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1.
以对磷酸三钙具有高效溶解作用且对玉米苗生长有促生效果的假单胞菌K3为模式菌株,采用NBRIP液体培养基研究了解磷菌K3的解磷机制及缓冲容量对其解磷量的影响。结果表明,解磷菌K3液体摇瓶培养7 d后,培养液中水溶性磷从6.54 μg/mL增加至655.23 μg/mL,pH从7.00降至3.99。高效液相色谱测定发现,K3菌液中的主要代谢产物为苹果酸、乳酸和草酸,浓度分别为47.39 mmol/L、25.67 mmol/L和1.89 mmol/L。人工模拟K3菌株产生的有机酸及调节培养基不同pH值对磷酸三钙溶解度影响的试验表明,有机酸的螯合作用是解磷细菌K3菌株解磷的主要机理,而调节培养基pH对解磷的作用有限。液体摇瓶和土培试验结果显示,土壤缓冲容量对K3解磷菌的解磷效应有显著的抑制作用。 相似文献
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优良解磷菌株诱变选育与解磷培养基优化 总被引：2，自引：1，他引：1
对解磷菌枯草芽孢杆菌X1055进行紫外线诱变选育,筛选到优良解磷突变株1055Y1。该菌株摇瓶培养72h发酵液中有效磷含量最高达到5.22mg/L,解磷能力比出发菌株提高17.8%。在单因子试验筛选最适碳源、氮源基础上,通过Plackett-Burman设计,确定接种量、pH、KCl浓度等因素对突变株1055Y1解磷能力具有显著影响。应用爬坡试验和Box-Behnken设计的响应面法,对突变株1055Y1解磷培养基进一步优化。结果表明:在250ml发酵培养基中,接种量10.21ml、pH6.75、KCl浓度0.12g/L时,突变株1055Y1解磷达到最佳水平,发酵液有效磷含量达到7.03mg/L。 相似文献
3.
本文运用构建成功的含有假单胞菌属自身启动子及绿色荧光蛋白(GFP)的质粒pTRGFP电转至假单胞菌解磷细菌K3中,通过激光共聚焦显微镜及质粒检测,获得了发光稳定的标记菌株K3GFP。7天液体摇瓶试验中,标记菌株K3GFP的可溶性P含量在第4天达到最高774μg/ml,而出发菌株K3在第3天时已经达到了最大含量780μg/ml,说明pTRGFP质粒的转入对K3菌株解P能力有一定的影响。标记菌株K3GFP施入自然土壤10天后数量维持在5.47×106CFU/g～2.40×106CFU/g,35天后降到5.0×103CFU/g,说明解磷细菌K3GFP可以在自然土壤中定殖。本试验为研究解磷细菌在根际及土壤中的生长动态等行为特征奠定了基础。 相似文献
4.
磷在植物生长发育过程中起着重要的作用。为获得优质的解磷促生菌,以北京平谷桃园根际土壤为原料,从中筛选具有较高解磷促生能力的菌株。通过平板试验初筛可产解磷圈的菌株,再进行液态培养,采用钼锑抗比色法测定其最大的解磷能力,并探索菌株解磷能力与pH变化的关系,同时借助盆栽试验研究其促生特性。从桃树根际土共分离筛选出3株解磷菌,编号分别为JP01、JP03和PG62,初步鉴定分别为黑曲霉、杰氏假单胞菌和苍白杆菌。其中,黑曲霉JP01解磷能力优异,其溶解磷酸三钙的能力为112.52 mg/L,对卵磷脂的转化量为145.50 mg/L,最大解磷矿粉能力为95.74 mg/L,在培养过程中,该解磷菌对应的培养液pH显著下降。盆栽试验表明,黑曲霉能增加玉米幼苗株高、茎粗和地下部干重,还可使植株全磷含量增加。本研究结果可为开发解磷微生物菌剂提供优良的菌株资源。 相似文献
5.
山西矿区复垦土壤中解磷细菌的筛选及鉴定 总被引：5，自引：1，他引：4
【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。 相似文献
6.
