Correlation of continuous ryegrass regrowth with cytokinin induced by root nitrate absorption

This study investigated the separate and combined effects of nitrate (NO₃ ^?) and cytokinin additions on continuous ryegrass regrowth after defoliation and the underlying mechanisms. Our results showed that frequent defoliation reduced the biomass of newly grown leaves and roots, the root soluble carbohydrate contents, the root vitality (an indicator of root absorption capacity), and the leaf contents of NO₃ ^?, zeatin and zeatin riboside (Z + ZR), and isopentenyl adenine and isopentenyl adenosine (IP + IPA). NO₃ ^?addition to the roots or leaves increased the biomass of newly grown leaves as well as the leaf contents of NO₃ ^?, Z + ZR, and IP + IPA without increasing the root-to-shoot delivery of endogenous cytokinin. Interestingly, cytokinin directly added to the leaves also increased the biomass of newly grown leaves and their Z + ZR and IP + IPA contents, suggesting that nitrate-induced leaf cytokinin production mediates the growth-promoting effects of nitrate. We also found that cytokinin had a direct whereas NO₃ ^? had an indirect effect on the biomass of newly grown leaves. Taken together, our results indicate that leaf cytokinin production induced by NO₃ ^? absorbed through the roots plays a key role in continuous ryegrass regrowth after defoliation.     

Role of roots in regulating the growth rate and cytokinin content in leaves

The role of roots in the enhancement of cytokinin content and leaf growth of Phaseolus vulgaris plants after decapitation and partial defoliation was investigated. Partial excision of the roots of plants which were decapitated above the primary leaf node resulted in a reduction of leaf growth and soluble proteins accumulation in the primary leaves. Roots excision was done at time of decapitation and repeated 8 days later. Endogenous cytokinins, known to be involved in enhancing shoot growth, accumulated in the leaves and stems of decapitated and partially defoliated plants. Lower levels of cytokinins were detected in the leaves of decapitated plants with only a partial root system. The level of cytokinins in the roots of decapitated plants was reduced by partial root excision. The growth and accumulation of cytokinins in leaves were, however, not totally suppressed by removing a large proportion of the roots. At the commencement of the experiment the stem had a higher cytokinin content than both the leaves and roots. This suggests that the stem could be an alternative source of cytokinins to the leaves. The cytokinin complement in the leaves of decapitated plants is not identical to that in the roots. It appears that cytokinins supplied by the roots are metabolized in the leaves, or that alternatively certain cytokinins are synthesized in the leaves themselves.     

Effect of exogenous application of sugars on the salt tolerance of perennial ryegrass protoplasts

Various sugars were introduced by electroporation into perennial ryegrass protoplasts, and the involvement of intracellular functional groups of the sugars in salt tolerance was investigated. The protoplasts were prepared from the young leaves of perennial ryegrass, and those into which sugars were introduced were treated with NaCl solution (250 mM, pH 7.0) for 6 h at 10°C. The survival rate of the protoplasts increased when xylitol, cellobiose, 1-kestose, maltose, maltotriose, raffinose and trehalose were introduced, while no changes occurred when fructose, fucose, galactose, glucose, inositol, mannitol, mannose, rhamnose, sorbitol, sorbose, fructobiose, lactose and sucrose were introduced. Cellobiose, 1-kestose, maltose, maltotriose, raffinose and trehalose possess a number of equatorial OH (e-OH) groups that promote the structuration of H₂O. Xylitol, however, structures H₂O even though it does not possess the e-OH groups. Hence, it is suggested that under conditions of NaCl stress, structured H₂O protects the structure of cell membranes and the activity of enzymes, and that e-OH groups are involved in enhancing salt tolerance.     

