Mode of high temperature injury to wheat. II. Comparisons of wheat and rice with and without inflorescences

High temperature injury to wheat ( Triticum aestivum L.) during grain development is manifested as acceleration of senescence. Experiments were conducted to elucidate the mode of senescence and site of high temperature responses. Wheat (cv. Chris) and rice ( Oryza sativa L. cv. Newbonnet), which have C₃ photosynthesis but different temperature responses, were grown with and without inflorescences under three temperature regimes after anthesis. Plant growth and constituents associated with senescence were measured weekly until physiological maturity. Increasing temperatures from 25°C/15°C to 35°C/25°C day/night after anthesis decreased growth, leaf viability, chlorophyll and protein concentrations, and RuBP carboxylase activity and increased protease and RNase activities in wheat. Inflorescence removal increased vegetative weights and slowed most senescence processes more in wheat than in rice, but did not alter the course of high temperature responses. Results are interpreted as indicating that diversion of nutrients from roots by inflorescence sinks at normal temperatures and by increased respiration at high temperatures caused similar responses. Source and sink activities appeared to be regulated jointly, probably by cytokinins from roots, during senescence at normal and elevated temperatures.     

Mediation of high-temperature injury by roots and shoots during reproductive growth of wheat

Abstract Previous studies suggest that high temperature stress on wheat (Triticum aestivum L.) involves root processes and acceleration of monocarpic senescence. Physiological changes in wheat roots and shoots were investigated to elucidate their relationship to injury from elevated temperatures after anthesis. Plants were grown under uniform conditions until 10 d after anthesis, when shoot/root regimes of 25°C/25°C, 25°C/35°C, 35°C/25°C and 35°C/35°C were imposed. Growth and senescence of shoots and grain were influenced more by root temperatures than by shoot temperatures. High root temperatures increased activities of protease and RNasc enzymes, and loss of chlorophyll, protein and RNA from shoots, whereas low root temperatures had opposite effects. High root temperatures appeared to induce shoot senescence directly. High shoot temperatures probably disrupted root processes, including export of cytokinins, and induced high leaf protease activity, senescence and cessation of grain development. The authors concluded that responses of wheat to high temperatures, whether of roots or shoots, are manifested as acceleration of senescence and may be mediated by roots during grain development.     

Response of carbonic anhydrase to polyethylene glycol-mediated water stress in wheat

Carbonic anhydrase (CA) activity in wheat leaves changed upon leaf dehydration: it decreased at mild stress (relative water content, RWC, 81 %), but increased at severe water stress (RWC 74 %). Phosphoenopyruvate carboxylase activity was not significantly affected by these stresses. This revised version was published online in June 2006 with corrections to the Cover Date.     

Changes in photosystem II function during senescence of wheat leaves

Analyses of chlorophyll fluorescence were undertaken to investigate the alterations in photosystem II (PSII) function during senescence of wheat ( Triticum aestivum L. cv. Shannong 229) leaves. Senescence resulted in a decrease in the apparent quantum yield of photosynthesis and the maximal CO₂ assimilation capacity. Analyses of fluorescence quenching under steady‐state photosynthesis showed that senescence also resulted in a significant decrease in the efficiency of excitation energy capture by open PSII reaction centers (F'_v/F'_m) but only a slight decrease in the maximum efficiency of PSII photochemistry (F'_v/F'_m). At the same time, a significant increase in non‐photochemical quenching (q_N) and a considerable decrease in photochemical quenching (q_P) were observed in senescing leaves. Rapid fluorescence induction kinetics indicated a decrease in the rate of Q_A reduction and an increase in the proportion of Q_B‐non‐reducing PSII reaction during senescence. The decrease in both F'_v/F'_m and q_P explained the decrease in the actual quantum yield of PSII electron transport ((φ_PSII). We suggest that the modifications in PSII function, which led to the down‐regulation of photosynthetic electron transport, would be in concert with the lower demand for ATP and NADPH in the Calvin cycle which is often inhibited in senescing leaves.     

