The Effect of Salicylic Acid on Peroxidase Activity in Wheat Calli Cocultured with the Bunt Pathogen Tilletia caries

The effect of salicylic acid (SA) on peroxidase activity in wheat (Triticum aestivum L.) calli cocultured with the bunt pathogen Tilletia caries was studied. Fungal infection was shown to activate cytoplasmic peroxidase. SA suppressed total peroxidase activity but did not inhibit the peroxidase with pI 9.8. A novel chitin-specific peroxidase with pI 3.5 appeared after the SA treatment. The infection of SA-treated cells with Tilletia caries activated the isoenzymes with pI 3.5, 4.8, and 7.5 and stimulated their secretion into the culture medium. The ability of SA to control wheat peroxidase activity during pathogenesis is discussed. The important role of this control in plant defense responses to the bunt pathogen is emphasized.     

Changes in the Levels of IAA,ABA, and Cytokinins in Wheat Seedlings Infected with Tilletia caries

The seedling growth and the content of endogenous phytohormones in wheat seedlings were estimated 3, 6, and 9 days after infection with the bunt pathogen (Tilletia caries) (DC.)TUL. The infection of a pathogen-susceptible species Triticum aestivum L. and a resistant species T. timopheevii Zhuk. resulted, respectively, in an increase and a decrease in the seedling growth and the IAA content as compared to the control. The cytokinin content increased in both species, and the increase in T. timopheevii was more rapid. The pathogen-induced increase in auxin content is suggested to enhance fungal invasion of plants. In the susceptible species, a high ABA concentration was retained for a longer period of time and could act as a factor of virulence. At the same time, in the resistant species, an increase in ABA content was transient and seems to trigger plant defense mechanisms.     

Effect of Increasing Concentrations of Heavy Metals on the Growth of Barley and Wheat Seedlings

The tolerance to increasing doses of lead and cadmium salts on the growth and survival of barley (Hordeum vulgare L.) and wheat (Triticum aestivum L.) seedlings were studied. Seedlings grown under controlled conditions were treated with dilute (0.001–0.005 mM) solutions of either lead nitrate or cadmium bromide for 1, 4, or 7 days. Subsequently, they were incubated for 7 days in solutions of the same compounds, but at sublethal or lethal concentrations (0.05–10 mM). Plant pretreatment with low concentrations of heavy metals induced an increase in their tolerance to the metals, because pretreated plants could tolerate heavy metals at high concentrations. It is concluded that plant tolerance to increasing concentrations of heavy metals is related to the activation of protective and adaptive processes in their tissues.     

Statistical Parameters Reflecting Morphogenetic Capacity of Soft Spring Wheat Calluses

Callus cultures of soft spring wheat were subcultured without separation into explants to follow the line one excised embryo–one callus. This approach revealed the following statistical correlations. Within every cultivar of Triticum aestivum L. and within a row of cultivars arranged in ascending order according to the frequency of embryogenic callus formation, positive correlations (at P = 95) were found between the proliferative activity of callus cells and the frequency of embryogenic callus formation. A reliable intraspecies correlation (significant at P = 95) between multiple regenerations of plants from calluses and the tillering trait (bushiness) of donor plants was also found. We assessed the importance of various statistical parameters of callus cultures for preliminary estimation of morphogenesis efficiency at early stages of culturing. Frequencies of callusogenesis and the growth curves for randomly selected calluses turned out to be noninformative characteristics, unless the morphogenetic activity of calluses was taken into account. The following statistical parameters were found to correlate with the morphogenetic capacity of wheat calluses: gradually increasing coefficients of variation in fresh weight of primary calluses, a larger callus size, and higher fresh weight gain in potentially morphogenetic calluses.     

