Detection of the regions of phytosiderophore release from barley roots

Abstract

We devised a method to detect regions of phytosiderophore release from barley (Hordeum vulgare cv. Minorimugi) roots. Plants were grown in Fe-sufficient (+Fe) or Fe-deficient (?-Fe) water cultures for 14 d in a phytotron. Intact or excised roots were sampled and put between two sheets of filter papers just after the onset of the light period. The filter papers with roots were wrapped with vinyl film to avoid drying out and covered with aluminum foil to shade. The roots between the filter paper were kept in the phytotron for 4 h and phytosiderophores (PS) released from roots were absorbed by the filter paper. Then, the shape of the roots was preserved by photocopy and PS released on the filter paper were visualized by the method employed for detecting PS on thin layer chromatography (TLC). Gelatinous Fe solubilizing activity was employed for detecting PS on the filter paper. Released PS were detected as white spots on the orange-colored background and the regions of PS release from the roots were visible on the filter paper. It was evident that the apical zones of roots were the main regions of PS release. It was apparent that a distinct primary root newly formed on the basal parts had higher activity to release PS than the other roots of ?Fe plants. It was also shown that apical root zones of +Fe plants released PS.     

Incorporation of 14C into methionine as an intermediate for phytosiderophore biosynthesis in barley

In order to verify the precursory role of methionine (Met) in the biosynthesis of the mugineic acid family of phytosiderophores (MAs), feeding experiments of ¹⁴C-Iabeled compounds to barley (Hordeum vulgare L. cv. Minorimugi) roots grown hydroponically were conducted. When both ^l4C-Glucose (Glc) and unlabeled Met were fed to segmented roots, ¹⁴C was incorporated into Met and MAs in the roots. Molar-radioactivity of Met was higher than that of the amino-butanoic-acid unit in MAs in the roots. When ^l4C-Glc and unlabeled homoserine (Hse) were fed to decapitated roots, l4C was incorporated into Met but not into Hse. Therefore, it was considered that Hse might not be a major precursor of MAs. In addition, ¹⁴C was incorporated into Met and MAs in the roots when both ¹⁴C-glycerol (Gol) and unlabeled Met were fed to segmented roots. It is suggested that MAs may be synthesized from Glc via Met, bypassing Hse, and that the MAs biosynthesis may involve an unknown pathway associated with Gol and leading to Met.     

抗氧化水平对小麦幼苗耐缺铁及铁累积能力的影响

以小麦品种"石4185"和"沧6001"为试验材料,采用水培方法,分析了抗氧化水平对小麦幼苗耐缺Fe和Fe累积能力的影响。结果表明:Fe缺乏引起小麦根部活性氧(ROS)累积,导致根部产生过氧化伤害,同时降低叶片中叶绿素含量。与"石4185"相比,"沧6001"具有相对较低的ROS水平和较高的叶绿素含量。缺Fe条件下,"沧6001"比"石4185"诱导出更多的根毛和侧根,并具有较高Fe累积。抗氧化酶活性分析结果表明,缺Fe条件下,"沧6001"比"石4185"具有更高的SOD、CAT和APX活性。在缺Fe条件下添加抗氧化剂NAC或AsA提高了幼苗中的Fe含量,表明抗氧化水平的提高促进了小麦幼苗对Fe的吸收和累积。本研究对于理解小麦对缺Fe胁迫响应的生理机制及筛选耐Fe缺乏和高Fe累积的小麦品种具有指导意义。     

Diurnal variations in absorption and translocation of a ferrated phytosiderophore in barley as affected by iron deficiency

The release of phytosiderophore (PS) from roots of Fe-deficient graminaceous plants follows a distinct diurnal rhythm with maximum release rates occurring usually 3 to 4 hours after the onset of light. However, it remains to be determined whether absorption of the PS-Fe³⁺ complex shows a diurnal rhythmicity similar to that of PS release, Barley plants grown with or without 10 µM FeEDTA for 7 days were fed with ferreted PS (10 µM labelled with ⁵⁹Fe) at 4-h intervals to study the diurnal variations in the absorption and transloca tion of ⁵⁹Fe, The absorption of ⁵⁹Fe, irrespective of the Fe nutritional status of the plants, was higher during the day and lower during the night but did not show any peak throughout the day-night cycle. On the other hand, the translocation of ⁵⁹Fe into shoots of Fe-deficient plants was lower than that of Fe-sufficient plants, while the Fe nutritional status of the plants did not affect the absorption of ⁵⁹Fe by roots, The formation of root apoplastic ⁵⁹Fe was lower during the day and higher during the night, regardless of the Fe nutritional status of plants. Our results showed that the absorption of the PS-Fe³⁺ complex by roots did not follow the PS release pattern.     

