磷细菌筛选及其对土壤无机磷转化的影响

从潮土、水稻土、砂姜黑土和石灰土等土壤的植物根际土壤和根中分离了86株磷细菌,通过液体摇瓶培养3d,培养液水溶磷含量为4.2～387.3mg/L,水溶磷含量与培养液pH呈显著负相关(R2=0.621 6)。用筛选出的1株磷细菌(HCW115)进行玉米盆栽试验,结果表明,磷细菌处理的玉米干物重和吸磷量与对照相比分别增加了37.5%和40.2%,达到显著差异。磷细菌对土壤Al-P、Fe-P和O-P转化无明显影响,但可以促进土壤Ca2-P、Ca8-P和Ca10-P向有效磷转化而被玉米吸收,与原土相比,Ca2-P、Ca8-P和Ca10-P含量分别减少了74.9%,12.3%和1.51%。     

解磷微生物及其在盐碱土中的应用研究进展

解磷微生物能够将土壤中难溶性磷酸盐转化为可供植物吸收利用的磷,在提高土壤有效磷含量、缓解植物盐碱胁迫损伤及修复盐碱土中表现出良好的应用前景.本文从解磷微生物的研究及应用现状入手,综述了盐碱土中解磷微生物的分离、解磷机制,同时阐述了盐碱耐受性和解磷能力关系及其在盐碱土修复中的应用进展,以期为利用耐盐碱解磷菌缓解植物盐胁迫...     

作物根际解磷真菌YTY的离子束诱变及其解磷机理探讨

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。     

Solubilization of Insoluble Inorganic Phosphate by Hyphal Exudates of Arbuscular Mycorrhizal Fungi

Increased phosphate (P) uptake in plants by arbuscular mycorrhizal (AM) fungi is thought to depend mainly on the extension of external hyphae into soil. On the other hand, it is known that the hyphae of some kinds of ectomycorrhizal fungi release organic acids into soil and that they dissolve the insoluble inorganic P. This study collected hyphal exudates of AM fungi within compartmentalized pot culture and clarified their ability to solubilize insoluble inorganic P. Sterilized Andisol was packed in pots that were separated into root and hyphal compartments with a nylon net of 30 μm pore size. Seedlings of Allium cepa inoculated with AM fungi, Gigaspora margarita, or Glomus etunicatum were grown. Control pots were not inoculated. Mullite ceramic tubes were buried in the soil of each compartment and soil solution was collected. The anionic fraction of the soil solution was incubated with iron phosphate (4 mg FePO₄ in 1 mL of 0.4 acetate buffer). Solubilized P was measured. The AM colonization of plants inoculated with G. margarita and G. etunicatum was 86% and 54%, respectively. Adhesion of external hyphae was observed on the surface of the mullite ceramic tubes buried in soil of the hyphal compartment. Colonization of both fungi increased shoot P uptake and growth. Soil solution collected from the hyphal compartments of both fungi solubilized more P than did that from uninoculated plants. It is suggested that hyphal exudates can contribute to increased P uptake of colonized plants.     

解磷菌剂对作物生长和土壤磷素的影响

利用课题组自行分离筛选的B2和B67解磷菌株研制的菌剂，在盆栽和大田试验过程中，菌剂在增加作物产量的同时，有提高土壤速效磷含量、培肥土壤的作用，而且是菌剂和化肥配合施用效果显著。施用了解磷菌剂或菌剂和化肥配合施用后，盆栽玉米的株高、鲜重和干重，较CK增加15．2％～89．2％，玉米单株吸养量较CK增加20％～146％；青菜的鲜重、干重和生物量较CK增加15．8％～41．6％，和CK比较差异显著；小麦的株高、有效小穗数、穗粒数、粒重以及生物量也较CK差异显著，大田试验的几种作物也有不同程度的增产。另外B2和B67接种到山西省典型土壤中，土壤速效磷含量较CK增加1．35～3．04倍，且发现其溶磷效果和土壤磷酸酶活性与有效活菌数相关，解磷菌剂在提高土壤速效磷含量的同时，也有提高土壤速效钾含量的作用。     