The Effect of Rapid Temperature Increase on the Growth Rate of Wheat Leaves

The growth rate of the first leaf of eight-day-old wheat plants was measured using a DLT-2 highly sensitive linear displacement transducer. Leaf extensibility was evaluated from the growth rate under the increase in the pulling force by 2 g. An increase in the air temperature resulted in the doubling of the transpiration rate and immediate slowing of the leaf growth followed by the leaf shrinkage. However, growth was later resumed almost completely. Heat treatment did not induce any changes in the leaf extensibility, indicating that cell-wall mechanical properties were not changed. Growth retardation was supposed to result from a decrease in the water content in the leaf tissues because the balance between water influx from roots and its loss through transpiration was shifted toward the water loss. An initial drop in the relative water content (RWC) indicates such a misbalance. Subsequent growth resumption coincided with a decreased water deficiency. Since the rate of transpiration was not reduced, RWC and growth rate restoring evidently occurred due to the activated water uptake by roots, which can be explained by the increased hydraulic permeability detected in our experiments.     

The Effects of Phytohormones and 5-Azacytidine on Apoptosis in Etiolated Wheat Seedlings

The development of etiolated wheat (Triticum aestivum L.) seedlings is necessarily accompanied by apoptosis in their coleoptiles and first leaves. Internucleosome DNA fragmentation, which is characteristic of apoptosis, was detected in the coleoptile as soon as six days after germination. After eight days of germination, DNA fragmentation was clearly expressed in the coleoptile and was noticeable in the apical part of the first-leaf blade. Growing of intact seedlings or incubation of their shoots in the presence of such phytohormones as benzyladenine, gibberellin A₃, fusicoccin C, and 2,4-D at the concentration of 10^–5 M did not essentially affect DNA fragmentation in the coleoptile. As distinct from antioxidants, none of the phytohormones used prevented apoptosis in wheat seedlings. In contrast, ABA (10^–5 M) and an ethylene producer, ethrel (2-chloroethylphosphonic acid, 10^–2–10^–3 M), stimulated sharply DNA fragmentation in the coleoptile. An inhibitor of DNA methylation, 5-azacytidine, was very efficient in the stimulation of DNA fragmentation in the coleoptiles of eight-day-old seedlings at its concentration of 100 g/ml. Thus, some phytohormones can regulate apoptosis, and DNA methylation is involved in this process. Our results indicate that apoptosis activation by some phytohormones may be mediated by their regulation of DNA methylation/demethylation, which is responsible for the induction of genes encoding apoptogenic proteins and/or the repression of antiapoptotic genes.     

胡桃醌对小麦种子萌发及幼苗生长的化感效应

以小麦种子为试验材料,采用培养皿药膜法、生化分析法研究了胡桃醌对小麦种子萌发和幼苗生长的化感效应。结果显示:(1)与对照相比,低浓度胡桃醌(4和20μmol.L-1)处理能促进小麦种子萌发和幼苗生长,种子α-淀粉酶活性增大,幼苗中可溶性蛋白含量升高,丙二醛(MDA)含量减少,超氧化物歧化酶(SOD)活性无显著变化。(2)当胡桃醌浓度大于100μmol.L-1时,对小麦种子萌发和幼苗生长开始呈现抑制作用,种子α-淀粉酶活力降低,幼苗相对含水量、可溶性蛋白含量降低,MDA含量升高,SOD活性先增大后减小。(3)在最大浓度胡桃醌(1 250μmol.L-1)处理下,小麦幼根的细胞形状不规则,排列混乱,层次不分明,维管束模糊不清,幼苗细胞质壁分离,叶绿体萎缩坏死,细胞核变形。研究表明,高浓度胡桃醌通过抑制受体小麦保护酶活性,加重膜脂过氧化程度,导致幼根、幼苗细胞结构的破坏,从而产生化感效应,且化感效应随浓度的增加不断增强。     

Hormonal Status in Wheat and Rice Seedlings under Anoxia

A comparative analysis of the effects of anoxia on growth, fresh weight gain, and phytohormones in wheat (Triticum aestivum L.) and rice (Oryza sativa L.) seedlings was performed. In both plant species, a total cessation of root growth occurred during the initial hours of anoxia. In an anaerobic environment, the fresh weight of wheat seedlings decreased. An increase in the shoot length and weight under the stress conditions was found only in rice seedlings. During the initial hours of anoxia, the level of free ABA in wheat and rice tissues increased manifold, and the accumulation of a free ABA form occurred at the expense of the hydrolysis of its bound forms. The IAA content in plant tissues also increased. In wheat, the accumulation of IAA was short, but in rice, a high hormone level was retained during the entire experiment, and, as a result, its concentration exceeded that of ABA. A level of cytokinins in the tissues of both plant species was affected by anoxia to a lesser extent than that of other phytohormones. This level somewhat decreased under anoxia similarly to the level in darkness under aeration. It is suggested that IAA accumulation in hypoxia-tolerant rice seedlings under anoxia favors maintenance of shoot growth and simultaneous inhibition of root growth. At the same time, in the hypoxia-sensitive wheat, an increase in the ABA level resulted in growth cessation.     