Amino acid metabolism associated with N-mobilization from the flag leaf of wheat (Triticum aestivum L.) during grain development

Field-grown winter wheat (Triticum aestivum L. cv. Castell) was used to study changes in the free amino acid pools of different plant parts and related enzyme activities in the flag leaf throughout the grain-filling period in three consecutive growing seasons. Amino acid analysis data indicated that, during senescence, the nitrogen flow in the flag leaf was directed towards the synthesis of glutamine as a specific nitrogen transport form. Of the enzymes involved, total glutamine synthetase (GS; EC 6.3.1.2) and especially ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1) activities declined continuously as senescence progressed. Unlike (chloroplastic) GS₂, (cytosolic) GS₁ was shown to be very persistent suggesting a special role for this isoenzyme in the N-reallocation process. Glutamate-oxaloacetate transaminase (GOT; EC 2.6.1.1), glutamate-pyruvate transaminase (GPT; EC 2.6.1.2) and isocitrate dehydrogenase (IDH; EC 1.1.1.42) showed a characteristic biphasic activity profile after anthesis. It is proposed that these enzymes, for each of which at least two isoenzymes were demonstrated, are involved in glutamate synthesis at the later stages of leaf senescence. Ammonium levels were fairly constant throughout the flag leafs life span, an ultimate rise often following peak values of glutamate dehydrogenase (GDH; EC 1.4.1.4) activity. The enzymology of flag leaf amino acid metabolism during grain development is further discussed in relation to observations of NH₃-volatilization from naturally senescing wheat plants.     

The influence of reduced photorespiration on long-term growth and development of wheat

The long-term role of photorespiration was investigated by comparing growth, development, gas exchange characteristics and mineral nutrition of a wheat crop ( Triticum aestivum L. cv. Courtot) cultivated in a culture chamber during a life cycle, either in 4% O₂ or in normal O₂ Low O₂ pressure reduced photorespiration, but CO₂ was controlled so that net photosynthesis remained the same as in the control crop. The growth and development of the low O₂ crop was slowed down. Ear appearance was 16 days late, but the rate of tillering was the same as in the control and was maintained longer so that the final number of tillers was doubled. Pigment, ribulose bisphosphate carboxylase (EC 4.1.1.39) and soluble sugar contents were similar. The response of photosynthesis to CO₂ and O₂ was not appreciably changed by the low O₂ treatment. There was almost no seed formation, and the senescence of the leaves was delayed. It appears that in non-stress conditions most of the photorespiration can be suppressed without damage to the photosynthetic apparatus. Retardation of development and inhibition of reproduction are likely due to other effects of O₂.     

Planting density effects on assimilation and partitioning of photosynthates during grain filling in the late-sown wheat

Leaf blades of the late-sown winter wheat produced the major portion, i.e., more than 60 %, of the total ¹⁴C-photosynthates at grain filling, but ear (rachis and glumes) only about 15 %, sheaths about 11 %, and stem internodes about 11 %. The change of plant density in this experiment had little influence on the ¹⁴CO2-photoassimilation of the ear (rachis and glumes), flag leaf lamina, sheaths and stem internodes, but markedly affected photosynthesis of the second, the third and lower leaves. The photosynthetic rate [expressed as specific radioactivity, s-¹ kg-¹(d.m.)] and the amount of ¹⁴CO₂ photosynthates decreased significantly in the second, the third and other lower leaves at a high plant density. Upon grain-filling of the late-sown wheat, the grain was the major importer of photosynthates. Yet partitioning to the stem internodes depended on the plant density. Stem was the importer of photosynthates at a low plant density, but the exporter at a high plant density. In plants at a low plant density a fairly large proportion of photosynthates was distributed into the roots. The middle and lower above-ground parts of the late-sown wheat at a high plant density decreased or lost their function early. As a result, the plant senesced earlier. However, the grain setting, filling and yielding were restricted. An appropriately low plant density was suitable for prolonging the function of the middle and lower organs, delaying the senescence of plant, increasing the source supply for grain filling, and improving the grain yield. This revised version was published online in August 2006 with corrections to the Cover Date.     

