两株解磷真菌的解磷能力及其解磷机理的初步研究

从不同处理的水稻土壤中分离筛选出两株高效解磷真菌HP2、P5,研究了不同碳源条件对溶磷效果的影响,以及解磷菌株在不同的碳源培养条件下,溶磷量与培养介质pH值之间的相关性。结果表明,HP2菌株解磷能力在不同的测定时间内均高于P5菌株;不同碳源培养基的溶磷量顺序为蔗糖〉葡萄糖〉纤维素,且彼此差异显著：测定时间内,菌株的溶磷量与介质pH值之间存在极显著相关性（P〈0．01）。     

两株解磷真菌的解磷能力及其解磷机理的初步研究*

从不同处理的水稻土壤中分离筛选出两株高效解磷真菌HP2、P5,研究了不同碳源条件对溶磷效果的影响,以及解磷菌株在不同的碳源培养条件下,溶磷量与培养介质pH值之间的相关性。结果表明,HP2菌株解磷能力在不同的测定时间内均高于P5菌株;不同碳源培养基的溶磷量顺序为蔗糖>葡萄糖>纤维素,且彼此差异显著;测定时间内,菌株的溶磷量与介质pH值之间存在极显著相关性(P<0.01)。     

溶磷菌和固氮菌溶解磷矿粉时的互作效应

采用4株溶磷菌（Lx81、Dm84、Jm92、Lx191）、和3株固氮菌（ChW5、ChW6、ChO6）单独和混合接种后测定培养液有效磷含量、pH值及总有机酸含量的方法,研究溶磷菌和固氮菌溶解磷矿粉时的互作效应。结果表明,相对于单独接种溶磷菌：Lx81与3株固氮菌分别混合培养能提高磷矿粉的溶解能力,4株溶磷菌与ChW6,Lx81、Dm84、Lx191与Ch06分别混合培养及Jm92＋ChW5组合溶磷量极显著增加（P〈0．01）;Dm84＋ChW5、Lxl91＋ChW5、Jm92＋Ch06组合的溶磷量下降（P〈0．01）。除Lx81＋ChW6、Lx81＋Ch06培养液pH值降低外,混合培养的其它组合培养液pH值均较单独接种溶磷菌时升高。有机酸测定结果表明,Lx81、Jm92与ChW5、Ch06分别混合培养、ChW6＋Lx81组合有机酸含量升高（P〈0．01）,其它7种组合的有机酸含量均较单独接种溶磷菌的值下降（P〈0．01）。溶磷菌和固氮菌单菌培养时溶磷量与pH值、溶磷量与总有机酸含量及pH值与总有机酸含量之间呈现线性相关;Dm84、Lx191与3株固氮菌分别混合培养溶磷量与pH值之间、Lx81与3株固氮菌分别混合培养溶磷量与总有机酸含量之间呈现线性相关,其它组合的溶磷量与pH值、总有机酸含量间没有相关性。溶磷菌和固氮菌混合培养对溶解磷矿粉既有协同作用也有拮抗作用。     

红树林溶磷菌的初步鉴定、溶磷能力测定及其优化培养

对分离来自华南红树林的溶磷菌进行16S rDNA或ITS等基因水平上的初步鉴定, 测定其溶解无机磷的能力, 并对溶磷菌的生长培养基条件进行优化。结果表明, 溶磷真菌的溶磷效果明显强于溶磷细菌, 且溶磷真菌的溶解无机磷能力与培养液的pH呈极显著负相关, 而溶磷细菌的溶磷能力与pH没显著相关关系。单因素实验表明, 对供试菌株生长的合适碳源为麦芽糖, 氮源为尿素。通过正交实验得到的优化培养基为麦芽糖5 g/L、尿素0.05 g/L、NaCl 5 g/L、pH 5, 在30°C下培养48 h菌落总数可达6.06×     

微生物溶解磷矿粉能力与pH及分泌有机酸的关系

从玉米根际和非根际土壤中分离得到的 74株溶解磷矿粉的微生物 ,发现它们的溶磷能力差异很大 ,主要决定于菌株本身的特性 ,与其来源无关 ,真菌普遍比细菌要强。真菌的溶磷能力与其培养介质的 pH之间呈显著的负相关 ,但细菌的这种关系非常弱。二者都产生多种有机酸 ,真菌主要分泌草酸、丙二酸和乳酸 ,而细菌主要分泌草酸、酒石酸、丙二酸、乳酸和乙酸 ,不同菌株分泌有机酸的数量和种类差异很大 ,但溶磷量与有机酸总量或单个有机酸浓度之间 ,没有发现显著的相关性 ,唯独柠檬酸与真菌的溶磷量之间存在显著的相关性。说明不同菌株有完全不同的溶磷机理 ,可能多种机理并存。     