Effect of Helminthosporic Root Rot on the Lipid Content in Wheat Seedlings

The concentration of dry matter and the content of esterified fatty acids in total lipids of roots and etiolated shoots of 3- to 10-day-old seedlings of wheat (Triticum aestivum L.) infected with Bipolaris sorokiniana (Sacc.) Schoemaker, the agent of helminthosporic root rot, were determined in the course of germination. At the onset of germination, fungal infection caused a considerable increase in the dry matter concentration in both roots and shoots due to the enhanced mobilization of seed reserves. However, after the 7th day of germination, dry matter concentration fell below the level of noninfected control seedlings as a result of infection. The content of total lipids rose immediately after infection and always exceeded the control index up to the end of germination, in spite of a continuous decrease in this index in both control and infected seedling. It is concluded that an increase in the content of cellular lipids is a characteristic response of both shoots and roots to the root rot infection of wheat seedlings.     

The Effect of Helminthosporic Root Rot on the Composition of Acyl Lipids of Wheat Seedlings

The changes of the molecular species composition of esterified fatty acids (FAs) of total and nonextractable lipids were determined in roots and etiolated shoots of 3- to 10-day-old wheat (Triticum aestivumL.) seedlings infected with the fungus Bipolaris sorokiniana, the agent of helminthosporic root rot. A novel technique of assessing the extent of the infection-induced deviation of FA composition, mol %, from the control value was developed. It consists in the quantitative determination of both the deviations in this composition and the extent of contribution of separate FA species to the deviations observed. The application of this technique has shown that, for the total lipids, the maximum of such a deviation, in accordance with the membrane theory of stress, directly coincided in time with the onset of a decrease in the dry matter content in both roots and shoots. In each of these, the deviation was primarily caused by the change in the content of those FA species that usually dominate in a specific group of membrane lipids prevailing in a given organ, viz., plastid glycolipids in shoots and extraplastidal membrane phospholipids in roots. In both cases, C₂₀–C₂₂FAs significantly contributed to the deviations observed. This fact seems to reflect an enhanced formation of epicuticular waxes rich in these FAs on the shoot and root surfaces as an adaptive response of plants to fungal infection. Nonextractable (annular) membrane lipids, because of their vital importance for the survival of plant cells, differed from the total lipids with a far greater stability of their quantitative FA composition under conditions of infection-induced metabolic disturbances.     

Effect of Growing Conditions on Wheat Hormonal Status and Productivity in Experimental Ecological System

The levels of free and bound forms of IAA, ABA, cytokinins (CK), and gibberellins, as well as growth characteristics and productivity were investigated in two wheat lines. The plants were grown under controlled conditions in an artificial ecosystem that allowed the irradiance, CO₂ concentration, and rhizosphere temperature to be changed. The main difference in the hormonal status of leaves of tall spring wheat, line 232, and dwarf wheat, line 95-3, was the absence of GA₉ gibberellins in the latter. It was found that the light intensity and temperature of rhizosphere insignificantly affected the balance of endogenous phytohormones and HI in wheat. The elevation of CO₂ concentration resulted in a considerable increase in the content of free IAA, an appearance of free GA₉, and a rise in the productivity of wheat, line 232. The concentration of CO₂ was shown to be a major parameter that determined HI in the experimental ecological system.     