Degradation of proteins from thylakoid membranes in senescing wheat leaves at high temperature

This investigation determined whether thylakoid proteins would be degraded more rapidly or not in senescing wheat (Triticum aestivum L. em. Thell.) leaves concurrently exposed to high temperatures. Excised leaves were pulse-labelled with [³⁵S]-methionine for a 12 h period, and then incubated at 22,32 or 42°C for 0, 1, 2, or 3 d, before extracting a thylakoid enriched membrane sample. After electrophoretic separation, two prominent [³⁵S]-labelled protein bands were chosen for further analyses. Band A contained the D-1 thylakoid protein and band B contained thylakoid proteins of the light harvesting complex (LHCII) associated with photosystem II (PSII). Total protein, [³⁵S]-labelled protein, band A protein, and band B protein within the thylakoid enriched membrane samples were measured. Unlabelled thylakoid enriched membrane samples, extracted from leaves given similar treatments, were used to measure uncoupled whole-chain and photosystem II (PSII) electron transport and chlorophyll fluorescence. Accentuated decline in whole-chain and PSII electron transport, increasing F_o values, and decreasing F_max values were a result of high temperature injury in leaves treated at 42°C. None of the thylakoid enriched membrane protein fractions were degraded more rapidly in high-temperature treated leaves. Degradation of the total [³⁵S]-labelled membrane proteins and band B was inhibited by the 42°C treatment. The results indicate that high temperature stress may disrupt some aspects of normal senescence.     

Protective and injuring action of visible light on photosynthetic apparatus in wheat plants during hyperthermia treatment

Illumination of wheat (Triticum aestivum L.) leaves during heat treatment produced either additional injury or protection of photosynthetic apparatus depending on irradiance and the heating dose. Furthermore, illumination of leaves during hyperthermia exerted differential impacts on thermal tolerances of photosynthesis and photosystem II-driven electron transport assessed from the reduction of 2,6-dichlorophenolindophenol (DCPIP). Measurements with infrared gas analyzer showed that mild heating of leaves in darkness (10 min at 38–40°C) had stronger inhibitory effect on CO₂ uptake than heating of leaves exposed to low and moderate complex irradiances (3–30 klx), as well as excessive irradiance (75–100 klx). When the leaves were heated at higher temperatures (42–44°C), the low and moderate irradiances had a protective action, while high-intensity light aggravated the inhibition of photosynthesis. Illumination of leaves with weak light during heat treatment mitigated the impairment of chloroplast ultrastructure, whereas irradiation with high-intensity light (100 klx) destroyed the sensitive population of chloroplasts. The heat-stimulated photoinhibition was stronger for leaf photosynthesis than for DCPIP reduction in chloroplasts isolated from heat-treated leaves. No correlation was observed between the extent of violaxanthin deepoxidation, zeaxanthin accumulation, and the protective effect of light on photosynthetic apparatus during heat treatments.     

Endogenous abscisic acid and wheat germ agglutinin content in wheat grains during development

Abscisic acid (ABA) and wheat germ agglutinin content of immature wheat grains and embryos was determined by immunoassay throughout the development of a field-grown wheat crop ( Triticum aestivum cv. Timmo). Wheat germ agglutinin accumulation in the embryo was not preceded by an increase in endogenous abscisic acid amount or concentration in either embryos or grains. At a later stage in development the endogenous concentration of abscisic acid in both embryos and grains was found to be two orders of magnitude lower than the endogenous levels required to inhibit precocious germination and promote wheat germ agglutinin accumulation in excised embryos cultured in vitro. These findings are discussed in the context of the control of embryo development in vivo by both ABA and the water status of the grain and embryo.     

A burst of CO2 from photosynthesizing leaves after a temperature decrease under constant light conditions

A transient CO₂ burst from seedlings of some plant species was observed after a rapid temperature decrease. The magnitude of the CO₂ release depended on initial temperature, oxygen concentration and light intensity. To obtain a maximal value of CO₂ release, the temperature had to decrease by more than 8°C. The phenomenon was detected only in the light, and was confined to C₃ species. It was inhibited by low oxygen concentration, indicating its possible connection with photorespiration.     