Crop growth and nitrogen transformations in wheat (Triticum aestivum L.) planted after wetland rice (Oryza sativa L.)

Summary A field study was undertaken to examine the effects of various management strategies on wheat (Triticum aestivum L.) performance and N cycling in an intensively cropped soil. Microplots receiving 100 kg N ha^–1 as¹⁵NH₄ ^{+ 15}NO₃ ^– at sowing, tillering or stem elongation were compared with unfertilized microplots. Stubble from the previous rice crop was either incorporated, burnt without tillage, burnt then tilled or retained on the surface of untilled soil. Wheat grain yield ranged from 1.5 to 5.1 t ha^– and was closely related to N uptake. Plant accumulation of soil N averaged 36 kg N ha^–1 (LSD 5% = 10) on stubble-incorporation plots and 54 kg N ha^–1 on stubble-retention plots. Fertilizer N accumulation averaged 18 kg N ha^–1 (LSD 51% = 6) on stubble-incorporation plots and 50 kg N ha^–1 on stubble-retention plots. Tillage had little effect on burnt plots. Delaying N application from sowing until stem elongation increased average fertilizer N uptake from 26 to 39 kg N ha^–1 (LSD 5% = 6), but reduced soil N uptake from 50 to 37 kg N ha^– (LSD 5% = 10).Immobilization and leaching did not vary greatly between treatments and approximately one-third of the fertilizer was immobilized. Less than 1% of the fertilizer was found below a depth of 300 mm. Incorporating 9 t ha^–1 of rice stubble 13 days before wheat sowing reduced net apparent mineralization of native soil N from 37 to 3 kg ha^–1 between tillering and maturity. It also increased apparent denitrification of fertilizer N from an average 34 to 53 kg N ha^–1 (LSD 5% = 6). N loss occurred over several months, suggesting that denitrification was maintained by continued release of metabolizable carbohydrate from the decaying rice stubble. The results demonstrate that no-till systems increase crop yield and use of both fertilizer and soil N in intensive rice-based rotations.     

Comparison of protein quality and mineral element concentrations in grain of spelt (Triticum spelta L.) and common wheat (Triticum aestivum L.)

The cultivation of Triticum spelta (spelt) has no tradition in Hungary. In recent years the interest towards this old species renewed in many countries. This high‐nutritional cereal, which has a high ash and fibre content, can be used in many health‐oriented grain‐based food products. Therefore, field experiments have been conducted for some years to test the performance of this species under home growing conditions. Here we report the results of analyses for some important quality parameters of grain samples from the 1996/97 season in comparison with those of older and new home‐grown bread wheat cultivars. Three common wheat cultivars and one advanced spelt line were grown on small plots fertilised with an NPK dose necessary to reach the highest yield and quality. Spikes were sampled weekly from the time of 70–77% grain moisture to full ripening. The grains were analysed for ash, N, P and K content and amino acid composition. Concentrations of 16 other macro + micro elements and in the ripe grains, baking quality parameters were also assessed. The grain development of spelt showed a remarkable time‐lag compared to that of the common wheat cultivars. However, the highest thousand‐grain‐masses, ash, N, and P concentrations were measured in this cultivar after milk ripening. The grains of spelt contained the macro‐nutrient Mg and four micro‐nutrients (Zn, Mn, Fe, Cu) in higher concentrations compared to those of the common wheat varieties. The total and essential amino acid concentrations measured in the ripe grains of spelt were also remarkably higher.

Although its wet gluten content (47.5%) was considerably higher than that of the bread wheat cultivars, its breadmaking quality was poor.     