两株解磷细菌的解磷活性及作用机制研究 总被引：4，自引：0，他引：4
解磷细菌在增加土壤可溶性磷含量、提高磷肥利用效率方面具有重要作用。为选筛高效解磷菌、探讨其解磷机制,本文利用平板溶磷圈法筛选解磷细菌,采用钼锑抗比色法研究其解磷活性,苯磷酸二钠法研究其磷酸酶活性,利用薄层层析分析其产生的有机酸,根据生理生化特征和16S r RNA基因序列系统发育分析,确定其分类学地位。结果表明,菌株JXJ-11和JXJ-15对植酸钙的降解活性很强,3 d后培养液中可溶性磷浓度分别增加219 mg·L~(-1)和216 mg·L~(-1);对磷酸钙降解活性较弱,最高可溶性磷浓度仅为植酸钙的21.79%~30.37%;解磷细菌可分泌酸性、中性和碱性磷酸酶,降解不溶性磷,可能产生丙酸和琥珀酸等有机酸,降低培养液p H,增加可溶性磷浓度。两株细菌均为革兰氏阴性杆菌,无芽孢,产生硫化氢,其中菌株JXJ-11的16S rRNA基因序列与Sphingomonas melonis DAPP-PG 224T和S.aquatilis JSS7T相似性最高(99.79%),菌株JXJ-15的16S rRNA基因序列与Klebsiella pneumoniae subsp.pneumoniae DSM 30104T相似性最高(99.73%),根据以上信息,确定菌株JXJ-11和JXJ-15分别是鞘氨醇单胞菌属和克雷白氏杆菌属的成员。菌株JXJ-11和JXJ-15的解磷机制包括分泌有机酸和磷酸酶,其中JXJ-11在微生物磷肥研制方面具有潜在应用价值。 相似文献
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8.
酸性土壤中磷易被固定,磷的生物有效性极低。解磷菌对土壤中难溶性磷具有重要的增溶作用。虽然已有不少解磷菌方面的研究,但是主要集中于中性和石灰性土壤中钙磷的解磷菌报道,而关于酸性土壤中高效溶解铝磷的微生物报道较少。采用培养基和土培试验,首先对酸性土壤上不同植物（胡枝子、大豆、水稻）根际土壤中的解磷菌进行了分离,然后比较了它们对不同磷源（磷酸钙和磷酸铝）的溶解能力,最后研究了它们对大豆生长和磷吸收的影响。通过使用难溶性磷源（磷酸钙和磷酸铝）的固体培养基,分离得到5株优势菌株L1、S1、S2、R1和R2,经16S rRNA序列鉴定,L1属于阮杆菌属（Nguyenibacter）,S1和S2分别属于假单胞菌属（Pseudomonas）和沙雷氏菌属（Serratia）,R1和R2分别属于伯克氏菌属（Burkholderia）和雷尔氏菌属（Ralstonia）。菌株S1、S2、R1和R2对难溶性磷酸钙有较强的溶解能力,对磷酸铝的溶解能力较弱;菌株L1对磷酸铝表现出较高的溶解能力,对难溶性磷酸钙的溶解能力弱。联合接种菌株L1+S1对大豆生长和磷吸收表现出良好的促进效果,而单独接种L1和S1效果不显著。... 相似文献
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盐碱地塔宾曲霉菌的解磷能力及其对小麦生长的影响 总被引：2，自引：0，他引：2
[目的]研发新型解磷生物菌肥,提高黄河三角洲盐碱障碍耕地作物产量。[方法]采用无机磷液体培养基培养的方法,从黄河三角洲盐碱化菜园根际土壤筛选得到一株解磷真菌CT1,即塔滨曲霉菌(Aspergillus tubingensis),对其解磷能力进行了深入研究。[结果]解磷菌CT1的解磷能力随发酵液盐浓度升高降低,当发酵液盐浓度在0.03%~6%时,发酵液中有效磷浓度可维持在523.5~338.5mg/L,且解磷菌的溶磷量与发酵液pH之间存在明显的负相关。解磷菌CT1在葡萄糖作为碳源的培养基上生长状况最好,在(NH_4)_2SO_4作为氮源的培养基上生长状况最好。接入解磷菌15d的小麦与未接菌的小麦相比,茎长增加了16.24%,茎鲜重增加了12.35%,根长增加了21.6%。[结论]解磷菌CT1对盐碱地小麦幼苗生长有一定的促进作用,可作为提高盐碱地作物产量的新型解磷生物菌肥利用。 相似文献
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为筛选并构建高效的作物根际解磷真菌,探讨解磷真菌的解磷机理,本研究从小麦和棉花根际土壤中分离筛选了一株高解磷能力的真菌YTY,对其离子束诱变的最佳条件进行探讨,进一步构建YTY的高效解磷突变菌,并从有机酸角度研究YTY的解磷机理。结果表明,从作物根际土壤分离的解磷真菌YTY具有较高的解磷能力,形态学和ITS鉴定表明该菌为草酸青霉;YTY的最佳诱变条件为30 keV和1×1015 ions·cm-2,利用该诱变条件获得了3株高效解磷突变株,即p-1-1、p-1-2、p-1-3,解磷能力分别较出发菌株YTY提高了56.88%、42.26%和32.15%;YTY培养液(5 d)的pH值为2.5,离子色谱法测得其中含有3种有机酸,即乳酸、乙酸和草酸,同时测得3株突变菌培养液的pH值为2.0,显著低于出发菌株YTY,且乳酸、乙酸、草酸和总酸含量均显著高于YTY,表明有机酸是YTY解磷的重要物质,且乳酸、乙酸、草酸是YTY解磷的主要有机酸。本研究结果为解磷真菌的离子束诱变选育提供了参考,并为真菌YTY解磷机理的探明及开发应用提供了理论依据和生物材料。 相似文献
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Influence of low-molecular-weight organic acids on the solubilization of phosphates 总被引：18，自引：0，他引：18
Dr N. S. Bolan R. Naidu S. Mahimairaja S. Baskaran 《Biology and Fertility of Soils》1994,18(4):311-319
A range of low-molecular-weight organic acids were identified in rhizosphere soil, leaf litter, and poultry manure compost. Laboratory and greenhouse experiments were carried out to examine the effects of seven low-molecular-weight organic acids on phosphate adsorption by soils, and the solubilization and plant uptake of P from soil pre-incubated with monocalcium phosphate and North Carolina phosphate rock. Acetic, formic, lactic (monocarboxylic), malic, tartaric, oxalic (dicarboxylic), and citric (tricarboxylic) acids were used in the study. The addition of organic acids decreased the adsorption of P by soils in the order tricarboxylic acid>dicarboxylic acid>monocarboxylic acid. The decreases in P adsorption with organic acid addition increased with an increase in the stability constant of the organic acid for Al (logK
