Additions of maize root mucilage to soil changed the structure of the bacterial community

The organic compounds released from roots (rhizodeposits) stimulate the growth of the rhizosphere microbial community. They may be responsible for the differences in the structure of the microbial communities commonly observed between the rhizosphere and the bulk soil. Rhizodeposits consists of a broad range of compounds including root mucilage. The aim of this study was to investigate if additions of maize root mucilage, at a rate of 70 μg C g⁻¹ day⁻¹ for 15 days, to an agricultural soil could affect the structure of the bacterial community. Mucilage additions moderately increased microbial C (+23% increase relative to control), which suggests that the turnover rate of microorganisms consuming this substrate was high. Consistent with this, the number of cultivable bacteria was enhanced by +450%. Catabolic (Biolog^® GN2) and 16S-23S intergenic spacer fingerprints exhibited significant differences between control and mucilage treatments. These data indicate that mucilage can affect both the metabolic and genetic structure of the bacterial community as shown by a greater catabolic potential for carbohydrates. We concluded that mucilage is likely to significantly contribute to differences in the structure of the bacterial communities present in the rhizosphere compared to the bulk soil.     

Impact of artificial root exudates on the bacterial community structure in bulk soil and maize rhizosphere

Bacterial densities, metabolic signatures and genetic structures were evaluated to measure the impact of soil enrichment of soluble organic carbon on the bacterial community structures. The exudates chosen were detected in natural maize exudates (glucose, fructose, saccharose, citric acid, lactic acid, succinic acid, alanine, serine and glutamic acid) and were used at a rate of 100 μg C g⁻¹ day⁻¹ for 14 days. Moreover two synthetic solutions with distinct carbon/nitrogen ratios (20.5 and 40.1), obtained by varying carboxylic and amino acids concentrations, were compared in order to evaluate the potential role of organic N availability. The in vitro experiment consisted of applying exudate solutions to bulk soil. In the case of the control, only distilled water was added. Both solutions significantly increased bacterial densities and modified the oxidation pattern of Biolog® GN2 plates with no effect of the C/N ratio on these two parameters. Genetic structure, measured by means of ribosomal intergenic spacer analysis (RISA), was also consistently modified by the organic amendments. N availability levels led to distinct genetic structures. In a second experiment, one of the previous exudate solutions (C/N 20.5) was applied to 15-day-old maize plants to determine the structural influence of exudates on the rhizosphere microbial community (in situ experiment). Bacterial densities were significantly increased, but to a lesser extent than had been found in the in vitro experiment. Metabolic potentials and RISA profiles were also significantly modified by the organic enrichment.     

微量元素亏缺对人参根系分泌物组成的影响

以3年生人参幼苗为试验材料,在室内用正常以及缺少硼、锰、锌、铁元素的1倍Hoagland营养液进行水培养试验,研究不同微量元素亏缺处理对人参根系分泌物主要成分的影响.结果表明:在包括对照在内的5个处理中分别检测到23～30种化合物,包括醇类、脂肪酸、脂肪酸酯、酚酸及其衍生物、醛类和其他类化合物.进一步研究发现,除20种化合物在上述5种不同处理中均检测到外,与对照相比,在微量元素亏缺胁迫下,人参根系分泌物中其他类化合物有所减少,有机酸及酚酸类化合物增多.试验结果表明,微量元素亏缺对根系分泌物组成有显著影响.     

连作对花生根系分泌物化感作用的影响

采用连续收集法提取连作5 年、3 年和轮作处理的花生结荚期根系分泌物, 研究其对土壤微生物及花生种子发芽、幼苗生长发育和细胞膜过氧化的化感作用及连作对花生根系分泌物化感作用的影响。结果表明,花生结荚期根系分泌物对花生根腐镰刀菌36194 菌丝的生长、叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性及丙二醛(MDA)含量存在促进作用, 对固氮菌14046 的生长, 花生种子胚根的伸长、幼苗的苗高、茎叶鲜重、根系鲜重、叶片叶绿素含量等有抑制作用, 促进和抑制作用均随根系分泌物添加浓度和连作年限的增加呈增强趋势。连作花生结荚期根系分泌物化感物质在土壤中的累积, 很可能是导致花生连作障碍的原因之一。     

