection of wheat seeds to osmotic stress under aerobic conditions lowered their O₂ uptake and CO₂ evolution but did not induce ethanol production. Exposure of pressurized seeds to NaCl stress did not alter their ethanol production beyond that which had been induced by pressure. Ethanol production by pressurized seeds increased following either the addition of sucrose or by excision of the embryos from the endosperms. More electrolytes leaked into the embedding solution from pressurized seeds than from control seeds. Exogenous ethanol was toxic to wheat seeds at concentrations as low as 343 m M . The effects of hydrostatic pressure and of the consequently induced ethanol production on the mortality of flooded seeds is discussed.