植物磷营养状况对丛枝菌根真菌生长及代谢活性的调控*

采用四室隔网培养装置,以玉米为宿主植物,通过在植物生长室设0、50、250和500 mgPkg-1 4个施磷水平,研究了植株体内的磷营养状况对AM真菌Glomus sinuosum和Glomus intraradices生长及活性的影响。研究发现在不施磷条件下,接种AM真菌G. intraradices显著促进了植物生长和磷的吸收;低磷条件(50 mgPkg-1)下,接种菌根真菌显著促进了植物对磷的吸收,但对植物生长没有明显的影响;而在高磷条件(250 mgPkg-1 和500 mgPkg-1)下,接种菌根真菌不但没有促进植物的生长和磷的吸收,反而对其有抑制作用。随着施磷水平的提高, AM真菌根内菌丝的碱性磷酸酶活性显著下降;与不施磷相比,低量(50 mgPkg-1)供磷增加了AM真菌土壤中根外菌丝的密度,高磷(250 mgPkg-1 和500 mgPkg-1)降低了土壤中根外菌丝的密度。上述结果说明:⑴ 给宿主植物施用磷肥引起的植物磷营养状况的改变,对AM真菌生长和代谢活性具有一定的调控作用;⑵ G. sinuosum和G. intraradices两种AM真菌的生长和代谢活性对施磷水平的响应程度无显著性差异;⑶ 高磷抑制AM真菌生长和代谢活性,使真菌吸磷量减少,可能是造成菌根效应降低的原因之一     

不同磷源对红三叶草根际和菌根际磷酸酶活性的影响

以红三叶草为研究对象,利用三室培养系统,在接种菌根真菌(Glomus mosseae)的条件下研究了不同磷源对根际和菌根际磷酸酶活性的影响,植株生长8周后收获并测定根室、菌丝室的土壤磷酸酶活性、植株干重及含磷量．结果表明,根室酸性磷酸酶活性比碱性磷酸酶活性更裔,接种条件下二者都稍有增加,特别是在供给有机磷(植酸钠)的条件下明显增加了菌丝室土壤磷酸酶活性．接种菌根真菌显著增加了植株干重、磷含量和总磷吸收．施用磷酸二氢钾(KH2PO4)时菌丝吸磷量占吸磷总量的43．1%,而施用植酸钠(Na-phytate)时菌丝吸磷量占吸磷总量的60．8%。     

干旱胁迫下覆膜和接种微生物对玉米生长与水分利用效率的影响

针对西北干旱半干旱地区土壤贫瘠与水分缺乏的问题,利用微生物与作物形成互惠互利的共生关系,本研究设置两个水分梯度:干旱胁迫(供试土壤最大持水量的35%)和正常水分(供试土壤最大持水量的75%),两个覆膜方式:无覆膜(NM)和覆膜(FM),4个接种微生物水平:单接AM真菌(AM)、单接解磷细菌(PSB)、联合接种AM真菌与解磷细菌(AM+PSB)以及对照(CK),研究不同水分和覆膜条件下4个接种微生物对玉米生长特性、地上养分吸收与水分利用效率的影响.结果表明: 与正常水分处理相比,干旱胁迫能够显著提高接种AM真菌处理的侵染率,但正常水分处理下土壤根外菌丝密度、总球囊霉素(T-GRSP)与易提取球囊霉素(EE-GRSP)含量明显提高.干旱胁迫下,单接AM真菌处理的促生作用和菌根效应表现最好,能够提高玉米生物量、水分利用效率和土壤有机碳含量,促进土壤N、P、K的吸收与运输,从而增加玉米地上部分N、P、K吸收量;而正常水分下,联合接种AM+PSB处理表现要好于单接AM和PSB处理,且其与覆膜的互作效果最好.相关分析结果表明,玉米生物量、叶片SPAD值和地上部分N、P、K吸收量均与土壤根外菌丝密度呈显著正相关,玉米水分利用效率与其呈显著负相关.     

