The Effects of Root Treatment with Various Stress Agents on Plant Cold- and Heat-Tolerance

The root systems of cucumber (Cucumis sativus L.), wheat (Triticum aestivum L.), and barley (Hordeum vulgare L.) plants were subjected to a short-term (6–7-h-long) treatment with low and high temperatures, sodium chloride, and lead nitrate, and the effect of these treatments on the changes in the cold- and heat-tolerance of leaf cells was determined. It was established that chilling of cucumber and wheat seedling roots at 10 and 2°C, respectively, or their heating at 38°C and 40°C, respectively, induced an increase in both cold- and heat-tolerance of leaf cells. An increase in the cold- and heat-tolerance was also observed in roots treated with sodium chloride at concentrations of 0.15 M (cucumber) and 0.2 M (wheat), as well as with lead nitrate at a concentration of 0.1 mM (barley, wheat). The tolerance increase induced by these stress agents was accompanied by a considerable increase in the ABA concentration in leaves. The effect of physical and chemical stress agents is suggested to induce the same nonspecific changes in the aboveground organs. These changes bring about, directly or indirectly, an increase in the cold- and heat-tolerance and are related to an increase in the ABA content.     

Effect of Whole Plant and Local Heating on the ABA Content in Cucumber Seedling Leaves and Roots and on Their Heat Tolerance

The effects of heating at 38°C of whole cucumber (Cucumis sativus L.) seedlings or local heating of their shoots and roots on ABA content and heat tolerance of leaves and roots were investigated. During the initial period of the high-temperature treatment of whole seedlings, the ABA concentration in leaves and roots increased considerably. Local heating of the shoot or root resulted in an increase in the ABA concentration not only in the heated organ, but also in unheated seedling parts. A high-temperature treatment of the whole seedlings and the local treatment of shoots or roots caused an increase in the heat tolerance of leaf cells. The heat tolerance of root cells virtually did not change after heating of the whole seedlings or shoots, but decreased after heating of roots. The possible role of ABA in changing the heat tolerance of leaf and root cells by local heating of the seedling is discussed.     

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.     

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.     

Changes in water relations and endogenous abscisic acid content of wheat and barley grains and embryos during development

Abstract. Immature cereal embryo development can be controlled by in vitro culture on media containing ABA, or by media of low osmotic potential. To assess the possible in vivo roles of these factors, endogenous ABA levels and water relations of embryos and grains of wheat ( Triticum aestivum L.) and barley ( Hordeum vulgare L.) were determined during development. ABA concentrations remained consistent with those required to inhibit precocious germination in vitro of early stage embryos but not of more mature embryos. With increasing maturity, a difference in water potential developed between grain and embryo, suggestive of an in vivo role for water status in controlling the development of the embryo.     

Apoptosis in developing anthers and the role of ABA in this process during androgenesis in Hordeum vulgare L.

Intra-nucleosomal cleavage of DNA into fragments of about 200 bp was demonstrated to occur in developing anthers, in which microspores had developed into the mid-late to late uni-nucleate stage in situ, i.e. at the verge of mitosis. The same was observed, but to a much larger extent, if these anthers were pre- treated by a hyper-osmotic shock. Pretreatment of anthers before the actual culture of microspores was required for optimal androgenesis of microspores. The use of the TUNEL reaction, which specifically labels 3 ends of DNA breaks, after intra-nucleosomal cleavage of DNA, revealed that DNA fragmentation mainly occurred in the loculus wall cells, tapetum cells and filament cells. TUNEL staining was absent or infrequently observed in the microspores of developing anthers in situ. Electron microscopy studies showed condensed chromatin in nuclei of loculus wall cells in the developing anthers. These observations at the chromatin and DNA level are known characteristics of programmed cell death, also known as apoptosis. Features of apoptosis were infrequently found in microspores from freshly isolated mature anthers. However, most tapetum cells had disappeared in these anthers and the remaining cell structures showed loss of cellular content. The viability of microspores in pre-treated anthers was comparable to those in freshly isolated anthers and almost four times higher than in anthers from control experiments. This observation was correlated with three to four times less microspores showing TUNEL staining and a two times higher level of ABA in the anther plus medium samples than in controls. Addition of ABA to the controls enhanced the viability and lowered the occurrence of apoptosis linked characteristics in the microspores. These data suggest that pre-treatment is effective in stimulating androgenesis because it leads to an increase in ABA levels which protects microspores from dying by apoptosis.     