Changes in bacterial populations along roots of wheat (Triticum aestivum L.) seedlings

Summary In this study the bacterial populations on root tips (1–2 days old) of wheat (Triticum aestivum L.) were compared with the populations on root segments about 1 week older (root base). The isolates were characterized with a set of physiological tests and the test results were used to group the bacteria by means of cluster analysis. Some clusters contained bacteria that occurred mainly on the root tips and were characterized by the ability to produce acid from different sugars and by the presence of the enzymes nitrate reductase, lipase, and oxidase; they were sensitive to high salt concentrations in the media. Another cluster included significantly more isolates from the root-base segments; these bacteria were characterized by a negative reaction to most of the physiological tests; the colonies formed by these bacteria had yellow pigmentation. Possiblemechanisms for the changes in the bacterial populations are discussed.     

气候变暖对春小麦籽粒痕量元素利用率的影响

为了回答温度升高是否会改变痕量元素在士壤中的溶解性,以及作物对痕量元素的生物利用率,在人工气候室模拟未来气候变化温度升高背景下,对种植在不同温度处理的3种春小麦,测定了籽粒中Cd,Cu,Fe和Zn含量.结果发现温度升高引起了土壤中Cd,Cu,Fe和Zn溶解性的显著变化,也显著地影响小麦籽粒中Cd,Cu,Fe和Zn的生物利用率.最高升温3℃处理使西旱1号、2号和3号小麦籽粒中Cd浓度相比对照分别下降43.4％、11.1％和13.4％,Cu浓度相比对照处理分别下降了30.4％、25.1％和10.8％.但Fe和Zn的情况却不同,1℃和2℃升温处理使西旱1号籽粒中Zn浓度比对照处理分别增加了28.9％和35.8％.根据未来气候变化两北地区温度升高1.9℃,估计到2050年,小麦籽粒中的质量分数范围分别在Cd(0.59-0.65)、Cu(5.91-7.64)、Zn(63.73-69.41)和Fe(185.23-202.70) mg/kg.     

Water stress effects on biochemical traits and antioxidant activities of wheat (Triticum aestivum L.) under In vitro conditions

Water stress is one of the major environmental stresses that affect agricultural production worldwide, especially in arid and semi-arid regions. This research investigated the effect of water deficit, induced by PEG-6000 on wheat genotypes (GA-2002, Chakwal-97, Uqab-2000, Chakwal-50 and Wafaq-2001) grown in modified MS medium solution. Osmotic stress caused a more pronounced inhibition in leaf relative water content and leaf membrane stability more sensitive (index in Wafaq-2001 and Uqab-2000) genotypes compared with the tolerant (Chakwal-50, GA-2002 and Chakwal-97) genotypes. Upon dehydration, an incline in proline, total soluble sugar, total soluble protein, superoxide dismutase, peroxidase, catalase and malondialdehyde activity content were evident in all genotypes, especially at osmotic stress of ?8 bars. The observed data showed that status of biochemical attributes and antioxidant enzymes could provide a meaningful tool for depicting drought tolerance of wheat genotypes. The present study shows that genotypic differences in drought tolerance could be likely attributed to the ability of wheat plants to induce antioxidant defense under drought conditions. In order to develop genotypes with stable, higher yields in dry farming conditions, it is necessary to characterise genetic resources based on drought adaptation, determine suitable genotypes, and then use them in breeding programmes.     

纳米氧化铜对小麦根系生理生化行为的影响

为阐明纳米金属材料暴露对植物生长的影响及其作用机制,采用模拟土壤的琼脂培养方法研究了纳米氧化铜(CuO NPs)对小麦(Triticum aestivum L.)根伸长及相关生理生化行为的影响。结果表明:小麦根伸长与CuO NPs暴露浓度之间存在指数相关关系,在低浓度CuO NPs(10 mg/L)暴露下得到一定程度促进,而在高浓度(100 mg/L)下受到强烈抑制。小麦根内超氧化物歧化酶(SOD)活性随CuO NPs暴露浓度的变化趋势与小麦根伸长具有一致性。另外,在1~100 mg/L范围内,随着CuO NPs暴露浓度的升高,小麦根内丙二醛(MDA)含量不断增加,但植物蛋白含量急剧降低。以上结果说明CuO NPs对根伸长抑制主要是由于纳米材料暴露造成植物细胞膜氧化损伤,小麦能通过提高根系活力对CuO NPs暴露作出适应性应激响应以减少纳米材料毒性的伤害。     

Investigating environmental factors that cause extreme gluten quality deficiency in winter wheat (Triticum aestivum L.)

