Response of winter wheat to cold: production of phenolic compounds and L-phenylalanine ammonia-lyase activity

The formation of soluble and polymeric (lignin) phenolic compounds, activity of L-phenylalanine ammonia lyase (PAL, EC 4.3.1.5), and content of free L-phenylalanine during cold hardening of winter wheat plants (Triticum aestivum L.) were studied. Cold treatment increased accumulation of soluble phenolic compounds in leaves while not affecting the content of lignin. The opposite was observed in tillering nodes. The activity of PAL was lower than in control plants in both tissues, and the content of free L-phenylalanine in tissues increased.     

冬小麦籽粒淀粉合成相关酶活性的日变化

蔗糖向淀粉的转化是决定小麦籽粒产量的重要因素.田间条件下研究了两个小麦(Triticum aestivum L.)品种"鲁麦22"和"鲁麦14"籽粒淀粉合成相关酶:蔗糖合酶(sucrose synthase,SS)、腺苷二磷酸葡萄糖焦磷酸化酶(ADP-glucose pyrophosphorylase,ADPGPPase)、可溶性淀粉合酶(soluble starch synthase,SSS)、束缚态淀粉合酶(starch granule-bound synthase,GBSS)的活性以及籽粒ATP含量的日变化.结果表明,上述酶活性呈现明显的昼夜变化特征,酶活性一般在白天较低,而在夜间呈现较高活性,而籽粒ATP含量趋势相反.相关分析表明,白天较低的酶活性可能与气温超过其适宜温度有关.对籽粒淀粉合成相关酶活性日变化的可能因子进行了讨论.     

冬小麦籽粒淀粉合成相关酶活性的日变化

蔗糖向淀粉的转化是决定小麦籽粒产量的重要因素。田间条件下研究了两个小麦(TriticumaestivumL.)品种“鲁麦22”和“鲁麦14”籽粒淀粉合成相关酶:蔗糖合酶(sucrosesynthase,SS)、腺苷二磷酸葡萄糖焦磷酸化酶(ADP-glucosepyrophosphorylase,ADPGPPase)、可溶性淀粉合酶(solublestarchsynthase,SSS)、束缚态淀粉合酶(starchgranule-boundsynthase,GBSS)的活性以及籽粒ATP含量的日变化。结果表明,上述酶活性呈现明显的昼夜变化特征,酶活性一般在白天较低,而在夜间呈现较高活性,而籽粒ATP含量趋势相反。相关分析表明,白天较低的酶活性可能与气温超过其适宜温度有关。对籽粒淀粉合成相关酶活性日变化的可能因子进行了讨论。     

Development of Frost Tolerance in Winter Wheat as Modulated by Differential Root and Shoot Temperature

Abstract: Winter wheat plants ( Triticum aestivum L. cv. Urban), grown in nutrient solution, were exposed to differential shoot/root temperatures (i.e., 4/4, 4/20, 20/4 and 20/20 C) for six weeks. Leaves grown at 4C showed an increase in frost tolerance from - 4C down to - 11 C, irrespective of root temperature. In 4/20 C plants, high root temperature decreased the rate of hardening of the leaves, but did not influence the final level of frost tolerance. In roots grown at 4C frost tolerance increased from - 3 C down to - 4 C, independently of shoot temperature. An accumulation of soluble sugars in the leaves was only observed when both shoot and root were grown at 4C and was not correlated with final frost tolerance achieved. However, the rate of hardening was correlated with the soluble sugar concentration. An increase in root soluble sugar concentration was exclusively observed in roots exposed to a temperature of 4C, irrespective of shoot temperature. Proline concentration only increased in plant parts exposed to a temperature of 4C. The present results indicate that the importance of root temperature in low-temperature hardening of winter wheat is limited, even though exposure to differential root and shoot temperatures brought about pronounced changes in growth, soluble sugar concentration, insoluble sugar concentration and proline concentration in roots and leaves.     

