Effect of Arbuscular Mycorrhizal Fungi and Phosphate-Solubilizing Bacteria Consortia Associated with Phospho-Compost on Phosphorus Solubilization and Growth of Tomato Seedlings (Solanum lycopersicum L.)

ABSTRACT

The exploitation of phosphate mines generates an important quantity of phosphate sludge that remains accumulated and not valorized. In this context, composting with organic matter and rhizospheric microorganisms offers an interesting alternative and that is more sustainable for agriculture. This work aims to investigate the synergetic effect of arbuscular mycorrhizal fungi (AMF), phosphate-solubilizing bacteria (PSB) and phospho-compost (PC), produced from phosphate-laundered sludge and organic wastes, and their combination on plant growth, phosphorus solubilization and phosphatase activities (alkaline and acid). Inoculated mycorrhizae and bacteria strains used in this study were selected from plant rhizosphere grown on phosphate-laundered sludge. Significant (p < .05) increases in plant growth was observed when inoculated with both consortia and PC (PC+ PSB+ AMF) similar to those recorded in plants amended with chemical fertilizer. Tripartite inoculated tomato had a significantly (p < .05) higher shoot height; shoot and root dry weight, root colonization and available P content, than the control. Co-inoculation with PC and AMF greatly increased alkaline phosphatase activity and the rate of mycorrhizal intensity. We conclude that PC and endophytic AMF and PSB consortia contribute to a tripartite inoculation in tomato seedlings and are coordinately involved in plant growth and phosphorus solubilization. These results open up promising prospects for using formulate phospho-compost enriched with phosphorus-solubilizing microorganisms (PSM) in crop cultivation as biofertilizers to solve problems of phosphate-laundered sludge accumulation.     

AM真菌对田间山银花生长、矿质营养及绿原酸含量的影响

Lonicera confusa, a traditional Chinese medicine herb for treating cold, flu, acute fever, and so forth, is often grown artificially in acidic soils and suffers from phosphorus (P) deficiency. A five-year field experiment was carried out to study the colonization rate, growth, nutrition, and chlorogenic acid content of Lonicera confusa seedlings inoculated with arbuscular mycorrhizal (AM) fungi, Glomus etunicatum and Glomus intraradices. Before transplanting into a field, both AM-inoculated and uninoculated control plants were cultured in nursery beds. In the plants inoculated with the AM fungi, the colonization rate decreased linearly with time and a greater decrease was observed in the plants inoculated with G. intraradices than with G. etunicatum, while the AM colonization increased from 0% to 12.1% in the uninoculated control plants 5 years after transplanting. Plant height, crown diameter, number of new branches, and flower yield increased significantly by AM inoculation as compared to the uninoculated control. Phosphorus concentrations in leaves and flowers increased, and plant uptake of nutrients, e.g., nitrogen (N), P, and potassium (K), was also enhanced significantly by AM inoculation. The Lonicera confusa seedlings had a better response to inoculation of G. intraradices than G. etunicatum in both growth and chlorogenic acid content in flowers. In contrast, both plant P uptake and P concentrations in leaves and flowers were similar between two fungal inoculations. The positive responses of Lonicera confusa to AM inoculation in growth, nutrient uptake, flowering, and chlorogenic acid content in flowers suggested that AM inoculation in nursery beds could promote the plant growth and increase chlorogenic acid content in flowers of Lonicera confusa when grown on acidic and P-deficient soils.     

棉花内生拮抗菌Paenibacillus xylanilyticus YUPP-1抗黄萎病研究

 在棉花现蕾期筛选到一个高抗黄萎病的内生菌菌株,根据16S rDNA序列同源性,将其命名为Paenibacillus xylanilyticus YUPP-1(解木聚糖类芽孢杆菌YUPP-1)。当把该菌标记后,每株棉花施用该菌株达到1×10⁶ cfu时,该菌能顺利定殖于棉花体内。室内盆栽抗病实验表明：当生防菌灌根处理的菌量为每株1×10⁶,1×10⁷,1×10⁸ cfu,其在结铃期的发病株率仅为8%,5%和2%,而此时对照的发病株率高达96%。2008－2009年的小区防治结果表明：当生防菌灌根处理的菌量为每株1×10⁸ cfu时,2008年处理区的棉花黄萎病发病株率和病指为14.8%和8.5,对照分别为53.3%和26.5,2009年处理区的棉花黄萎病发病株率和病指为9.4%和6.5,对照为37.8%和18.8。这些结果表明,解木聚糖类芽孢杆菌YUPP-1对防治棉花黄萎病具有巨大的应用潜力。     

