Transformation of rock phosphate during composting and the effect of humic acid

Summary Phosphorus from Mussoorie rock phosphate (MRP) was solubilized and transformed into available forms when MRP was incorporated during composting of organic wastes. Clusterbean and redgram utilized phosphorus efficiently from the phosphorus enriched compost containing 3.1% P when added in the soil of pH 7.6 to 7.8. The solubilization of phosphorus during composting has been attributed to the formation of humic substances.     

香蕉根际土壤解磷细菌的筛选、鉴定及解磷能力

【目的】以磷矿粉为难溶态磷,以期从香蕉根际土壤筛选出高效的解磷细菌。【方法】采用透明圈法和钼锑抗比色法分离筛选解磷细菌,通过形态学特征、生理生化试验结合16S rRNA基因序列分析及系统发育树比对鉴定其种属,并利用单因素试验方法研究不同碳源、氮源及C/N比值(40:1、20:1和8:1)对菌株溶解磷矿粉能力的影响。研究不同菌株解磷能力和培养介质pH值的变化关系。【结果】分离具有解磷能力的细菌8株,筛选出具有代表性的3个菌株B3-5-6、M-3-01和T1-4-01,初步鉴定菌株B3-5-6为嗜气芽孢杆菌(Bacillus aerophilus),M-3-01为虫内生沙雷氏菌(Serratia nematodiphila),T1-4-01为艾博丽肠杆菌(Enterobacter asburiae)。B3-5-6解磷能力与介质pH值之间存在线性负相关性(|r|=0.949 66>0.735),其相关性达到极显著水平;B3-5-6在碳源为蔗糖、氮源为(NH4)2SO4、C/N为40:1,M-3-01在碳源为葡萄糖、氮源为(NH4)2SO4、C/N为20:1,T1-4-01在碳源为乳糖、氮源为蛋白胨、C/N为20:1条件下解磷效果较好。解磷效果与初筛相比分别提高了1.12、1.17、2.55倍。【结论】不同的碳氮源、C/N值会直接影响磷细菌的解磷能力;筛选出一株解磷能力与培养介质pH之间存在着极显著相关性的细菌,其解磷机理有待进一步研究。     

Factors determining rock phosphate solubilization by microorganisms isolated from soil

Forty two soil isolates (31 bacteria and 11 fungi) were studied for their ability to solubilize rock phosphate and calcium phosphate in culture medium. Eight bacteria and 8 fungi possessed solubilizing ability. Pseudomonas cepacia and Penicillium purpurogenum showed the highest activity. There was a correlation between final pH value and titratable acidity (r=–0.29 to –0.87) and between titratable acidity and soluble phosphate (r=0.22 to 0.99). Correlation values were functions of insoluble phosphate and of the group of microorganisms considered. A high correlation was observed between final pH and soluble phosphate only for the rock phosphates inoculated with the highest concentration of solubilizing bacteria (r=–0.73 to –0.98).     

不同碳源对三种溶磷真菌溶解磷矿粉能力的影响

通过液体培养法 ,对 3种溶磷真菌利用葡萄糖、果糖、蔗糖、麦芽糖、淀粉和纤维素等碳源溶解宜昌产磷矿粉的试验 ,结果表明 ,菌株P2 3在供给葡萄糖时的溶磷能力最高 ,并在一定程度上能够利用长链碳源淀粉和纤维素为营养而溶磷 ;而高效溶磷菌株P6 6和P39溶磷的最佳碳源是果糖和麦芽糖 ,该两菌株利用淀粉和纤维素的溶磷效果很小 ,甚至不溶磷。 3种溶磷真菌培养滤液 pH值、可滴定酸含量与其溶磷量之间的相关性因菌株而异 ,差别很大。菌株P2 3培养滤液pH值、可滴定酸含量与其溶磷量之间相关性很低 ,但菌株P6 6和P39培养滤液pH值、可滴定酸含量与其溶磷量之间相关性却达到极显著水平 (P <0 0 1)。结果表明 ,不同碳源对溶磷菌溶解磷矿粉能力影响很大 ,分析推断 3种菌株产生的有机酸活化磷矿粉能力为P6 6>P39>P2 3。     

The influence of rhizosphere in rice crop on resin-extractable phosphate in flooded soils at various levels of phosphate applications

