Localisation of phosphomonoesterase activity in ectomycorrhizal fungi grown on different phosphorus sources

Phosphorus (P) is a major limiting nutrient for plants in boreal forest ecosystems where a substantial part of the total P is sequestered in organic compounds. Some ectomycorrhizal (ECM) fungi are known to produce phosphomonoesterases, enzymes that degrade organic P sources. Here, we test 16 ECM species for this enzymatic activity by growing them on media containing orthophosphate, phytic acid or apatite. A method with an overlay gel that determined both phosphomonoesterase activity and its spatial distribution was developed. The phosphomonoesterase activity was not significantly higher when growing on organic P; conversely some isolates only produced measurable enzyme activity when grown on apatite. Species-specific variations with respect to phosphomonoesterase activity as well as growth responses to different substrates were found. The production of phosphomonoesterases was found to be widespread in ECM fungi and the enzyme activity did not need induction by organic P. The enzyme activity was highest in the central parts of the mycelia, potentially reflecting breakdown and recycling of phospholipids from old hyphae or potentially higher mycelial density.     

氮添加对毛竹林土壤酸性磷酸单酯酶动力学参数的影响

土壤磷酸酶在有机磷矿化和磷循环过程中发挥着重要作用,然而,土壤磷酸酶响应氮(N)沉降的动力学机制仍不清楚。本研究在亚热带毛竹林中设置对照(0)、20(低氮)、40(中氮)和80 g N·hm^-2·a^-1(高氮)4种不同氮添加处理,在氮添加满3年、5年和7年时采集0～15 cm土层土壤样本,测定了土壤化学性质、微生物生物量,并分析了酸性磷酸单酯酶(ACP)的最大反应速率(V_m)、半饱和常数(K_m)和催化效率(K_a)。结果表明: 氮添加显著降低了土壤可溶性有机碳、有效磷和有机磷含量,显著增加了土壤铵态氮、硝态氮含量和V_m,且V_m与有效磷、有机磷和可溶性有机碳含量存在显著相关关系;总体上,氮添加显著提高了K_a;除了在氮添加满5年时高氮处理下K_m显著高于对照外,氮添加对K_m无显著影响,且K_m与有效磷和有机磷含量有显著负相关关系。中、高氮处理对ACP动力学参数的影响大于低氮处理。方差分解分析表明,土壤化学性质的变化而非微生物学性质的变化主导了V_m(47%)和K_m(33%)的变化。总之,氮添加显著影响了毛竹林土壤的基质有效性,通过调控ACP动力学参数(尤其是V_m)进而影响了土壤磷循环。本研究有助于了解氮素富集下土壤微生物调节土壤磷循环的潜在机制,并为全球变化下土壤磷循环模型优化提供重要参数。     

间伐和凋落物处理对华北落叶松人工林土壤磷形态的影响

土壤磷在维持生态系统功能稳定性中发挥重要作用,研究间伐和凋落物处理下的土壤磷组分特征及转化机理,对森林生态系统磷素管理和可持续发展具有重要意义。采用Tiessen改良的Hedley分级方法,探究了不同间伐强度(未间伐、轻度间伐、中度间伐、重度间伐)和凋落物处理(对照、加倍、去凋、切根去凋)下土壤磷形态的变化特征及其驱动因子。结果显示:随着间伐强度的增大,土壤活性磷(Resin-Pi、NaHCO_3-Pi和NaHCO_3-Po)、土壤微生物量磷和酸性磷酸酶活性呈先增加后降低的趋势,且在中度间伐最高。凋落物加倍(DL)显著增加了土壤活性磷(Resin-Pi、NaHCO_3-Pi和NaHCO_3-Po)、土壤微生物量磷和酸性磷酸酶活性。稳定态磷(HCl-Pi、浓HCl-Pi和浓HCl-Po)、残留态磷(Residual-P)不受间伐和凋落物处理的影响。冗余分析(RDA)显示,土壤微生物量磷、酸性磷酸酶活性和土壤有机碳是引起华北落叶松人工林表层土壤磷组分变化的重要因子。研究表明,适度的间伐和增加凋落物能够显著提高华北落叶松人工林表层土壤磷素的活化能力。本研究为华北落叶松人工林的可持续经营提供依据。     