绿色荧光蛋白标记解淀粉芽孢杆菌Bam22在油菜体内的定殖

研究GFP标记的解淀粉芽孢杆菌Bam22在油菜植株上的定殖规律,为其开发利用提供科学依据.采用自然转化法将携带绿色荧光蛋白基因的质粒pGFP4412导入Bam22细胞中,构建GFP荧光标记菌株;对比野生型菌株Bam22与标记菌株Bam22-GFP的生长曲线和抑菌能力;通过灌根法,观察标记菌株在油菜体内不同组织部位的定殖...     

Population size and specific potential of P-mineralizing and -solubilizing bacteria under long-term P-deficiency fertilization in a sandy loam soil

The effects of organic manure, mineral fertilizer (NPK), and P-deficiency fertilization (NK) on the individual biomass of young wheat plants, arbuscular mycorrhizal (AM) colonization in wheat root systems, population sizes of soil organic phosphorus mineralizing bacteria (OPMB) and inorganic phosphate solubilizing bacteria (IPSB) as well as soil P-mineralization and -solubilization potential were investigated in a long-term (18-year) fertilizer experiment. The experiment included five treatments: organic manure, an equal mixture of organic manure and mineral fertilizer, fertilizer NPK, fertilizer NK, and the control (without fertilization). Plant biomass, population sizes of soil OPMB and IPSB were greatly increased (P<0.05) by the application of organic manure and slightly increased by the balanced application of mineral fertilizer, while undiminished AM colonization in wheat root system was only observed in the case of the NK treatment. Compared to balanced fertilization, P-deficiency fertilization resulted in a significant increase (P<0.05) of OPMB-specific mineralization potential (soil P-mineralization potential per OPMB cell) and highest IPSB-specific solubilization potential (soil P-solubilization potential per IPSB cell), suggesting that OPMB and IPSB are likely more metabolically active in P-deficiency fertilized soils after long-term fertilizer management, and mycorrhizal plants are more dependent on AM in P-poor soils than in P-fertilized soils. Our results also showed the different effects of mineral fertilizer versus organic manure on soil P-mineralization and -solubilization potentials, as well as specific potentials of OPMB and IPSB in arable soils.     

棉花黄萎病拮抗细菌HMB-1005鉴定及定殖

以大丽轮枝菌(Verticillium dahliae)为指示菌,对从棉田土壤中已经分离筛选到的细菌HMB-1005进行形态、19项生理生化特征及16S rDNA试验,结果表明,HMB-1005为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);通过灭菌土和非灭菌土进行盆栽试验,拮抗菌HMB-1005芽孢液浸种、土壤全部混菌和部分混菌3种处理,经过5个月的盆栽试验表明,拮抗细菌HMB-1005能够在灭菌土和非灭菌土中定殖,且定殖数量达106cfu/g土左右,并仍能保持较高的抑菌活性。通过对棉苗根内细菌的分离,证实拮抗细菌HMB-1005能在棉花根内定殖,数量达到103cfu/g。实验结果显示,灭菌土的拮抗菌数高于非灭菌土,并且3种处理方式差异不明显,在实际生产应用中,建议使用浸种方法。     

短小芽孢杆菌BX-4抗生素标记及定殖效果研究

为研究短小芽孢杆菌BX-4在作物根际的定殖及防病效果,通过浓度梯度法用氨苄青霉素对其进行了抗性标记,并通过番茄盆栽试验研究了其在根际土壤中的定殖规律。结果表明,筛选出的突变体菌株BX-4＇能够耐受浓度为200 μg·mL^-1的氨苄青霉素,并且具有耐药和遗传双重稳定性;应用试验显示该突变体菌株能成功在番茄根际定殖,接种20 d后根际土壤中存活数量达到最高值1.34×10^8 cfu·g^-1干土,以后逐渐下降,到50 d时趋于稳定;筛选的突变体菌株对番茄青枯病具有明显的防治效果,防效达37.9%-50.9%。短小芽孢杆菌BX-4在作物根部的定殖规律为揭示其生防机理及应用该菌提供了科学根据。     