Drought stress-induced changes in the composition and physical state of phospholipids in wheat

Seedlings of winter wheat ( Triticum aestivum L. cv. Jubilejnaja 50) were grown under normal and dry conditions. Frost resistance (LT₅₀) of 10-day-old control seedlings was −6°C. LT₅₀ of the subsequently drought-stressed leaves shifted to −16°C. In plants of the same physiological age (28 days) but grown without stress, LT₅₀ was −12°C. Phosphatidylcholine accumulated and phosphatidylethanolamine decreased in drought-stressed leaves. Fatty acid unsaturation of these phospholipids increased with leaf age, independently of water supply. Both ageing and drought stress produced a decrease in the apparent phase separation temperature of isolated total phospholipids as determined by electron spin resonance. The possible role of dehydration-induced structural changes in the bilayer matrix in triggering adaptive alterations in membrane composition, similar to those observed during cold hardening, is discussed.     

Carbohydrate levels among winter wheat cultivars varying in freezing tolerance and snow mold resistance during autumn and winter

The LT₅₀ values and soluble carbohydrate levels in wheat ( Triticum aestivum L.) crowns and leaves were monitored throughout autumn and winter in cultivars varying in freezing tolerance and snow mold resistance during 1993/1994 and 1994/1995 in the field at Sapporo, Japan. During the first stage of hardening, from sowing to mid‐November, the pattern of accumulation of mono‐ and disaccharides was similar for all cultivars. During the second stage of cold hardening, from mid‐November to mid‐December, the greatest accumulation of mono‐ and disaccharides, without a corresponding increase in fructan, was observed among the freezing‐tolerant cultivars; and levels of simple saccharides rapidly decreased under snow cover. Conversely, levels of mono‐ and disaccharides in snow mold‐resistant cultivars were less than 70% of those in freezing‐tolerant cultivars before snow cover and maintained low levels throughout winter, while polysaccharide levels in snow mold‐resistant cultivars were about 120% of those in freezing‐tolerant cultivars in December. Sugar metabolism during the winter was examined using 18 cultivars in 1994/1995. LT₅₀ values were correlated to the greatest extent with total mono‐ and disaccharide and fructan content among wheat cultivars excluding snow mold‐resistant cultivars in December. Snow mold‐resistant cultivars tended to metabolize carbohydrates more slowly until the end of the snow cover period. This result suggested that the enzymatic metabolism of the synthesis of sugars and the conversion of fructan to cryoprotective sugars in response to low temperatures, especially subzero ones, might be different between the two contrasting types in resistance to winter stress.     

Adenine Nucleotide Changes during Cold Acclimation of Winter Rape Plants

During the first stage of hardening of winter rape plants (Brassica napus L. var. oleifera L., cv. Górczański), marked increase of ATP content in leaves was observed. Lowering the temperature from 5 to 0 C (the second stage of hardening) had no further effect on ATP content. In roots, not capable of hardening, pronounced decrease of ATP content was noted after prolonged exposure to cold. It was found that increased ATP content and higher energy charge in cold-treated leaves were due to light and dark processes.     

Factors contributing to enhanced freezing tolerance in wheat during frost hardening in the light

The interaction between light and temperature during the development of freezing tolerance was studied in winter wheat (Triticum aestivum L. var. Mv Emese). Ten-day-old plants were cold hardened at 5 degrees C for 12 days under normal (250 micromol m(-2)s(-1)) or low light (20 micromol m(-2)s(-1)) conditions. Some of the plants were kept at 20/18 degrees C for 12 days at high light intensity (500 micromol m(-2)s(-1)), which also increased the freezing tolerance of winter wheat. The freezing survival rate, the lipid composition, the antioxidant activity, and the salicylic acid content were investigated during frost hardening. The saturation level of hexadecanoic acid decreased not only in plants hardened at low temperature, but also, to a lesser extent, in plants kept under high light irradiation at normal growth temperature. The greatest induction of the enzymes glutathione reductase (EC 1.6.4.2.) and ascorbate peroxidase (EC 1.11.1.11.) occurred when the cold treatment was carried out in normal light, but high light intensity at normal, non-hardening temperature also increased the activity of these enzymes. The catalase (EC 1.11.1.6.) activity was also higher in plants grown at high light intensity than in the controls. The greatest level of induction in the activity of the guaiacol peroxidase (EC 1.11.1.7.) enzyme occurred under cold conditions with low light. The bound ortho-hydroxy-cinnamic acid increased by up to two orders of magnitude in plants that were cold hardened in normal light. Both high light intensity and low temperature hardening caused an increase in the free and bound salicylic acid content of the leaves. This increase was most pronounced in plants that were cold treated in normal light.     