Proteolysis and cell death in clover leaves is induced by grazing

Summary.  Programmed plant cell death is a widespread phenomenon resulting in the formation of xylem vessels, dissected leaf forms, and aerenchyma. We demonstrate here that some characteristics of programmed cell death can also be observed during the cellular response to biotic and abiotic stress when plant tissue is ingested by grazing ruminants. Furthermore, the onset and progression of plant cell death processes may influence the proteolytic rate in the rumen. This is important because rapid proteolysis of plant proteins in ruminants is a major cause of the inefficient conversion of plant to animal protein resulting in the release of environmental N pollutants. Although rumen proteolysis is widely believed to be mediated by proteases from rumen microorganisms, proteolysis and cell death occurred concurrently in clover leaves incubated in vitro under rumenlike conditions (maintained anaerobically at 39 °C) but in the absence of a rumen microbial population. Under rumenlike conditions, both red and white clover cells showed progressive loss of DNA, but this was only associated with fragmentation in white clover. Cell death was indicated by increased ionic leakage and the appearance of terminal deoxynucleotidyl transferase-mediated dUTP-nick-end-labelled nuclei. Foliar protein decreased to 50% of the initial values after 3 h incubation in white clover and after 4 h in red clover, while no decrease was observed in ambient (25 °C, aerobic) incubations. In white clover, decreased foliar protein coincided with an increased number of protease isoforms. Received June 24, 2002; accepted August 15, 2002; published online March 11, 2003     

Protein accumulation in leaves and roots associated with improved drought tolerance in creeping bentgrass expressing an ipt gene for cytokinin synthesis

Cytokinins (CKs) may be involved in the regulation of plant adaptation to drought stress. The objectives of the study were to identify proteomic changes in leaves and roots in relation to improved drought tolerance in transgenic creeping bentgrass (Agrostis stolonifera) containing a senescence-activated promoter (SAG12) and the isopentyl transferase (ipt) transgene that increases endogenous CK content. Leaves of SAG12-ipt bentgrass exhibited less severe senescence under water stress, as demonstrated by maintaining lower electrolyte leakage and lipid peroxidation, and higher photochemical efficiency (F(v)/F(m)), compared with the null transformant (NT) plants. SAG12-ipt plants had higher root/shoot ratios and lower lipid peroxidation in leaves under water stress than the NT plants. The suppression of drought-induced leaf senescence and root dieback in the transgenic plants was associated with the maintenance of greater antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase). The SAG12-ipt and NT plants exhibited differential protein expression patterns under well-watered and drought conditions in both leaves and roots. Under equivalent leaf water deficit (47% relative water content), SAG12-ipt plants maintained higher abundance of proteins involved in (i) energy production within both photosynthesis and respiration [ribulose 1,5-bisphosphate carboxylase (RuBisCO) and glyceraldehyde phosphate dehydrogenase (GAPDH)]; (ii) amino acid synthesis (methionine and glutamine); (iii) protein synthesis and destination [chloroplastic elongation factor (EF-Tu) and protein disulphide isomerases (PDIs)]; and (iv) antioxidant defence system (catalase and peroxidase) than the NT plants. These results suggest that increased endogenous CKs under drought stress may directly or indirectly regulate protein abundance and enzymatic activities involved in the above-mentioned metabolic processes, thereby enhancing plant drought tolerance.     

Glycolipid changes in tissues of wheat leaves induced by exogenous hydrogen peroxide

The results of the study of H2O2 treatment of wheat leaf glycolipid composition are presented. The fraction changes depended upon H2O2 concentration, way of treatment and exposition. The most typical reaction was galactolipid (MGDG, DGDG) content increase while lower sulpholipid variations at low doses of the root treatment and at leaf spraying.     