Summary Poor or lack of response of lowland rice to P fertilization is a well-known fact. Several studies were conducted in this direction however, our understanding regarding the underlying mechanism has been far from clear. A remarkable influence of rice plants on P transformation in submerged soil is identified in this experiment which may shed light on this problem. Accordingly, in presence of rice plants P was mobilized during the initial growing period followed by immobilization. The increased microbial activity in presence of physiologically active roots was responsible for P mobilization, while capacity of rice plants to reoxidize the rhizosphere, by secretion of oxygen from roots, during later growing period was responsible for P immobilization.The extent of P mobilization decreased while that of immobilization increased with increasing P levels in different soils. At a given P level the ratio of P mobilization to immobilization was higher in a soil where crop growth was better and P uptake was higher as compared to another soil where crop growth was poor with lower P uptake.Thus, lowland rice plants appear to possess an unique physiological mechanism, to regulate the contrasting changes in P availability in the rhizosphere depending on P requirement by the plants or P availability in soil, which in turn is responsible for the poor or lack of response to P fertilization.The experiment was conducted in a growth chamber. Two soils with widely varying properties were used.     

原生动物与解磷微生物协同解磷作用

磷是农业生产重要的限制因素之一,土壤生物包括原生动物和微生物在土壤磷素转化过程中起着重要的作用。通过纯培养和土壤培养试验发现,供试的原生动物能够提高磷矿粉的有效磷,土壤有效磷含量也显著增加。所溶解出来的磷很大一部分以无机磷酸盐的形式贮藏在细胞内,熏蒸时才释放出来,少部分磷转化为生物量磷。原生动物与解磷菌之间没有显著的相互促进作用。     

Rhizosphere mediated growth enhancement using phosphate solubilizing rhizobacteria and their tri-calcium phosphate solubilization activity under pot culture assays in Rice (Oryza sativa.)

Phosphate solubilizing rhizobacteria are considered as an important alternative to increase the availability of accumulated phosphates through solubilization. These increase the growth of plant by enhancing the efficiency of fixing biological nitrogen. This was studied through a pot experiment involving two Phosphate Solubilizing Rhizobacteria (PSRB) isolates, Pseudomonas aeruginosa and Bacillus subtilis along with Tri-calcium phosphate (TCP) on availibity of nutrients, biological composition of soil and yield attributes of rice crop at its growth stages. Experiment was laid in factorial completely randomized design (CRD) comprising of eight treatments replicated thrice with two factors viz. factor 1 with or without TCP (1 g⁻¹soil) and factor 2 with single or combined inoculation of PSRB isolates. Considerable enhancement in available content of potassium (K), phosphorous (P), nitrogen (N) in soil was found with TCP 1 g⁻¹soil (P₁) and consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture at crop growth stages. Highest increase in available N (17.13% and 19.1%), available P (232% and 265%), available K (19.6% and 29.2%) over control were recorded in B₃ (consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture). Similarly, maximum nutrient uptake N (6.4%), P (15.8%) and K (8.9%) were recorded with same treatment. A considerable growth in soil microbial biomass carbon and dehydrogenase activity at crop growth stages was recorded on application of TCP 1 g⁻¹soil (P₁) and consortium of PSRB isolates'' Pseudomonas aeruginosa and Bacillus subtilis (B₃). Highest increase in microbial biomass carbon (16.4% and 16.5%) and dehydrogenase activity 34.7% and 43.8% over control were recorded in B₃ (consortium of PSRB isolates Pseudomonas aeruginosa and Bacillus subtilis) and was found best among all treatments in terms of yield (63.2%) and yield attributes; number of panicles⁻¹plant (54.8%), number of grains⁻¹panicle (156%) and average panicle length (63.9%).     

Extracellular phosphate solubilization by the cyanobacteriumAnabaena ARM310

Endogenous polyphosphate depletedAnabaena ARM310, solubilized extracellular tricalcium phosphate through increased phosphatase activity.     

