香蕉根系丛枝菌根(AM)真菌染色方法比较

AM真菌侵染情况观察是AM真菌研究的重要基础。本研究比较了5种不同染色剂(5%醋酸墨水,酸性品红,苏丹红Ⅳ,台酚蓝,苯胺蓝)对香蕉根系AM真菌的染色效果。研究结果表明:5%醋酸墨水和台酚蓝染色液的染色效果最佳,根皮层细胞内的AM真菌的菌丝、丛枝、泡囊及内含物等结构清晰可见,且能够明确分辨出AM真菌与其他未知真菌。但综合考虑操作难易程度、价格成本和毒性等因素,5%醋酸墨水染色液更适用于香蕉根系AM真菌的染色和制片观察。     

丛枝菌根观察与侵染率测定方法的比较

菌根生长状况观察与侵染率测定是菌根学研究中一项重要的基础性工作。综述了丛枝菌根（AM）染色观察与侵染率测定方法研究概况,并对其进行比较和评价。认为采用醋酸墨水染色观察AM生长状况与采用根段侵染率加权法和放大交叉法测定AM真菌侵染率是目前较为科学、准确、易行的方法。根据不同需要也可选择其他适宜的方法,如要了解丛枝发育状况,可采用放大交叉法;如要了解泡囊和侵入点数量,可采用直接计数法,从而使其研究结果具有可比性。有必要建立基于分子生物学技术和脂肪酸定量分析技术测定一种或数种AM真菌侵染状况,这将有力推动AM真菌生理、生态功能研究的发展。     

濒危药用植物桃儿七根的显微结构及其菌根真菌分布研究

本文研究了桃儿七Sinopodophyllum hexandrum根的显微结构及其真菌分布。结果表明,桃儿七的根为根状茎,节状,不定根形成的须根系发达。根的结构主要由表皮、皮层、维管柱三部分构成,其中,皮层所占比例最大,超过80%。根的木质部有四原型和五原型两种类型,五原型较为常见;四原型的根和五原型的根在皮层细胞形态上存在一定差异。在桃儿七的不定根和其上的侧根观察到真菌菌丝分布,其数量和种类与根的直径有关,在不定根较细(先端)的部位真菌以暗色有隔内生真菌(DSE真菌)为主,侵染率为77.9%;而较粗根中真菌菌丝为无隔菌丝为主,分布很少且仅存在于皮层细胞的一至二层,不侵染皮层深部和维管柱。不定根侧根中真菌以丛枝菌根真菌为主,丛枝菌根常常占据大部分的皮层细胞,侵染率高达90%以上。桃儿七根中没有发现根毛存在,因此,侧根中共生的丛枝菌根真菌可能是桃儿七养分和水分吸收的主要途径。     

植酸钠对菌根真菌根内菌丝碱性磷酸酶活性及根外菌丝生长的影响

采用三室隔网培养装置,以玉米为宿主植物,接种丛枝菌根真菌(AM)(Glomus intraradices),研究了不同用量的植酸钠对AM真菌生长和代谢活性的影响．研究发现,接种AM真菌的植株地上部和根系的P浓度和吸P量,比非菌根植物的提高了1～2倍．外源植酸钠的存在,显著降低了AM真菌根内菌丝的碱性磷酸酶活性,增加了AM真菌在土壤中的菌丝密度．结果表明,外源植酸钠对根内AM真菌碱性磷酸酶活性和真菌根外菌丝的生长具有调控(增减)作用,并且AM真菌提高了植物对土壤固有养分和外源植酸钠中P的吸收和利用．     