土壤水分对花生根际分泌物中低分子量有机酸的释放及根质外体中铁的积累的影响

A three-compartments rhizobox was designed and used to study the low-molecular-weight organic acids in root exudates and the root apoplastic iron of “lime-induced chlorosis“ peanut grown on a clacareous soil in realtion to different soil moistrue conditions.Results showed that chlorosis of peanuts developed under condition of high soil mositure level(250 g kg^-1),while peanuts grew well and chlorosis did not develop when soil moisture was managed to a normal level(150 g kg^-1).The malic acid maleic acid and succinic acid contents of chlorotic peanut increased by 108.723,0.029,and 22.446ug cm^-1 ,respectively,compared with healthy peanuts.The content of citric acid and fumaric acid also increased in root exudates of chlorotic peanuts.On Days 28 and 42 of peanut growth,the accumulation of root apoplastic iron in chlorotic peanuts was higher than that of healthy peanuts.From Day 28 to Day 42,the mobilization percentages of chlorotic peanuts and healthy peanuts to root apoplastic iron were almost the smae,being 52.4% and 52.8%,respectively,indicating that the chlorosis might be caused by the inactivation of iron within peanut plant grown on a calcareous soil under soil moisture conditions.     

Effect of soil drying on mucilage exudation and its water repellency: a new method to collect mucilage

Despite the importance of mucilage for soil–plant relations, little is known about the effect of soil drying on mucilage exudation. We introduce a method to collect mucilage from maize growing in wet and dry soils. Mucilage was collected from brace roots. The amount of mucilage exuded did not change with soil water content and transpiration rate. Mucilage exuded in dry soils had a higher degree of hydrophobicity, suggesting that the wetting properties of mucilage change in response to soil drying.     

Effect of maize root mucilage on phosphate adsorption and exchangeability on a synthetic ferrihydrite

 This study was conducted to determine the influence of root mucilage (RM) from Zea mays, polygalacturonic acid (PGA), and galacturonic acid (GA), on the adsorption, desorption, and exchangeability of orthophosphate (P) on a synthetic ferrihydrite in deionized water. The adsorption rate and affinity on the oxide decreased in the order P>GA>PGA≥RM. The results suggest that P and GA were adsorbed by ligand exchange on Fe-OH surface sites, while RM and PGA might in addition also be adsorbed by hydrogen and van der Waals bonds to the oxide surface. Whereas P and GA could be adsorbed at sites located inside the micropores of ferrihydrite aggregates, PGA and RM probably remained on the external surfaces because of their large molecular weight. The preliminary adsorption of organic compounds decreased the subsequent P adsorption. The larger decrease of P adsorption occurred when dry ferrihydrite powder was directly added to the RM/water suspension. This effect was due to the flocculation of ferrihydrite aggregates, which limited the transport of P to the adsorption sites. Preliminary adsorption of GA also decreased P adsorption, because both compounds competed for the same Fe-OH sites. The ability of the organic compounds to desorb P from the ferrihydrite was not very pronounced and decreased in the order GA>PGA≥RM. The preliminary adsorption of organic compounds on the oxide resulted in an increase in isotopically exchangeable P in 300 min compared to the treatment without organic compounds. However, because of its higher affinity for the oxide surface, P exchangeability was not modified when P was added before the organic compounds. Received: 23 July 1999     

Organic acids in root exudates and soil solution of Norway spruce and silver birch

