球囊霉素相关土壤蛋白与根系形态的相关性研究

球囊霉素相关土壤蛋白(GRSP)是丛枝菌根(AM)真菌菌丝分泌的糖蛋白,能够促进土壤团聚体的形成。多种因素影响GRSP的产量,植物根系形态是否通过碳素竞争机制影响GRSP产量目前尚不清楚。以Glomus mosseae的两个生态型菌株和红三叶草Trifolium repense为试材,通过砂培试验探讨宿主的根系形态与GRSP产量的相关性。发现接种AM真菌导致宿主细根的比例降低,粗根的比例增加,但对总根长和根表面积没有影响;侵染率和菌丝长度随着时间的延长而增加,菌株之间存在显著差异;接种AM真菌导致GRSP产量显著提高;GRSP产量与根系形态没有显著的相关性,但是与侵染率和菌丝长度显著相关。结果表明,尽管宿主根系形态建成和AM真菌都对碳素具有竞争关系,但是前者并没有抑制GRSP的形成,可能存在根系碳素分配的自调控机制。     

球囊霉素相关土壤蛋白根际环境功能研究进展

球囊霉素(glomalin)是丛枝菌根真菌产生的一种含有金属离子的耐热糖蛋白, 能够改善土壤结构, 固定土壤中的重金属, 近期被更名为球囊霉素相关土壤蛋白(glomalin-related soil protein)。该文从球囊霉素的定义、性质与环境功能等方面对相关文献进行了综述, 认为目前对球囊霉素的共识仍停留在理论假设蛋白的程度上, 包括: 1)该蛋白可能是热激蛋白60 (HSP 60)的同系物; 2)该蛋白所携带的阳离子可能随着土壤性质的改变而不同。目前还没有清楚确切地定义球囊霉素的真实分子结构与理化性质。今后需从分子层面对球囊霉素予以深入研究。同时, 需要不断改进球囊霉素的提取和测定方法, 以便进一步探讨球囊霉素固定重金属离子的机理, 提高植物的重金属抗性。     

球囊霉素相关土壤蛋白的分布及环境功能研究进展

球囊霉素相关土壤蛋白(GRSP)是由丛枝菌根真菌(AMF)在土壤中产生的一种糖蛋白,其在土壤中大量存在,可分为总球囊霉素、易提取球囊霉素、免疫反应性总球囊霉素、免疫反应性易提取球囊霉素.土地利用方式、施肥条件、AMF及宿主类型、外界环境条件等均会影响土壤中GRSP的含量及分布.GRSP能改善土壤团聚体的水稳定性、降低陆地生态系统土壤中CO₂排放、促进土壤中碳贮存、降低土壤中重金属的有效性、减弱重金属的植物毒害.GRSP的提取及定量表征技术仍是限制人们深入了解其在土壤中分布及环境功能的瓶颈.今后有关GRSP的研究应重视以蛋白及其编码该蛋白的基因为基础,阐释GRSP在土壤生态系统中的分子生物学作用及机制,以及GRSP对土壤中有机污染物环境行为的影响.     

赤红壤地区森林土壤球囊霉素相关蛋白测定方法

球囊霉素是丛枝菌根真菌菌丝产生的一种糖蛋白,被证明具有改善土壤结构、固定土壤中重金属、保护有机碳等重要的生态功能。球囊霉素目前尚无法纯化,一般通过测定球囊霉素相关蛋白(GRSP)来对其定量。由于GRSP测定方法的非专一性,目前尚无适宜于各种陆地生态系统的标准测定方法。本文针对GRSP测定结果可能受到提取时间、离心力以及测定前再次离心时离心力的大小等因素的影响,设置对比试验,对亚热带赤红壤地区土壤GRSP提取测定,以期获取适合于该地区的GRSP测定方法。结果表明:当提取时离心时间为10 min,10000×g的离心力提取的易提取GRSP(EE-GRSP)是5000×g提取的1.15~1.82倍,但离心力大小对总GRSP(T-GRSP)影响不显著;当提取时离心力为5000×g时,离心时间由10 min延长至15 min能够显著提高EE-GRSP含量;测定之前对上清液再次离心,若使用较大的离心力(≥10000×g),将显著影响GRSP测定值;经过6次连续提取,能够提取T-GRSP理论最大含量的97.7%~99.8%;因此,在亚热带酸性赤红壤森林生态系统中,GRSP的提取建议采用10000×g 10 min的离心力和离心时间组合,测定前对上清液再次离心时,应以≥10000×g的离心力为宜,研究结果为该地区GRSP的标准化测定提供了理论依据。     