Changes in the Phytohormone Levels in Wheat Calli as Affected by Salicylic Acid and Infection with Tilletia caries,a Bunt Pathogenic Agent

The role of salicylic acid (SA) in growth regulation and the change in the levels of phytohormones (IAA, ABA, and cytokinins) were studied in the wheat calli co-cultured with bunt pathogen Tilletia caries. Calli infection with T. caries resulted in the hypertrophied callus growth and simultaneous increase in phytohormone level. The addition of SA to the nutrient media decreased the callus growth induced by the pathogen, whereas the level of investigated phytohormones was not affected. In the SA-treated infected calli, the formation of necrotic lesions was observed in the zones of contact of the fungal mycelium with callus cells that limited pathogen growth. The authors suggest that the stabilization of the hormonal balance of plant cells at pathogenesis is one of the possible mechanisms of the SA protective action in vitro and in vivo. Hence, co-culturing wheat calli and T. caries fungus appeared to be a convenient model for assessing SA protective action.     

The Effect of Local Cooling of Cucumber and Wheat Seedlings on Various Kinds of Stress Resistance of Their Leaves and Roots

Shoots and roots of wheat (Triticum aestivum L., cold-resistant species) and cucumber (Cucumis sativus L., cold-sensitive species) were chilled at 2°C or 10°C, respectively, for 7 h. The changes in cold, heat, and salt resistance in treated leaf and root cells were recorded. Local cooling of the leaf resulted in an increase of its cold and salt tolerance, but its heat tolerance remained unchanged. At the same time, cold tolerance of the root slightly increased as a result of local cooling, but its heat and salt tolerance decreased. Cooling of the shoot did not affect the cold and heat tolerance of root cells but caused a decrease in their salt tolerance. Finally, in the leaf maintained at a normal temperature, there was an increase in all kinds of stress resistance as a result of root cooling. We discuss the possibility of an unspecific change in stress resistance caused by metabolic shifts. These shifts are induced by a signal, which is transmitted inside the plant into plant organs located at a considerable distance from the chilled ones.     

Molecular Characteristics of Transgenic Wheat and the Effect on Transgene Expression

A population of R0 transgenic wheat plants, generated by particle bombardment, was analyzed to define molecular, genetic and phenotypic properties resulting from transformation with a cointegrate vector, or cotransformation with two separate plasmids. By evaluating the progeny of 70 independently-derived transgenic plants, we also identified rare events such as chimerism and transgene elimination, which provide valuable information concerning the development of transgenic cereal plants following bombardment experiments. The frequency of chimerism in our transgenic wheat plants was very low. Furthermore, while transgene elimination did occur, this was also a very rare event. We determined the copy numbers of integrated transgenes and the levels of transgene expression. Comparisons to transgenic rice plants generated in the same manner demonstrated some similarities, but also important differences in transgene behavior. Whereas in rice there is no evidence for any direct relationship between transgene copy number and transgene expression or stability, multicopy populations in wheat demonstrated a bias towards higher levels of expression for the two genes and the maize ubiquitin promoter evaluated in the present study.     

一氧化氮对小麦叶片镍毒害的缓解作用

采用溶液培养法研究了重金属镍(Ni)对扬麦158(Triticum aestivumL)幼苗生长的影响及外源一氧化氮(NO)对Ni毒害的缓解作用。结果表明,100μmol/LNi处理显著抑制小麦幼苗生长,导致叶绿素含量下降,丙二醛(MDA)含量显著升高,且叶片过氧化物酶(POD)、超氧化物歧化酶(SOD)和谷胱甘肽转硫酶(GST)等抗氧化酶活性升高。400μmol/L硝谱钠(SNP,NO供体)预处理2d,能够明显减轻Ni毒害,使叶绿素和MDA含量基本恢复至对照水平。NO可能是通过提高APX和GR等抗氧化酶的活性及谷胱甘肽含量而增强植株抗氧化能力,显著减轻由Ni导致的叶片Ca和Fe含量下降而增强小麦幼苗抵御Ni毒害的能力。     

中国普通小麦初选核心种质的产生

对中国普通小麦种质资源构建了初选核心种质。地方品种和选育品种分别构建。按栽培区(地理生态区)分组。地方品种按亚区分为28组,选育品种按大区分为10组。各组内在21个表型性状聚类的基础上,按平方根法取样,并依遗传多样性指数与遗传丰富度加以调整。提出在生产上或育种中起过重要作用的品种为必选材料。初步选定的材料经种植核对,淘汰错杂后,产生初选核心种质。地方品种全部供试材料11694份,初选核心种质3283份,取样比例为28．18％。选育品种全部11441份,初选核心种质1684份,取样比例为14．9％。计划经分子标记分析,最后核心种质的比例占全部种质的10％左右。根据全部材料21个性状遗传多样性指数测验,初选核心种质,除芒和壳两性状外,与全部种质的遗传差异均未达到显水平。讨论了初选核心种质的构建方法。指出陕南部西山地和汾渭谷地是中国小麦地方品种遗传变异多样性的富集地。育成品种多样性程度以西南冬麦区和黄淮冬麦区为最高。     