Here we report on low molecular weight organic acids in root exudates and soil solutions of Norway spruce and silver birch grown in rhizoboxes, sterile microcosms and the field. Monocarboxylic acids dominated in all three experimental systems. Formic, shikimic and oxalic acids were found in both spruce and birch microcosms. Fumaric acid was exclusive for spruce, while lactic, malonic, butyric and phthalic acids were only found in the birch microcosms. In spruce rhizoboxes oxalic, lactic, formic, butyric and pthalic acids were found. In addition, citric, adipic, propionic, succinic and acetic acids were observed in the rhizosphere of birch. Behind root windows in the field, only oxalic and lactic acids were found in the rhizosphere of spruce fine roots, whereas also formic and phthalic were observed close to birch fine roots, all at low concentrations. The rhizosphere of mycorrhizal short roots of birch contained butyric acid along with the acids observed for birch fine roots. Our results emphasise that characteristics of both the trees e.g. species, developmental stage, root density, mycorrhizal status, and the experimental system, i.e. growth conditions are important for the composition and the amount of organic acids. We conclude that the rhizosphere of birch contains more organic acids at higher concentrations than spruce.     

Maize root mucilage decreases adsorption of phosphate on goethite

 This work was carried out to determine the influence of root exudates on phosphorus solubility in the presence of a strong phosphate sorbant. Root mucilage was collected from maize grown under field conditions. Polygalacturonic acid (PGA) was also used as a model of exudates. Adsorption of phosphate by synthetic goethite was measured in the absence of either (1) root mucilage collected from maize in the field, or (2) PGA. The ranking order of the adsorption on goethite was phosphate at low concentration >PGA >mucilage >phosphate at high concentration. Then, competitive adsorption between phosphate ions and PGA or mucilage was studied. Increasing the concentration of PGA or mucilage added to the goethite suspension decreased the amount of phosphate adsorbed on goethite. It is likely that PGA, mucilage and phosphate compete for the same adsorption sites on goethite: the effect of the coating of the mineral by the mucilage is another possible explanation. Received: 18 April 2000     

Pruned tea bushes secrete more root exudates to influence microbiological properties in soil

Pruning is adopted at 3–4 years interval as an agronomic practice during tea cultivation. It was hypothesized that biomass loss during pruning will imply stress on tea bushes. The aim of this study was to quantify changes in different parameters (labile organic carbon fractions, phosphatase activity, microbial biomass and microbial respiration) in tea rhizosphere due to pruning by collecting soil samples from the rhizosphere of ten of each pruned and un-pruned tea bushes. Hot-water extractable and dissolved organic C contents in rhizosphere soil of pruned tea were significantly (P ≤ 0.05) higher than those in the soil of un-pruned tea bushes. Analysis of phospholipid fatty acids (PLFA) revealed that the rhizosphere of pruned tea plants had higher population of Gram (+) and Gram (-) bacteria, fungi, actinomycetes and lower denitrifying bacterial population as compared to un-pruned tea plants. Activity of acid phosphatase enzyme in soil was also increased due to pruning. A separate study revealed that de-centering may induce production of up to 50% more labile organic carbon compounds by young tea as compared to un-pruned plants. Therefore, it could be concluded that pruned tea bushes secrete more root exudates to influence microbiological and biochemical properties in rhizosphere.     

An efficient method for the collection of root mucilage from different plant species—A case study on the effect of mucilage on soil water repellency

Root mucilage may play a prominent role in understanding root water uptake and, thus, there is revived interest in studying the function of root mucilage. However, mucilage research is hampered by the tedious procedures of mucilage collection. We developed a mucilage separator which utilizes low centrifugal forces (570 rpm) to separate the mucilage from seminal roots without the need of handling individual seeds or removing the germinated seeds from the tray/mesh to a centrifuge tube. For the different plant species, between 1 and 3.7 mL tray^?1 of hydrated mucilage could be produced, with 6 trays being handled successively within 45 min. For Triticum aestivum, which showed a dry matter content of 0.5%, this was equivalent to 98.6 mg mucilage dry matter. The lowest total production was found for Zea mays with just 34 mg dry matter. The amounts of mucilage produced normalized to root tip agree well with literature data. The mucilage obtained by the new method was used to measure its effect on repellency of soil as this property directly relates to the phenomenon of lower rhizosphere soil water content during rewetting. It could be shown that repellency of the rhizosphere is affected by the quantity as well as by species‐dependent quality of mucilage in the rhizosphere. Among the species tested (Lupinus albus, Vicia faba, Zea mays, Triticum aestivum), the largest differences were observed between the two legumes. For Zea mays seminal root mucilage obtained with the new system was compared to mucilage of air born brace roots. The differences between these two mucilages, representing different root orders, indicate clearly that there is still a need for methods which enable the investigation of roots from older plants.     