Al). Organic acids extracted greater amounts of P from soils meubated with both monocalcium phosphate and phosphate rock than water did. Although more phosphate was extracted by the organic acids from monocalcium phosphate — than from phosphate rock — treated soils in absolute terms, when the results were expressed as a percentage of dissolved phosphate there was little difference between the two fertilizers. The amount of P extracted by the organic acids from both fertilizers increased with an increase in logK
Al values. The addition of oxalic and citric acids increased the dry matter yield of ryegrass and the uptake of P in soils treated with both fertilizers. The agronomic effectiveness of both fertilizers increased in the presence of organic acids and the increase was greater with the phosphate rock than with the monocalcium phosphate. The results indicated that organic acids increase the availability of P in soils mainly through both decreased adsorption of P and increased solubilization of P compounds. 相似文献
12.
Biosolubilization of phosphorus from rock phosphate and other P fertilizers in response to phosphate solubilizing bacteria and poultry manure in a silt loam calcareous soil
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下载免费PDF全文 ‘Phosphate solubilizing bacteria' (PSBs) are able to release unavailable P from native and applied P sources into plant‐available soil pool through their solubilizing and acidifying effects. The effects of three indigenous and one exotic PSBs on P solubilization from different P sources, plant biomass production, and P‐uptake efficiency of maize (Zea mays L.) were examined in an incubation and greenhouse study. For incubation study, surface (0–15 cm) soil was collected from an arable field (Inceptisols) and amended with rock phosphate (RP), single superphosphate (SSP), poultry manure (PM), and RP+PM with and without PSBs. The amended soil was incubated in the control environment at 25 ± 2°C for a total of a 100‐d period to establish relative potential rate of P solubilization of added P sources. A complementary greenhouse experiment was conducted in pots by growing maize as a test crop. Growth characteristics, P‐uptake, and P‐utilization efficiency (PUE) were determined. Phosphate solubilizing bacteria generated a solubilization effect on different P sources by releasing more P into plant‐available soil pool, i.e., 14.0–18.3 µg g?1 in RP, 5.0–9.9 µg g?1 in SSP, 1.4–4.4 µg g?1 in PM, and 4.5–7.8 µg g?1 in RP+PM compared to their sole application without PSBs. The available P from inorganic SSP declined continuously from the mineral pool (after day 30) and at the end 40% of applied P was unaccounted