4株溶磷细菌和真菌溶解磷矿粉的特性

溶磷微生物广泛地分布在土壤,根际等生态环境中,了解这些微生物溶解难溶性磷酸盐如磷矿粉的特性,对于开发利用这些微生物,提高磷素利用效率具有重要作用,研究发现;真菌比细菌溶解磷矿粉的能力要强得多,培养基中的铁,镁,锰,钠等成分可以提高真菌的溶磷量,但降低了细菌的溶磷量,培养基中磷矿粉用量越高,溶磷量越低;碳源物质浓度高于3%将显地降低溶磷量,微生物能够破坏磷矿粉的结构。使其中的磷在以后的培养过程中更加容易释放出来,可见利用微生物活化磷矿粉中的磷,具有良好前景。     

牡丹根际溶磷放线菌的筛选及其溶磷特性

通过从牡丹根际土壤中分离筛选溶磷放线菌,得到一株具有较强溶磷能力的菌株PSPSA1,根据形态特征、生理生化特性以及16S rDNA序列分析对菌株进行鉴定,并研究其溶磷遗传稳定性及溶磷特性.菌株PSPSA1被鉴定为白网链霉菌,具有较好的溶磷遗传稳定性.在不同磷源培养液中溶磷量依次为磷酸钙(158.5 mg·L^-1)>磷酸铝(139.9 mg·L^-1)>磷酸铁(127.7 mg·L^-1)>卵磷脂(45.6 mg·L^-1),在无机磷培养液中的溶磷量均与pH呈现显著负相关性,在有机磷培养液中的溶磷量与pH没有显著相关性.在不同碳源条件下的溶磷量依次为乳糖>葡糖糖>麦芽糖>果糖>蔗糖>淀粉>纤维素,在不同氮源条件下的溶磷量依次为蛋白胨>硝酸铵>硫酸铵>硝酸钾>尿素,以葡萄糖为碳源、蛋白胨为氮源时,菌株的溶磷量最高可达202.6 mg·L^-1.土培60 d,单施菌株土壤有效磷含量比对照增加68.2%,菌株与有机肥混施土壤有效磷含量比单施有机肥增加76.7%.表明菌株PSPSA1能够溶解多种难溶磷,在土壤中溶磷效果显著,与有机肥混施其溶磷能力明显提高,有望成为高效生物磷肥的优良菌种.     

4株溶磷细菌和真菌溶解磷矿粉的特性*

溶磷微生物广泛地分布在土壤、根际等生态环境中 ,了解这些微生物溶解难溶性磷酸盐如磷矿粉的特性 ,对于开发利用这些微生物 ,提高磷素利用效率具有重要作用。研究发现 :真菌比细菌溶解磷矿粉的能力要强得多 ,培养基中的铁、镁、锰、钠等成分可以提高真菌的溶磷量 ,但降低了细菌的溶磷量。培养基中磷矿粉用量越高 ,溶磷量越低 ;碳源物质浓度高于 3 %将显著地降低溶磷量。微生物能够破坏磷矿粉的结构 ,使其中的磷在以后的培养过程中更加容易释放出来。可见利用微生物活化磷矿粉中的磷 ,具有良好前景。     

一株节杆菌溶解磷矿粉的动态

磷矿粉用量明显地影响节杆菌1TCRi7菌株对其溶解,随着磷矿粉用量的增加,溶磷率急剧下降,超过5 g/L,溶磷率不到0.1%,培养液的pH也随磷矿粉用量增加而升高,但菌体生长繁殖几乎不受磷矿粉用量的影响。振荡培养时,第1 d菌体繁殖量就达到高峰,pH也降至最低,同时表现出强烈的溶磷活力,但菌株的溶磷量并不与其生长量吻合,溶磷量第9 d才达到高峰,以后逐渐下降。培养过程中有机酸分泌发生交替变化,主要分泌乙酸和丙二酸。第1 d、第9 d和第15 d出现3个峰值,说明微生物利用了所分泌出来的有机酸,并发生了有机酸代谢的变化。     

