Ammonium and amino acid metabolism in excised leaves of wheat (Triticum aestivum) senescing in the dark

Several parameters of amino acid metabolism were studied in detached primary leaves of wheat (Triticum aestivum L. cv. Castell) during a 14 day incubation period in the dark. Protein loss was accompanied by a 5-fold increase in the total amount of free amino acids during the first 4 days of the incubation period with asparagine being the most important. Beyond this stage a pronounced intracellular accumulation of ammonium occured. A gradual decrease in the levels of free amino acids and ammonium at the later stages of senescence could in part be accounted for by leakage from the leaves. Additionally, some nitrogen was lost due to ammonia volatilization. The rapid decay of the glutamine synthetase (GS; EC 6.3.1.2)-glutamate synthase (Fd-GOGAT; EC 1.4.7.1) system and the fast decline of glutamate-pyruvate transaminase (GPT; EC 2.6.1.2) activity appear to be predominant features of senescence in the dark. Decreasing Fd-GOGAT activity was slightly compensated by a small and temporary increase in the activity of NADH-GOGAT (EC 1.4.1.14). Glutamateoxalocetate transaminase (GOT: EC 2.6.1.1) activity, although declining continuously, proved to be much more persistent. Changes in glutamate dehydrogenase (GDH; EC 1.4.1.3) activity closely resembled the profile of ammonium evolution in the leaves and NADP-isocitrate dehydrogenase (IDH; EC 1.1.1.42) activity revealed a temporary maximum during the period of rapid increase in GDH activity. Increased activity of GDH could also be induced by exogenous ammonium. Ammonium accumulation could, at least partly, be caused by increased asparaginase (EC 3.5.1.1) activity which accompanied the rapid conversion of asparagine to aspartic acid. Asparagine aminotransferase (EC 2.6.1.14) activity declined sharply from the beginning of the senescence period. Although the activity profile of glutaminase (EC 3.5.1.2) was similar to that of asparaginase, glutamine was of little importance quantitatively and an analogous relationship between glutamine and glutamic acid could not be detected.     

Sucrose metabolism during endosperm development in wheat (Triticum aestivum)

This work reports changes in sucrose synthase and invertase activities throughout endosperm development in wheat, together with the associated substrates and metabolites, sucrose, UDP, glucose, fructose and UDP-glucose. Throughout endosperm development, sucrose synthase had consistently higher activity than invertase and indeed invertase activity did not change appreciably. The observed variation in pattern and amounts of glucose and fructose present during the mid- and late stages of endosperm development confirmed the suggestion that invertase was not the preferred pathway of sucrose catabolism. Kinetic parameters for sucrose synthase were determined in crude extracts. Estimates of UDP and sucrose concentrations suggest that sucrose synthase is unlikely to achieve its potential maximum velocity. This limitation may however be overcome in part by the apparent excess catalytic activity measured during endosperm development.     

Gibberellic acid (GA) increases fibre cell differentiation and secondary cell-wall deposition in spring wheat (Triticum aestivum L.) culms

The role of gibberellic acid (GA) in differentiation and secondary cell-wall deposition of fibre cells of spring wheat (Triticum aestivum) culms was studied using applications of GA and chlormequat (a GA biosynthesis inhibitor). In certain genotypes, higher GA levels may increase the number of cortical fibre cell files by changing cell fate from parenchyma to fibre, and induce thicker secondary cell-walls.     