Acid phosphomonoesterase and phytase activities of wheat (Triticum aestivum L.) roots and utilization of organic phosphorus substrates by seedlings grown in sterile culture

Wheat seedlings exhibited a differential ability to utilize P from a range of organic P substrates when grown in agar culture under sterile conditions. Plants showed limited ability to obtain P from inositol hexaphosphate (IHP), whereas other monoester substrates such as glucose 1‐phosphate (G1P), were equivalent sources of P for plant growth as compared with inorganic phosphate (P_i). Poor utilization of IHP was exemplified by significantly lower rates of dry matter accumulation and reduced P content of tissues, which were generally not significantly different to control plants that were grown in the absence of added P. The inability of wheat seedlings to obtain P from IHP was not associated with poor substrate availability but was due to either insufficient root phytase activity or inappropriate localization of phytase within root tissues. Phytase activities of 4 and 24 mU g ^{? 1} root fresh weight (FW) were determined for crude root extracts prepared from plants that were grown with either adequate P or under deficient conditions, respectively. Similar levels of phytase activity (approximately 12 mU g ^{? 1} FW) were observed in assays using intact roots, although no secreted activity was detected. By comparison, a secreted acid phosphomonoesterase activity was observed, and activities of between 466 and 1029 mU phosphomonoesterase g ^{? 1} root FW were measured for intact roots. On the basis of the differences in enzyme activity, and the observed differences in the ability of wheat seedlings to utilize G1P and IHP, it is evident that low intrinsic levels of phytase activity in wheat roots is a critical factor that limits the ability of wheat to obtain P from phytate when supplied in agar under non‐limiting conditions. This hypothesis was further supported by the observation that the ability of wheat to obtain P from IHP was significantly improved when the seedlings were inoculated with a soil bacterium (Pseudomonas sp. strain CCAR59) that possesses phytase activity.     

中亚热带森林转换对土壤磷积累的影响

磷是植物生长的必需元素之一,是维持亚热带森林生态系统生产力的关键因子。研究森林转换后土壤因素对磷素的影响,对生态系统的稳定和森林经营具有重要意义。选取由亚热带常绿阔叶林转换而成的米槠次生林(SF)、米槠人促林(AR)和杉木人工林(CF)为研究对象,测定了土壤理化性质、铁铝氧化物、各形态磷含量以及酸性磷酸酶活性,旨在探究土壤磷对森林转换的响应和驱动土壤磷变化的影响因子。结果显示:米槠人促林土壤的全磷、有机磷和微生物生物量磷显著高于米槠次生林和杉木人工林;冗余分析(RDA)发现,土壤含水量、总氮和无定型铁是影响淋溶层土壤磷的主要因子,而在淀积层,则是酸性磷酸酶、游离型铁和总氮起主要作用;土壤生物化学属性和微生物特性都会影响着不同形态土壤P的积累,其中土壤中的水分和酸性磷酸酶活性是调控土壤磷的关键因子。研究表明,中亚热带地区天然林转换为人促更新林更有利于森林土壤磷的储存和供应,有助于维持本区域森林生态系统的稳定。     

Barley genotypes differ in activity of soluble extracellular phosphatase and depletion of organic phosphorus in the rhizosphere soil

In the present investigation we studied the extent of variation among barley genotypes (Hordeum vulgare L. cv. Alexis, Canut, Digger, Etna, Peel) in their ability: i) to induce activity of soluble extracellular phosphatase in rhizosphere soil. ii) to withdraw bicarbonate extractable organic phosphorus (NaHCO₃-P₀). All the genotypes induced 3–4 times higher phosphatase activities in rhizosphere soil as compared to bulk soil. Among the genotypes, there were significant (p>0.01) differences in soluble extracellular and non-soluble phosphatase activities and depletion of NaHCO₃-P₀ in soil near their root mats. Etna induced highest phosphatase activities and depleted most NaHCO₃-P₀ from the rhizosphere soil. A high correlation (r=0.79) was found between the activity of soluble extracellular phosphatase and the quantity of NaHCO₃-P₀ withdrawn from the rhizosphere soil by the barley genotypes.     