Effects of chlorosubstituted indoleacetic acids on root growth, ethylene and ATP formation

7-Chloroindoleacetic acid and dichloroindoleacetic acids with a Cl in the 7 position showed anti-auxinic activity and promoted root growth in wheat ( Triticum aestivum L. cv. Diamant II). In contrast, 4-, 5- and 6-chloroindoleatetic acids acted as strong auxins inhibiting the growth of wheat roots. Flax ( Linum usitatissimum L. cv. Concurrent) and cucumber ( Cucumis sativus L. cv. Favör) roots showed similar, but less clear-cut responses. 7-Chloroindoleacetic acid and 4,7-dichloroindoleacetic acid alleviated root growth inhibition in wheat caused by IAA, monochloroindoleacetic acids and benzyladenine. 2,4-D, 4- and 6-chloroindoleacetic acids strongly induced ethylene formation in cucumber seedlings; 4,7- and 6,7-dichloroindoleacetic acids did not, except at high concentrations. The more lipid-soluble dichloroindoleacetic acids were stronger inhibitors of ATP formation in cucumber mitochondria than monochloroindoleacetic acids, while IAA itself had only a very slight effect.     

Waterlogging tolerance in the tribe Triticeae: the adventitious roots of Critesion marinum have a relatively high porosity and a barrier to radial oxygen loss

Nine species from the tribe Triticeae – three crop, three pasture and three ‘wild’ wetland species – were evaluated for tolerance to growth in stagnant deoxygenated nutrient solution and also for traits that enhance longitudinal O₂ movement within the roots. Critesion marinum (syn. Hordeum marinum) was the only species evaluated that had a strong barrier to radial O₂ loss (ROL) in the basal regions of its adventitious roots. Barriers to ROL have previously been documented in roots of several wetland species, although not in any close relatives of dryland crop species. Moreover, the porosity in adventitious roots of C. marinum was relatively high: 14% and 25% in plants grown in aerated and stagnant solutions, respectively. The porosity of C. marinum roots in the aerated solution was 1·8–5·4‐fold greater, and in the stagnant solution 1·2–2·8‐fold greater, than in the eight other species when grown under the same conditions. These traits presumably contributed to C. marinum having a 1·4–3 times greater adventitious root length than the other species when grown in deoxygenated stagnant nutrient solution or in waterlogged soil. The length of the adventitious roots and ROL profiles of C. marinum grown in waterlogged soil were comparable to those of the extremely waterlogging‐tolerant species Echinochloa crus‐galli L. (P. Beauv.). The superior tolerance of C. marinum, as compared to Hordeum vulgare (the closest cultivated relative), was confirmed in pots of soil waterlogged for 21 d; H. vulgare suffered severe reductions in shoot and adventitious root dry mass (81% and 67%, respectively), whereas C. marinum shoot mass was only reduced by 38% and adventitious root mass was not affected.     

The effect of Ca2+-stress on fluxes of Ca2+ and K+ in wheat and cucumber

Betula papyrifera Marsh, seedlings adapted very poorly to flooding for up to 60 days. Responses to flooding included increased ethylene production; stomatal closure; leaf senescence; drastic inhibition of shoot growth, cambial growth, and root growth; decay of roots, and death of many seedlings. Flooding inhibited growth of leaves that formed prior to flooding, inhibited formation of new leaves, and induced abscission of old leaves. As a result of extensive leaf abscission, fewer leaves were present after flooding than before flooding was initiated. The drastic reduction in leaf area was associated with greatly decreased growth of the lower stem and roots. No evidence was found of adaptive morphological changes to flooding. The data indicate that intolerance of B. papyrifera seedlings to flooding is an important barrier to regeneration of the species on sites subject to periodic inundation.     

Interaction Between Root and Leaf Disease Development in Barley Cultivars after Inoculation with Different Isolates of Bipolaris sorokiniana

The relationship between root and leaf infection in 11 cultivars of barley ( Hordeum vulgare ) by different isolates of Bipolaris sorokiniana was investigated in young plants. Roots of 10-day-old seedlings, grown in filterpaper rolls, and the third leaf of 17-day-old seedlings were inoculated with the different isolates and a Disease Development Index (DDI) was calculated.
The rate of lesion development in leaves was higher than in roots, indicated by generally higher DDI after leaf inoculation than after root inoculation. Significant differences in resistance were found among the barley cultivars. Inoculation with different isolates of B. sorokiniana caused significant differences in DDI for both roots and leaves. In the leaves, but not in the roots, a significant cultivar–isolate interaction was found. No significant correlations, neither in isolate aggressiveness nor in cultivar reaction between root and leaf, were observed.     