Effects of exogenous polyamines on nitrate tolerance in cucumber

Putrescine (Put), spermidine (Spd), and spermine (Spm) are the major polyamines (PAs) in plant, which are not only involved in the regulation of plant developmental and physiological processes, but also play key roles in modulating the defense response of plants to diverse environmental stresses. In this study, Cucumis sativus L. seedlings were cultivated in nutrient solution and sprayed with three kinds of PAs (Put, Spd, and Spm). The effects of PAs were investigated on excess nitrate stress tolerance of C. sativus by measuring growth and nitrogen (N) metabolism parameters. The contents of NO_3-^?N, NH_4-⁺N, proline and soluble protein in leaves were increased; while plant height, leaf area, shoot fresh and dry weight, root fresh weight were decreased under 140 mM NO₃^? treatment for 7 d. In addition, the activities of nitrate reductase (NR), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH) were significantly inhibited under 140 mM NO₃^? treatment for 7 d. With foliar treatment by 1 mM Spd or Spm under stress treatment, the contents of Spm, Put, and Spd in leaves increased significantly, except that Spm content decreased under Spd treatment. The activities of NR, glutamine synthetase (GS), GOGAT and GDH and plant height, leaf area, shoot fresh and dry weights were significantly increased. The contents of proline and soluble protein in leaves were significantly enhanced. In contrast, the accumulation of NO_3-^?N and NH_4-⁺N were significantly decreased. However, there were minor differences in activities of N metabolism enzymes and the content of osmotic adjustment substances under 1 mM Put treatment. These findings suggest that 1 mM exogenous Spm or Spd could enhance the capacity of N metabolism, promote growth and increase resistance to high concentrations of NO₃^?. The ameliorating effect of Spd was the best, and that of Put the worst.     

A stronger coordination of litter decomposability between leaves and fine roots for woody species in a warmer region

Key message

There is a positive correlation between leaf and root decomposition across species, both in a warm-temperate forest in Japan, as well as globally.

Abstract

Evaluating the effects of plant species traits on litter decomposition would increase our understanding of plant–soil feedbacks in forest ecosystems. Currently, an assessment of a possible coordination between leaf and root decomposition across different species is required. However, previous studies have generated conflicting results. We hypothesized that such inconsistencies may be attributed to differences in local climatic effects on the decomposition process. We focused on the linkages between leaf and fine-root decomposition of woody species in a warm-temperate forest, which have not been addressed in previous studies. We found a significant positive correlation between leaf and root decomposition, and this linkage may be attributed to a wider range of decomposition rates across the species in our study forest. Additionally, we combined our data with those of previous studies of woody species to infer a global linkage in the decomposition process between leaves and roots. We found a positive correlation in decomposition rates between leaves and roots at the global scale, as well as a relatively strong correlation in warmer regions. These results support the importance of litter quality on biogeochemical processes and suggest that synergetic interactions between climate and plant communities could be amplified in a warmer future.

     

Effects of exogenous proline and glycinebetaine on the salt tolerance of rice cultivars

Salinity significantly increased trisodium-8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS) uptake and decreased the K(+)/Na(+) ratio in salt-sensitive rice (Nipponbare) but did not markedly in salt-tolerant rice (Pokkali). Proline and glycinebetaine (betaine) suppressed the increase in PTS uptake and the decrease in the K(+)/Na(+) ratio in Nipponbare, but did not affect PTS uptake or the K(+)/Na(+) ratio in Pokkali.     

Effects of proline and carbohydrates on the metabolism of exogenous proline by excised bean leaves in the dark

Proline was metabolized when vacuum infiltrated into starved bean (Phaseolus vulgaris L.) leaves from plants previously in the dark for 48 hours, but an equivalent increase in protein proline was not observed. When ¹⁴C-proline was infiltrated into starved leaves, a large percentage of the ¹⁴C was recovered in other amino acids, organic acids, and CO₂, in addition to that recovered as protein proline. However, extensive oxidation of proline was observed only if enough proline was added to increase substantially the endogenous concentration of proline. Increasing the endogenous concentration did not affect the amount of proline that was incorporated into protein.     