施用生物炭基肥对喀斯特石灰土磷元素特性的影响

以贵州省喀斯特山地石灰土为研究对象,采用盆栽试验方法,研究施用生物炭(稻壳炭)、猪粪堆肥和NPK肥3种肥料制成的生物炭基肥,测定土壤中不同形态磷含量、碱性磷酸酶活性及刺槐幼苗生物量。试验共设10个处理,分别为CK、M、F、MF、RH1MF、RH2MF、RH4MF、RH8MF、RH4M、RH4F(其中CK代表对照,M代表堆肥,F代表化肥,RH代表稻壳炭,数字代表生物炭按炭土质量比计算在生物炭基肥中的配比)。结果表明,施用生物炭基肥可显著提高喀斯特石灰土中总磷、有效磷、有机磷、微生物量磷含量及刺槐幼苗生物量,生物炭高施用量下处理(RH8MF)的效果更好,且4种不同形态的磷含量相互之间呈极显著正相关(P0.01);生物炭中等用量配比下生物炭基肥处理(RH2MF、RH4MF)的土壤碱性磷酸酶活性最高,分别比CK提高82.7%、63.4%。综上所述,施用生物炭基肥,尤其在生物炭较高施用量下,可以显著改善喀斯特石灰土中磷素含量,可改善石灰土中磷素含量较低状况,提高喀斯特山地人工幼林地生态恢复的成效。     

不同林分土壤磷形态与磷酸酶特征

以湖南省平江县国有芦头林场的次生林以及经人工翻垦种植的油茶(Camellia oleifera)、黄桃(Amygdalus persica)、杨梅(Myrica rubra)和杉木(Cunninghamia lanceolata)四种人工林为研究对象,比较了不同林分土壤理化性质、磷酸酶活性与磷形态特征,分析了三者之间的相关性,探讨了次生林转变为人工林后,土壤磷形态和磷酸酶的变化特征以及驱动土壤磷素形态变化的关键因子。结果表明：(1)次生林土壤有机碳(SOC),全氮(TN)、铵态氮(NH⁺₄-N)含量与磷酸酶活性显著高于其他四种林分。(2)五种林分中土壤残余磷(Residual-P)含量最高,是林地土壤主要的磷素存在形态。林分转变后,黄桃林与杉木林树脂提取态无机磷(Resin-Pi)显著增加,黄桃林与油茶林NaHCO₃提取态磷(NaHCO₃-Pi、NaHCO₃-Po)含量显著增加,而四种人工林的NaOH提取态有机磷(NaOH-Po)含量均显著降低。可利用磷、中等可利用磷与稳定态磷...     

磷肥输入对稻田土壤剖面胶体磷含量的影响

磷肥管理与土壤磷素赋存息息相关,选取杭嘉湖地区典型稻田定位实验,研究磷肥输入下稻田土壤剖面胶体磷含量的变化,分析不同施肥管理下胶体磷在土壤中的分布规律。结果表明,(1)胶体磷是磷素在土壤中赋存的重要形式,能够占到土壤胶体溶液(1μm)总磷的85%以上,土壤全磷的0.1%—2%;(2)磷肥施用增加了土壤胶体磷含量,特别是在有机肥处理下0—5 cm土壤胶体磷含量达到了8.0 mg/kg;(3)随着土壤深度的增加,胶体磷含量减少,有机肥输入下对深层土壤胶体磷含量的影响较无机肥明显;(4)水稻收割后表层土壤胶体磷含量与油菜收割后相比减少明显,0—5 cm土壤胶体磷含量减少了90%左右,但有机肥施用下胶体磷含量减少较小;(5)水稻收割后30—60 cm土壤胶体磷含量有所增加,可能与胶体磷发生的纵向迁移有关。了解磷肥输入对稻田土壤剖面胶体磷含量的影响,为研究土壤磷素赋存形态及迁移变化提供科学依据,对有效评估胶体磷的环境风险起到指导作用。     