庐山常绿阔叶林常见树种根际球囊霉素相关土壤蛋白分布特征及其影响因素

庐山森林具有明显的纬度地带性特征,常绿阔叶林与常绿落叶阔叶混交林是庐山中部区域常见的森林类型,对于庐山生物多样性及生态服务功能有重要意义.球囊霉素相关土壤蛋白(GRSP)的动态变化是评价和指示森林土壤质量的重要指标之一,探明庐山森林GRSP分布特征及其影响因素,对于提高森林土壤质量具有指示意义.以庐山自然保护区常绿阔叶...     

丛枝菌根真菌产球囊霉素研究进展

球囊霉素（Glomalin）是由丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）产生的一种含金属离子的糖蛋白,由于丛枝菌根真菌在自然和人工陆生生态系统中广泛分布,丛枝菌根真菌在生态系统中的生态学功能一直是菌根生物学研究中诱人的问题。自1996年球囊霉素被发现以来,球囊霉素在土壤生态系统中的生态学功能、生态学地位日益受到重视。本文对球囊霉素作为土壤主要有机源和超级胶的功能作了简介,综述了球囊霉素的研究现状,并对其研究前景作了展望。     

黄土高原紫穗槐丛枝菌根真菌与土壤因子和球囊霉素空间分布的关系

在黄土高原的陕西省甘泉、绥德、米脂、榆林4县(市)选取4个样地,研究紫穗槐不同深度土层丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)侵染率、孢子密度、球囊霉素和土壤因子之间的关系。结果表明:(1)AMF侵染率平均值绥德样地最大(100%),米脂样地最小(75.02%);孢子密度平均值米脂样地最大(5.91个/g),榆林样地最小(1.57个/g);4个样地孢子密度最大值均在0～10cm土层,且随土层加深而降低;AMF侵染率与孢子密度在各样地间差异显著,同一样地侵染率与孢子密度变化规律不一致。(2)榆林样地的有机碳、铵态氮、速效磷、脲酶和碱性磷酸酶活性平均值显著高于其他3个样地;除绥德、米脂样地的蔗糖酶和甘泉样地的碱性磷酸酶外,其他样地的土壤酶活性和球囊霉素含量均随土层加深而降低,且各土层之间差异显著,最大值均在0～10cm土层。(3)AMF侵染率与pH、总球囊霉素含量、易提取球囊霉素含量呈极显著正相关,与有机碳、硝态氮、速效磷含量呈显著正相关;孢子密度与速效磷、蔗糖酶、脲酶活性呈显著正相关,与铵态氮含量呈显著负相关。(4)第1、2主成分的累积方差贡献率达到56.4%,第1主成分主要综合了球囊霉素、AMF侵染率和孢子密度,第2主成分综合了pH和部分土壤因子信息。因此,球囊霉素、AMF侵染率、孢子密度、pH、部分土壤因子对决定土壤生态起主要作用。     

库布齐沙漠白沙蒿根际丛枝菌根及其球囊霉素的时空异质性

为探明流动沙地先峰植物白沙蒿(Artemisia sphaerocephala)根际丛枝菌根真菌产球囊霉素时空分布, 在库布齐沙漠选设白沙蒿样地, 于春、夏和秋季分0-10, 10-20, 20-30, 30-40和40-50 cm土层采集土壤样品, 测定其根际丛枝菌根真菌的菌丝侵染率、孢子密度、球囊霉素含量和土壤理化因子, 并系统分析了两两间的相互关系。结果表明: ①白沙蒿和丛枝菌根真菌具有良好的共生关系, 夏季和秋季的真菌菌丝侵染率略高于春季, 夏季平均为89.75%, 秋季平均达到92.37%, 两季的最大值都出现在20-30 cm土层。②白沙蒿根际丛枝菌根真菌活性有明显的空间异质性。真菌孢子密度为1.21-12.31 个·g土-1, 最大值出现在夏季的0-10 cm土层。孢子密度在不同季节有显著差异, 夏季>秋季>春季, 各季都随土层加深而递减。③白沙蒿根际土壤中总球囊霉素含量范围为0.37-1.27 mg·g-1, 易提取球囊霉素含量范围为0.19-0.81 mg·g-1, 两者在各季节最大值都出现在0-10cm土层, 呈现明显的表层土富集性。④球囊霉素与土壤中真菌孢子密度呈极显著正相关, 并和土壤养分及大多土壤酶活性呈显著正相关, 可作为评价土壤丛枝菌根真菌活性和土壤肥力的重要指标。     