内生菌BS-2对蔬菜立枯病的抑制效果

抗利福平标记的菌株内生定殖测定结果表明,分离自辣椒叶片的内生枯草芽孢杆菌BS-2菌株既可在辣椒体内定殖,又可进入茄子、蕃茄、白菜、黄瓜、西瓜、丝瓜、甜瓜和豇豆等多种植物体内定殖;拮抗和防病作用测定发现,该菌株对多种植物病原真菌具有较强拮抗作用.菌液浸种处理后对黄瓜、甜瓜和蕃茄苗期立枯病有78.96%以上的抑制效果,并对这些蔬菜有较明显的促生作用.     

我国南方桉树人工林菌根合成及其多样性研究(英文)

对我国南方部分地区桉树人工林菌根真菌资源和菌根类型进行了调查 .对采自广东、云南 15个桉树林地的根系样品进行了显微观察 ,结果证实了在自然土壤中桉树能形成不同类型的菌根 ,检查的根样在不同程度上均有菌根菌的感染 .其中 ,有 4个人工林的根系样品AM感染率在 50 %以上 ,另有 4个根样ECM感染根段长超过 50m .调查发现林分年龄大小对菌根形成及菌根类型有一定的影响 ,表现在幼林及过熟林菌根感染率均较低 ,而 4～ 7年生按树林菌根感染率相对较高 ,在同一根系上AM与ECM菌间存在一定的竞争性 .在菌根菌资源方面 ,从总体上来说 ,我国南方菌根菌资源较少 ,尤其是ECM菌 .调查结果表明 ,我国南方桉树人工林根际土壤中有AM菌 2 8种 ,隶属于 4个属 ,其中以球囊霉属真菌最多 ,占 6 4 % ;地球囊霉在所采集的土样中出现的频度最高 .南方 15个桉树人工林中发现有 17种ECM菌 ,隶属 10个属 ,其中彩色豆马勃、多根硬皮马勃、光硬皮马勃等菌种在两广地区许多桉林中较常见 ,云南地区则多以黄须腹菌、硬皮马勃等菌较多 .这些ECM菌通常生长于松树林中 ,能否与引进的桉属树种形成较好的菌根还有待于进一步研究 .本文对自然条件下混合菌根的形成及真菌间的竞争关系进行了讨论     

群体感应信号物质对吡咯伯克霍尔德氏菌JK-SH007在杨树内定殖的影响

为了明确杨树溃疡病生防菌吡咯伯克霍尔德氏菌JK-SH007的群体感应系统是否与其内生定殖相关,本文通过群体感应指示菌紫色杆菌CV026和液相色谱串联四级杆飞行时间质谱技术(HPLC-Q-TOF-MS)确定其群体感应信号物质种类,并利用结晶紫染色、菌体回收、GFP标记等技术,研究其群体感应信号物质对该菌株生物膜及在杨树苗内的定殖能力的影响。结果表明,菌株JK-SH007产生的群体感应信号物质为含8个碳原子酰基侧链的辛酰基-L-高丝氨酸内酯(C8-HSL)。群体感应信号物质C8-HSL的添加对该菌株生物膜及在杨树苗内的定殖能力有影响,并且呈现低浓度促进,较高浓度抑制的规律。C8-HSL的添加与菌株JK-SH007生物膜的OD值和菌体量具有显著相关性,相关系数分别为0.923和0.756。当添加1.0%C8-HSL终浓度达到5×10^-8 g/L时,菌株JK-SH007生物膜的形成量达到峰值。荧光显微镜镜检发现,当添加100μL C8-HSL时,杨树组培苗根和茎中GFP荧光标记的菌株JK-SH007的定殖数量明显较CK多。菌株JK-SH007的群体感应与其在杨树内的内生定殖能力密切相关,群体感应信号物质C8-HSL具有增强菌株JK-SH007内生定殖的能力,同时也表明该物质有利于该菌株生防效果的发挥。