The phosphate uptake mechanism

The slow rate of diffusion of phosphate in soil results in a zone of depletion of phosphate ions in solution around the roots of plants in low phosphate soils. Transfer of phosphate to the site of uptake into the root symplasm limits phosphate uptake in such soils. This transfer involves movement across the depletion zone and through the root apoplasm. The apoplasm is made up of the cell walls of epidermal and cortical cells, together with the associated intercellular spaces. Although the pores in the open latticework of these cell walls permit movement of nutrients around cells, they increase the path length across which phosphate ions have to diffuse. The structural components and net negative charges of the cell walls also influence the effective concentrations of phosphate in the apoplasm. This concentration may be further modified by excreted organic compounds around cell walls and the presence of micro-organisms that use such compounds as carbon sources. A membrane on the inner surface of the cell wall, the plasmalemma, separates the apoplasm from the symplasm. Uptake of nutrients into the root symplasm occurs through transporter proteins embedded in this membrane. Understanding of the mechanisms by which phosphate is transported across the plasmalemma into the plant symplasm has advanced considerably over the past 4 years due to the application of molecular techniques. Genes encoding the transporters involved in this process have been isolated from a number of plant species. These transporters belong to a family of membrane proteins characterized by having 12 membrane-spanning domains arranged in a '6+6' configuration. H₂PO₄ ^– ions, together with protons, are transported through this protein. This transport process is driven by the potential across the membrane maintained by the action of a H⁺-ATPase, the `proton pump', that extrudes protons to the outer surface of the membrane. The expression of genes encoding high-affinity root phosphate transporters is regulated by the phosphorus (P) status of the plant. The transduction pathway involved in this regulation is not known at present. It is a systemic response rather than a localized response, however, the overall phosphate status of the plant being the controlling factor. Under phosphate stress, the expression of genes encoding these phosphate transporters is up-regulated. This results in a greater number of transporter proteins in the plasmalemma and enhanced phosphate uptake rates, if phosphate is available at the membrane surface. Uptake occurs around the root tip, into epidermal cells with their associated root hairs and into cells in the outer layers of the root cortex. Further back along the root axis, phosphate can also be taken up by transfer from mycorrhizal fungi to root cortical cells.Strategies for increasing nutrient uptake by overexpressing genes encoding high-affinity phosphate transporters are likely to be mainly applicable to situations where a reasonable phosphate concentration can be maintained at the outer surface of the plasmalemma. Maintaining such a concentration is a major problem in the phosphate deficient soils of the semi-arid tropics (SAT), so emphasis in these soils is on strategies to improve the movement of phosphate to the surface of the plasmalemma. There may be scope, however, for manipulating the expression of genes involved in the internal mobilisation of phosphate within the plant, thereby improving phosphate utilisation.     

The solubilization of low-grade rock phosphate with anaerobic digester slurry

The effects of biogas slurry on the solubilization of low-grade rock phosphate was studied by digestion in compost pits. Biogas slurry had solubilizing power for up to 60 days while molasses addition prolonged it throughout a digestion period of 120 days. In this case solubilization reached a maximum of 42.6% of added rock phosphate. Pyrite incorporation depressed drastically the rate of solubilization in spite of decreasing the pH and increasing the citrate-soluble P of the digesting biogas slurry.The concentration of total P increased with increase the loss of organic matter during digestion. The maximum weight loss was recorded in rock phosphate-containing treatment, where release of water-soluble P increased continuously with time and organic P decreased after 30 days of incubation. On the other hand, incorporation of molasses both in presence and absence of pyrite, decreased the water-soluble P but increased the organic P content of the digesting material.     

盐碱地柠条根围土中黑曲霉的分离鉴定及解磷能力测定

在盐碱滩地的改良过程中,柠条具有提升土壤供氮、供磷、供钾的潜力.以盐碱滩地上建植的柠条灌木林为研究对象,以柠条根围土壤为培养基质,采用无机磷培养基筛选,用平板溶菌圈法分离获得1株具有溶磷能力的真菌.将测得的ITS基因序列在NCBI上进行同源性检索,结果表明,所测序列与黑曲霉(Aspergillus niger)同源性为100％.综合形态特征和ITS基因序列同源性两方面分析,该菌株鉴定为黑曲霉(Aspergillus niger).168h连续监测无机磷培养液pH值、速效磷含量、菌丝重量和菌体吸磷量,研究该菌株的解磷能力.研究结果表明:随着培养时间的延长,培养液pH值从7.0下降到2.0左右,溶液中速效磷含量逐渐增加到4.7 mg,菌体自身吸磷量由5.4 mg下降到0.5mg,在36-48h后各项指标达到稳定状态.可见,黑曲霉菌体可以有效利用难溶性磷源,并将其转化成可被植物吸收利用的有效磷.     

Mechanism of insoluble phosphate solubilization by Pseudomonas fluorescens RAF15 isolated from ginseng rhizosphere and its plant growth-promoting activities

Aims:  To investigate the mechanism of insoluble phosphate (P) solubilization and plant growth-promoting activity by Pseudomonas fluorescens RAF15.
Methods and Results:  We investigated the ability of Ps. fluorescens RAF15 to solubilize insoluble P via two possible mechanisms: proton excretion by ammonium assimilation and organic acid production. There were no clear differences in pH and P solubilization between glucose-ammonium and glucose-nitrate media. P solubilization was significantly promoted with glucose compared to fructose. Regardless of nitrogen sources used, Ps. fluorescens RAF15 solubilized little insoluble P with fructose. High performance liquid chromatography analysis showed that Ps. fluorescens RAF15 produced mainly gluconic and tartaric acids with small amounts of 2-ketogluconic, formic and acetic acids. During the culture, the pH was reduced with increase in gluconic acid concentration and was inversely correlated with soluble P concentration. Ps. fluorescens RAF1 showed the properties related to plant growth promotion: pectinase, protease, lipase, siderophore, hydrogen cyanide, and indoleacetic acid.
Conclusion:  This study indicated that the P solubility was directly correlated with the organic acids produced.
Significance and Impact of the Study:  Pseudomonas fluorescens RAF15 possessed different traits related to plant growth promotion. Therefore, Ps. fluorescens RAF15 could be a potential candidate for the development of biofertilizer or biocontrol agent.     