Availability of phosphorus and sulfur of insecticide origin by fungi

Thirteen fungal species isolated from soil treated with pesticides were tested for their ability to mineralize and degrade three organophosphate insecticides currently used in Egypt (Cyolan®, Malathion® and Dursban®) in liquid media free from phosporus (P) and sulfur (S). All fungal species grew successfully on the culture media treated with the three used doses of insecticides (10, 50 and 100 ppm active ingredient) but the growth rate varied with the species, the insecticide and the doses. At 10 ppm level, insecticide degradation expressed in term of organic P mineralization (calculated as % of applied P) was the highest with all fungi tested. Organic P mineralization from pesticides was decreased by increasing the dose used to 50 and 100 ppm. The highest amount of P mineralized was observed with Cyolan® followed by Malathion® whilst P mineralization from Dursban® proceeded very slowly. Aspergillus terreus showed the greatest potential to mineralize organic P followed by A. tamarii, A. niger, Trichoderma harzianum and Penicillium brevicompactum whilst the remaining fungi only moderately mineralized the organic P component of the insecticides tested. Organic sulfur mineralization by the used fungal species paralleled, to some extent, organic P mineralization. The extracellular protein content of culture filtrates in the presence of various doses of insecticides was also decreased by increasing insecticide concentrations. The extracellular protein was significantly correlated with P and S mineralization (r = 0.89** and 0.64**, respectively) whilst correlation with cell dry mass was not significant (r = 0.03 and 0.003) suggesting a direct relationship between pesticide degradation and microbial protein production. The addition of P or S to the growth media enhanced extracellular protein excretion, and increased organic P and S mineralization by the most potent species tested (A. niger, A. tamarii, A. terreus and T. harzianum). This increment was significant in most cases, especially at the higher application rates. The relationship between extracellular protein excretion and organic P and S mineralization from insecticides was highly significant with the addition of inorganic phosphorus (r = 0.96** and 0.83**, respectively) or sulfur (r = 0.85** and 0.89**, respectively) to the growth media.     

菌根真菌磷酸酶活性对红三叶草生境中土壤有机磷亏缺的影响

利用PVC分室培养装置研究了菌根际和菌丝际磷酸酶活性变化与土壤有机磷亏缺间的关系,结果表明,施用有机磷（植酸钠）能促进菌根根系侵染、提高土壤磷酸酶尤其是酸性磷酸酶的活性,使菌丝际范围变宽。菌丝际的存在使土壤有机磷亏缺范围加大,与非菌根植物相比,由于菌根真菌的作用,植物能更容易地从有机磷中获得磷营养以满足植物生长的需要,从而使其干物重和磷吸收量更高。     