细菌溶磷作用与磷矿粉伴生性重金属元素的释放

以4株溶磷细菌和5种磷矿粉为材料,对细菌溶磷作用与磷矿粉伴生性重金属元素释放间的关系进行了研究。结果表明,溶磷细菌在促进磷矿粉中磷溶出的同时,还促进了磷矿粉中伴生性重金属的释放,在测定的6种伴生性金属元素(Zn、Pb、Cu、Co、Ni、Cd)中,Zn最容易随细菌溶磷作用而释放,最多可占该磷矿粉中总Zn量的97.0%,Pb和Cd随细菌溶磷释放率皆低于10%;细菌溶磷作用强度与磷矿粉中各伴生性重金属释放量间的关系因磷矿粉产地和伴生性重金属种类而异。细菌作用于湖南石门磷矿粉和四川马边磷矿粉的溶磷量与Zn、Cu、Co、Ni释放量间(四川马边磷矿粉中Co除外)正相关,但未达到显著水平(r=0.50~0.95,P>0.05),但在溶解贵州开阳磷矿粉时,Zn、Co、Ni的释放量都与溶磷量显著正相关(r>0.88,P<0.05)。     

Investigation on Phosphate Solubilization Potential of Agricultural Soil Bacteria as Affected by Different Phosphorus Sources,Temperature, Salt,and pH

Ten phosphate-solubilizing bacterial strains belonging to genera Pseudomonas, Burkholderia, Enterobacter, Serratia, Klebsiella, and Aeromonas were tested for mineral phosphate solubilization activity in Pikovskaya's broth using different phosphate sources at four temperatures (15, 25, 35, and 45 ^οC). Dicalcium and tricalcium phosphate were solubilized more effectively (≥1000 mg L^?1) than ferric and rock phosphate (≥100 mg L^?1) and 35 °C was found to be the optimum temperature. Although Klebsiella and Aeromonas spp. are well known for their dinitrogen (N₂)–fixing ability, to the best of our knowledge, this is the first report of inorganic phosphate solubilization by Klebsiella terrigena and Aeromonas vaga. Interestingly, A. vaga BAM-77 is the most efficient strain at solubilizing inorganic phosphorus (P) even in the presence of 8% sodium chloride (NaCl) at pH 10. These findings indicate that all four strains are efficient P solubilizers under variable conditions of temperature, pH, and P source and thus can be recommended for P fertilization in different soils.     

Cd2+ 对无机磷细菌生长及溶磷作用的影响

无机磷细菌(phosphorus solubilization bacteria,PSB)是重要的土壤有益微生物,它们将植物不能吸收的难溶性无机磷转化为有效磷,对土壤磷素供应十分重要。镉是污染土壤的常见重金属,研究Cd~(2+)胁迫下PSB的生长和溶磷能力,可揭示镉污染对土壤无机磷转化供应的影响,为改善植物磷素供应提供有益信息。试验以重庆市缙云山国家森林公园毛竹林地分离的4株假单胞菌为供试菌株,比较它们的溶磷能力,研究了不同浓度Cd~(2+)对PSB生长和溶磷作用的影响。结果表明:供试菌株溶磷能力PSB05PSB09PSB07PSB01,溶磷指数和溶磷率最高值是最低值的2.08倍和4.14倍。摇瓶培养120 h后,培养液pH表现为PSB01PSB07PSB09PSB05,并与培养液中的无机磷含量(y)呈显著负相关(y=–83.148 pH+581.96,R2=0.9052,n=25)。在固体培养时,PSB05的溶磷圈和菌落直径随培养基中Cd~(2+)浓度提高而降低。在Cd~(2+)浓度0.5~2.0 mg/L液体培养基中,PSB05数量较无Cd~(2+)对照减少22.09%~68.18%,草酸、乙酸、柠檬酸和氢离子分泌量显著下降,溶磷量降低5.27%~38.45%。供试菌株的溶磷能力差异显著;高浓度Cd~(2+)抑制PSB05生长、有机酸和氢离子分泌及溶磷作用,不利于土壤供给磷素。     

Phosphate Solubilizing Ability and Phylogenetic Diversity of Bacteria from P-Rich Soils Around Dianchi Lake Drainage Area of China

