Organic phosphorus compounds as a phosphorus source for higher plants through the activity of phosphatases produced by plant roots and microorganisms

Summary The efficiency of phosphatases produced by clover, barley, oats and wheat was investigated in soils treated with sodium glycerophosphate, lecithin and phytin. Root exudates of aseptically grown clover were also examined for the breakdown of different organic P compounds in order to test the efficiency of plant-produced phosphatases. In general, the plants were able to use P from all the organic sources used in the study almost as efficiently as inorganic sources. Dry-matter yield, P uptake, acid and alkaline phosphatase activity and microbial population were increased in all the P treatments. Organic P enhanced alkaline phosphatase activity. Lecithin increased fungal, and phytin bacterial growth. There was no alkaline phosphatase activity in the asepticallly grown clover root exudates. Phosphatase released in aseptic culture after 4 weeks of clover growth was able to efficiently hydrolyse sodium glycerophosphate, lecithin and phytin. The amount of organic P hydrolysed in this and in the soil experiment surpassed plant uptake by a factor of 20. This suggests that the limiting factor on plant utilization of organic P is the availability of hydrolysable organic P sources.     

Influence of organic and inorganic phosphorus supply on the maximum secretion of acid phosphatase by plants

Three cereals (wheat, pearl millet, sorghum), three legumes (mung bean, moth bean, clusterbean) and three oil seed crop species (groundnut, sesame, mustard) were grown in solution culture under conditions of P deficiency and with additions of inorganic P or organic P (phytin, lecithin, glycerophosphates). The plants started secreting acid phosphatase as soon as their roots emerged (24-96 h). The activities of the secreted acid phosphatases increased with plant age and were at a maximum under P-deficient conditions. Phytin increased acid phosphatase secretion to a greater extent than lecithin and glycerophosphate. The increased activities of acid phosphatase secreted under P-deficient conditions when compared with those under P-sufficient (inorganic P) conditions ranged from 2.7 times (glycerophosphate) to 5 times (negligible P, phytin) higher. Legumes secreted the maximum amount of acid phosphatase within 2 weeks after germination, then activity decreased gradually. Acid phosphatase secretion by oilseeds started at a low level and increased with the age of the plants. Only a small increase in acid phosphatase secretion by cereals was observed during the first 3 weeks of growth. The amount of acid phosphatase secreted by legumes was 22% higher than by oilseeds and 72% higher than by cereals. The results showed that an organic P concentration of 250 mg L^-1 and above, and an inorganic P concentration <50 mg L^-1 provide the most suitable conditions for plants to secrete a maximum amount of acid phosphatase.     

Relative efficiency of fungal intra‐ and extracellular phosphatases and phytase

A comparison of intra‐ and extracellular acid phosphatase, alkaline phosphatase and phytase activity in six fungi is reported. A strong linear relationship between intra versus extracellular fungal acid phosphatase (R² = 0.94), alkaline phosphatase (R² = 0.96), and phytase (R² = 0.97) is observed. Three‐fourth of acid phosphatase were generally present inside the fungal cells and only 25 % were released extracellularly after a three weeks period. Phytase shows the reverse trend where thirty nine times higher extracellular phytase activity was noticed than present inside the fungal cells. The extracellular enzymes are found 60 % more efficient in the hydrolysis of phytin than their intracellular counterpart but they are at par in the hydrolysis of glycerophosphate. The results clearly demonstrated that phytase types of phosphatases mostly occur outside the fungal cells whereas most of the acid phosphatase and alkaline phosphatase are located inside the cells.<?show $6#>     

Phytase and phosphatase producing fungi in arid and semi-arid soils and their efficiency in hydrolyzing different organic P compounds