The regulation of nitrogen accumulation in the grain of wheat plants (Triticum aestivum)

Nitrogen accumulation in the ear of wheat plants ( Triticum aestivum L. cv. Klein Chamaco) during ear growth was studied under 4 experimental conditions. Plants were grown in pots with Perlite or soil, and fertilized with nutrient solutions. In one experiment the plants were grown in a greenhouse and supplied with high (16m M ) or low (1.6 m M ) N in the nutrient solutions until anthesis, and then with or without nitrogen supply until ripening. In a second experiment the plants were grown with high N supply until anthesis, and then for half of the plants light intensity was decreased by 50%, and at the same time. N supply was terminated for half of the plants within each light treatment. A third experiment was similar to the previous one, but was carried out in a growth cabinet under 20% of the maximal irradiance in the greenhouse. In a fourth experiment half the ear was excised at anthesis in half of the plants, and these plants were then supplied with or without nitrogen.
In all experiments there was a linear relation between the rate of N accumulation and the rate of ear growth. A wide range of final individual grain weights and N concentration was observed among the experiments. The same maximum N concentration was observed for all grain sizes, although the N concentration could be different between grains of the same size. The grain N concentration correlated with the rate of N accumulation per unit of ear weight increase during ear growth. It is suggested that in wheat plants there is a dependence of nitrogen transport on carbon transport to the ear, and to the ear, and that the final grain N concentration is determined by the N/C ratio exported from the vegetative tissues.     

Diversity of root-associated bacteria associated with field-grown canola (Brassica napus L.) and wheat (Triticum aestivum L.)

Little is known about the composition and diversity of the bacterial community associated with plant roots. The purpose of this study was to investigate the diversity of bacteria associated with the roots of canola plants grown at three field locations in Saskatchewan, Canada. Over 300 rhizoplane and 220 endophytic bacteria were randomly selected from agar-solidified trypticase soy broth, and identified using fatty acid methyl ester (FAME) profiles. Based on FAME profiles, 18 bacterial genera were identified with a similarity index >0.3, but 73% of the identified isolates belonged to four genera: Bacillus (29%), Flavobacterium (12%), Micrococcus (20%) and Rathayibacter (12%). The endophytic community had a lower Shannon-Weaver diversity index (1.35) compared to the rhizoplane (2.15), and a higher proportion of Bacillus, Flavobacterium, Micrococcus and Rathayibacter genera compared to rhizoplane populations. Genera identified in the endophytic isolates were also found in the rhizoplane isolates. Furthermore, principal component analysis indicated three clusters of bacteria regardless of their site of origin, i.e., rhizoplane or endophytic. In addition, the rhizoplane communities of canola and wheat grown at the same site differed significantly. These results indicate that diverse groups of bacteria are associated with field-grown plants and that endophytes are a subset of the rhizoplane community.     

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     

In vitro development of wheat (Triticum aestivum L.) from zygote to plant via ovule culture

Ovules of the wheat breeding line Veery #5 were excised and transferred to culture within 24 h after pollination. When ovules were cultured on Phytagel-solidified medium, and the pericarp removed exclusively at the micropylar tip and the abaxial side, zygotes from up to 79.2% of the ovules underwent embryogenesis with the same developmental pattern as found in planta. Embryos from more than 50% of the cultured ovules germinated when transferred to regeneration medium. More than 100 plantlets were randomly chosen for transfer to soil, all of which developed to phenotypically normal and fertile plants. With this system, the entire process of zygotic embryogenesis can be studied using living material. Furthermore, the method could be used as an embryo rescue technique for plant breeding purposes. Received: 17 June 1996 / Revision received: 22 October 1996 / Accepted: 15 December 1996     

Enzyme activities associated with developing wheat(Triticum aestivum L.) grain amyloplasts

Intact amyloplasts from endosperm of developing wheat grains have been isolated by first preparing the protoplasts and then fractionating the lysate of the protoplasts on percoll and ficoll gradients, respectively. Amyloplasts isolated as above were functional and not contaminated by cytosol or by organelles likely to be involved in carbohydrate metabolism. The enzyme distribution studies indicated that ADP-glucose pyrophosphorylase and starch synthase were confined to amyloplasts, whereas invertase, sucrose synthase, UDP-glucose pyrophosphorylase, hexokinase, phosphofructokinase-2 and fructose-2,6-P₂ase were absent fro the amyloplast and mainly confined to the cytosol. Triose-P isomerase, glyceraldehyde-3-P dehydrogenase, phosphohexose isomerase, phosphoglucomutase, phosphofructokinase, aldolase, PPi-fructose-6-P-1 phosphotransferase, and fructose-l,6-P₂ase, though predominantly cytosolic, were also present in the amyloplast. Based on distribution of enzymes, a probable pathway for starch biosynthesis in amyloplasts of developing wheat grains has been proposed.     