转基因植物对土壤污染相关研究

随着转苏云金芽孢杆菌基因（Bacillus thuringiensis, Bt）作物（后面全用转Bt基因作物代替）的推广应用,有关转Bt基因作物对土壤中微生物的生态风险尚无统一的结论。丛枝菌根(Arbusular Mycorrhiza, AM）真菌在土壤中分布广泛,是监测转Bt基因作物生态风险评估的重要微生物群落。Bt杀虫蛋白的表达及释放,可能会使AM真菌共生过程中一些重要阶段的生态共生效应受到影响。转Bt基因作物对AM真菌的影响可作为转Bt基因作物生态风险评价的重要指标。文章以紫云英为宿主植物,通过盆栽实验,人为添加提取的Bt蛋白,检测Bt蛋白对AM真菌侵染率、酶的活性以及生物量的影响,以探索Bt蛋白对AM真菌侵染性及共生效应的影响,从而为转Bt基因作物的生态风险评估提供参考。     

缩球囊霉孢子萌发及细胞核在孢子和芽管中分布的研究

本文研究了从本校分离的丛枝菌根真菌缩球囊霉Glomus constrictum的孢子萌发和萌发孢子的细胞核DAPI(4’6-diamidino-2-phenylindol)染色。结果显示,G．constrictum孢子直径为179．5-198．7μm,顶生于产孢菌丝上。经表面消毒处理后,孢子在水琼脂平板上7天开始萌发。DAPI染色后,经稀释荧光计数,单个孢子细胞核数目约为5300,在孢子中无序分布,细胞核直径约为9．9-11．2μm。孢子萌发过程中,细胞核总数无明显变化,只是部分细胞核从孢子流向了萌发伸长的菌丝。     

两种土壤类型草本植物根系丛枝菌根真菌多样性

将分属12个科的17种宿主植物分别种植于石灰土和紫色土上,120 d后收获取样,通过CTAB法提取共生菌根内AM真菌的DNA,用特异引物U1/U2扩增编码核糖体28S大亚基的rDNA部分序列,并利用聚丙烯酰胺凝胶电泳(PAGE)和银染显影技术显色, 比较了石灰土和紫色土上草本植物根系内丛枝菌根真菌群落的物种多样性.结果表明：生长于石灰土上的17种宿主植物根系内共发现AM真菌条带29种,平均每种宿主植物内8.29种;生长于紫色土上的17种宿主植物根系内发现条带24种,平均每种宿主植物内9.47种.根系内的AM真菌条带包括特异条带和共有条带.经聚类分析发现,AM真菌对宿主植物的侵染有一定的科属专一性,且受土壤环境等外在条件的影响.同时探讨了AM真菌应用于石灰岩地区生态恢复的可能性.     

Influence of arbuscular mycorrhizal fungi on soil structure and aggregate stability of a vertisol

The influence of arbuscular mycorrhizal (AM) fungi on aggregate stability of a semi-arid Indian vertisol was studied in a pot experiment in which Sorghum bicolor (L.) was grown as test plant for 10 weeks. Pasteurized soil inoculated with AM fungi was studied with pasteurized and unpasteurized soils as references. A part of the soil in each pot was placed in nylon mesh bags to separate effects of roots and hyphae. The sorghum plants were planted outside the mesh bags which permitted AM hyphae to enter while excluding roots. Aggregate stability of the soil was determined by wet-sieving and turbidimetric measurements. Development of the AM fungi was quantified as colonized root length and external hyphal length. Soil exposed to growth of roots and hyphae (outside mesh bags) showed aggregates with larger geometric mean diameter (GMD) in pasteurized soil inoculated with AM fungi than in pasteurized uninoculated soil. There was no significant difference in GMD of the inoculated, pasteurized soil and the unpasteurized soil. No significant effects of inoculation or plant growth were found in pasteurized soil exposed to hyphal growth only (inside the mesh bags). However, the unpasteurized soil had significantly higher GMD than the pasteurized soil, irrespective of plants and inoculum. Turbidimetric measurements of soil exposed to roots and hyphae (outside mesh bags) showed the highest aggregate stability for the inoculated pasteurized soil. These results demonstrate that AM fungi contribute to the stabilization of soil aggregates in a vertisol, and that the effect is significant after only one growing season. The effect was associated with both AM hyphae and the stimulation of root growth by AM fungi. The contribution from plant roots and AM hyphae to aggregate stability of different size fractions is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Depth-dependent effects of ericoid mycorrhizal shrubs on soil carbon and nitrogen pools are accentuated under arbuscular mycorrhizal trees