Seven most efficient phytase and phosphatases producing fungi were isolated from the soils of arid and semi-arid regions of India and tested for their efficiency on hydrolysis of two important organic P compounds: phytin and glycerophosphate. The native soil organic P may be exploited after using these organisms as seed inoculants, to help attain higher P nutrition of plants. The identified organisms belong to the three genera: Aspergillus, Emmericella and Penicillium. Penicillium rubrum released the most acid into the medium during growth. Aspergillus niger isolates were found to accumulate biomass the fastest. A significant negative correlation (r=−0.593,n=21, p<0.01) was observed between the development of fungal mat and pH of the media. The extracellular (E) phosphatases released by different fungi were less than their intracellular (I) counterpart, but the trend was reversed in case of phytase production. The E:I ratio of different fungi ranged from 0.39 to 0.86 for acid phosphatase, 0.29 to 0.41 for alkaline phosphatases and 9.4 to 19.9 for phytase. The efficiency of hydrolysis of different organic P compounds of different fungi varied from 2.12-4.85 μg min⁻¹ g⁻¹ for glycerophosphate to 0.92-2.10 μg min⁻¹ g⁻¹ for phytin. The trend of efficiency was as follows: Aspergillus sp.>Emmericella sp.>Penicillium sp. The results indicated that the identified fungi have enough potential to exploit native organic phosphorus to benefit plant nutrition.     

Efficiency of Bacillus coagulans as P biofertilizer to mobilize native soil organic and poorly soluble phosphates and increase crop yield

Bacillus coagulans, a phosphatase- and phytase-producing bacterium was isolated and tested under greenhouse conditions and in the field in a loamy sand soil. Bacterial population build-up and efficiency was compared under sterilized and non-sterilized soil conditions. Exploitation of plant unavailable (poorly soluble) P was higher in sterilized soil, mainly due to an increased bacteria population. A gradual increase in microbial build-up of up to 21 times the inoculated population was observed over a 4-week period under the sterilized soil condition. Clusterbean influenced acid phosphatase and phytase activity. The depletion of organic P was much higher than the depletion of mineral and phytin P. The microbial contribution to the hydrolysis of the different P fractions was significantly higher than the plant contribution. The maximum effect of inoculation on different enzyme activities (acid phosphatase, alkaline phosphatase, phytase and dehydrogenase) was observed in pants between 5 and 8 weeks of age. A significant improvement in plant biomass (25%), root length (28%), plant P concentration (22%), seed (19%) and straw yield (28%) resulted from inoculation. The results suggested that B. coagulans produces phosphatases and phytase, which mobilized P from unavailable native P sources and enhanced the production of clusterbean.     

根际pH对玉米利用磷酸单酯和双酯盐的影响

【目的】土壤有机磷在土壤全磷中占有很大比重,是植物潜在的有效磷源,但必须通过磷酸酶的水解作用释放出无机磷才能被植物利用。土壤中有机磷的主要形式为磷酸单酯和磷酸双酯。本研究中,我们探讨了无菌条件下不同形态的氮源引起的根际pH变化如何影响植物对这两种有机磷的活化利用。【方法】采用琼脂无菌培养体系种植玉米,向玉米植株供应两种形态的氮源和磷源, 氮源为硝态氮和铵态氮,磷源为植酸钙和卵磷脂,植酸钙属于磷酸单酯盐,卵磷脂属于磷酸双酯盐。不同的供氮形态会导致根际pH变化,进而研究根际pH变化对磷酸单酯盐和磷酸双酯盐的活化利用所产生的影响。【结果】当给玉米供应铵态氮时,根际pH从5.5降至4.0; 供应硝态氮时,根际pH升至6.6。测定玉米根际的琼脂中根系分泌的磷酸单酯酶和磷酸双酯酶活性发现,磷酸单酯酶活性在pH 6.0~7.0之间最高,磷酸双酯酶活性在pH 7.0~8.0之间达到最高。无论以植酸钙还是卵磷脂为有机磷源,相对于铵态氮处理,硝态氮处理能够使根际保持较高的磷酸单酯酶或磷酸双酯酶活性。有机磷的水解过程由磷酸酶活性和底物有效性两个因素控制,而植酸钙的水解受根际pH影响很大,在一定pH范围内,植酸钙的溶解度随根际pH值降低而升高,有效态磷浓度的增加,使得磷酸酶的底物有效性提高。在供应铵态氮时,根际pH值降低,玉米对植酸钙的利用效率高于硝态氮处理,尽管供硝态氮时磷酸单酯酶活性更高。同时,在供应铵态氮条件下,植株对植酸钙的利用率要显著高于卵磷脂,原因在于虽然磷酸双酯酶和磷酸单酯酶活性较低,由于植酸钙的溶解度较大,它的底物有效性更高。因此,植酸钙处理中植株的磷含量更高。相反,在供应硝态氮时,植酸钙溶解度减小而两种磷酸酶活性较高,卵磷脂处理中植株的磷含量更高。【结论】土壤中有机磷的水解过程由磷酸酶活性和有机磷底物有效性两个因素控制,酶活性与根际pH密切相关。本研究说明土壤有机磷的活化必须首先转化为溶解于水溶液中的状态,才能作为磷酸酶的底物被催化水解。我国长期施用化肥导致北方土壤大范围酸化,这种酸化无疑对土壤固有或随有机物料进入农田的有机磷的活化利用是具有重要贡献的,应该在北方土壤养分管理中应加以考虑。     