Elevated CO<Subscript>2</Subscript> and temperature differentially affect photosynthesis and resource allocation in flag and penultimate leaves of wheat

Differences in acclimation to elevated growth CO₂ (700 μmol mol⁻¹, EC) and elevated temperature (ambient +4 °C, ET) in successive leaves of wheat were investigated in field chambers. At a common measurement CO₂, EC increased photosynthesis and the quantum yield of electron transport (Φ) early on in the growth of penultimate leaves, and later decreased them. In contrast, EC did not change photosynthesis, and increased Φ at later growth stages in the flag leaf. Contents of chlorophyll (Chl), ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO), and total soluble protein were initially higher and subsequently lower in penultimate than flag leaves. EC decreased RuBPCO protein content relative to soluble protein and Chl contents throughout the development of penultimate leaves. On the other hand, EC initially increased the RuBPCO:Chl and Chl a/b ratios, but later decreased them in flag leaves. In the flag leaves but not in the penultimate leaves, ET initially decreased initial and specific RuBPCO activities at ambient CO₂ (AC) and increased them at EC. Late in leaf growth, ET decreased Chl contents under AC in both kinds of leaves, and had no effect or a positive one under EC. Thus the differences between the two kinds of leaves were due to resource availability, and to EC-increased allocation of resources to photon harvesting in the penultimate leaves, but to increased allocation to carboxylation early on in growth, and to light harvesting subsequently, in the flag leaves.     

Nitrogen redistribution during grain growth in wheat (Triticum aestivum L.)

The flag leaf of wheat was examined for changes in quantity and activity of ribulose-bisphosphate carboxylase (RuBPCase; EC 4.1.1.39), in the proteolytic degradation of RuBPCase and other native proteins, and in the ultrastructure of the leaf cells during grain development. Proteolytic degradation of RuBPCase at pH 4.8 increased until 8–10 d after anthesis, then declined, and increased again 16–18 d after anthesis. The second peak coincided with the onset of a preferential loss of immunologically recognizable RuBPCase. The specific activity and number of active sites per molecule of RuBPCase did not change during senescence. Examination of ultrastructure with the electron microscope showed little change in the appearance of the mitochondria as the flag leaf aged. Prominent cristae were still evident 35 d after anthesis. In contrast, the chloroplasts showed a progressive disruption of the thylakoid structure and an increasing number of osmiophilic glubules. The double membrane envelope surrounding the chloroplast appeared intact until at least 20 d after anthesis. The tonoplast also appeared intact up to 20 d. At later stages of senescence of the leaf the outer membrane of the chloroplast adjacent to the tonoplast appeared to break but the inner membrane of the envelope appeared intact until at least 35 d after anthesis.Abbreviation RuBPCase ribulose-1,5-bisphosphate carboxylase (EC. 4.1.1.39) I=Waters et al. 1980     

Response of photosynthetic apparatus to moderate high temperature in contrasting wheat cultivars at different oxygen concentrations

The photosynthetic responses to moderately high temperatures (38 degrees C, imposed at 21% or 2% O(2) in air and 1500 mumol m(-2) s(-1)) were compared in wheat (Triticum aestivum L.) cultivars grown in northern regions of Ukraine and expected to be relatively sensitive to high temperatures ('North' cultivars) and in cultivars grown in southern regions and expected to be relatively heat-tolerant ('South' cultivars). Heating intact leaves in 21% O(2) for 1 h decreased CO(2) assimilation by c. 63% in 'North' cultivars and only c. 32% in 'South' cultivars, with a decrease in PSII activity being observed in only one of the 'North' cultivars. Carboxylation efficiency was decreased by about 2.7-fold in 'North' cultivars with no significant effect in 'South' cultivars. The maximum rates of carboxylation by Rubisco in vivo, V(cmax), estimated from Farquhar's model, increased more than 2-fold in 'South' cultivars and remained unchanged in 'North' cultivars while the maximum rate of RuBP regeneration, J(max), decreased by 53% and 21% in 'North' and 'South' cultivars, respectively. Where the heat treatment was imposed in 2% O(2) this increased (as compared with treatment at 21% O(2)) the inhibitory effect on CO(2) assimilation in tolerant cultivars, but decreased it in sensitive ones. The results suggested that differences in tolerance of moderately high temperatures in wheat relate to the stability of the Rubisco function and to RuBP regeneration activity rather than to the effects on PSII activity or stomatal control.     