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

The technique of EDTA-enhanced phloem exudation (King and Zeevaart, 1974: Plant Physiol. 53, 96–103) was evaluated with respect to the collection and identification of amino acids exported from senescing wheat leaves. Whilst the characteristics of the exudate collected conform with many of the accepted properties of phloem exudate, unexpectedly high molar proportions of phenylalanine and tyrosine were observed. By comparing exudation into a range chelator solutions with exudation into water, the increased exudation of phenylalanine and tyrosine relative to the other amino acids occurring when ethylene-diaminetetracetic acid was used, was considered to an artefact.In plants thought to be relying heavily on mobilisation of protein reserves to satisfy the nitrogen requirements of the grain, the major amino acids present in flag-leaf phloem exudate were glutamate, aspartate, serine, alanine and glycine. Only small proportions of amides were present until late in senescence when glutamine became the major amino acid in phloem exudate (25 molar-%). Glutamine was always the major amino acid in xylem sap (50 molar-%).The activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 1.4.7.1), glutamate dehydrogenase (EC 1.4.1.3) and asparagine synthetase (EC 5.3.5.4) were measured in the flag leaf throughout the grain-filling period. Glutamine synthetase and glutamate-synthase activities declined during this period. Glutamate-dehydrogenase activity was markedly unchanged despite variation in the number of multiple forms visualised after gel electrophoresis. The activity of the enzyme reached a peak only very late in the course of senescence of the flag leaf. No asparagine-synthetase activity could be detected in the flag leaf during the grain-filling period.II. Peoples et al. (1980)     

Effect of source-sink relations and nitrogen nutrition on senescence and N remobilization in the flag leaf of wheat

The relation between the source-sink ratio and nitrogen nutrition on grain yield of wheat ( Triticum aestivum L. cv. Klein Chamaco) was studied in a greenhouse experiment. Plants were grown until anthesis in pots with soil fertilized with 0.16 mmol N per plant twice a week. At anthesis, all leaves but the flag leaf were excised in a group of plants. In another group the treatment consisted in a similar defoliation plus the longitudinal excision of half the ear, while a third group was left untouched as a control. At the same time, the N supply to half of the plants in each group was interrupted, while the other half continued receiving 16 m M N. The defoliated plants showed a longer functional life of the flag leaf than the control, retaining the chlorophyll, soluble proteins and total reduced nitrogen for a longer time. The ear-excised plants showed an intermediate behavior. The plants with the interrupted N supply showed a faster leaf senescence than the N supplied ones, and this coincided with an increase in the proteolytic activity and nitrogen transport to the ear. However there were no differences in ear weight between the two nitrogen treatments. It is concluded that leaves and ear compete for the nitrogen, and that a low level of carbohydrates in the flag leaf, due to a low source-sink ratio, delays leaf senescence.     

Salicylic acid changes the properties of extracellular peroxidase activity secreted from wounded wheat (Triticum aestivum L.) roots