大豆根系分泌物中氨基酸对根腐病菌生长的影响

采用砂培和室内模拟方法,研究了两个抗病性不同的大豆品种水溶性根系分泌物中氨基酸组分随作物生长的变化;同时检测了培养基中添加大豆根系分泌物和纯品氨基酸对大豆根腐病菌菌落生长的影响。结果表明,添加大豆苗期和花荚期根分泌物均显著促进尖镰孢菌菌丝生长,添加成熟期根分泌物显著促进腐皮镰孢菌菌丝生长。易感根腐病大豆品种合丰25号花荚期以后根分泌物中氨基酸种类多于抗根腐病大豆品种绥农10号。感病大豆品种根系分泌的氨基酸总量随生育时期增加,在鼓粒期达到最高;抗病大豆品种根系分泌的氨基酸总量在花荚期最高。感病大豆品种根系分泌的主要氨基酸为精氨酸,抗病大豆品种根系分泌的氨基酸主要为天冬氨酸。氨基酸纯品培养中,添加精氨酸和酪氨酸处理的尖镰孢菌菌落直径显著高于不加氨基酸的对照菌落直径;添加丝氨酸和天冬氨酸的处理菌落直径则显著低于对照处理。同时,添加天冬氨酸的培养基上腐皮镰孢菌菌落直径显著低于不加氨基酸的对照。可见,不同大豆品种根系分泌物中氨基酸组分对病原菌生长起着一定的作用,其表现的作用受根际氨基酸种类和氨基酸浓度影响较大,对于不同病原菌的作用存在差异。     

Effects of root exudates and humic substances on weathering kinetics

The effect of complex natural organic ligands on the weathering kinetics of aluminum oxide was investigated in laboratory experiments. A peat-derived humic substance and root exudates obtained from ectomycorrhizal (Picea abies — Hebeloma crustuliniforme) and non-mycorrhizal Norway Spruce trees; and γ-Al₂O₃ were used as a model system. The experimental weathering rates are in accordance with a surface-controlled dissolution mechanism. The effect of the humic material on dissolution rates appears to depend on the degree of protonation of the humic (macro)molecules: we observed dissolution-enhancement or -inhibition at pH 3 and 4, respectively. Ectomycorrhizal exudates proved to be effective weathering agents at pH 4, as opposed to humic material and non-mycorrhizal exudates. Our results suggest that (i) the role of humic materials in mineral weathering and podzolization is different from what is commonly thought, and (ii) mineral weathering rates in the rhizosphere may be higher than in the bulk soil.     

Effect of root exudates on growth of newly isolated nitrifying bacteria from barley rhizoplane

An ammonia-oxidizing bacterium, strain PJA1, and nitrite-oxidizing bacterium, strain PJN1, were newly isolated from the rhizoplane of barley. The cells of strain PJA1 were lobate, compartmentalized, and showed characteristics of the genus Nitrosolobus. The similarity of the 16S rRNA gene to Nitrosolobus multiformis ATCC25196 was 99.04%. The cells of strain PJN1 were rod-shaped. The similarity of the 16S rRNA gene to Nitrobacter agilis ATCC14123 was 98.57%. These newly isolated bacteria were identified as Nitrosolobus sp. PJA1 and Nitrobacter sp. PJN1, respectively. The exudates prepared from barley roots of both the pre-heading and post-heading stages promoted the growth of strain PJA1 (15–20% increase). The growth of the non-rhizoplane strains like Nitrosomonas europaea ATCC25978 was remarkably inhibited by the exudates. The growth of strain PJN1 was also remarkably promoted by the root exudates at the pre-heading stage (6 time increase), and by the root exudates at the post-heading stage (2.5 time increase). The root exudates did not effect the growth of Nitrobacter winogradskyi IFO14297, isolated from the non-rhizoplane.     