丝氨酸内肽酶在黄瓜叶片衰老中的作用

采用丝氨酸内肽酶抑制剂和植物生长调节剂处理离体黄瓜叶片,研究了黄瓜叶片暗诱导衰老过程中丝氨酸内肽酶的作用。结果表明,6-BA50μmol／L与丝氨酸内肽酶抑制剂AEBSF能抑制叶片内肽酶活性的升高,延缓蛋白质降解,而ABA50μmol/L则促进了内肽酶活性的升高：其作用效果与AEBSF相反。活性电泳结果显示,黄瓜叶片中检测到6条内肽酶同工酶,其中4条（CEP2、3、4、6）为丝氨酸类型内肽酶,而ABA使丝氨酸内肽酶CEP2、3、4、6的活性明显增强,提示了丝氨酸类型内肽酶在黄瓜叶片衰老过程中具有重要作用。     

胚乳和蔗糖对大麦成熟胚无菌苗生长和叶片切段愈伤组织诱导及分化的影响

大麦无菌苗的苗高。苗干重和单位长度、叶片切段的干重均依胚培养时所带胚乳的增多而递增。但叶片切段培养时的愈伤组织诱导率和再分化率并不与胚乳多少正相关。成熟胚培养时蔗糖的供应明显影响叶片切段培养时的愈伤组织诱导率。培养带1/2胚乳的胚,用萌发率高的新鲜种子培养无菌苗时,以不供给蔗糖的有较高的叶段愈伤组织诱导率,用萌发率显著降低的陈种子时,则以育苗时供给6％蔗糖为好。     

Stress-dependent redistribution of abscisic acid (ABA) in Hordeum vulgare L leaves: the role of epidermal ABA metabolism, tonoplastic transport and the cuticle

When ¹⁴C-labelled abscisic acid ([¹⁴C]ABA) was supplied to isolated protoplasts of the barley leaf at pH 6, initial rates of metabolism were about five times higher in epidermal cell protoplasts than in mesophyll cell protoplasts if equal cytosolic volumes were considered. In spite of the fact that epidermal cells make up only about 35% of the total water space in barley leaves, and despite the small cytosolic volume of these cells, in intact leaves all epidermal cells would thus metabolize half as much ABA per unit time as the mesophyll cells (0–27 and 0–51 mmol h^?1 m^?3 leaf water). Therefore, under these conditions epidermal cells seem to be a stronger sink than mesophyll cells for ABA that arrives via the transpiration stream. However, at an apoplastic pH of 7–25, which occurs in stressed leaves, the proportion of total metabolized ABA would be much smaller in epidermal than in mesophyll cells (0–029 and 0–204 mmolh^?l m^?3 leaf water). Our results indicate that under conditions of slightly alkaline apoplastic pH the epidermis may serve as the main source for fast stress-dependent ABA redistribution into the guard cell apoplast. This is partly the result of ABA transport across the epidermal tonoplast, which is dependent on the apoplastic pH and possibly on the cytosolic calcium concentration. The cuticle seems to be of no particular importance in stress-induced apoplastic ABA shifts and cannot be regarded as a significant sink for high ABA concentrations under stress.     

An analysis of the light-induced unrolling of the grass leaf

The light-induced unrolling of primary leaves of dark-grown wheat ( Triticum aestivum L. cv. Weibull's Folke) and barley ( Hordeum vulgare L. cv. Weibull's Ida) does not follow the reciprocity law but shows maxima and minima like the phototropic dose-response curve. These fluctuations of the unrolling can be explained by differences and non-synchronizations of the growth in width of the two sides of the leaves. As the thickness of the leaves is small, even minute differences between the widths of the two sides will give rise to great changes in the unrolling and thus in the projection values.     