Summary.  Wheat (Triticum aestivum L.) roots released proteins showing peroxidase activity in the apoplastic solution in response to wound stress. Preincubation of excised roots with 1 mM salicylic acid at pH 7.0 enhanced the guaiacol peroxidase activity of the extracellular solution (so-called extracellular peroxidase). The soluble enzymes were partially purified by precipitation with ammonium sulfate followed by size exclusion and ion exchange chromatography. Despite an increase in the total activity of secreted peroxidase induced by pretreatment of excised roots with salicylic acid, the specific activity of the partially purified protein was significantly lower compared to that of the control. Purification of the corresponding proteins by ion exchange chromatography indicates that several isoforms of peroxidase occurred in both control and salicylic acid-treated samples. The activities of the extracellular peroxidases secreted by the salicylic acid-treated roots responded differently to calcium and lectins compared with those from untreated roots. Taken together, our data suggest that salicylic acid changes the isoforms of peroxidase secreted by wounded wheat roots. Received June 10, 2002; accepted September 24, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Institute of Biochemistry and Biophysics, Russian Academy of Sciences, P.O. Box 30, Kazan 420111, Russia.     

Accelerated production of transgenic wheat (Triticum aestivum L.) plants

We have developed a method for the accelerated production of fertile transgenic wheat (Triticum aestivum L.) that yields rooted plants ready for transfer to soil in 8–9 weeks (56–66 days) after the initiation of cultures. This was made possible by improvements in the procedures used for culture, bombardment, and selection. Cultured immature embryos were given a 4–6 h pre-and 16 h post-bombardment osmotic treatment. The most consistent and satisfactory results were obtained with 30 g of gold particles/bombardment. No clear correlation was found between the frequencies of transient expression and stable transformation. The highest rates of regeneration and transformation were obtained when callus formation after bombardment was limited to two weeks in the dark, with or without selection, followed by selection during regeneration under light. Selection with bialaphos, and not phosphinothricin, yielded more vigorously growing transformed plantlets. The elongation of dark green plantlets in the presence of 4–5 mg/l bialaphos was found to be reliable for identifying transformed plants. Eighty independent transgenic wheat lines were produced in this study. Under optimum conditions, 32 transformed wheat plants were obtained from 2100 immature embryos in 56–66 days, making it possible to obtain R3 homozygous plants in less than a year.     

Contribution of the wheat (Triticum aestivum L.) flag leaf to grain yield in response to plant growth regulators

In West-Europe, intensive cereal management uses plant growth regulators (PGRs) especially for wheat. A green-house experiment compared the effects of two PGRs on flag leaf characteristics and yield of winter wheat. Chlormequat chloride + choline chloride (CCC) and chlormequat chloride + choline chloride + imazaquin (CCC+I) were applied to winter wheat at growth stage 5 (Feekes Large scale). CCC and CCC+I significantly increased flag leaf surface area at anthesis. Both treatments also enhanced chlorophyll content of the main stem flag leaf. The grain filling period was extended with PGR application by 2 days. CCC and CCC+I significantly increased net CO₂ assimilation rates during the flag leaf life. No effects of PGR spraying were observed on the pattern of ¹⁴C labelled assimilate distribution. Increased grain yield was due to the increase in average grain weight. The results indicate that PGR treatments increased flag leaf contribution to grain filling. The addition of imazaquin (I) to chlormequat (CCC) improved the effects of CCC.     

Effects of foliar applied benzyladenine on grain yield and grain protein in wheat (Triticum aestivum L.)

The effects of foliar applications of nitrogen and benzyladenine (BA) on grain yield and grain protein of wheat grown under field conditions were studied over 2 years with 5 cultivars at 2 locations. Nitrogen (N) at 20 kg.ha^–1, and BA at 100 or 800 mg.l^–1 were applied alone or combined at pre and post-anthesis; applications of BA at 8 mg.l^–1 were also made on individual ears in order to study the effect on cell number. Weekly determinations of the chlorophyll content of the flag leaf were conducted after anthesis to study leaf senescence. At harvest, yield, yield components and grain protein percentage were determined. N and BA applications delayed chlorophyll loss in the flag leaf, but modified neither yield nor yield components. Foliarly applied BA increased grain protein in four of the five cultivars tested. It is concluded that delay of the senescence induced by BA might allow more energy to be available for N uptake by the crop leading to an increase in grain protein.Research supported by a CAFPTA grant 1656/86 and by CONICET, PID 30017700/85.CONICETComisión de Investigaciones Cientificas de la Provincia de Buenos AiresInstituto de Fisiologia Vegetal     