一株红壤溶磷菌的分离、鉴定及溶磷特性

【目的】为了提高红壤磷素利用率,探讨溶磷菌溶磷机理。【方法】利用难溶性无机盐培养基从花生根际土壤样品中分离到一株溶磷菌C5-A,结合菌落形态特征、生理生化和16S rRNA序列确定该菌株的系统发育地位;通过菌株C5-A在NBRIP液体培养基培养过程中培养液pH变化确定其溶磷能力;利用液体发酵实验测定不同的碳源、氮源对菌株C5-A溶磷的影响;通过高效液相色谱检测C5-A在不同氮源培养液中有机酸的种类和浓度。【结果】菌株C5-A鉴定为洋葱伯克霍尔德氏菌(Burkholderia cepacia),遗传稳定性较好。在FePO4和AlPO4培养液中,菌株C5-A的溶磷量和pH变化呈显著负相关;菌株C5-A对磷酸三钙、磷酸铝、磷酸铁、磷矿粉均有较强的溶解能力,最高溶磷量分别为125.79、227.34、60.02和321.15 mg/L;菌株C5-A对不同浓度的两种磷矿粉有较强的溶解能力;分别以麦芽糖和草酸铵为碳源和氮源时溶磷量最高。高效液相色谱检测出10种有机酸,分别为草酸(葡萄糖酸)、乙酸、苹果酸、琥珀酸和5种未知有机酸,然而,乙酸而非草酸似乎是影响C5-A溶磷的重要有机酸。【结论】从红壤花生根际土壤中筛选到一株对难溶性无机盐具有较强溶解能力溶的菌株C5-A,有望为开发高效红壤微生物磷肥提供种质资源。     

Stress induced phosphate solubilization in bacteria isolated from alkaline soils

Phosphate solubilizing bacteria NBRI0603, NBRI2601, NBRI3246 and NBRI4003 were isolated from the rhizosphere of chickpea and alkaline soils. All four strains demonstrated diverse levels of phosphate solubilization activity under in vitro conditions in the presence of various carbon and nitrogen sources. Acid production may have contributed to phosphate solubilization, but was not the only reason for phosphate release into the medium. Among the four strains, NBRI2601 was the most efficient strain in terms of its capability to solubilize phosphorus in the presence of 10% salt, pH 12, or 45 degrees C. The strains showed varied levels of phosphate solubilization when the effects of different sources of nitrogen were examined during growth. The presence of low levels of Ca(2+) and EDTA in the medium enhanced phosphate solubilization.     

一株高效解磷真菌新菌株的筛选鉴定及解磷特性

从辽宁省辽中县多年耕种的日光温室番茄根际土壤中筛选出一株解磷真菌,通过菌落形态特征和ITS rDNA序列对比,鉴定该菌株为草酸青霉菌的一株新菌株,将其命名为PSF1.该菌株能利用葡萄糖、蔗糖、乳糖、半乳糖、可溶性淀粉等多种碳源和硫酸铵、氯化铵、硝酸铵、硝酸钾、尿素等多种氮源进行生长代谢并表现出较强的解磷能力,在C/N 10∶1~60∶1、初始pH 7~8的条件下生长情况较好且解磷能力较高.该菌株有很强的产酸能力,在培养过程中培养液pH由7.00~7.50下降至2.06~4.87;在4种磷源培养液中的最高解磷量分别为磷酸三钙(869.62 mg·L^-1)>磷矿粉(233.56 mg·L^-1)>磷酸铝(44.77 mg·L^-1)>磷酸铁(28.42 mg·L^-1).Pearson相关分析表明,菌株在磷酸三钙、磷矿粉和磷酸铁培养液中的解磷量与pH的变化之间呈极显著负相关关系,在磷酸铝培养液中无显著相关关系.菌株PSF1解磷能力强,生长条件广,推测其在土壤中有较强的解磷能力.     

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.     