Response of cowpea (Vigna unguiculata) to inoculation with VA-mycorrhizal fungi and to rock phosphate fertilization in some unsterilized Nigerian soils

Summary The effects on cowpea of inoculation with vesicular-arbuscular (VA) mycorrhizal fungi and rock phosphate (RP) fertilization were studied in pots using Alagba and Araromi series soils and in the field on Alagba, Apomu and Egbeda series soils. Inoculation of the plants with VA-mycorrhizal fungi caused very rapid infection of the roots. A higher per cent mycorrhizal infection was maintained during subsequent plant growth in the field. RP application reduced the degree of infection without affecting plant growth in the field and in pot experiments. Nodulation, nitrogen fixation and utilization of RP were increased by inoculation with mycorrhizal fungi in the pot experiments but not in the field experiments. In the pot experiments, inoculated plants supplied with RP flowered earlier, and took up more phosphorus than either inoculated plants without RP or uninoculated plants. The largest response to inoculation in terms of shoot dry matter, nodule yield and nitrogen content of shoots was obtained in Alagba soil under both pot and field conditions.IITA Journal Series Paper No. 136.     

不动杆菌JL-1菌株的解磷机理

【背景】不动杆菌(Acinetobacter indicus) JL-1菌株可将土壤中难溶性磷转化为植物可吸收利用的可溶性磷,但其解磷机理尚不明确。【目的】探讨不动杆菌JL-1菌株的解磷机理。【方法】采用钼蓝比色法测定菌株在不同难溶性磷源中的解磷效果,确定最佳磷源;通过粒径测定法、超声破碎法、高效液相色谱法等多种方法,检测不动杆菌JL-1菌株在解磷过程中对磷酸钙的溶解作用、对磷的固化作用及产有机酸和磷酸酶酶活情况。【结果】不动杆菌JL-1菌株在磷酸钙液体培养基中解磷效果最佳,解磷量在48 h达到最高值118.04μg/mL。在解磷过程中,JL-1菌株对磷酸钙具有溶解作用,菌株自身固化了部分磷;可溶性磷的释放是发酵过程中产生的葡萄糖酸、丙酸、乙酸、乳酸等多种有机酸与磷酸酶共同作用的结果,有机酸中丙酸含量最高可达118.11 mg/mL;酸性磷酸酶酶活最高为22901.32μmol/(L·h),碱性磷酸酶酶活最高为23826.02μmol/(L·h)。【结论】不动杆菌JL-1菌株在解磷过程中对磷酸钙颗粒显示出一定的溶解作用,通过分泌有机酸和磷酸酶使可溶性磷释放,同时菌体固化了一部分磷,本研...     

Phosphate supplying power of rock phosphates in an oxisol

Summary The P-supplying power of triple superphosphate, three apatitic rock phospates and a calcined aluminum rock phosphate were tested by measuring the quantities of fertilizer P recovered in soybean and in four chemical extractants, after 3-day and 75-day periods of contact between soil and fertilizer.The triple superphosphate supplied the highest amounts of P, but it lost efficiency during the longer incubation period. The rock phosphates maintained their original efficiencies, probably as a result of a balance between P released from the fertilizer and P converted into non-labile forms.The following coefficients of correlation between P uptake by soybean from an acid oxisol and P extracted by chemical extractants, after the two incubation periods, were found: 0.902^** for 0.01M CaCl₂; 0.823^** for anion-exchange resin; 0.720^** for 0.03N NH₄F+0.025N HCl; –0.037 (n.s.) for 0.025N H₂SO₄+0.050N HCl.The acid NH₄F solubilized residual calcined aluminum phosphate particles, and double acid extracted P from residual apatite particles, thus accounting for their poorer performances in predicting availability of fertilizer P.The relative efficiencies of the rock phosphates could largely be predicted after an incubation period of only three days. This finding attests to the presence in these rock phosphates of an easily soluble fraction of P which is not indicative of the degree of reactiveness of the phosphate as a whole.on leave at the Agricultural University during 1977.     