for. However, P losses were reduced to 28 and 27% when PSBs (PSB1 and PSB3) were applied with superphosphate treatments. In the absence of PSBs, the recoveries of applied P (in soil) from RP, SSP, PM and RP+PM were 4, 25, 9, and 12%, respectively, those had been increased to 14, 30, 12 and 15% in the presence of PSBs. Similarly, the plant biomass in RP+PSBs treatments compared to the RP without PSBs increased between 12–30% in first sampling (30 DAG) and 13–30% in the second sampling (60 DAG). The P utilization efficiency (PUE) in plants supplemented with PSBs was 20–73% higher compared to those without PSBs. The detection of oxalic and gluconic acids in culture medium treated with PSBs (7.8–25.0 and 25–90 mg L?1, respectively) confirmed the production of organic acids by the indigenous bacterial isolates. This study indicate that low P recovery both in plant and soil can likely be improved by using indigenous PSBs and organic amendment poultry manure, which allowed a more efficient capture of P released due to P solubilization. 相似文献
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从石灰性土壤作物根际土样筛选出2株具有较强解P能力的优势菌株“B2”和“B67”,平板培养溶P圈D/d值分别为5.1和3·8,液体摇床培养过程中产生苹果酸、丙二酸、草酸和乳酸等多种有机酸,产酸总量分别为对照的28·25和19.78倍,pH分别较对照下降2.3和1.1,将“B2”和“B67”的培养液分别接种到4个不同地区土壤中恒温恒湿培养20d后测定结果表明,所有土壤中速效磷含量均有所提高,且接种“B2”处理分别为对照的2.64、3.04、1.71和2.20倍,接种“B67”处理分别为对照的2.12、1.90、1.35和1.78倍,且发现其溶P效果和土壤磷酸酶活性与有效活菌数变化趋势基本一致。 相似文献
14.
To confirm whether endophytes are members that play important roles in phosphorus (P) solubilization in red acidic soil, five endophytes that have the potential to dissolve insoluble P were isolated from cassava (Manihot esculenta Crantz) root. Based on the maximum amount of soluble P in Ca3(PO4)2, AlPO4, and FePO4 liquid NBRIP, the strain Pantoea dispersa was selected to investigate the ability to solubilize phosphate over time. Our results showed that the solubilizing process of P. dispersa was accompanied by acid production. Beside succinate, oxalic acid, and citric acid, two special organic acids, salicylic acid, and benzeneacetic acid were found during microbial P solubilization. Based on PCR-DGGE (denaturing gradient gel electrophoresis) analysis, soil application of P. dispersa triggered natural soil microbial activity. This phenomenon could be maintained up to 25 days, suggesting that the endophyte P. dispersa would be a suitable candidate for optimizing agro-microecological systems via soluble P release in red acidic soil. 相似文献
15.
从滇池富磷区的89份土样中筛选出48株溶磷真菌,采用钼蓝法测定这些真菌溶解Ca3(PO4)3的能力。结果表明培养液中可溶性磷含量在14.45～64.87?mg/L,其中菌株SPF46、SPF47和Mo-Po的溶磷能力最强,其培养液中可溶性磷含量分别达到55.44、59.78和64.87?mg/L。结合形态特征及ITS rDNA 系统亲缘关系分析,将PSF46鉴定为黄暗青霉（Penicillium citreonigrum）,PSF47菌株鉴定为黑曲霉（Aspergillus niger）,Mo-Po菌株鉴定为草酸青霉（Penicillium oxalicum）。在Ca3(PO4)3、FePO4.4H2O和AlPO4 3种磷源中,这3种溶磷真菌对Ca3(PO4)2的溶磷效果最好。通过盆栽试验表明这3种溶磷真菌对油菜具有较好的促生效果,单一菌株处理,根系增长率在10.16%～294.7%,株高增长率在32.19%～134.5%;叶片直径增长率在35.53%～170.5%;鲜质量和干质量增长率分别在30.71%～189.5%,56%～224%。3种溶磷真菌混合处理,根系增长129.4%,株高增加60.41%,叶片直径增加170.5%;鲜质量和干质量分别增加246.1%、272.2%。 相似文献
16.