一氧化氮参与调节盐胁迫下脱落酸诱导的小麦幼苗叶片脯氨酸的累积

不同浓度(0.01～5.00mmol/L)的外源一氧化氮(NO)供体硝普钠(SNP)以浓度依赖性的性式诱导150mmol/LNaCl胁迫下小麦(Triticum aestivum L.cv.Yangmai 158)幼苗叶片脯氨酸的累积.其中0.1 mmol/L的SNP效果最明显,而结合采用NO清除剂c-PTIO和血红蛋白的处理均分别逆转了该效应.研究结果还发现:0.1 mmol/L SNP诱导的脯氨酸累积还可能有利于盐胁迫下小麦幼苗的保水性;0.1 mmol/L的SNP显著激活了内源ABA的合成,而结合血红蛋白的处理则证实,在外源ABA诱导脯氨酸累积的过程中NO可能作用于ABA信号分子的下游,但NO和ABA信号分子在此诱导反应中不存在累积效应.进一步研究脯氨酸合成和降解的酶促反应途径,发现外源NO处理前4天内可能主要是通过提高△'-吡咯啉-5-羧酸合成酶(P5CS)的活性来促进脯氨酸的合成,以后直至第8天主要是通过抑制脯氨酸脱氢酶(ProDH)的活性来抑制脯氨酸的降解;ABA对于P5CS和ProDH活性的调节能力弱于NO.此外,Ca2 在NO诱导的盐胁迫下小麦叶片脯氨酸累积的信号分子途径中起重要的介导作用.     

Role of TaALMT1 malate-GABA transporter in alkaline pH tolerance of wheat

Malate exudation through wheat (Triticum aestivum L) aluminium-activated malate transporter 1 (TaALMT1) confers Al³⁺ tolerance at low pH, but is also activated by alkaline pH, and is regulated by and facilitates significant transport of gamma-aminobutyric acid (GABA, a zwitterionic buffer). Therefore, TaALMT1 may facilitate acidification of an alkaline rhizosphere by promoting exudation of both malate and GABA. Here, the performance of wheat near isogenic lines ET8 (Al⁺³-tolerant, high TaALMT1 expression) and ES8 (Al⁺³-sensitive, low TaALMT1 expression) are compared. Root growth (at 5 weeks) was higher for ET8 than ES8 at pH 9. ET8 roots exuded more malate and GABA at high pH and acidified the rhizosphere more rapidly. GABA and malate exudation was enhanced at high pH by the addition of aluminate in both ET8 and transgenic barley expressing TaALMT1. Xenopus laevis oocytes expressing TaALMT1 acidified an alkaline media more rapidly than controls corresponding to higher GABA efflux. TaALMT1 expression did not change under alkaline conditions but key genes involved in GABA turnover changed in accordance with a high rate of GABA synthesis. We propose that TaALMT1 plays a role in alkaline tolerance by exuding malate and GABA, possibly coupled to proton efflux.     

Polyribosome metabolism, growth and water status in the growing tissues of osmotically stressed plant seedlings

Relationships between growth of osmotically stressed intact seedlings and polyribosome levels and water status of growing tissues were examined. Sudden exposure of barley (Hordeum vulgare L. cv. Arivat) roots to a solution of ?0.8 MPa polyethylene glycol caused leaf growth to stop almost immedately, but growth resumed at a much lower rate after 0.5–1 h. In the growing region of leaves, the polyribosome: total ribosome ratio of free (non-membrane-bound) ribosomes was significantly reduced after 15 min stress, but a decrease in the large polyribosome:total polyribosome ratio occurred only after 1–2 h. Membrane-bound and free polyribosome levels both decreased to 70% of unstressed control values after 4 h stress. Recovery of total polyribosomes occurred within 1 h after relief of 4 h stress, but required 3 h after relief of 24 h stress. Stress detectably reduced the water potential and osmotic potential of growing tissue within 0.5–1.0 h, and osmotic adjustment continued for up to 10 h. Recovery of water status was incomplete after 1 h relief of a 4 h stress. In contrast, expanded blade tissues of stressed plants underwent minor changes in water status and slow decreases in polyribosomes levels. These results confirm that growing tissues of barley leaves are selectively responsive to stress, and suggest that changes in growth, water status and polyribosome levels may be initiated by the same signal. Measurements of seedling growth, polyribosome levels and water status of growing tissues of barley and wheat (Triticum aestivum L. cv. Zaragoza) leaves, etiolated pea (Pisum sativum L. cv. Alaska) epicotyl and etiolated squash (Cucurbita pepo L. cv. Elite) hypocotyl stressed with polyethylene glycol solutions of ?0.3 to ?0.8 MPa for 12 h or more showed that polyribosome levels were highly correlated with seedling growth rate as well as with tissue water and osmotic potentials, while turgor remained unchanged. These results suggest that long-term growth of osmotically stressed plants may be limited by a reduced capacity for protein synthesis in growing tissues and is not dictated by turgor loss.