Phosphate solubilizing microorganisms isolated from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome

Many soil microorganisms are able to transform insoluble forms of phosphorus to an accessible soluble form, contributing to plant nutrition as plant growth-promoting microorganisms (PGPM). The objective of this work was to isolate, screen and evaluate the phosphate solubilization activity of microorganisms in maize rhizosphere soil to manage soil microbial communities and to select potential microbial inoculants. Forty-five of the best isolates from 371 colonies were isolated from rhizosphere soil of maize grown in an oxisol of the Cerrado Biome with P deficiency. These microorganisms were selected based on the solubilization efficiency of inorganic and organic phosphate sources in a modified Pikovskaya's liquid medium culture containing sodium phytate (phytic acid), soybean lecithin, aluminum phosphate (AlPO₄), and tricalcium phosphate (Ca₃(PO₄)₂). The isolates were identified based on nucleotide sequence data from the 16S ribosomal DNA (rDNA) for bacteria and actinobacteria and internal transcribed spacer (ITS) rDNA for fungi. Bacteria produced the greatest solubilization in medium containing tricalcium phosphate. Strains B17 and B5, identified as Bacillus sp. and Burkholderia sp., respectively, were the most effective, mobilizing 67% and 58.5% of the total P (Ca₃(PO₄)₂) after 10 days, and were isolated from the rhizosphere of the P efficient L3 maize genotype, under P stress. The fungal population was the most effective in solubilizing P sources of aluminum, phytate, and lecithin. A greater diversity of P-solubilizing microorganisms was observed in the rhizosphere of the P efficient maize genotypes suggesting that the P efficiency in these cultivars may be related to the potential to enhance microbial interactions of P-solubilizing microorganisms.     

Arbuscular mycorrhizal fungal hyphae mediating acidification can promote phytate mineralization in the hyphosphere of maize (Zea mays L.)

Mycorrhizal and non-mycorrhizal maize (Zea mays L.) plants were grown in two-compartment rhizoboxes, in order to study the effect of mycorrhizal hyphae-mediated acidification on organic P mineralization in the hyphosphere. The soil in the two compartments was supplemented with either KNO₃ or (NH₄)₂SO₄, and phytin (0 or 75 mg P kg⁻¹) was added to the hyphal compartment. P content in the shoots was significantly higher for the NH₄⁺ treatment than for the NO₃⁻ treatment, but only in the combined presence of phytin and AM fungal mycelium (Rhizophagus intraradices). NH₄⁺ treatment under these conditions also led to a decrease in hyphosphere pH, enhanced phosphatase activity in the hyphosphere and accelerated mineralization of phytin compared to the NO₃⁻ treatment. The results show that hyphosphere acidification induced by absorption of NH₄⁺ by the AM fungal mycelium leads to an increase in phosphatase activity, and consequently enhances mineralization of phytin and improves maize uptake of P from phytin-P.     

Kinetic characterization of O-phospho-L-tyrosine phosphohydrolase activity of two fungal phytases

Fungal phytases belonging to "histidine acid phosphatase" or HAP class of phosphohydrolases that catalyze the hydrolysis of phytic acid could also hydrolyze O-phospho-L-tyrosine, which is also called phosphotyrosine. Two phytases from Aspergillus niger and Aspergillus awamori with pH optima 2.5 were tested for phosphotyrosine hydrolase activity; both enzymes cleaved the phosphomonoester bond of phosphotyrosine efficiently at acidic pH. The Km for phosphotyrosine ranged from 465 to 590 microM as opposed to 135 to 160 microM for phytate. The Vmax, however, is 2-4 times higher for phosphotyrosine than it is for phytate. The catalytic efficiency of phytase for phosphotyrosine is on the same order as it is for phytate (3.5 x 10(6) to 1.6 x 10(7) M(-1) s(-1)); the pH versus activity profile for phosphotyrosine is, however, different from what it is for phytate. The temperature optima shifted 5 degrees C higher to 70 degrees C when phosphotyrosine was used as the substrate. Taken together, the kinetic data show that fungal HAPs that are known as PhyB are capable of cleaving the phosphomonoester bond in phosphotyrosine. This is the first time that phosphotyrosine phosphatase (PTPase) activity has been reported for the subgroup of HAP known as phytase.     