Complex phytohormone responses during the cold acclimation of two wheat cultivars differing in cold tolerance, winter Samanta and spring Sandra

Hormonal changes accompanying the cold stress (4°C) response that are related to the level of frost tolerance (FT; measured as LT50) and the content of the most abundant dehydrin, WCS120, were compared in the leaves and crowns of the winter wheat (Triticum aestivum L.) cv. Samanta and the spring wheat cv. Sandra. The characteristic feature of the alarm phase (1 day) response was a rapid elevation of abscisic acid (ABA) and an increase of protective proteins (dehydrin WCS120). This response was faster and stronger in winter wheat, where it coincided with the downregulation of bioactive cytokinins and auxin as well as enhanced deactivation of gibberellins, indicating rapid suppression of growth. Next, the ethylene precursor aminocyclopropane carboxylic acid was quickly upregulated. After 3-7 days of cold exposure, plant adaptation to the low temperature was correlated with a decrease in ABA and elevation of growth-promoting hormones (cytokinins, auxin and gibberellins). The content of other stress hormones, i.e., salicylic acid and jasmonic acid, also began to increase. After prolonged cold exposure (21 days), a resistance phase occurred. The winter cultivar exhibited substantially enhanced FT, which was associated with a decline in bioactive cytokinins and auxin. The inability of the spring cultivar to further increase its FT was correlated with maintenance of a relatively higher cytokinin and auxin content, which was achieved during the acclimation period.     

玉米赤霉烯酮在冬小麦短日春化中的作用

应用酶联免疫技术,研究了冬小麦品种燕大１８１７ 在不同光周期条件下,体内内源玉米赤霉烯酮（ＺＥＮ） 和ＡＢＡ 含量的变化。结果表明,冬小麦品种燕大１８１７ 具有短日春化性,苗期经４ 周ＳＤ 处理后转到ＬＤ 下能正常抽穗。ＳＤ 诱导春化的幼苗茎尖和叶片中ＺＥＮ 含量高于未经诱导的ＬＤ 幼苗,而在两种光周期条件下生长的幼苗中ＡＢＡ 含量变化趋势并无明显差异。马拉硫磷（ ＭＡＬ） 抑制ＳＤ 幼苗体内ＺＥＮ 含量高峰的出现,也部分抑制抽穗;外源ＺＥＮ 可部分抵消ＭＡＬ对植株的影响,降低茎尖内源ＡＢＡ 含量,并有促进小麦拔节的作用。说明在冬小麦短日春化过程中,ＺＥＮ 具有促进作用,而ＡＢＡ 没有直接影响。     

Response of Winter Wheat to Cold: Production of Phenolic Compounds and L-Phenylalanine Ammonia Lyase Activity

The formation of soluble and polymeric (lignin) phenolic compounds, activity of L-phenylalanine ammonia lyase (PAL, EC 4.3.1.5), and content of free L-phenylalanine during cold hardening of winter wheat plants (Triticum aestivum L.) were studied. Cold treatment increased accumulation of soluble phenolic compounds in leaves while not affecting the content of lignin. The opposite was observed in tillering nodes. The activity of PAL was lower than in control plants in both tissues, and the content of free L-phenylalanine in tissues increased.     