大兴安岭落叶松林丛枝菌根真菌多样性

对大兴安岭落叶松林17个科26种植物丛枝菌根(AM)定居情况和丛枝菌根真菌(AMF)多样性进行了调查。在26种植物根样中形成典型AM定居的有23种,占88.46%。菌根侵染率为0～78.7%,平均为21.69%。在26种植物根样中,4种植物形成重楼型(Par-is-type)AM,占15.38%;10种植物形成疆南星型(Arum-type)AM,占38.46%;9种植物形成中间型(Intermediatetype)AM,占34.62%,3种植物未能形成AM菌根,占11.54%。从23种植物根围土样中分离到50种AMF,分属于5个属,其中无梗囊霉属(Acaulospora)24种占48%;球囊霉属(Glomus)19种占38%;内养囊霉属(Entrophospora)5种占10%;原囊霉属(Archaeospora)1种占2%;巨孢囊霉属(Gigaspora)1种占2%。Acaulospora和Glomus2属真菌在大兴安岭落叶松林土壤中占绝对优势;尼氏无梗囊霉(A.nocolsonii)和幼套球囊霉(G.etunicatum)是优势种。植物根围土壤中AMF的孢子密度平均为68.94个.100g-1风干土;AM...     

西双版纳地区龙脑香科植物AM真菌的初步研究

 对云南省西双版纳地区17种龙脑香科树种根系丛枝菌根（Arbuscular mycorrhiza, AM）真菌的定居情况进行了调查,并对根围土壤中AM真菌进行了分离和鉴定。结果表明,调查根样均有不同程度的菌根感染,感染率最高可达40%,调查揭示了西双版纳地区龙脑香科植物在自然条件下可形成丛枝菌根。初步从龙脑香科植物根际土壤中分离、鉴定出32种AM真菌,隶属于无梗囊霉属（Acaulospora）、球囊霉属（Glomus）、原囊霉属（Achaeospora）、拟球囊霉属（Paraglomus）和盾巨孢囊霉属（Scutellospora）,其中,无梗囊霉属和球囊霉属真菌为西双版纳地区龙脑香科植物AM真菌优势类群。     

墨兰菌根的结构及酸性磷酸酶定位研究

利用光学显微镜、电子显微镜及细胞化学方法,对墨兰菌根的结构和酸性磷酸酶定位进行了初步研究。结果表明墨兰具有典型的兰科植物根结构,发现该兰花的根的外皮层不具薄壁通道细胞,菌根真菌通过破坏部分根被和外皮层细胞而侵入根的皮层细胞并在细胞内形成菌丝结,侵入的菌丝被染菌皮层细胞质膜和电子透明物质包围,进一步被消化并聚集成衰败菌丝团块。酸性磷酸酶在染菌皮层细胞及包围菌丝的皮层细胞质膜和衰败菌丝细胞壁上有强烈的酶反应,衰败菌丝周围分布有许多单层膜的含酶小泡,它们可相互愈合形成大的含酶泡或与包围菌丝的质膜融合,类似于兰科植物共生原球茎中观察到的现象。说明皮层细胞可主动释放水解酶参与对菌丝的消化     

珠状巨孢囊霉的单主寄生营养液培养及宿主根分泌物特性*

为研究丛枝菌根真菌对宿主植物根分泌物的影响,在营养液培养条件下成功地建立了菌根真菌Gigaspora margarita与转移Ri T-DNA胡萝卜根器官之间的无菌单主寄生培养方法。接种发芽的孢子四个月后,对根段染色观测,发现萌发孢子的菌丝能够入侵到根段的皮层细胞内,根细胞内有菌丝的折叠卷曲现象,并开始形成了新的孢子。利用高压液相色谱测定各处理营养液中根分泌物的成分,发现菌根分泌的苹果酸和乙酸含量高于对照处理,菌根营养液的pH升高幅度也大于未接种的对照根。     

密花石斛等六种兰科植物菌根的显微结构研究

本文对密花石斛等六种兰科植物根的显微结构进行了比较观察。结果表明,它们具有典型的兰科植物根,具根被和发达的皮层组织,皮层细胞内分布有针状结晶和菌根真菌形成的菌丝结,发现菌根真菌通过外皮层薄壁通道细胞或破坏根被组织和外皮层细胞侵入皮层细胞,形成内生菌根。     