不同种类解磷微生物的溶磷效果及其磷酸酶活性的变化

比较了不同种类的微生物菌株对不同种类难溶性磷酸盐及磷矿粉的溶解能力。结果发现,细菌、酵母、霉菌在解磷方面均有一定作用,发挥着不同优势。磷酸钙、磷酸铝、磷酸铁等难溶性磷酸盐容易被酵母菌、霉菌溶解,而磷矿粉容易被巨大芽孢杆菌溶解,显示不同微生物与不同磷源的亲和溶解能力不同。不同种类磷酸盐或磷矿粉对微生物磷酸酶活力的影响不同,贫磷条件可以促进酸性和碱性磷酸酶活性的增加     

土壤磷酸酶活性对施肥-种植-耕作制度的响应

植物所需的磷素营养,主要来源于土壤内源、外源无机磷。而大量存在的有机磷类化合物在土壤磷酸酶的作用下催化水解为无机磷,不仅是土壤内源无机磷的主要来源,也是土壤磷素循环的一个重要的环节。土壤磷酸酶在土壤中主要以吸附态的形式存在,其活性受到施肥、种植和耕作制度的影响。本文对国内外不同施肥方式、种植及耕作制度下土壤磷酸酶(酸性磷酸酶、中性磷酸酶、碱性磷酸酶、磷酸二酯酶、磷酸三酯酶、焦磷酸酶)活性响应进行综述,拟为不同农业管理制度下土壤磷酸酶活性的调节提供依据。     

Characterization of extracellular phosphatase activities in periphytic biofilm from paddy field

Periphytic biofilms exist widely in paddy fields, but their influences on the hydrolysis of organic phosphorus(P) have rarely been investigated. In this study,a periphytic biofilm was incubated in a paddy soil solution, and hydrolysis kinetic parameters(half-saturation constant(Km) and maximum catalytic reaction rate(Vmax)), optimal environmental conditions, substrate specificity, and response to different P regimes of the phosphatase activities in the periphytic biofilm were determined, in order to characterize extracellular phosphatase activities in periphytic biofilms from paddy fields. The results indicated that the periphytic biofilm could produce an acid phosphomonoesterase(PMEase), an alkaline PMEases, and a phosphodiesterase(PDEase). These three phosphatases displayed high substrate affinity, with Km values ranging from 141.03 to 212.96 μmol L^-1. The Vmax/Km ratios for the phosphatases followed the order of alkaline PMEase > acid PMEase > PDEase, which suggested that the PMEases, especially the alkaline PMEase, had higher catalytic efficiency. The optimal pH was 6.0 for the acid PMEase activity and 8.0 for the PDEase activity, and the alkaline PMEase activity increased with a pH increase from 7.0 to 12.0. The optimal temperature was 50℃ for the PMEases and 60℃ for the PDEase. The phosphatases showed high catalytic efficiency for condensed P over a wide pH range and for orthophosphate monoesters at pH 11.0, except for inositol hexakisphosphate at pH 6.0. The inorganic P supply was the main factor in the regulation of phosphatase activities. These findings demonstrated that the periphytic biofilm tested had high hydrolysis capacity for organic and condensed P,especially under P-limited conditions.     