Biotechnological solubilization of rock phosphate on media containing agro-industrial wastes

Rock phosphate (RP) is an important natural material traditionally used for the production of phosphorus (P) fertilizers. Compared with chemical treatment, microbial solubilization of RP is an alternative environmentally mild approach. An overview of biotechnological techniques, mainly based on solubilization processes involving agro-industrial residues, is presented. Potential advantages of composting, solid-state fermentation, and liquid submerged fermentation employing free and immobilized microorganisms that produce organic acids and simultaneously solubilize RP are discussed. Subsequent introduction of the final fermented products into soil-plant systems promotes plant growth and P acquisition.     

Optimizations of particle size and pulp density for solubilization of rock phosphate by a microbial consortium from activated sludge

Microbial solubilization of rock phosphate is getting more and more attention recently. However, the microorganisms used in previous studies were mostly single or known species, and seldom studies focused on the mixed microorganisms or microbial consortia from natural environments. In this study, a microbial consortium taken from activated sludge was used to solubilize two different mid-low-grade rock phosphates. The results showed that the microbial consortium could effectively solubilize the rock phosphates in National Botanical Research Institute’s phosphate growth medium and released soluble phosphorus in the broth. The biomass increased gradually, whereas the pH decreased sharply during the solubilizing process. The maximum phosphorus solubilization was recorded at particle size of 150?µm. Higher or lower than this optimal particle size, the phosphorus solubilization decreased. The phosphorus solubilization gradually decreased with a larger pulp density from 1 to 5%, and the optimal pulp density was 1%. The solubilization level of microbial consortium varied with different rock phosphates. The results revealed that the soluble phosphorus released from high-silicon ore was higher than which from high-magnesium ore. A strong positive correlation between biomass and phosphorus solubilization in the broth was observed from regression analysis results, and the phosphorus solubilization also had a significant negative correlation with pH in the broth.     

Microbial solubilization of rock phosphate on media containing agro-industrial wastes and effect of the resulting products on plant growth and P uptake

Four agro-industrial wastes were assayed as substrates for microbial solubilization of rock phosphate (RP). Sugar beet wastes (SB), olive cake (OC) and olive mill wastewaters (OMWW) were treated by Aspergillus niger, and dry olive cake (DOC) was treated by Phanerochaete chrysosporium. In conditions of solid-state fermentation 46% of SB and 21% of OC were mineralized by A. niger while 16% of DOC was mineralized by P. chrysosporium. Repeated-batch mode of fermentation was employed for treatment of OMWW by immobilized A. niger, which resulted in conversion of 80% of the fermentable sugars. Acidification of all media treated by A. niger was registered with a simultaneous solubilization of 59.7% (SB), 42.6% (OC), and 36.4% (OMWW) of the total P present in the RP. The same mechanism of RP solubilization was observed in DOC-based medium inoculated with P. chrysosporium but other mechanisms were probably involved during the process. A series of microcosm experiments were then performed in the greenhouse to evaluate the effectiveness of the resulting fermented products. All amendments improved plant growth and P acquisition, which were further enhanced by mycorrhizal inoculation. The level of all studied parameters including the root mycorrhizal colonization depended on the substrate characteristics. The reported biotechnological schemes offer a potential application particularly for degraded soils.     

Enhanced solubilization of rock phosphate by Penicillium bilaiae in pH-buffered solution culture

Little is known about how pH-buffering capacity affects phosphorus (P) solubilization by Penicillium bilaiae. This study compared solubilization of rock phosphate (RP) by P. bilaiae in nonbuffered (pH 5.0) and buffered (pH 7.0) media. Fungal growth reached the stationary phase around day 12 and was slightly enhanced in the buffered medium. The fungus reduced solution pH from 5.0 to 4.1 in the nonbuffered medium and from 7.0 to 4.9 in the buffered medium by day 12. Phosphorus concentrations increased after day 9 more in the buffered than in the nonbuffered media (53 and 5 mg P x L(-1), respectively, on day 12). On day 12, higher concentrations of citric and oxalic acids were detected in the buffered (2.0 and 1.2 g x L(-1), respectively) than nonbuffered media (0.5 and 0.04 g x L(-1), respectively). Solubilization of RP was simulated without P. bilaiae in solutions equivalent to the nonbuffered and buffered cultures of P. bilaiae grown with RP. After a 24 h incubation, the P concentrations were of similar magnitudes to those observed in the P. bilaiae culture (18 and 47 mg P x L(-1), respectively, in the nonbuffered and buffered media). Under increased pH-buffering conditions, the enhanced production of citric and oxalic acids led to significant RP solubilization.