The phylogenetic diversity of phosphate solubilizing bacteria(PSB)distributed in P-rich soils in the Dianchi Lake drainage area of China was characterized,and the tricalcium phosphate(TCP)solubilizing activities of isolated PSB were determined.Among 1 328 bacteria isolated from 100 P-rich soil samples,377 isolates(28.39%of the total)that exhibited TCP solubilization activity were taken as PSB.These PSB showed different abilities to solubilize TCP,with the concentrations of solubilized P in bacterial cultures varying from 33.48 to 69.63 mg L^-1.A total of 123 PSB isolates,with relatively high TCP solubilization activity(>54.00 mg L^-1),were submitted for restriction fragment length polymorphism(RFLP)analysis,which revealed 32 unique RFLP patterns.Based on these patterns,62 representative isolates,one to three from each RFLP pattern,were seffected for 16S rRNA sequencing.Phylogenetic analysis placed the 123 PSB into three bacterial phyla,namely Proteobacteria,Actinobacteria and Firmicutes.Members of Proteobacteria were the dominant PSB,where 107 isolates represented by 26 RFLP patterns were associated with the genera of Burkholderia,Pseudomonas,Acinetobacter,Enterobacter,Pantoea,Serratia,Klebsiella,Leclercia,Raoultella and Cedecea.Firmicutes were the subdominant group,in which 13 isolates were affiliated with the genera of Bacillus and Brevibacterium.The remaining 3 isolates were identified as three species of the genus Arthrobacter.This research extends the knowledge on PSB in P-rich soils and broadens the spectrum of PSB for the development of environmentally friendly biophosphate fertilizers.     

AM真菌菌丝际细菌具有固氮解磷双重功能

为了认识和理解丛枝菌根真菌(AM真菌)根外菌丝表面定殖的细菌是否同时具有固氮和解磷能力,从田间生长的玉米菌根根外菌丝表面分离鉴定了固氮菌,评价了其固氮和解磷能力.从AM真菌根外菌丝表面分离出23株可在无氮培养基中生长的细菌,分属于变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和放线菌门(Ac...     

溶磷细菌Enterobacteria sp EnHy-401对盐渍土壤中小麦生长的影响

盆栽试验结果表明,溶磷菌Enterobacteria sp EnHy-401能较好地在盐渍土壤中定植,并对小麦的生长和矿质营养的吸收有促进作用。与对照相比,生长在经接菌处理盐渍土中的小麦,其地上部分和根系的干重增加了28.1%和14.6%,植株中对磷、钙、钾和镁的吸收率分别提高了34.4%3、6.3%、31.5%6、.3%。该菌株所表现出来的对盐渍土壤中磷素有效性的提高、对小麦吸收K+、Ca2+、Mg2+、P等矿质营养的促进作用以及小麦生长与矿质营养元素吸收之间高度的相关性表明,溶磷菌株Enterobacteria sp.EnHy-401对盐胁迫下小麦生长的促进效应主要是通过改善盐渍土中部分矿质营养的供应,增强植株对P、Ca,Mg、K等营养元素的选择性吸收而实现,该效应受到土壤盐分和营养基质的影响。     

Absorption of Ammonia Released from Poultry Manure to Soil and Bark and the Use of Absorbed Ammonia in Solubilizing Phosphate Rock

Composting systems were designed to utilize ammonia (NH₃) released during composting of poultry manure to solubilize phosphate rock (PR). The NH₃ released from decomposing manure was allowed to pass through columns containing soil or bark materials mixed with North Carolina phosphate rock (NCPR) at a rate of 1 mg P g^?1. After eight weeks of incubation, the columns were dismantled and the forms of P and N in PR/soil or PR/bark mixtures were measured. The dissolution of PR was determined from the increases in the amount of soluble and adsorbed P (resin plus NaOH extractable P) or from the decreases in the residual apatite P (HC1 extractable P).

The amounts of NH₄^+-N in the soil and bark columns increased due to absorption of the NH₃ released from poultry manure. No nitrification of absorbed NH₃ occurred, however, unless the soil or bark were reinoculated with a fresh soil solution and incubated for further six weeks.

In the absence of NH₃ absorption, soil and bark materials dissolved approximately 33 percent and 82 percent of NCPR, respectively. The higher dissolution of NCPR in bark was attributed to its higher exchangeable acidity and Ca sink size. There was no increase in NCPR dissolution during the initial NH₃ absorption phase (36 percent and 85 percent dissolution in soil and bark respectively), which may be due to the absence of nitrification. However, during subsequent reincubation when nitrification occurred, the final dissolution of NCPR in the NH₃ treated soil and bark was slightly higher (41 percent and 100 percent, respectively). Protons (H⁺) are released during the oxidation of NH₄⁺ to NO_3? (nitrification) which promote the dissolution of PR. However, most of the H⁺ released during nitrification was involved with soil and bark pH buffering reactions. Only five to 10 percent was involved in PR solubilization in PR/soil mixtures whereas about 50 percent was involved in PR/bark systems.

Bark covers for poultry manure and poultry manure compost heaps have the potential to reduce NH₃ loss and conserve N and may be useful for other purposes such as PR solubilization.     