珠状巨孢囊霉的单主寄生营养液培养及宿主根分泌物特性

为研究丛枝菌根真菌对宿主植物根分泌物的影响,在营养液培养条件下成功地建立了菌根真菌Gigaspora margarita与转移Ri T-DNA胡萝卜根器官之间的无菌单主寄生培养方法。接种发芽的孢子四个月后,对根段染色观测,发现萌发孢子的菌丝能够入侵到根段的皮层细胞内,根细胞内有菌丝的折叠卷曲现象,并开始形成了新的孢子。利用高压液相色谱测定各处理营养液中根分泌物的成分,发现菌根分泌的苹果酸和乙酸含量高于对照处理,菌根营养液的pH升高幅度也大于未接种的对照根。     

Rapid methods for quantifying VAM fungal infections in maize roots

Roots of maize (Zea mays cv W64A × W182E) infected by vesicular arbuscular mycorrhizal (VAM) fungi (Glomus versiforme (Karst) Berch or a Glomus species isolated from an alfalfa soil) exhibit a bright yellow pigmentation. The percentage of pigmented roots can be quantified by a rapid visual estimate or by a grid intersect method. Both methods gave similar estimates of VAM infection to those obtained using a grid intersect count on cleared roots stained with chlorazol black E. Thus for experimental or field evaluation where speed and quantity are important, the rapid visual estimate (less than one minute for each washed root system) yields reliable results. The yellow root intersect method takes longer (5–15 minutes per root system) but gives more reproducible results. The yellow root pigmentation is light sensitive However, root systems can be reliably assayed after 1 week when stored at 5°C in the dark or after 1 year if dried.     

A Standardized Cotton Blue Stain for Pollen Germination and Growth in Andropogoneae Grasses

Styles along with ovaries were dissected from spikelets of sorghum and other grasses and fixed in 1:3 acetic alcohol at 0.5 hr intervals for 3 hr after pollination. The material was hydrated and then macerated in 50% HCl for 10 min at 56 C, washed in running water for 30 min, cleared in lactophenol for 35 min at 60 C, stained in 1% cotton blue for 50 min at 50 C, and destained for 1 hr in lactophenol at room temperature. Styles were dissected from the ovary in a drop of lactophenol. Slight pressure on the coverslip with the blunt end of a needle and a little warming spreads the stylar tissue and reveals the darkly stained pollen tubes.     

丛枝菌根真菌对植物根尖分生区和根冠细胞的侵染*

一般说来,从枝菌根（ＡＭ）真菌大多数是从植物根系根毛区（成熟区）侵入和扩展的,在显微镜下往往看不到根尖分生区和根冠表皮细胞被ＡＭ真菌侵染的特征。这就很容易给人们造成一种假象,似乎ＡＭ真菌不能侵染根尖分生区和根冠表皮细胞,即它们对ＡＭ真菌是免疫的。然而笔者多次于显微镜下看到ＡＭ真菌侵染根尖分生区和根冠表皮细胞,并形成典型的泡囊、丛枝、菌丝等结构。这一现象导致作者在温室盆栽和大田条件下研究了玫瑰红巨孢囊霉（ Ｇｉｇａｓｐｏｒａ　ｒｏｓｅａ Ｎｉｃｏｌ　＆ Ｓｃｈｅｎｃｋ）、珠状巨孢囊霉（Ｇｉｇａｓｐｏｒａ　ｍａｒｇａｒｉｔａ　Ｂｅｃｋｅｒ　＆　Ｈａｌｌ）、根内球囊霉（Ｇｌｏｍｕｓ　ｏｍｔｒａｒａｄｉｃｅｓ　ｓｃｈｅｎｃｋ　＆　Ｓｍｉｔｈ、摩西球囊霉（Ｇｌｏｍｕｓ ｍｏｓｓｅａｅ （Ｎｉｃｏｌ　＆ Ｇｅｒｄ．） Ｇｅｒｄｅｍａｎｎ　＆ Ｔｒａｐｐｅ）、地表球囊霉（ Ｇｌｏｍｕｓ ｖｅｒｓｉｆｏｒｍｅ（ Ｋａｒｓｔｅｎ）Ｂｅｒｃｈ）和弯丝硬囊霉（ Ｓｃｌｅｒｏｃｙｓｔｉｓ　ｓｉｎｕｏｓａ　Ｇｅｒｄｅｍａｎｎ　＆ Ｂａｋｈｉ）对棉花（Ｇｏｓｓｙｐｉｕｍ　ｈｉｒｓｕｔｕｍ　Ｌ．）、烟草（Ｎｉｃｏｔｉａｎａ　 ｔａｂａｃｕｍ Ｌ．）和白     