不同间伐强度下华北落叶松人工林土壤磷组分特征及其影响因素

间伐是人工林经营的重要措施,磷是森林生态系统的限制性养分元素之一,但间伐对土壤磷素的影响尚不明确.本研究采用Tissen改良的Hedley磷分级体系对土壤样品进行连续浸提,研究不同间伐强度[对照,未间伐;轻度间伐,15%;中度间伐,35%;重度间伐,50%]下山西太岳山好地方林场华北落叶松人工林表层土壤(0～10 cm)磷组分特征及其影响因素.结果表明: 与对照相比,中度间伐显著增强了土壤总无机磷含量;轻度和中度间伐显著增强了Resin-Pi、NaHCO₃-Pi、NaHCO₃-Po、NaOH-Pi、微生物生物量磷和土壤酸性磷酸酶活性,显著降低了NaOH-Po含量;间伐对土壤全磷、总有机磷、稳定态磷(HCl-Pi、浓HCl-Pi和浓HCl-Po)和闭蓄态磷(Residual-P)的影响不显著.土壤有机碳含量、含水率、土壤微生物和酸性磷酸酶活性是影响土壤磷素有效性的重要因子.中度间伐能提高华北落叶松人工林土壤磷素的有效性.     

土壤解磷微生物促进植物磷素吸收策略研究进展

随着全球磷肥需求增加和磷矿资源储量短缺矛盾逐渐加重,土壤解磷微生物在促进磷循环方面的重要性日益突显,因此有必要对其种类和促进植物磷素吸收策略进行全面梳理总结。荟萃分析了土壤解磷微生物种类并构建了系统进化树,重点论述了土壤解磷微生物促进植物磷素吸收主要策略。土壤解磷微生物主要通过矿化和溶解作用直接活化难溶性磷,但其也能与植物根系互作间接活化磷素。其间接途径主要包括解磷微生物与根系分泌物和丛枝菌根真菌互作,它们通过碳磷交换间接活化磷素;其次包括一些解磷微生物可以通过固氮作用使植物生长受到磷的限制,从而调节植物磷酸盐转运系统间接活化土壤磷;解磷微生物也能通过分泌植物激素和生物防治剂促进植物根系生长间接促进植物磷素吸收。解磷微生物还能通过磷素固定减少磷流失,也可以通过加速自身磷素周转促进植物磷素吸收。对完善和发展解磷微生物主导的土壤磷循环和植物磷素吸收利用理论体系具有重要意义。     

Interactions among nitrogen fixation and soil phosphorus acquisition strategies in lowland tropical rain forests

Paradoxically, symbiotic dinitrogen (N₂) fixers are abundant in nitrogen (N)‐rich, phosphorus (P)‐poor lowland tropical rain forests. One hypothesis to explain this pattern states that N₂ fixers have an advantage in acquiring soil P by producing more N‐rich enzymes (phosphatases) that mineralise organic P than non‐N₂ fixers. We assessed soil and root phosphatase activity between fixers and non‐fixers in two lowland tropical rain forest sites, but also addressed the hypothesis that arbuscular mycorrhizal (AM) colonisation (another P acquisition strategy) is greater on fixers than non‐fixers. Root phosphatase activity and AM colonisation were higher for fixers than non‐fixers, and strong correlations between AM colonisation and N₂ fixation at both sites suggest that the N–P interactions mediated by fixers may generally apply across tropical forests. We suggest that phosphatase enzymes and AM fungi enhance the capacity of N₂ fixers to acquire soil P, thus contributing to their high abundance in tropical forests.     

模拟氮沉降对低磷胁迫下马尾松不同家系根系分泌和磷效率的影响

近年来大气氮(N)沉降的增加, 导致森林土壤中有效N含量增加、N:P发生改变, 研究N沉降对低磷(P)胁迫下林木根系分泌和P效率的影响具有重要意义。该文以马尾松(Pinus massoniana)家系作为试验材料, 设置模拟N沉降与同质低P (介质表层与深层均缺P)、异质低P (介质表层P丰富、深层缺P)耦合的二年生盆栽实验, 系统研究了模拟N沉降对低P胁迫下马尾松根系分泌性酸性磷酸酶(APase)活性、有机酸分泌以及P效率的影响。结果表明: (1)同质低P和异质低P下, 模拟N沉降均显著提高了植株N:P化学计量比、增加了P素的相对匮乏程度, 从而诱导根系增加了APase和有机酸的分泌, 而同质低P比异质低P下增加幅度更大, 其中有机酸分泌均与马尾松生长呈正相关关系, 而APase活性与P效率相关性较小; (2)同质低P下, N沉降虽然增加了根系分泌, 但未提高马尾松P素吸收和生长量, 其原因在于, 同质低P下植株N:P过高, 因而植株对N沉降敏感性低; 在异质低P下, 植株表现为N、P共同限制, 因而对N敏感性较高, N沉降增加了根系分泌, 同时提高了N和P吸收效率、增加了生物量; (3)马尾松根系分泌对模拟N沉降的响应存在较大的家系差异。同质低P下, 家系71×20的有机酸分泌和生物量对N沉降的响应幅度较大; 异质低P下, 家系36×29、71×20和73×23对N沉降的响应幅度较大。     