Effects of leaf zeatin and zeatin riboside induced by different clipping heights on the regrowth capacity of ryegrass

The effect of clipping height on ryegrass regrowth was investigated by examining the roles of several plant hormones. Our study consisted of three treatment conditions: (1) darkness over whole plants, (2) darkness only over stubble leaf sheaths, and (3) light over whole plants. Results showed that under darkness over whole plant, low stubble height resulted in low leaf regrowth biomass. Similar leaf regrowth biomass was observed under conditions of darkness only over stubble leaf sheaths as well as light over whole plants. Each unit weight of stubble at different clipping heights has relatively similar potential of providing stored organic substance for leaf regrowth. Therefore, regrowth index, calculated as newly grown leaf biomass divided by unit stubble weight, was used to evaluate regrowth capacity at different clipping heights under minimal influence of organic substances stored in stubbles. Under light over whole plants and single clipping, low stubble height and high stubble height with root thinning resulted in low leaf biomass and high regrowth index. On the other hand, under light over whole plants and frequent clipping high leaf biomass and regrowth index were observed in high stubble height. In addition, we found that leaf zeatin and zeatin riboside (Z + ZR) affected ryegrass regrowth and that roots regulated leaf Z + ZR concentration. Thus, our results indicate that root-derived cytokinin concentration in leaves influences ryegrass regrowth at different clipping heights.     

Effects of the exogenous application of auxin and cytokinin on carbohydrate accumulation in grains of rice under salt stress

Phytohormones, such as auxin and cytokinin, are known to be involved in the regulation of plant responses to salinity stress and counteract the adverse effect of stress conditions. This work investigated the effects of the exogenous spraying of indole-3-acetic acid (IAA) and kinetin (KIN) during the reproductive phase on grain yield by examining the 1000-grain weight and filled-grain percentage as well as the changes in starch, total soluble sugars, sucrose, glucose and fructose concentrations in the grains of two rice cultivars under salt stress. The results indicated that the applied IAA and KIN led to an increased grain yield, 1000-grain weight and filled-grain percentage for both rice cultivars under salt stress. The storage starch content in the grain of the salt-sensitive cultivar was more than that in the salt-tolerant cultivar under IAA application compared with KIN, whereas a decrease in the total soluble sugar content was observed with both IAA and KIN treatments, in comparison to the non-hormone treatment. Interestingly, this study showed that IAA led to a much higher increase in the sucrose content in grain, as compared to the KIN. Furthermore, this experiment suggests that glucose and fructose may play important roles during salt stress because there were clearly higher concentrations of these sugars in the grain of the stressed cultivars under IAA and KIN application: it appears that their accumulation was the earliest response detected during the grain-filling period in rice. Finally, this work indicated that an increase in the rice grain yield, 1000-grain weight and filled-grain percentage are associated with an increase in the contents of starch, sucrose, glucose and fructose in grain caused by the application of IAA and KIN.     

Aluminum inhibition of shoot lateral branches ofGlycine max and reversal by exogenous cytokinin

Aluminum effects on the morphological development of soybean (Glycine max (L.) Merr.) were characterized in greenhouse and growth chamber experiments. An Al-sensitive cultivar, ‘Ransom’, was grown in an acid soil (Aeric Paleudult) adjusted to 3 levels of exchangeable Al. Lateral shoot development at the nodes of the main stem was extensive in the limed soil containing 0.06 cmol(+) Alkg⁻¹. However, lateral shoot length and weight were severely inhibited in the unlimed soil containing 2.19 cmol(+) Alkg⁻¹, and in the unlimed soil amended to 2.63 cmol(+) Alkg⁻¹ with AlCl₃. This inhibition by the high Al/low pH condition was reversed by the exogenous application of a synthetic cytokinin 6-benzylaminopurine (BA). The daily application of 20 μg mL⁻¹ BA applied locally to the lateral meristems of plants grown in the unlimed soil stimulated lateral shoot growth substantially, such that it was either comparable to or greater than that observed in the limed treatment without BA. Accumulation of K, Ca, and Mg in lateral shoot branches was also stimulated by the local application of BA. The inhibitory effects of Al on lateral shoot development were confirmed in solution culture. In addition, differential sensitivity to Al was evident among the primary root, first order lateral roots, and second order lateral roots. The length of the primary root was only slightly decreased by increasing concentrations of Al up to 30 μM. In contrast, the length of basipetally located first order lateral roots was restricted to greater extent; up to 50% by 30 μM Al. Second order lateral lengths were inhibited even more severely; up to 86% by 30 μM Al. Substantial evidence in the literature indicates that the root apex is a major site for the biosynthesis of cytokinin that is supplied to shoots, and cellular function and development in this region of the root are impaired during Al toxic conditions. This suggests that one mode of action by which Al may affect shoot growth is by inhibiting the synthesis and subsequent translocation of cytokinin to the meristematic regions of the shoot. The present observation of a reversal of Al-inhibited lateral shoot development by exogenously applied cytokinin supports this hypothesis. However, the inability of applied cytokinin to counter the restriction imposed by Al on total shoot dry matter production implies the impairment by Al toxicity of other root functions, such as ion and water transport, also played an important role in altering shoot morphology.     