Phosphorus and phosphate solubilizing bacteria: Keys for sustainable agriculture

Abstract

Phosphorus (P) is one of the most important minerals required for plant growth occupying a strong position among soil macro nutrients. Soil P deficiency is often fulfilled by phosphate fertilizers. P deficiency in soils is due to less total P contents in the soil and fixation of added P from chemical fertilizers as well as other organic sources like manures. The response of plant under P stress or even when it is present in adequate amount is very mild. The basic constraint in the availability of P is its solubilization as it gets fixed both in acidic and alkaline soil. Soil fixed P can only be solubilized by phosphate solubilizing microorganisms (PSMs).These bacteria released different types of organic acids in the soil which make P soluble and available to plants. The potential of these PSMs to solubilize P varies and mainly depends upon mechanism adopted for solubilization, their molecular genetics as well as their ability to release P in soil. The PSMs, having all the characteristics of phytohormone production, nitrogen fixation, as well as, heavy metal decontamination and creating salt stress tolerance in plants, are quite rare for sustainable agriculture. Application of this environment friendly approach for increasing crop productivity as well as its impact on soil and plant health is discussed in this review which will not only open new avenues of research but also provide fruitful information about phosphate solubilizing microbes for sustainable agriculture development.     

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.     

Rock phosphate solubilization by psychrotolerant Pseudomonas spp. and their effect on lentil growth and nutrient uptake under polyhouse conditions

Psychrotolerant Pseudomonas isolates (RT5RP2 and RT6RP) isolated from the rhizoplane of wild grass at 3,100 and 3,800 m above mean sea level, respectively, from Rudraprayag district of Uttarakhand (India), were found to solubilize Udaipur rock phosphate (URP). Both isolates grew at temperatures ranging from 4 to 30 °C. Kinetics of phosphate solubilization by the bacterial strains showed a nonlinear regression of the rate of P solubilization, which fitted best in the power model, and showed a declining trend across three different temperatures. Under pot culture conditions, bacterization of lentil seeds (cv. VL Masoor 507) with the psychrotolerant Pseudomonas strains when combined with URP as a sole source of phosphorus resulting in significant enhancement in P uptake of the plants, compared to the application of rock phosphate alone.     

Medium pH, carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of Penicillium purpurogenum through organic acid production

Aims: To study phosphate solubilization in Penicillium purpurogenum as function of medium pH, and carbon and nitrogen concentrations. Methods and Results: Tricalcium phosphate (CP) solubilization efficiency of P. purpurogenum was evaluated at acid or alkaline pH using different C and N sources. Glucose‐ and (NH₄)₂SO₄‐based media showed the highest P solubilization values followed by fructose. P. purpurogenum solubilizing ability was higher in cultures grown at pH 6·5 than cultures at pH 8·5. Organic acids were detected in both alkaline and neutral media, but the relative percentages of each organic acid differed. Highest P release coincided with the highest organic acids production peak, especially gluconic acid. When P. purpurogenum grew in alkaline media, the nature and concentration of organic acids changed at different N and C concentrations. A factorial categorical experimental design showed that the highest P‐solubilizing activity, coinciding with the highest organic acid production, corresponded to the highest C concentration and lowest N concentration. Conclusions: The results described in the present study show that medium pH and carbon and nitrogen concentrations modulate the P solubilization efficiency of P. purpurogenum through the production of organic acids and particularly that of gluconic acid. In the P solubilization optimization studies, glucose and (NH₄)₂SO₄ as C and N sources allowed a higher solubilization efficiency at high pH. Significance and Impact of the Study: This organism is a potentially proficient soil inoculant, especially in P‐poor alkaline soils where other P solubilizers fail to release soluble P. Further work is necessary to elucidate whether these results can be extrapolated to natural soil ecosystems, where different pH values are present. Penicillium purpurogenum could be used to develop a bioprocess for the manufacture of phosphatic fertilizer with phosphate calcium minerals.     

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).     

Solubilization of inorganic phosphate byRhizobium

A large number of strains ofRhizobium were able to solubilize the insoluble phosphate compound, hydroxy-apatite, in liquid culture. Solubilization of hydroxyapatite byRhizobium was not mediated by an enzyme but acidity developed in the cultures was involved in the process. An inverse relationship between the level of soluble phosphate and medium pH was evident. The ability to solubilize hydroxyapatite varied among the strains. In a medium without NH ₄ ⁺ , some of the strains showed better activity than when NH ₄ ⁺ was present, suggesting involvement of different mechanisms for phosphate solubilization.R. meliloti SU 47 produced 2-ketogluconic acid along with an unidentified acid in the medium containing NH ₄ ⁺ . 2-Ketogluconic acid was identified as the major factor in inorganic phosphate solubilization. Initial presence of soluble phosphate in the medium had no discernible influence on the extent of hydroxyapatite solubilization. Initial presence of calcium reduced solubilization of phosphate and addition of EDTA to stationary phase cultures caused an increase in the level of soluble phosphate.