Plant mycorrhizal associations influence the accumulation and persistence of soil organic matter and could therefore shape ecosystem biogeochemical responses to global changes that are altering forest composition. For instance, arbuscular mycorrhizal (AM) tree dominance is increasing in temperate forests, and ericoid mycorrhizal (ErM) shrubs can respond positively to canopy disturbances. Yet how shifts in the co-occurrence of trees and shrubs with different mycorrhizal associations will affect soil organic matter pools remains largely unknown. We examine the effects of ErM shrubs on soil carbon and nitrogen stocks and indicators of microbial activity at different depths across gradients of AM versus ectomycorrhizal (EcM) tree dominance in three temperate forest sites. We find that ErM shrubs strongly modulate tree mycorrhizal dominance effects. In surface soils, ErM shrubs increase particulate organic matter accumulation and weaken the positive relationship between soil organic matter stocks and indicators of microbial activity. These effects are strongest under AM trees that lack fungal symbionts that can degrade organic matter. In subsurface soil organic matter pools, by contrast, tree mycorrhizal dominance effects are stronger than those of ErM shrubs. Ectomycorrhizal tree dominance has a negative influence on particulate and mineral-associated soil organic matter pools, and these effects are stronger for nitrogen than for carbon stocks. Our findings suggest that increasing co-occurrence of ErM shrubs and AM trees will enhance particulate organic matter accumulation in surface soils by suppressing microbial activity while having little influence on mineral-associated organic matter in subsurface soils. Our study highlights the importance of considering interactions between co-occurring plant mycorrhizal types, as well as their depth-dependent effects, for projecting changes in soil carbon and nitrogen stocks in response to compositional shifts in temperate forests driven by disturbances and global change.     

The arbuscular mycorrhizal fungal protein glomalin is a putative homolog of heat shock protein 60

Work on glomalin-related soil protein produced by arbuscular mycorrhizal (AM) fungi (AMF) has been limited because of the unknown identity of the protein. A protein band cross-reactive with the glomalin-specific antibody MAb32B11 from the AM fungus Glomus intraradices was partially sequenced using tandem liquid chromatography-mass spectrometry. A 17 amino acid sequence showing similarity to heat shock protein 60 (hsp 60) was obtained. Based on degenerate PCR, a full-length cDNA of 1773 bp length encoding the hsp 60 gene was isolated from a G. intraradices cDNA library. The ORF was predicted to encode a protein of 590 amino acids. The protein sequence had three N-terminal glycosylation sites and a string of GGM motifs at the C-terminal end. The GiHsp 60 ORF had three introns of 67, 76 and 131 bp length. The GiHsp 60 was expressed using an in vitro translation system, and the protein was purified using the 6xHis-tag system. A dot-blot assay on the purified protein showed that it was highly cross-reactive with the glomalin-specific antibody MAb32B11. The present work provides the first evidence for the identity of the glomalin protein in the model AMF G. intraradices, thus facilitating further characterization of this protein, which is of great interest in soil ecology.     

Influence of arbuscular mycorrhizal fungi on soil structure and soil water characteristics of vertisols