Effects of exogenous nitric oxide on cadmium toxicity, element contents and antioxidative system in perennial ryegrass

The effects of sodium nitroprusside (SNP, a donor of NO) on cadmium (Cd) toxicity in ryegrass seedlings (Lolium perenne L.) were studied by investigating the symptoms, plant growth, chlorophyll content, lipid peroxidation, H⁺-ATPase enzyme and antioxidative enzymes. Addition of 100???M CdCl₂ caused serious chlorosis and inhibited the growth of ryegrass seedlings, and dramatically increased accumulation of Cd in both shoots and roots, furthermore, the absorption of macro and micronutrients were inhibited. Addition of 50, 100, 200???M SNP significantly decreased the transport of Cd from roots to shoots, alleviated the inhibition of K, Ca, Mg and Fe, Cu, Zn absorption induced by Cd, reduced the toxicity symptoms and promoted the plant growth. The accumulation of reactive oxygen species (ROS) significantly increased in ryegrass seedlings exposed to Cd, and resulted in the lipid peroxidation, which was indicated by accumulated concentration of thiobarbituric acid-reactive substances. Addition of 50, 100, 200???M SNP significantly decreased the level of ROS and lipid peroxidation. Activities of antioxidant enzymes also showed the same changes. Addition of 50, 100, 200???M SNP increased activities of superoxide dismutase, peroxidase, catalase and ascorbate peroxidase in ryegrass seedlings exposed to Cd. Addition of 100???M SNP had the most significant alleviating effect against Cd toxicity while the addition of 400???M SNP had no significant effect with Cd treatment.     

Effects of salicylic acid and cold on freezing tolerance in winter wheat leaves

The effects of salicylic acid (SA) (0.01, 0.1 and 1 mM) and cold on freezing tolerance (freezing injury and ice nucleation activity) were investigated in winter wheat (Triticum aestivum cv. Dogu-88) grown under control (20/18 °C for 15, 30 and 45-day) and cold (15/10 °C for 15-day, 10/5 °C for 30-day and 5/3 °C for 45-day) conditions. Cold acclimatisation caused a decrease of injury to leaf segments removed from the plants and subjected to freezing conditions. Exogenous SA also decreased freezing injury in the leaves grown under cold (15/10 °C) and control (15 and 30-day) conditions. Cold conditions (10/5 and 5/3 °C) caused an increase in ice nucleation activity by apoplastic proteins, which were isolated from the leaves. For the first time, it was shown that exogenous SA caused an increase in ice nucleation activity under cold (15/10 and 10/5 °C) and control conditions. These results show that salicylic acid can increase freezing tolerance in winter wheat leaves by affecting apoplastic proteins.     