条锈病对小麦(Triticum aestivum L.)叶片光合功能及光合功能蛋白D1表达的影响

测定了小麦(Triticum aestivum L.)感染小麦条锈病后的光合常数,以及叶绿素含量、类囊体膜光合电子传递速率和光合反应中心D1蛋白的变化.实验显示,条锈病侵染导致感病小麦叶片净光合速率与叶绿素含量降低;抗病小麦经侵染后净光合速率却有恢复过程,叶绿素含量先降后升.此外,感病小麦叶片被侵染后全链电子传递速率受到抑制,PSII电子传递速率的变化与全链电子传递速率的变化趋势相似,但PSI电子传递速率受到的影响较小;抗病小麦小麦叶片被侵染后电子传递速率所受影响较小.同时发现,病程中,感病和抗病小麦PSII的光合反应中心D1蛋白含量变化总是与PSII电子传递速率的变化类似,推测D1蛋白的表达量变化是引起PSII电子传递活性与全链电子传递速率变化的主要因素之一.     

基于全基因组的河北省小麦品种遗传多样性分析

丰富的遗传变异对于提高作物的环境适应性和遗传改良进度至关重要。小麦是重要的粮食作物,河北省作为我国第三大小麦产区,在保障国家粮食安全中占有重要地位。建国以来,河北省审定了大量小麦品种,然而有关其遗传基础的研究却相对缺乏。本研究以1949年至2012年的169份河北省小麦品种为材料,利用覆盖小麦全基因组的SSR标记分析了供试小麦品种的遗传多样性。结果表明,供试河北省小麦品种的3个染色体组中,以B染色体组具有最高的遗传多样性,A组最低;7个同源群中,以第5、4群具有最高的多样性水平,而第7群最低;在21条染色体上的遗传多样性变化较大,以4A、2D具有较高的多样性水平,1A染色体最低。从品种的更新换代角度看,自1949年以来,尽管品种的等位基因频率在下降,河北省小麦品种的多样性水平基本呈现上升趋势。在此基础上,根据遗传相似系数,供试品种可聚为5大类,聚类结果既反映出河北省小麦品种多样性水平的复杂多样,也反映出品种的地域性分布特征。     

PFCA uptake and translocation in dominant wheat species (Triticum aestivum L.)

The comparative uptake of four perfluorinated carboxylic acids (PFCAs) by wheat (Triticum aestivum L.) grown in nutrient solution was investigated. Wheat is the main food crop in northern China and may become a potential pathway of human exposure to these PFCAs. The uptake of four PFCAs from water at a fixed concentration (1 μg/mL) increased over time, approaching a steady state, and except for the short-chain perfluorobutanoic acid, most of the total mass of each of the PFCAs taken up by wheat was found to be at the root. The root concentration factor (RCF) and shoot/root concentration factor (SRCF) were calculated, and with the increase in carbon chain length, the RCFs increased but SRCFs decreased, which indicated that long-chain PFCAs had stronger root uptake and weaker translocation capacities than short-chain PFCAs. In addition, pH could obviously impact the uptake of four PFCAs in the roots and shoots of wheat, and the highest concentrations were found at pH = 7 when the pH increased from 4 to 10.     

低磷和干旱胁迫对小麦生长发育影响的研究初探

研究了低磷和干旱胁迫对小麦（Triticum aestivum L.）生长发育的影响。结果表明,低磷胁迫能显著降低小麦的分蘖数、叶片相对含水量和叶绿素含量,进而抑制小麦的生长发育,降低其生物产量和经济产量,不耐低磷品种中国春受影响的程度要大于耐低磷品种烟中144。在相同条件下,干旱能够强化磷胁迫效应,表现出明显的胁迫叠加现象。