Endogenous cytokinins mediate growth-stimulating and protective action of the HUMI preparation on wheat plants inoculated with Tilletia caries

Humic substances (HS) and preparations created on their basis are known to be effective in growth stimulation and improvement of resistance and productivity of different plant species. We have revealed that the presowing semidry treatment of wheat seeds with preparation HUMI (NVP ‘BashIncom’, Ufa, Bashkortostan, Russia) promoted seedling growth and increased wheat yield. This effect is likely to be due to the shifts induced by HUMI in the hormonal system of plants associated with a 1.5-fold persistent accumulation of cytokinins (CKs) without any changes in the contents of indoleacetic acid (IAA) and abscisic acid (ABA). Meanwhile, presowing treatment with HUMI had a stabilizing effect on the state of the hormonal system in the course of ontogenesis of wheat inoculated with teliospores Tilletia caries. Obtained data about prevention of the decline in the cytokinin content in HUMI-treated infected wheat is fundamental in the implementation of immuno-stimulatory effect of HUMI resulting in the suppression of the growth and development of pathogens.     

Activity of Lectin-Like Proteins of the Cell Walls and the Outer Organelle Membranes as Related to Endogenous Ligands in Cold-Adapted Seedlings of Winter Wheat

The hemagglutinating activity (HA) of lectin-like components in the cell walls and the outer organelle membranes was studied in freezing-tolerant winter wheat (Triticum aestivum L., cv. Mironovskaya 808) plants in the course of hardening at 2°C, in parallel with the effects of endogenous ligands from the soluble fraction on HA. Low hardening temperature divergently changed HA of the lectin-like components in the cell walls, the outer membranes of nuclei, plastids, and mitochondria, and the microsomal membranes: HA increased in the cell walls, nuclei, and plastids and decreased in the mitochondria and microsomal membranes. Under hardening conditions, with plant growth slowed down, HA of the lectin-like proteins from the outer organelle membranes was inhibited in the presence of the soluble fraction components (soluble ligands); such inhibition was not observed in the case of actively growing nonhardened seedlings. The authors put forward a hypothesis that the lectin-like proteins from both peripheral (cell walls) and intracellular (outer organelle membranes) compartments are essential for developing freezing tolerance. HA of the cell walls and the outer membranes of nuclei and plastids enhanced by hardening manifested positive correlation with freezing tolerance and negative correlation with the growth rate. In contrast, HA of the outer membranes of mitochondria and microsomes was positively related to plant growth and negatively, to freezing tolerance. As negative and positive effectors of membrane-dependent processes, the lectin-like components of the outer organelle membranes seem to control membrane functional activities in the course of cold adaptation.     

Effect of increased irradiance on the hormone content,water relations,and leaf elongation in wheat seedlings

In wheat (Triticum durum Desf., cv. Bezenchukskaya 139) seedlings, an increase in irradiance from 20 to 400 μmol/(m² s) PAR enhanced transpiration and increased stomatal conductance by three times on the background of reduced relative water content (RWC). After this treatment, leaves quickly ceased to grow and became even shrunk later. In 40 or 50 min, leaf growth was resumed. At this period, we observed an increase in hydraulic conductivity and RWC and also in leaf extensibility. As soon as 10 min after treatment, some changes in hormone content were noted. In the zones of leaf growth and its mature part, zeatin and zeatin riboside were accumulated, whereas ABA accumulation was observed in the zone of leaf growth and in the roots. The results obtained indicate that leaf expansion at increased irradiance was related to changes in cell-wall extensibility and hydraulic conductivity. The first effect could be due to cytokinin accumulation, whereas the second one, to ABA accumulation.