Unstable breadmaking quality of wheat due to environmental influence has been a problem for Norwegian milling industries. Large variation in gluten quality was observed from field trials with Norwegian winter wheat conducted in several locations between 2005 and 2013. Moreover, extremely poor gluten quality was observed in several locations in the 2007 and 2011 season, and indicated almost complete loss of breadmaking quality. To investigate the environmental factors which cause extremely weak gluten, gluten proteins were characterized in samples selected within the 2011 season. The results revealed that the proportion of large glutenin polymers decreased in wheat samples with extremely weak gluten. Moreover, re-polymerization of large glutenin polymers, which normally occur during the resting period of a dough, did not take place in gluten prepared from these samples. Incubation of total proteins extracted from these samples in an in vitro system showed a drastic degradation of gluten proteins indicating protease activities. The origin of the proteases remains unclear; however, exogenous proteases derived from Fusarium spp. seem to play a key role for protein degradation, and thus causing severe quality deficiency. A genotypic difference was found between the two cultivars and one of them had higher resistance against the factors influencing gluten quality in negative way.     

小麦TaWRKY51基因的克隆、表达分析和转基因功能鉴定

WRKY是植物中特有的锌指型转录因子,其广泛参与植物对生物及非生物胁迫的响应过程.本研究从小麦(Triticum aestivum L.)中分离出一个新的WRKY转录因子基因TaWRKY51,其全长cDNA序列长度为1295 bp,其中开放阅读框(ORF)为942 bp,编码一个由313个氨基酸组成的多肽.用半定量RT-PCR进行表达谱分析,结果显示,TaWRKY51基因在分蘖节、叶和根系中的表达水平较高,并且受干旱胁迫诱导上调表达.在拟南芥(Arabidopsis thaliana)中过量表达TaWRKY51基因导致转基因株系侧根数目明显增多,并且对ABA、干旱和盐等胁迫处理的敏感性增加,表明该基因可能在植物响应非生物逆境胁迫信号传导过程中起负调控作用.本研究有助于揭示TaWRKY51基因调控植物侧根发育及响应非生物逆境胁迫的分子机制.     

Copper induced iron deficiency in wheat (Triticum aestivum L)

Abstract

A pot experiment was conducted to study the interaction effects of phosphorus and copper on wheat. The soils used were calcareous loamy sand (ls) and non calcareous sandy loam (sl). Four levels of Cu (0, 5, 10 and 20?mg Cu kg^?1 soil) and six levels of P (0, 25, 50, 100, 200 and 400?mg P kg^?1 soil) were applied in all possible combinations with three replications. Soil pH decreased with Cu application while Olsen P increased with P application in both soils. Growth and yield of wheat improved significantly with graded levels of applied P. However, when any level of P was combined with 20?mg Cu kg^?1 soil, severe iron chlorosis of leaves, a drastic reduction in growth and chlorophyll content was observed in calcareous ls only. The results indicated that it was Cu and not P that induced Fe deficiency in wheat grown in alkaline calcareous soil and the Cu requirement of the crop seemed to be much lower in the calcareous ls. Root dry matter, grain and straw yield decreased with increasing levels of applied Cu in ls but in sl maximum increase of 62.5, 74.3 and 63.7 per cent in root, grain and straw yield was observed with a combined application of 400?mg P and 5?mg Cu kg^?1 soil over control. Accumulation of Cu in roots decreased the Fe absorption by roots which indicated that Fe chlorosis of wheat leaves is expected when Cu: Fe concentration ratio in root is > 0.30.     

Priming effects of 15N-labelled ammonium nitrate on uptake of soil N by wheat (Triticum aestivum L.) under field conditions

Summary The uptake of labelled and unlabelled N by wheat was measured in a field experiment using ¹⁵N-labelled ammonium nitrate fertilizer. The dry matter yield and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The uptake of applied N by wheat ranged between 25 and 34%. Fertilizer N application increased the uptake of unlabelled soil N which was attributed to a positive priming effect or added N interaction. The added N interaction observed by applying 20, 60, and 120 kg fertilizer N was 11.4, 19.1, and 27.9 kg, corresponding to 26, 44 and 64%, respectively of the N taken up from unfertilized soil. The A values did not alter with the increase in fertilizer N application. The observed added N interaction may have been the result of pool substitution whereby added labelled fertilizer N stood proxy for unlabelled soil N. A significant correlation coefficient (r=0.996^**) between the uptake of soil N and the dry matter yield showed that soil N was more important than fertilizer N in wheat production.     