有机酸对土壤无机态磷转化和速效磷的影响

土壤有效磷含量低是影响作物生产的重要限制因素之一.作物根分泌活化难溶性磷的有机酸对改善其磷素营养具有重要意义.采用张守敬和Jackson无机磷分级方法,以湖北省3种pH值土壤为材料,加入不同磷源和有机酸,经过室温培养后,测定速效磷含量和无机磷组分的变化.结果表明:施磷显著提高了土壤中速效磷含量,中性土、酸性土Fe-P和Al-P含量大幅上升, Fe-P占增加量的50%以上,而碱性土Ca-P含量显著增加.加施有机酸使中性土速效磷含量增多,除苹果酸处理的变幅较小外,草酸和柠檬酸的加入速效磷显著增加.由于有机酸的作用,中性土Al-P含量下降,Ca-P含量上升,变幅大小依次为草酸＞柠檬酸＞苹果酸;酸性土中Al-P含量呈下降趋势,碱性土中Ca-P含量有不同程度的减少,3种土壤中O-P含量均有所增加.说明有机酸活化的磷主要来源于中性土和酸性土Al-P、Fe-P及碱性土Ca-P中的磷,同时有机酸能够促进土壤中闭蓄态磷(O-P)的形成与积累.     

低分子量有机酸对红壤无机态磷转化及酸度的影响

以鄂南、赣北两红壤样品为材料,加入不同有机酸并经室温培养后,测定不同P组分、pH及活性Al含量的变化。结果表明,供试有机酸均使土壤Ca2－P含量增高,增幅大小依次为柠檬酸＞苹果酸＞琥珀酸＞乙酸;2种土壤的Ca8－P和Ca10－P含量无明显变化规律,Fe-P、Al-P和O－P含量有所下降,除乙酸处理的土壤pH值无显著变化外,其它有机酸的加入使pH下降0．65－1．96;有机酸引起活性Al量增多,除乙酸处理的变化较小外,其它有机酸或混合物的加入使土壤中0．02mol.L^-1CaCl2提取Al增加4．7－50．3倍,1mol.L^-1提取Al增加4．0－67．3倍。可见,有机酸具有双重作用,既增加P的有效性,又增加土壤酸度和Al毒。     

Partitioning of soil phosphorus among arbuscular and ectomycorrhizal trees in tropical and subtropical forests

Partitioning of soil phosphorus (P) pools has been proposed as a key mechanism maintaining plant diversity, but experimental support is lacking. Here, we provided different chemical forms of P to 15 tree species with contrasting root symbiotic relationships to investigate plant P acquisition in both tropical and subtropical forests. Both ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) trees responded positively to addition of inorganic P, but strikingly, ECM trees acquired more P from a complex organic form (phytic acid). Most ECM tree species and all AM tree species also showed some capacity to take up simple organic P (monophosphate). Mycorrhizal colonisation was negatively correlated with soil extractable P concentration, suggesting that mycorrhizal fungi may regulate organic P acquisition among tree species. Our results support the hypothesis that ECM and AM plants partition soil P sources, which may play an ecologically important role in promoting species coexistence in tropical and subtropical forests.