山西矿区复垦土壤中解磷细菌的筛选及鉴定

【目的】矿区复垦土壤贫瘠、 有效磷含量低。解磷细菌能够将有机磷和难溶性无机磷转化为可溶性磷,促进植物对磷素的利用。因此筛选和鉴定具有解磷能力的菌株,可为解决矿区生态恢复使用的微生物肥料提供菌种资源。【方法】采用平板分离法初筛菌株,得到D/d1.5的菌株,然后以磷酸钙为磷源,通过液体发酵试验复筛菌株,挑选出解磷率高于巨大芽孢杆菌(Bacillus megaterium)As1.223的菌株。以磷矿粉和卵磷脂为磷源,液体发酵试验测定菌株的解磷能力及磷酸酶活性。进行菌株的生长试验以测定菌株温度适宜性、 耐盐性及耐酸碱性。通过形态学、 基因序列分析及脂肪酸组成分析综合进行菌株鉴定。 菌落形态观察用营养琼脂平板培养基培养;菌体形态即细胞形态及其大小采用扫描电镜观察;基因序列分析采用16S rDNA序列测定,基因在线比对采用EzTaxon数据库;使用美国MIDI公司的Sherolock全自动细菌鉴定系统对菌株进行脂肪酸组成分析。【结果】利用无机磷和有机磷平板培养基,从山西省矿区复垦区土壤样品中筛选出19株解磷微生物,其中D/d1.5的有7株。在以磷酸钙为磷源的液体培养试验中,4株菌的解磷率高于巨大芽孢杆菌As1.223,解磷率为7.89%～12.61%,最高的为菌株Y14。4株菌对磷矿粉的解磷率为0.81%～1.21%,最高的为菌株Y14。在以卵磷脂为磷源的液体培养试验中,4株菌的解磷率与酸性磷酸酶活性分别为1.79%～3.07%和24.3～28.4U/L,均高于巨大芽孢杆菌As1.223; 碱性磷酸酶活性为11.9～50.2U/L;菌株Y14的解磷率与磷酸酶活性均最高。4株菌均有较强的环境适应能力,以Y14的适应性最强。H22、 Y11和Y34与假单胞菌属(Pseudomonas sp.)同源性在99%以上,Y14与泛菌属(Pantoea sp.)有99.79%的同源性; H22、 Y11和Y34的细胞脂肪酸组成特征峰与假单胞菌属(Pseudomonas sp.)相一致,Y14与泛菌属(Pantoea sp.)相一致;H22、 Y11和Y34被鉴定为假单胞菌(Pseudomonas sp.),Y14为泛菌属(Pantoea sp.)。【结论】分离、 筛选到4株高效解磷菌,对于磷酸钙和卵磷脂的解磷率均高于巨大芽孢杆菌As1.223。4株菌分别隶属于假单胞菌属(Pseudomonas sp.)和泛菌属(Pantoea sp.)。菌株Y14无机磷与有机磷平板的D/d值分别为3.28与1.59,降解磷酸钙、 磷矿粉、 卵磷脂的解磷率分别为12.61%、 1.21%、 3.07%,酸性与碱性磷酸酶活性分别为28.4 U/L和50.2 U/L,均为4株菌里最高的,且环境适应能力最强,生长温度为20～60℃,能耐受pH 4～11的酸碱梯度和2%～7%的盐分梯度,Y14被鉴定为泛菌属(Pantoea sp.)。4株菌均具有良好的解磷能力及较强的环境适应能力,可望进一步研发成为微生物肥料生产菌种。综合D/d值、 解磷率、 磷酸酶活性和生长试验,本试验最终确定适合山西矿区复垦农田推广的高效解磷菌菌株为Y14。     

可利用的碳和氮对丛枝菌根真菌萌发孢子氨基酸代谢的影响

The effects of carbon (C) and nitrogen (N) sources on N utilization and biosynthesis of amino acids were examined in the germinating spores of the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith after exposure to various N substrates,CO2,glucose,and/or root exudates.The N uptake and de novo biosynthesis of amino acids were analyzed using stable isotopic labeling with mass spectrometric detection.High-performance liquid chromatography-based analysis was used to measure amino acid levels.In the absence of exogenous N sources and in the presence of 25 mL L-1 CO2,the germinating AM fungal spores utilized internal N storage as well as C skeletons derived from the degradation of storage lipids to biosynthesize the free amino acids,in which serine and glycine were produced predominantly.The concentrations of internal amino acids increased gradually as the germination time increased from 0 to 1 or 2 weeks.However,asparagine and glutamine declined to the low levels;both degraded to provide the biosynthesis of other amino acids with C and N donors.The availability of exogenous inorganic N (ammonium and nitrate) and organic N (urea,arginine,and glutamine) to the AM fungal spores using only CO2 for germination generated more than 5 times more internal free amino acids than those in the absence of exogenous N.A supply of exogenous nitrate to the AM fungal spores with only CO2 gave rise to more than 10 times more asparagine than that without exogenous N.In contrast,the extra supply of exogenous glucose to the AM fungal spores generated a significant enhancement in the uptake of exogenous N sources,with more than 3 times more free amino acids being produced than those supplied with only exogenous CO2.Meanwhile,arginine was the most abundant free amino acid produced and it was incorporated into the proteins of AM fungal spores to serve as an N storage compound.     