Many phosphate solubilizing microorganisms (PSM) require external pyrroloquinoline quinone (PQQ) for strong phosphorus (P)
solubilization in vitro. The objective of this study was to isolate efficient and PQQ-independent PSM. A total of 21 PSM were
isolated from the rhizosphere soil of wheat and maize grown in the pots. Acinetobacter strains were the only PQQ-independent and most effective solubilizers of tricalcium phosphate containing agar. The mean P
dissolved in liquid cultures of Acinetobacter strains in a 5-day incubation ranged from 167 to 888 μg/ml P. The pH dropped to below 4.7 from 7.8 in six isolates, which
produced gluconic acid in concentrations ranging between 27.5 and 37.5 mM. There was a linear regression between soluble P
and gluconic acid concentrations in the bacterial cultures (P < 0.05; R
2 = 0.59). Inoculation with Acinetobacter sp. WR922 significantly (P < 0.05) increased wheat (Triticum aestivum L.) P content by 27% at 15 days after emergence (DAE) and dry matter by 15% at 30 DAE compared to the control. The plant
P content in inoculated plants at 30 DAE was linearly correlated with soluble P of the bacterial cultures (P < 0.05; R
2 = 0.69). Gluconic acid production directly affected phosphate solubilization in vitro, which in turn influenced plant P content
of inoculated plants in PQQ-independent P-solubilizing Acinetobacter strains. 相似文献
17.
Jose Martin Scervino Milton Prieto Mesa Ivana Della Mónica Marina Recchi Nubia Sarmiento Moreno Alicia Godeas 《Biology and Fertility of Soils》2010,46(7):755-763
Phosphorus availability is a major limiting factor for yield of most crop species. The objective of this study was to compare
the solubilization of three sources of phosphorus (P) by different fungal isolates and to determine the possible mechanisms
involved in the process. Talaromyces flavus (S73), T. flavus var flavus (TM), Talaromyces helicus (L7b) and T. helicus (N24), Penicillium janthinellum (PJ), and Penicillium purpurogenum (POP), fungal strains isolated from the rhizosphere of crops, are known to be biocontrol agents against pathogenic fungi.
The P solubilization efficiency of these fungal strains in liquid media supplemented either with tricalcium phosphate (Ca3(PO4)2; PC), aluminum phosphate (AlPO4; AP), or phosphorite (PP) depended on the source of P and the fungal species. The type and concentration of organic acids
produced by each species varied according to the source of available P. In the medium supplemented with PC, the highest proportion
was that of gluconic acid, whereas in the media supplemented with the other P sources, the highest proportion was that of
citric and valeric acids. This suggests that the release of these organic compounds in the rhizosphere by these microorganisms
may be important in the solubilization of various inorganic P compounds. Results also support the hypothesis that the simultaneous
production of different organic acids by fungi may enhance their potential for solubilizing insoluble phosphate. 相似文献
18.
绿肥腐解液中有机酸组成对铝磷和铁磷活化能力的影响 总被引：1，自引：0，他引：1
19.
磷细菌P17对不同来源磷矿粉的溶磷作用及机制 总被引：22，自引：1，他引：22
比较了一株磷细菌P17对来自多种产地磷矿粉的溶解能力。通过摇瓶试验、扫描电镜观察提出磷细菌P17对不同来源磷矿粉生物风化的证据 ,选出了P17最适合作用的磷矿粉类型。从摇瓶试验看出 ,磷细菌P17对来源于黄麦岭、黄金卡黄的变质岩型磷矿粉有较好的溶解能力。经过P17长达 70d的溶解 ,磷矿粉的难溶磷逐渐被P17菌株溶解下来。连续 10次接种培养后 ,P17菌株能溶解黄麦岭磷矿粉全磷的81 0 2 % ;而对于黄金卡黄磷矿粉 ,P17溶解了全磷总量的 78 97%。试验结果表明磷细菌P17能够提高黄麦岭磷矿粉和黄金卡黄磷矿粉的持续利用率。另外 ,培养 7d后 ,磷细菌P17能够在以黄麦岭磷矿粉、黄金卡黄磷矿粉为唯一磷源的发酵液中产生 3 10mmolL-1的挥发性有机酸 ,同时能分别产生 4 0 5mmolL-1、5 1 1mmolL-1的难挥发性有机酸。经过气相色谱检测 ,磷细菌P17能够产生柠檬酸、琥珀酸、乳酸以及乙酸等有机酸 ,可能螯合磷矿粉中的金属离子 ,使磷游离出来。初步探讨了磷细菌P17的溶磷机制 相似文献