Involvement of abscisic acid and cytokinins in the senescence and remobilization of carbon reserves in wheat subjected to water stress during grain filling

This study investigated the possibility that abscisic acid (ABA) and cytokinins may mediate the effect of water deficit that enhances plant senescence and remobilization of pre‐stored carbon reserves. Two high lodging‐resistant wheat (Triticum aestivum L.) cultivars were field grown and treated with either a normal or high amount of nitrogen at heading. Well‐watered (WW) and water‐stressed (WS) treatments were imposed from 9 d post‐anthesis until maturity. Chlorophyll (Chl) and photosynthetic rate (Pr) of the flag leaves declined faster in WS plants than in WW plants, indicating that the water deficit enhanced senescence. Water stress facilitated the reduction of non‐structural carbohydrate in the stems and promoted the re‐allocation of prefixed ¹⁴C from the stems to grains, shortened the grain filling period and increased the grain filling rate. Water stress substantially increased ABA but reduced zeatin (Z) + zeatin riboside (ZR) concentrations in the stems and leaves. ABA correlated significantly and negatively, whereas Z + ZR correlated positively, with Pr and Chl of the flag leaves. ABA but not Z + ZR, was positively and significantly correlated with remobilization of pre‐stored carbon and grain filling rate. Exogenous ABA reduced Chl in the flag leaves, enhanced the remobilization, and increased grain filling rate. Spraying with kinetin had the opposite effect. The results suggest that both ABA and cytokinins are involved in controlling plant senescence, and an enhanced carbon remobilization and accelerated grain filling rate are attributed to an elevated ABA level in wheat plants when subjected to water stress.     

Endonuclease activities in the coleoptile and the first leaf of developing etiolated wheat seedlings

DNase activity in coleoptiles and the first leaf apices of winter wheat (Triticum aestivum L., cv. Mironovskaya 808) etiolated seedlings was found to increase significantly during seedling growth, peaking on the eighth day of plant development. The maximum of DNase activity was coincident with apoptotic internucleosomal DNA fragmentation in these organs. Wheat endonucleases are capable of hydrolyzing both singleand double-stranded DNA of various origins. The leaf and coleoptiles were found to exhibit nuclease activities that hydrolyzed the lambda phage DNA with N⁶-methyladenine and 5-methylcytosine more actively compared to the hydrolysis of similar unmethylated DNAs. Thus, the endonucleases of wheat seedlings are sensitive to the methylation status of their substrate DNAs. The leaves and coleoptiles exhibited both Ca²⁺/Mg²⁺- and Zn²⁺-dependent nuclease activities that underwent differential changes during development and senescence of seedling organs. EDTA at a concentration of 50 mM fully inhibited the total DNase activity. Electrophoretic heterogeneity was observed for DNase activities operating simultaneously in the coleoptile and the first leaf at different stages of seedling development. Proteins exhibiting DNase activity (16–80 kD mol wt) were revealed in the first leaf and the coleoptile; these proteins were mostly nucleases with the pH optimum around 7.0. Some endonucleases (mol wts of 36, 39, and 28 kD) were present in both organs of the seedling. Some other DNases (mol wts of 16, 56, and about 80 kD) were found in the coleoptile; these DNases hydrolyzed DNA in the nucleus at terminal stages of apoptosis. Different suites of DNase activities were revealed in the nucleus and the cytoplasm, the nuclear DNase activities being more diverse than the cytoplasmic ones. Thus, the cellular (organspecific) and subcellular heterogeneity in composition and activities of DNases has been revealed in wheat plants. These DNases undergo specific changes during seedling development, serving at various stages of programmed cell death in seedling tissues.     

Glutamate dehydrogenase in the first leaf of wheat

Glutamate dehydrogenase extracted from wheat leaves ( Triticum aestivum L. cv. Capitole) taken at two different physiological stages was analysed by electrophoretic and immune-chemical techniques. Two NAD-dependent antigens were identified which bear the balk of the glutamate dehydrogenase activity in the two extracts. The first enzyme was found in much larger amounts in young than in senescent leaves and the reverse situation was observed for the second antigen. The possible relationships between this antigenic polymorphism and the heterogeneity detected by isoelectric focusing from the two extracts were investigated. A charge heterogeneity (isoelectric points about 5.7 and 4.8) was found for the first antigen in both extracts. The second antigen appeared homogeneous (isoelectric point about 5.7) at least in senescent leaves. The last result indicates that two quite different antigens appear in the same isoelectric focusing zone.