Comparative Effects of Two Forage Species on Rhizosphere Acidification and Solubilization of Phosphate Rocks of Different Reactivity

ABSTRACT

Dissolution of phosphate rocks (PR) in soils requires an adequate supply of acid (H⁺) and the removal of the dissolved products [calcium (Ca^{2 +}) and dihydrogen phosphate (H₂PO₄ ^?)]. Plant roots may excrete H⁺ or OH^? in quantities that are stoichiometrically equal to excess cation or anion uptake in order to maintain internal electroneutrality. Extrusion of H⁺ or OH^? may affect rhizosphere pH and PR dissolution. Differences in rhizosphere acidity and solubilization of three PRs were compared with triple superphosphate between a grass (Brachiaria decumbens) and a legume (Stylosanthes guianensis) forage species at two pH levels (4.9 and 5.8) in a phosphorus (P)-deficient Ultisol with low Ca content. The experiment was performed in a growth chamber with pots designed to isolate rhizosphere and non-rhizosphere soil. Assessment of P solubility with chemical extractants led to ranking the PRs investigated as either low (Monte Fresco) or high solubility (Riecito and North Carolina). Solubilization of the PRs was influenced by both forage species and mineral composition of the PR. The low solubility PR had a higher content of calcite than the high solubility PRs, which led to increased soil pH values (> 7.0) and exchangeable Ca, and relatively little change in bicarbonate-extractable soil P. Rhizosphere soil pH decreased under Stylosanthes but increased under Brachiaria. The greater ability of Stylosanthes to acidify rhizosphere soil and solubilize PR relative to Brachiaria is attributed to differences between species in net ion uptake. Stylosanthes had an excess cation uptake, defined by a large Ca uptake and its dependence on N₂ fixation, which induced a significant H⁺ extrusion from roots to maintain cell electroneutrality. Brachiaria had an excess of anion uptake, with nitrate (NO₃ ^?) comprising 92% of total anion uptake. Nitrate and sulfate (SO₄ ^{2 ?}) reduction in Brachiaria root cells may have generated a significant amount of cytoplasmic hydroxide (OH^?), which could have increased cytoplasmic pH and induced synthesis of organic acids and OH^? extrusion from roots.     

解钾菌及其释钾微观机制的研究进展

解钾菌主要通过黏附到矿物表面并释放酸性物质,将难溶性钾、磷和硅等营养元素释放出来进而供植物吸收利用。研究解钾菌释钾机制,对土壤肥力改善和绿色农业发展具有重要作用,并促使其成为国内外学者关注的热点问题。前人有关解钾机制（酸解、酶解、多糖络合溶解等）和室内或田间施用效果等宏观方面做了大量的研究和总结,然而,从微观尺度揭秘解钾菌和矿物间的生物物理相互作用机制很可能是剖析解钾过程黑匣子的关键和新思路。本文简要概述了解钾菌的种类和施用效果,重点剖析了单细胞水平上解钾菌在含钾矿物表面生物膜形成微观过程及影响因素,并详细介绍了研究解钾菌相关的前沿科学技术。     

Investigation on Co-sorption and Desorption of Fluoride and Phosphate in a Red Soil of China

Agricultural soils high in both fluoride (F) and phosphate (P) are common due to long-term accumulation of F from multi-sources and extensive application of phosphate fertilizers. Iron (Fe) and aluminum (Al) (hydro)oxides in acidic soils serve as main geochemical sinks of both P and F, influencing their transport and bioavailability. Though sorption of P and F in their single-ion system has been extensively investigated, studies on co-sorption of F and P on soils are very limited. In this study, the batch technique was used to investigate mutual effects of F and P on their co-sorption/desorption in an acidic red soil with high contents of Fe and Al (hydro)oxides. Results indicate that, in F–P coexisting system, a decrease in pH enhances the sorption of both F and P. An increase in F concentration suppresses P sorption due to competitive effect. However, F sorption can be improved in the presence of P due to surface precipitation of (Al,Fe)–F–P. Sorption of F and P follows both the Langmuir and Freundlich isotherms. Different orders of F and P addition into the soil have no appreciable effect on P sorption, but exert significant impact on F sorption. The presence of F has no measurable effect on P desorption, while the stability of F in the presence of P can be significantly diminished in comparison with that in the absence of P, which would lead to an improvement of F mobility.     