Cold acclimation of wheat (Triticum aestivum): Properties of enzymes involved in proline metabolism

Two cultivars of wheat ( Triticum aestivum L.), a winter wheat, Kharkov, and a spring wheat, Glenlea, were acclimated under controlled conditions at 2 temperatures, 5°C and 25°C with a 12-h photoperiod. Water content, protein and proline concentrations were determined. Enzymatic properties (activity and apparent energy of activation) were investigated for enzymatic systems involved in 2 pathways of proline metabolism, the glutamic acid and ornithine pathways. Four enzymes were studied, proline dehydrogenase (PDH, EC 1.5.1.2), glutamate dehydrogenase (GDH, EC 1.4.1.2-4), glutamine synthetase (GS, EC 6.3.1.2) and ornithine transaminase (OT, EC 2.6.1.13). Cold acclimation led to an accumulation of proline, a decrease in water content and an increase in soluble protein, especially in winter wheat. For both cultivars, cold acclimation modulated enzyme properties of PDH and GDH. Increased activities of GS and OT were observed as a result of cold acclimation in both cultivars, with the greatest increase in Kharkov. The apparent energy of activation of these 2 enzymes decreased, particularly for Kharkov, which accumulated proline in cold conditions.     

Involvement of cell-wall invertase in low-temperature hardening of tobacco plants

Specific features of low-temperature hardening (6 days at 8°C) of cold-sensitive tobacco plants (Nicotiana tabacum, cv. Samsun) related to changes in the cell-wall invertase activity were studied. During cold hardening, oppositely directed changes in this enzyme activity occurred in tobacco leaves and roots. In the leaves, cell-wall invertase was activated (approximately by 30%), the content of sugars increased (approximately by 25%), and the content of sucrose, the main transport form of sugars, in the apoplast reduced by three times; all these changes indicate that assimilate outflow from leaves to roots was inhibited. In contrast, in the root system, enzyme activity was decreased almost twice and the content of sugars in them was essentially unchanged. It is suggested that a strategy of low-temperature adaptation of cold-sensitive tobacco plants aimed at creating the high cold tolerance of aboveground parts, even at the expense of the root system, which, under conditions of native vegetation, is not practically exposed to damaging low temperatures.     

Accumulation of Free Proline in Citrus Leaves during Cold Hardening of Young Trees in Controlled Temperature Regimes

Free proline increased in leaves of orange (Citrus sinensis [L.] Osb. cv. Valencia) and grapefruit (Citrus paradisi Macfad. cv. Star Ruby) trees on a wide range of citrus rootstocks during cold hardening. Increases in sugars accompanied proline accumulation. During cold hardening, the rate of proline accumulation was greater in old than in young leaves. In leaves of grapefruit trees kept in the dark during cold hardening, neither proline nor sugars increased and the degree of cold hardiness was less than in trees exposed to light. Like sugar accumulations, proline accumulation does not reflect specific degrees of cold hardiness in citrus cultivars.     

Accumulation of a High Molecular Weight Protein during Cold Hardening of Wheat (Triticum aestivum L.)

Electrophoretic patterns of soluble protein fractions from cold-tolerantwinter wheats (Triticum aestivum L. cv. Frederick and cv. Norstar)and cold-sensitive spring wheat (T. aestiaum L. cv. Glenlea)were analysed in hardened and unhardened plants. One and two-dimensionalgel electrophoresis analysis reveals that cold hardening conditionsinduce changes in the soluble protein patterns. The most importantis the accumulation of a high molecular weight protein in therange of 200 kDa. This protein accumulated at higher concentrationin cold-tolerant cultivars compared to the coldsensitive onesuggesting a correlation between the degree of freezing toleranceand the accumulation of this specific protein. In addition,the intensity of three protein bands (mol wt 48, 47 and 42 kDa)increased while that of five others (mol wt 93, 89, 80, 67 and63 kDa) decreased during hardening. These changes occured inthe three cultivars suggesting that they are part of the metabolicadjustments in response to low temperature rather than a specificchange associated with the development of cold hardiness. (Received April 23, 1987; Accepted June 5, 1987)     

Sucrose and fructan metabolism of different wheat cultivars at chilling temperatures

Four wheat ( Triticum aestivum L.) varieties cultivated in different climates from subtropics to North Patagonia were used to study sucrose and fructan metabolism in plants when submitted to a cold period. Higher levels of sugars were found in the more cold tolerant cultivars. Sucrose synthase (EC 2.4.1.13) and sucrose phosphate synthase (EC 2.4.1.14) activities showed a 2–3 fold increase when plants were grown at 4°C for 10 days. The more cold-tolerant wheat cultivars also showed the higher levels of enzyme activities. These metabolical changes were not due to anatomical or morphological differences produced during growth at 4°C     