Variation of morpho-physiological traits in geographically diverse pigeonpea [Cajanus cajan (L.) Millsp] germplasm under different phosphorus conditions

Genotypic variation for morpho-physiological parameters, phosphorus (P) content and root acid phosphatase activity was studied in 52 pigeonpea genotypes. Data related to shoot (length, dry weight, number of leaves, and leaf area), root (volume, length, dry weight, area, perimeter, and number of root tips), acid phosphatase activity, and P content (root, stem, and leaf) were recorded at 60 days after sowing (DAS). The P use efficient genotypes showed high root length, root area, root perimeter, root dry weight, P content in leaves, and root to shoot dry weight ratio under the P not added condition. Significant variation was found among genotypes for root- and shoot-associated characteristics under both P treatments. The P use efficient genotypes with improved root morphological phenes have potential to acquire and utilize more P from immobile soil bound P sources may be of additional factor for increasing efficiency of acquisition and utilization of supplied P fertilizer.     

植物对不同形态磷响应特征研究进展

磷是植物生长发育所必需的大量营养元素之一,参与植物体内许多重要化合物的合成与代谢。土壤中磷素具有多种形态,且不同形态磷的植物有效性差异较大;植物在不同形态磷环境下,体内会形成相应的适应性机制。植物吸收积累磷通常与根形态、根系分泌物、体内磷转运等因素有关,受到特异基因表达的调控。了解植物对磷的吸收积累特性是筛选磷高效植物或磷富集植物的前提,也是充分利用土壤磷素资源、修复磷过剩环境的关键。根据国内外研究现状,本文从磷素吸收积累、根系形态特征、磷酸酶与植酸酶的变化以及磷营养高效的分子机制,综述了植物对不同形态磷的响应特征,并对未来该领域的研究进行了展望。     

Phosphatase activity in earthworm faeces

Wormcasts of Allolobophora caliginosa have been shown to have higher phosphatase activity than occurs in uningested soil, resulting in an increase in inorganic P released by mineralization of organic P. This paper describes similar observations on organic wastes which may be used for vermiculture. Besides A. caliginosa, three species which will feed on dung or other organic substrates were investigated, Eisenia foetida, Dendrobaena veneta and Lumbricus rubellus. When cow dung was used as a substrate, background phosphatase activity was too high for the effects of earthworm activity to be detected and a culture medium was therefore developed from sterilized paper waste sludge with added phytin. Phosphatase activity, assayed by a modified Hoffman method, was higher in the presence of all four earthworm species than in controls. It showed two peaks in relation to pH, at 3–5 and 9–10. The former is attributed to microbial activity and the latter to microbial activity or earthworm alkaline phosphatase. After 1 month, cultures of paper waste sludge with phytin contained about twice as much water soluble P when worms were present than in their absence.     

Organic and Inorganic P Sources Interacting with Applied Rhizosphere Bacteria and Their Effects on Growth and P Supply of Maize

Plant growth–promoting rhizobacteria (PGPR) may enhance the plant availability of phosphorus (P) in soil. A greenhouse pot experiment was conducted cultivating maize (Zea mays L.) on a P-deficient soil. Three bacterial treatments (control without PGPR and application of either Enterobacter radicincitans sp. nov. strain DSM 16656 or Pseudomonas fluorescens strain DR54) were tested in conjunction with three P treatments [no P addition, inorganic P as triplesuperphosphate (TSP), and organic P as phytin] at two different growth stages of maize (V6 and V9). Amendment with TSP enhanced growth, P uptake, and highly bioavailable P pools in soil to a greater extent than phytin. In contrast, arbuscular mycorrhiza (AM) formation of maize roots after phytin application doubled those for the TSP treatment or the control without P. Application of PGPR was also able to increase AM formation and P uptake of maize, especially when no P source was added. Furthermore, P. fluorescens inoculation resulted in an increase of highly soluble soil P pools at the early growth stage. Greater impacts of phytin on P nutrition of maize may exist in a longer term as a result of slow P release and promotion of AM fungi. Benefits to maize P nutrition derived from PGPR application can be expected under P deficiency.     