丛枝菌根真菌通过上调根系及自身水孔蛋白基因表达提高玉米抗旱性

在模拟干旱条件下, 研究了接种丛枝菌根(AM)真菌Glomus intraradices对玉米(Zea mays)根部13种质膜水孔蛋白基因表达的影响, 同时观测了AM真菌自身水孔蛋白基因的表达情况。结果表明, 干旱条件下, 除Zm PIP1;3、Zm PIP1;4、Zm PIP1;5和Zm PIP2;2之外的接种处理能显著提高根部其他8种质膜水孔蛋白基因的表达(Zm PIP2;7表达量未检测出), 并且AM真菌菌丝中水孔蛋白基因GintAQP1表达也显著增强。与此同时, 接种处理明显改善了植物水分状况, 提高了叶片水势。AM真菌增强宿主植物根部及自身的水孔蛋白基因的表达对于提高植物抗旱性具有潜在的重要贡献。     

Phosphorus uptake by a community of arbuscular mycorrhizal fungi in jarrah forest

Communities of indigenous arbusuclar mycorrhizal (AM) fungi are expected to alter phosphorus uptake and biomass productivity of plants according to characteristics of the life cycles of the fungi present and the way they interact with each other inside roots and with host plants. Differences in the relative abundance of AM fungi inside roots could influence P uptake if the fungi present differ in effectiveness at accessing P and transferring it to the plant. However, it is difficult to assess the contribution of AM fungi under field conditions. We investigated P uptake, from point sources of P placed 2, 4 and 6 cm from roots, by plants colonised by a community of AM fungi in jarrah forest soil. Roots were retained within a mesh bag to prevent them from growing towards the point source of P. The relative abundance of morphotypes of fungi inside roots and the P status of plants were assessed after 12 and 16 weeks. First, a bioassay was carried out in undisturbed forest soil cores using two host plants, a forest understorey plant Phyllanthus calycinus Labill and the annual pasture species subterranean clover (Trifolium subterraneaumL.), to assess the infectivity of the indigenous community of AM fungi. Roots of both bioassay host plants were colonised in similar proportions by morphotypes of AM fungi resembling Glomus, Acaulospora, Scutellospora and fine endophytes. In this bioassay, there were positive correlations between the proportion of root length colonised and plant biomass and P uptake for P. calycinus, but not for subterranean clover. In the experiment assessing the capacity of P. calycinus to access P placed at increasing distances from the root, shoot P content and concentration in P. calycinus were greater when P was placed 2 cm compared with 4 and 6 cm from roots. The length of hyphae in the vicinity of the point source of P decreased with increasing distance from the plant. The extent to which the individual AM fungi were involved in P uptake is not known. The Glomus morphotype was dominant at both times of sampling.     