Effect of zinc deficiency in wheat on the release of zinc and iron mobilizing root exudates

The effect of Zn deficiency in wheat (Triticum aestivum L. cv. Ares) on the release of Zn mobilizing root exudates was studied in nutrient solution. Compared to Zn sufficient plants, Zn deficient plants had higher root and lower shoot dry weights. After visual Zn deficiency symptoms in leaves appeared (15–17 day old plants) there was a severalfold increase in the release of root exudates efficient at mobilizing Zn from either a selective cation exchanger (Zn-chelite) or a calcareous soil. The release of these root exudates by Zn deficient plants followed a distinct diurnal rhythm with a maximum between 2 and 8 h after the onset of light. Re-supply of Zn to deficient plants depressed the release of Zn mobilizing root exudates within 12 h to about 50%-, and after 72 h to the level of the control plants (Zn sufficient plants). The root exudates of Zn deficient wheat plants were equally effective at mobilizing Fe from freshly precipitated Fe^III hydroxide as Zn from Zn-chelite. Furthermore, root exudates from Fe deficient wheat plants mobilized Zn from Zn-chelite, as well as Fe from Fe^III hydroxide. Purification of the root exudates and identification by HPLC indicated that under Zn as well as under Fe deficiency, wheat roots of the cv. Ares released the phytosiderophore 2′-deoxymugineic acid. Additional experiments with barley (Hordeum vulgare L. cv. Europa) showed that in this species another phytosiderophore (epi-3-hydroxymugineic acid) was released under both Zn and Fe deficiencies. These results demonstrate that the enhanced release of phytosiderophores by roots of grasses is not a response mechanism specific for Fe deficiency, but also occurs under Zn deficiency. The ecological relevance of enhanced release of phytosiderophore also under Zn deficiency is discussed.     

Effect of soil structure on soil water status

Although the need for research into soil structural features responsible for water loss from soil is recognised, the research has been hampered by the lack of suitable techniques for measuring in situ soil macrostructure. Therefore, in a field experiment, structural parameters obtained from eight differently tilled (loam) soils in winter and spring were correlated with gravimetric soil water content (at 5 and 10-cm depths) and evaporative water loss during daylight hours. Percentage of voids and aggregates of different sizes, mean aggregate size, mean void size and macroporosity percentage were all calculated from structural data obtained from sectioned blocks of soil that were impregnated with paraffin wax in the field.Significant negative correlations were observed between soil water content, and percentage of 4–8 and 8–16-nm voids, mean aggregate size and macroporosity percentage, especially in winter when there are long periods without precipitation. However, inverse relationships were detected between evaporative water loss during daylight hours on selected days and mean aggregate size, mean void size and macroporosity percentage. But evaporative water loss during daylight hours apparently did not influence water content of bare soil with coarse structure at the surface.     

根系分泌物与重金属的化学行为研究

通过毛细管电泳测定根系分泌物的组成发现,根系在受到重金属铅、镉或铅-镉共同作用下分泌物种类与对照都有不同程度的变化;透析袋平衡透析方法则证实了根系分泌物与重金属之间存在着络合反应,络合能力的大小与重金属本身的性质有关,小麦根系分泌物对铅的络合能力大于镉。溶解试验表明,小麦分泌物对红壤中铅、镉的溶出没有显著的影响。小麦无菌根系分泌物、非无菌分泌物、非无菌过0.45m分泌物均促进红壤对铅、镉的吸附。其中铅吸附量较对照分别增加了12%,16%,21%;镉吸附量较对照分别增加了66%,102%,72%。     