Fluctuations in bacterial populations on the root surface of wheat (Triticum aestivum L.) grown under different soil conditions

Summary Populations of several bacterial groups on the root surface of wheat and in root-free soil were investigated in volcanic ash soil and non-volcanic ash soil throughout a series of predetermined intervals. Over time, the populations changed similarly both on the root surface and in root-free soil. The numbers of total bacteria, fluorescent Pseudomonas spp., phosphate-solubilizing bacteria, and NH _inf+ ^sup4 -oxidizing bacteria, were consistently lower in the plots with volcanic ash soil than with nonvolcanic ash soil, but the numbers of cellulose-decomposing bacteria were opposite to those of the other groups. Superphosphate application improved the growth of wheat in the volvanic ash soil. It did not, however, bring about any significant changes in the bacterial populations among the volcanic ash soils supplemented with three different levels of superphosphate, though there were some variations with plant age.     

Molecular characterization of salinity tolerance in wheat (Triticum aestivum L.)

Random amplified polymorphic DNA (RAPD) markers were used to estimate the genetical variability of three salt-resistant genotypes, SARC-1, SARC-5 and S-24, exposed to saline environment. High-yielding and salt-sensitive variety MH-97 was used as standard for comparison. The behavior of these genotypes under saline environment was analyzed by using the hydroponics screening methods at the seedling stage. One hundred and fifty primers were tested of which 52 primers revealed differences between SARC-1 and SARC-5, 54 revealed differences between SARC-1 and S-24 and 61 revealed differences between SARC-5 and S-24. Polymorphism differences between MH-97 and SARC-1, MH-97 and SARC-5 and MH-97 and S-24 were 53%, 64% and 42%, respectively. Four primer pairs amplified special fragments, which were located in all the three salt-resistant genotypes but none on the salt-sensitive genotype MH-97. Primer GLD-15 (5?-CCGTGGCATT-3?) generated a prominent fragment of length 1460 bp; primer GLF-18 (5?-ACCCGGAACC-3?) produced a fragment of length nearly 980 bp in the salt-resistant genotype; the primer pair GLE-5 (5?-TTCAAGCCCG-3?) located one polymorphic amplified band of 1290 bp and the primer GLH-9 (5?-ATCCAGGTCA-3?) performed as a weak polymorphic band of 640 bp, respectively.     

Potential for nitrate reduction in wheat (triticum aestivum L.)

Following the prevalent agronomic practice of applying N fertilizer in two splits at optimum levels recommended for maximum yield viz. 120 kg N ha, to two wheat (Triticum aestlvum L.) cultivars, which differ in in vivo nitrate reductase (NR) activity, it was observed that the activity is high in the first formed leaf blades and declines in the successively formed ones. Enhancement in the activity subsequent to incubation of excised leaf blades in NO₃ ^‐ suggests that the substrate (NO^‐ ₃) is limiting and that the leaf blades, particularly the upper ones, have the potential to reduce additional amounts of NO₃ ^‐. High NR cultivar has greater potential than the low NR cultivar. The studies suggest that it may be possible to increase the NO₃ ^‐ moles reduced and thus enhance the reduced N content in case the nitrogen is available at later stages of growth.     

Physiological basis of iron chlorosis tolerance in rice (Oryza sativa) in relation to the root exudation capacity

Micronutrient deficiency in cultivable soil, particularly that of iron (Fe) and zinc (Zn), is a major productivity constraint in the world. Low Fe availability due to the low solubility of the oxidized ferric forms is a challenge. An experiment was, thus, executed to assess the performance of eight genetically diverse rice genotypes on Fe-sufficient (100 µM) and Fe-deficient (1 µM) nutrient solution, and their ability to recover from Fe deficiency was measured. Fe efficiency under Fe deficiency in terms of biomass production showed a significant positive correlation with the root release of phytosiderophore (PS) (R² = 0.62*). This study shows that the Fe deficiency tolerance of Pusa 33 was related to both a high release of PS by the root and an efficient translocation of Fe from the root to the shoot as the Fe–PS complex, which could be useful for improving the Fe nutrition of rice particularly under aerobic conditions.