施磷对吉林省西部盐碱土水田土壤无机磷组分的影响

为探明磷肥施用对盐碱土水田土壤无机磷组分的影响,以吉林省西部地区盐碱土水田土壤为研究对象,通过田间试验结合室内分析的方法,研究了不同磷肥施用量的情况下,土壤不同形态无机磷在水稻生育期内含量的动态变化。结果表明:施用磷肥可以在整个水稻生育期间提高盐碱土水田土壤中的有效磷源及潜在磷源的含量,包括Ca_2-P、Ca_8-P、Al-P和Fe-P等不同形态的无机磷含量;O-P是土壤无机磷组分中含量较高且极稳定的磷,在一个水稻生育期内,施肥不会引起O-P含量的明显变化;虽然以往认为Ca_(10)-P是土壤中不可利用的磷,但本研究中发现,Ca_(10)-P在水稻生育期间仍可发生一定的转化,较高磷肥施用量有利于土壤中Ca_(10)-P的积累。研究表明,盐碱土水田磷肥的施用,在水稻生育前期可以形成可被水稻吸收的有效态磷源和潜在磷源,水稻生育后期则开始向无效态磷转化并在土壤中积累。     

溶磷细菌对不同性质磷矿粉的溶磷效果研究

以磷矿粉为唯一磷源,在液体摇床培养条件下,研究了溶磷细菌Jm170对不同性质磷矿粉的溶磷作用。结果表明,在本试验条件下,细菌Jm170对四种供试磷矿粉均有较好的溶磷作用,且溶磷能力表现出明显选择性:品位愈高,溶磷能力愈强;磷矿中钙含量愈高,铁、铝含量愈低,溶磷能力愈强。     

长期施肥下红壤旱地解钾菌变化及其驱动因子

在南方红壤区,研究长期施肥下土壤解钾菌的变化规律及其驱动因素,可为该地区土壤钾素资源管理和钾肥合理施用提供理论依据。基于红壤旱地长期定位施肥试验(始于1986年),选取不施肥(CK)、施氮磷肥(NP)、氮磷钾肥(NPK)和氮磷钾肥配施有机肥(NPKM)处理,分析玉米开花期根际土壤的解钾菌类型和解钾能力及其有机酸和激素含量,并结合玉米根系特性和根际土壤理化性质探讨影响红壤旱地解钾菌变化的关键因子。结果表明:与NPK处理相比,NPKM处理的根长、根表面积、根直径和根体积分别增加了112.3%、174.4%、32.43%和291.5%,根际土壤pH提高了0.67个单位,有机质、非交换性钾和交换性钾分别增加了29.50%、19.34%和53.89%。各施肥处理根际土壤中均存在解钾菌,CK和NP处理的解钾菌均为类芽孢杆菌属,NPK和NPKM处理则均为纤维菌属。与CK处理相比,NP、NPK和NPKM处理下根际土壤解钾菌的解钾效率分别增加了162.4%、139.0%和105.6%,其中NP处理解钾菌的解钾效率最高。偏最小二乘路径模型的结果显示,根系和施肥可同时调控解钾菌的解钾效率。冗余分析(RDA)...     

Isolation of a Phytase-Producing Bacterial Strain from Agricultural Soil and its Characterization and Application as an Effective Eco-Friendly Phosphate Solubilizing Bioinoculant

In the present study, we isolated from agricultural soils, a Bacillus strain designated SD01N-014, expressing high-level extracellular phytase activity and investigated its potential to be used as a soil phosphate-solubilizing bioinoculant to improve soil phosphorus (P) availability and thus enhance plant growth. Soil inoculation test showed that being as an inoculant, SD01N-014 substantially enhanced soil P nutrition. Pot experiments showed that when applied to soils, SD01N-014 is capable of colonizing maize seedling roots, increasing plant-available P levels in the rhizosphere and promoting seedling growth. High tunnel trials with different vegetable plants demonstrated that SD01N-014 as a soil inoculant greatly promoted plant growth and was positively correlated with inoculum sizes. Together, these results indicate that SD01N-014 holds promising potential as a cheap, viable and eco-friendly alternative to chemical P fertilizers for increasing soil fertility and crop production in sustainable farming.     

坡地赤红壤不同利用模式对土壤磷和酶的影响

试验采用广东东莞花岗岩发育的坡地赤红壤为样品进行研究表明 :在坡地赤红壤的 4种利用模式中 ,旱地种植花生 ,荔枝可以有效地改善土壤的酸性 ,提高土壤肥力 ;提高土壤Al-P和Fe -P的含量 ,减少O -P的所占的比例 ,提高坡地赤红壤磷酸盐的有效性 ;增加土壤表层的脲酶和蛋白酶的活性 ,可能使土壤的磷酸酶的活性降低