黄瓜砧木对南方根结线虫生长发育的影响及其与根系分泌物的关系

黄瓜是设施主栽蔬菜,长期连作导致根结线虫病发生和危害加重;嫁接是提高黄瓜对根结线虫抗性的有效手段,但不同砧木品种间存在差异。为探明嫁接提高黄瓜抗根结线虫能力的机制,试验选用不同抗性的砧木品种‘云南黑籽南瓜’(低感)和‘甜砧南瓜’(高感),以‘新泰密刺’自根黄瓜为对照,采用人工接种,研究了黄瓜砧木对南方根结线虫生长和繁殖的影响,并探讨了根系分泌物与线虫卵孵化的关系。结果表明:线虫侵染初期,3个品种根内的二龄幼虫(J2)数差异显著,‘云南黑籽南瓜’最少,‘甜砧南瓜’次之,‘新泰密刺’最多;侵入根内的J2均能正常发育为成虫,根内线虫总数以‘云南黑籽南瓜’最少,‘新泰密刺’最多;根际J2数量以‘新泰密刺’最多,‘云南黑籽南瓜’最少;发生二次侵染前,根际卵粒数呈现出相同的特点。黄瓜及其砧木根系分泌物明显影响根结线虫卵的孵化,孵化率以‘云南黑籽南瓜’最低,‘甜砧南瓜’次之,‘新泰密刺’最高;接种根结线虫后,根系分泌物影响下的卵孵化率较未接种时明显增加,‘新泰密刺’增幅最大,‘云南黑籽南瓜’增幅最小;随着生育时间延长,三者根系分泌物对卵孵化的促进作用逐渐增强。低感砧木根系分泌物对南方根结线虫卵孵化具有明显的抑制作用,可显著降低线虫在其根内及根际的发育,可能是抗线虫的机制之一。     

Effect of nitrogen forms on reduction of manganese oxides in an Oxisol by plant root exudates

Reductive dissolution of soil manganese (Mn) oxides increases potential toxicity of Mn²⁺ to plants. In order to examine the effect of nitrogen forms on reduction of Mn oxides in rhizosphere soil, a rhizobox experiment was employed to investigate the reduction of Mn oxides due to the growth of soybean and maize in an Oxisol with various contents of NO₃^–-N and NH₄⁺-N and a total N of 200 mg kg^?1. The results showed that exchangeable Mn²⁺ in rhizosphere soil was 9.6–32.7 mg kg^?1 higher than that in bulk soil after cultivation of soybean and maize for 80 days, which suggested that plant root exudates increased reduction of soil Mn oxides. Application of ammonium-N promoted reduction of Mn oxides in rhizosphere soil compared to application of nitrate and nitrate together with ammonium. Soybean cultivation led to a higher reduction in soil Mn oxides than maize cultivation. Application of single ammonium enhanced Mn uptake by the plants and led to more Mn accumulating in plant leaves, especially for soybean. Therefore, application of ammonium-based fertilizer can promote reduction of soil Mn oxides, while application of nitrate-based fertilizer can inhibit reduction of soil Mn oxides and thus reduce Mn²⁺ toxicity to plants.     

Metabolic profiling of root exudates of Arabidopsis thaliana

In addition to accumulating biologically active chemicals, plant roots continuously produce and secrete compounds into their immediate rhizosphere. However, the mechanisms that drive and regulate root secretion of secondary metabolites are not fully understood. To enlighten two neglected areas of root biology, root secretion and secondary metabolism, an in vitro system implementing root-specific elicitation over a 48-day time course was developed. After roots of Arabidopsis thaliana had been elicited with salicylic acid, jasmonic acid, chitosan, and two fungal cell wall elicitors, the secondary metabolites subsequently secreted were profiled. High-performance liquid chromatography was used to metabolically profile compounds in the root exudates, and 289 possible secondary metabolites were quantified. The chemical structures of 10 compounds were further characterized by (1)H and (13)C NMR: butanoic acid, trans-cinnamic acid, o-coumaric acid, p-coumaric acid, ferulic acid, p-hydroxybenzamide, methyl p-hydroxybenzoate, 3-indolepropanoic acid, syringic acid, and vanillic acid. Several of these compounds exhibited a wide range of antimicrobial activity against both soil-borne bacteria and fungi at the concentration detected in the root exudates.