A conceptual framework and an empirical test of complementarity and facilitation with respect to phosphorous uptake by plant species mixtures

Plant species have different traits for mobilizing sparingly soluble phosphorus (P) resources, which could potentially lead to overyielding in P uptake by plant species mixtures compared to monocultures due to higher P uptake as a result of resource (P) partitioning and facilitation. However, there is circumstantial evidence at best for overyielding as a result of these mechanisms. Overyielding (the outcome) is easily confused with underlying mechanisms because of unclear definitions. We aimed to define a conceptual framework to separate outcome from underlying mechanisms and test it for facilitation and complementarity with respect to P acquisition by three plant species combinations grown on four soils. Our conceptual framework describes both mechanisms of complementarity and facilitation and outcomes (overyielding of mixtures or no overyielding) depending on the competitive ability of the species to uptake the mobilized P. Millet/chickpea mixtures were grown in pots on two calcareous soils mixed with calcium-bound P (CaP) and phytate P (PhyP). Cabbage/faba bean mixtures were grown on both acid and neutral soils mixed with P-coated iron (hydr)oxide (FeP) and PhyP. Wheat/maize mixtures were grown on all four soils. Rhizosphere carboxylate concentration and acid phosphatase activity (mechanisms) as well as plant P uptake and biomass (outcome) were determined for monocultures rhizosphere and species mixtures. Facilitation of P uptake occurred in millet/chickpea mixtures on one calcareous soil. We found no indications for P acquisition from different P sources, neither in millet/chickpea, nor in cabbage/faba bean mixtures. Cabbage and faba bean on the neutral soil differed in rhizosphere acid phosphatase activity and carboxylate concentration, but showed no overyielding. Wheat and maize, with similar root exudates, showed overyielding (the observed P uptake being 22% higher than the expected P uptake) on one calcareous soil. We concluded that although differences in plant physiological traits (root exudates) provide necessary conditions for complementarity and facilitation with respect to P uptake from different P sources, they do not necessarily result in increased P uptake by species mixtures, because of the relative competitive ability of the mixed species.     

Inoculation with Phosphate Solubilizing Mesorhizobium Strains Improves the Performance of Chickpea (Cicer aritenium L.) Under Phosphorus Deficiency

The use of phosphate-solubilising bacteria as inoculants increases plant phosphorus (P) uptake and thus crop yield. Strains from the genus Mesorhizobium are among the most powerful phosphate solubilizing microorganisms. In order to study efficiency in P uptake and N₂ fixation in chickpea (Cicer aritenium), forty-two rhizobia strains natively from Tunisian soils were studied in symbiosis with the chickpea variety “Béja1” which is frequently cultivated in Tunisia. Plants were inoculated separately with these strains under controlled conditions in perlite under two sources of P i.e. soluble (KH₂PO₄) and insoluble P (Ca₂HPO₄). At flowering stage, growth, nodulation, P uptake and N₂ fixation were assessed in all symbiotic combinations. The results showed that the S27 strain efficiently mobilized P into plants, observed as a significant increase of plant P content when insoluble P (Ca₂HPO₄) was supplied to the soil. This was associated with a significant increase in plant biomass, nodule number and N content under insoluble P conditions. Additionally, inoculation with the Mesorhizobium strain S27 significantly increased the root acid phosphatase activity under insoluble P. This study also shows significant correlations found between plant P content and acid phosphatase activity under low P conditions which may highlight the contribution of acid phosphatases in increasing P use efficiency. A field experiment also showed that most of the chickpea analyzed parameters were improved when plants inoculated with two selected rhizobia strains (S26 and S27) and supplied with P2O5. Overall, these findings postulate that rhizobial inoculation should not only be based on the effectiveness of strains regarding N fixation, but also to other traits such as P solubilisation potential.