拉杰沙希大学校园药用植物菌根研究

Forty different medicinal plants were investigated for arbuscular mycorrhizal association in the Rajshahi University Campus in Bangladesh. The results indicated that 35 different plants were infected by AM (arbuscular mycorrhizal) fungi as found by trypan blue staining procedure. The percentage of root colonization by AM fungi varied from 13.3% to 100%. Mangifera indica and Morus indica have maximum percentage of colonization (100%). The intensity of root colonization were abundant in the plants belonging to the families Anacardiaceae, Asclepiadaceae, Moraceae, Leguminosae and Apocynaceae whereas the intensity of colonization of crop roots were moderate and poor belonging to Gramineae and Leguminosae. The presence of greater number of spore in soil was always associated with the incidence of abundant mycelia. In plant roots the formation of spore and mycelia was restricted by low pH. Number of mycorrhizal fungus spores ranged between 35 to100 per 100g air dried soil in different family respective soils. The frequency of mycorrhizal fungus infection showed positive correlation with soil pH, moisture, water holding capacity, texture, total nitrogen, organic carbon, phosphorus, calcium, potassium, and magnesium. Especially phosphorus and nitrogen in the soil greatly influenced the plant root infection by AM fungi.     

Staining human lymphocytes and onion root cell nuclei with madder root

We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5–10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials.     

Staining human lymphocytes and onion root cell nuclei with madder root.

We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5-10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials.     

Atypical morphology of dark septate fungal root endophytes of<Emphasis Type="Italic"> Bouteloua</Emphasis> in arid southwestern USA rangelands

Native grasses of semi-arid rangelands of the southwestern USA are more extensively colonized by dark septate endophytes (DSE) than by traditional mycorrhizal fungi. Roots of dominant grasses ( Bouteloua sp.) native to arid southwestern USA rangelands were prepared and stained using stains specific for fungi (trypan blue) and for lipids (sudan IV). This revealed extensive internal colonization of physiologically active roots by atypical fungal structures that appear to function as protoplasts, without a distinguishable wall or with very thin hyaline walls that escape detection by methods staining specifically for fungal chitin. These structures were presumed to be active fungal stages that progressed to form stained or melanized septate hyphae and microsclerotia characteristic of DSE fungi within dormant roots. The most conspicuous characteristic of these fungi were the unique associations that formed within sieve elements and the accumulation of massive quantities of lipids. This interface suggests a biologically significant location for carbon transfer between the plant and fungus. The continuous intimate association with all sieve elements, cortical and epidermal cells as well as external extension on the root surface and into the soil indicates that they are systemic and considerably more prevalent than previously thought. A fungal network associated with a mucilaginous complex observed on the root surface and its potential role in root function in dry soil is discussed. It is suggested that those fungi that non-pathogenically and totally colonize plant cells be classed as systemic endophytic fungi (SEF). This would refine the broad designation of DSE fungi. The potential mutualistic benefit of SEF for native plants in arid ecosystems based on the extent of lipid accumulation and its apparent distribution is discussed.     

不同土壤类型下甘蔗根系AM真菌多样性及与土壤因子关系

甘蔗是广西重要的糖料作物,本研究通过建立克隆文库、土壤养分分析和根样染色等方法测定了33个采样点3种土壤类型(赤红壤,红壤,砖红壤)下甘蔗根系AM真菌多样性及其与土壤因子的关系。结果表明,3种土壤类型的甘蔗根系共鉴定出6科6属11种AM真菌,AM真菌各属频度存在明显差异,其中球囊霉属的频度值最高,在33个根系样品中有32个存在该属,为广西甘蔗根系AM真菌的优势属,而类球囊霉属、无梗囊霉属、近明球囊霉属、多样孢囊霉属和盾巨孢囊霉属5个属为稀有属。3种土壤类型的甘蔗根系均发现有球囊霉属和盾巨孢囊霉属;近明球囊霉属、无梗囊霉属和类球囊霉属仅在赤红壤的甘蔗根系中出现;而多样孢囊霉属仅在在赤红壤和砖红壤的甘蔗根系中出现。土壤的pH与AM菌根侵染率呈显著正相关,而有机质、总N、有效P、交换性Mg2+与AM菌根侵染率均呈负相关。本研究表明,自然条件下甘蔗根系具有相对丰富的AM真菌类群,这些AM真菌可能在甘蔗生长过程中发挥着重要的生态功能。