Competition for ammonium between plant roots and nitrifying and heterotrophic bacteria and the effects of protozoan grazing

The competition for limiting amounts of ammonium between the chemolithotrophic ammonium-oxidizing species Nitrosomonas europaea, the heterotrophic species Arthrobacter globiformis and roots of Plantago lanceolata (Ribwort plantain) was studied in a series of model systems of increasing complexity, i.e. energy-limited continuous cultures, non-water-saturated continuously percolated soil columns and pots with γ-sterilized soil planted with axetic P. lanceolata seedlings. The effects of bacterial grazing by the flagellate species Adriamonas peritocrescens on the competition for ammonium were also investigated in the three model systems. It was found that N. europaea was a weaker competitor for ammonium than either A. globiformis or plant roots of P. lanceolata. It is assumed that the heterotrophic bacteria have a higher affinity for ammonium than the nitrifying bacteria, whereas growing plant roots have a greater capacity to exploit the soil for ammonium than the immobile nitrifying bacteria. It is not very likely that allelochemicals were involved in suppressing the nitrification process. Four reasons are given for this assumption. Presence of the flagellates strongly stimulated the potential nitrification rate in all model systems. It is assumed that there is a more even distribution over the soil of either nitrifying bacteria or their substrate ammonium in the presence of flagellates. In addition to the distribution effect, there is a stimulation of the potential ammonium oxidation rate. The results are discussed in the light of the function of nitrate as nitrogen sink in the biogeochemical nitrogen cycle.     

Isolation and purification of cytokinin binding protein from tobacco leaves by affinity column chromatography

Cytokinin binding protein from tobacco leaves was isolated and purified to a single protein by means of affinity chromatography on benzyladenine-linked Sepharose column combined with polyacrylamide gel electrophoresis. In vitro binding of this protein to [¹⁴C] benzyladenine was inhibited remarkably by cold benzyladenine and kinetin and slightly by adenine, but not adenosine. The molecular weight of the protein was determined to be about 4,000 daltons by gel filtration and SDS polyacrylamide gel electrophoresis.     

Differential accumulation of proteins in leaves and roots associated with heat tolerance in two Kentucky bluegrass genotypes differing in heat tolerance

To understand the mechanisms of heat stress responses in perennial grasses, differential proteins in leaves and roots of two genotypes of Kentucky bluegrass (Poa pratensis), including heat-tolerant ‘Midnight’ and heat-sensitive ‘Brilliant’, were analyzed with two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Plants were exposed to heat stress for 28 days in growth chambers. Under 7–28 days of heat stress, leaf photochemical efficiency declined significantly while electrolyte leakage increased in leaves and roots, and to a lesser extent for heat-tolerant ‘Midnight’ than for heat-sensitive ‘Brilliant’. Compared with leaves, cell membrane damage due to heat stress was more severe in roots. The 2-DE and MS analysis identified 37 heat-responsive proteins in leaves, 28 heat-responsive proteins in roots; 14 proteins in leaves and 9 proteins in roots exhibited differential expression between the two genotypes. The results indicate that proteins involved in metabolism and energy in leaves and those in antioxidant defense in roots are associated with heat tolerance in Kentucky bluegrass. The differential accumulation of these proteins might be the reason for different heat tolerance in two Kentucky bluegrass genotypes in aerial and underground parts.     

水溶性有机物对黑麦草吸收铜的影响

采用水培试验,研究蚓粪及蚯蚓培养载体牛粪中水溶性有机物(DOM)对不同Cu2+浓度下(0、5、10 mg·L-1)黑麦草吸收Cu2+的影响.结果表明:随着Cu2+浓度的增加,黑麦草地上部、根干质量,以及根系的长度、表面积、体积和根尖数均逐渐下降;DOM显著增加了Cu2+处理下黑麦草地上部及根系生物量,促进了其根系的长度、表面积、体积和根尖数的增长.DOM降低了黑麦草地下部Cu2+浓度,促进了Cu2+从地下部向地上部的运输,显著增加了地上部Cu2+积累量.蚓粪DOM对黑麦草的影响优于牛粪DOM,并且供试高浓度DOM效果优 于低浓度.