The influence of inoculation with arbuscular mycorrhizal fungi (AM fungi) on soil water characteristics of fast and slowly wetted vertisol samples was studied. Vertisols characteristically have a low stability to wetting, and the disruption of their larger pores when they swell leads to reduced water infiltration and thereby to runoff. The degree of aggregate breakdown determines the ability of the soil to drain. A vertisol was used in this pot experiment with four treatments: T₁: Pasteurized soil, T₃: Pasteurized soil, with plants, T₄: Inoculated, pasteurized soil, with plants, T₅: Unpasteurized soil, with plants. A treatment using inoculated, pasteurized soil (T₂) was included in a related study (Bearden and Petersen, 2000) comparing aggregate stability, and the present study follows the same numbering to aid in comparison of experiments. After fast, disruptive wetting, the soil inoculated with AM fungi (T₄) was found to have a lower soil water content than did the soils from the other treatments at matric potentials lower than –3.92 kPa. This indicates greater drainage from pores smaller than 75 m for the soil inoculated with AM fungi, and the greater drainage appears to be directly related to a characteristic pore range between 67 and 75 m. The soil without plants (T₁), when wetted fast, had a lower soil water content at matric potentials higher than –3.92 kPa than soils from the other treatments, which indicates less pore volume due to pores larger than 75 m in the treatment without plants. The pore indexes, calculated as the ratio between the slope of the fast and the slope of the slowly-wetted water characteristics, generally had the highest values for the soil inoculated with AM fungi (T₄) from matric potential 0.00 to –0.29 kPa. In this matric potential range, the pore indexes were less than one. The unpasteurized soil with naturally present AM fungi (T₅) generally had the highest pore indexes from matric potential –0.49 to –3.92 kPa, and the pore indexes in this matric potential range were above one. These results indicate the smallest loss of very large pores in the soil inoculated with AM fungi (T₄) and the largest gain of smaller sized pores in the unpasteurized soil (T₅). This suggests that the resistance to breakdown of the largest pores is related to the presence of roots, and that the gain of groups of smaller pores is related to the presence of hyphae.     

盐渍化土壤根际微生物群落及土壤因子对AM真菌的影响

为探明盐渍化土壤影响下AM真菌与根际土壤间的关系,试验选取宁夏碱化龟裂土、草甸盐土、盐化灌淤土3种类型4个样地上典型植被群落,测定了植物根际土壤养分含量、微生物群落结构、AM真菌侵染率以及孢子密度。结果显示:盐渍化土壤根际微生物碳源利用类型显著不同,对芳香类化合物的代谢能力整体较弱;红寺堡草甸盐土上微生物优势群落为氨基酸代谢类群,惠农盐化灌淤土为多聚化合物代谢群,西大滩碱化龟裂土为碳水化合物代谢群。AM真菌孢子密度与微生物碳源代谢群间的关系比较复杂。其中,惠农样点根际土壤孢子密度与多聚化合物微生物代谢群呈显著正相关,西大滩地区孢子密度与碳水化合物微生物代谢群呈显著正相关。土壤有机质、全盐、全氮、碱解氮等土壤肥力因子及土壤中的HCO-3、Na+、Cl-等盐基离子含量能解释AM真菌孢子密度与土壤环境因子之间相互关系的大部分信息。较高的HCO-3浓度促进了AM真菌侵染率的提高,但高盐浓度下Na+和Cl-降低了菌根侵染率。土壤对AM真菌孢子密度、侵染率的影响因土壤盐分组成类型的不同而异。研究结果为深入了解AM真菌多样性,促进宁夏盐碱地的合理开发与利用提供了理论依据。     

Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots

Material on the surface of hyphal walls of arbuscular mycorrhizal fungi (AMF) during active colonization of plant roots was detected by a monoclonal antibody. Pot-cultured isolates of Glomus, Acaulospora, Gigaspora, Scutellospora, and Entrophospora had immunofluorescent material (IM) on younger, thinner, intact hyphae, but IM was scant to absent on thicker, melanized or lysing hyphae. Colonization of corn (Zea mays L.), Sudangrass (Sorghum sudanense (Piper) Staph.) or red clover (Trifolium pratense L.) was examined during 5 months of plant growth by removing cores and performing an indirect immunoassay on roots with attached hyphae. Fresh spores of some Glomus spp. had IM on the outer layer of the spore wall. Abundant IM was seen on root hairs of plants colonized by some isolates, and some IM was detected on root surfaces of all plants examined even during early colonization. After cultures were dried, hyphae, roots and spores had little to no IM. Uninoculated control roots had very rare, small patches of IM. An immunoreactive protein was extracted from hyphae of Gigaspora and Glomus isolates by using 20mM citrate (pH 7.0) at 121°C for 90 min. Gel electrophoresis profiles indicated that all isolates tested had the same banding patterns. Lectin-binding of extracted protein is suggestive of a glycoprotein. The immunofluorescence assay can be used to examine root sections for active colonization by AMF, and the potential use of the protein to quantify AMF activity in soil is discussed.