Early Developments in RNA, Protein, and Sugar Levels during Cold Stress in Winter Rye (Secale cereale) Leaves

Changes in freezing tolerance of winter rye (Secale cerealeL. cv. Voima) were determined for leaf tissues during a 1-weekcold stress, which was performed by transferring the 7-d-oldseedlings from a greenhouse (25°C, long day) to 3°Cand short day conditions. The development of cold hardeningwas shown by using an ion leakage test and by determining theamounts of carbohydrates, soluble proteins and RNA. The firstevidence of the development of freezing resistance was foundafter 1 d at low temperature, i.e. an LT₅₀ value increased from-5 to -7°C. Plants cold treated for 7 d reached an LT₅₀value of -9°C. This increase in freezing tolerance was foundto be associated with the increased levels of soluble carbohydrates,total RNA and soluble proteins. These metabolic changes indicatethe association with adjustment of growth and cell metabolismto low temperatures at the beginning of cold acclimation ofwinter rye.Copyright 1994, 1999 Academic Press Secale cereale L., winter rye, cold stress, proteins, RNA, sugars     

The effect of hypothermia on the content of 310 kD stress protein in seedlings of winter rye and wheat

The effect of hypothermia on the content of 310 kD stress protein in seedlings of winter rye and wheat was studied by rocket-immunoelectrophoresis and radioactive label. The 1-h low-temperature stress was found to result in an increase in the content of this protein at both above- and below-zero temperatures. It was found that the increase in the relative content of the protein with mol. wt 310 kD, under the effect of short-term low-temperature stress, occurs due to induction of its synthesis. It has been found that during cold hardening of winter wheat the content of this protein decreases up to 64% compared to the control during the first day of hardening but starts to rise by the third day and reaches 179% by the seventh day, which is well correlated with the increase in cold resistance of winter wheat plants during cold hardening.     

不同施肥水平对旱地冬小麦水分利用效率的影响

1987—1988年,研究了旱地施肥对冬小麦(Triticum aestivum cv．Shanhe No．6)水分利用效率的影响,初步探讨了“以肥调水”的生理机制。施肥不仅提高了旱地土壤含水量,更重要的是提高了土壤水势和土壤水的有效性,从而增加了有效水分利用。施肥增大旱地冬小麦绿叶面积,延缓叶片衰老,从而降低土壤蒸发,增加蒸腾用水潜势和光合潜势,但净同化率不一定提高。施肥增加旱地冬小麦总的水分利用(ET,即蒸散量)和蒸腾(T)用水,增加地上部生物产量,提高了经济产量和水分利用效率。施肥使冬小麦同时具有耗水和节水以抵御干旱的能力,对植株具有调节作用,使之更好地适应干旱环境。     

Electron Microscopic Cytochemical Studies on the Glycoproteins in Cellular Membranes of Winter Wheat Seedlings with Concanavalin A-Horseradish Peroxidase Method

The localization and distribution of the glycoproteins in cellular membranes of strong cold hardy winter wheat (Triticum aestivum L. cv. Yanda 1817) seedlings were studied employing the modified cytochemical method using Concanavalin A-Horseradish peroxidase conjugates. The results obtained showed that the reaction products of Horseradish peroxidase activity which indicated the presence of glycoproteins appeared as individual particles distributed at plasmalemma, endoplasmic reticulmn, nuclear envelope and tonoplast in the leaflet cells of the seedlings grown under optimum temperature 25℃ day/20℃ night. After cold acclimation of the seedlings at low temperature during late autumn and early winter, the quantity of glycoproteins distributed in endoplasmic reticulum and nuclear envelope was significantly increased, and glycoproteins were transported into almost all of the plasmodesmata. At some sites of mitochondria and plastides newly formed glycoproteins were observed. However, no distribution of glycoproteins was for,nd in Golgi bodies during the whole observations. The role of glycoproteins for the development and stability of cold resistance in plant through blocking up the plasmodesma passage in the overwintering period of winter wheat seedlings, and the possible differences of the synthetic site and transportation pathway of glycoproteins between animal and plant systems were discussed.