黄土高原天然和人工油松林根际土壤解磷细菌群落特征及其功能

为了揭示油松-根系微生物的互作关系及其对油松林分稳定性的影响,本研究选择陕西省黄龙县天然和人工油松林,采集油松根际与非根际土壤,测定非根际土壤化学特性,分离纯化根际土壤解磷(有机磷和无机磷)细菌,通过DNA基因测序鉴定解磷细菌,并测定解磷细菌的解磷能力。结果表明: 天然油松林非根际土壤中全碳(TC)、全氮(TN)含量以及C/N、N/P极显著高于人工油松林。2种油松根际土壤中共鉴定出20属65种解磷细菌,其中以芽孢杆菌属、链霉菌属和假单胞菌属为优势菌群;天然油松林根际解磷细菌多样性、丰富度和均匀度指数均高于人工油松林,而优势度指数低于人工油松林。链霉菌属与非根际土壤TC、TN和C/N、N/P呈正相关,而芽孢杆菌属和假单胞菌属与非根际土壤硝态氮、铵态氮、有效磷及全磷含量呈正相关。2种油松林根际土壤不同解磷细菌的解磷能力存在差异,其中天然和人工油松林根际共有的解磷细菌为假单胞菌Pseudomonas sp.34-5,其对磷酸钙的解磷能力最高,为11.40 μg·mL^-1;天然油松林根际独有的解磷细菌蕈状芽胞杆菌BF1-5对卵磷脂的解磷能力最高,为4.58 μg·mL^-1。该林区2种油松林根际解磷细菌菌群结构存在显著差异。与人工油松林相比,天然油松林根际土壤解磷细菌群落多样性更丰富且分布更均匀,解磷能力普遍高于人工林。     

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．     

不同磷浓度对丛枝菌根真菌吸收同化外源氮能力的影响

以高粱（ Sorghum bicolor）为宿主植物,丛枝菌根（ arbuscular mycrohiza,AM）真菌根内球囊霉（ Glomous intraradices）为接种菌剂,三室隔离培养盒为培养容器,通过在菌丝室添加不同浓度梯度磷素及外源氮NH4 NO3、Gln,研究磷浓度对AM真菌同化吸收不同外源氮能力的影响。实验结果显示：AM真菌能够侵染于高粱植物根系,但菌根侵染率差异不显著;在高磷浓度下孢子数量显著高于低磷浓度下孢子数量;菌丝室内根外菌丝（ ERM）干重在低磷浓度下含量最高,且以Gln为外源氮时含量比不加氮源和NH4 NO3为氮源时高;低磷浓度促使高粱地上茎叶和地下菌根干重显著提高,叶绿素含量在不同处理下没有显著差异。茎叶总氮含量均在以NH4 NO3为外源氮时最高,不同磷浓度下其总氮含量为P30＆gt;P120＆gt;P0＆gt;P60,菌根精氨酸含量在Gln为外源氮时含量比其他氮源下高,且在低磷（P30）浓度下含量最高。研究表明AM真菌对于吸收同化外源氮的能力与其生长环境中磷浓度高低有关,在低磷浓度下更利于AM真菌根外菌丝同化吸收外源氮,且对NH4＋形式氮源吸收能力最强。     

丛枝菌根真菌对玉米秸秆降解的影响及其作用机制

为了比较菌根、菌丝、植物根系对玉米秸秆降解的影响,采用4室分根装置即土壤室(S)、根室(R)、菌根室(M)和菌丝室(H),分室间用400目尼龙网和有机板分隔,尼龙网袋包埋玉米秸秆于不同分室内,以玉米为宿主植物,接种丛枝菌根(AM)真菌Glomus mosseae。试验分别在移栽后第20、30、40、50、60天时取样,通过测定接种AM真菌后玉米秸秆的碳、氮释放,土壤中3种常见酶活性、微生物量碳和氮及土壤呼吸的动态变化,探讨AM真菌降解玉米秸秆可能的作用机制。研究结果表明:经60 d的培养后,与未接种S室相比,接种AM真菌的M室和H室玉米秸秆降解量提高了27.72%和8.07%;另外,M室玉米秸秆碳素释放显著增加,而氮素的释放减少,致使碳氮比显著低于其他3室,较初始值降幅达8.72%,有利于秸秆进一步降解。在试验条件下,M室中土壤酸性磷酸酶、蛋白酶、过氧化氢酶活性较其他3室都有显著提高,并增加了微生物量碳、氮和土壤呼吸作用,形成了明显有别于根际的微生物区系。这一系列影响都反映出AM直接或间接作用于玉米秸秆的降解过程,是导致玉米秸秆降解加快的重要原因。     

丛枝菌根菌丝精氨酸双向运转并分解为鸟氨酸

通过^15N和^13C同位素标记证明了在丛枝菌根（arbuscular mycorrhiza,AM）菌丝内精氨酸（arginine,Arg）双向运转并分解为鸟氨酸（omithine,Ore）。离体菌根真菌Glomus intraradices和Ri T-DNA转化的胡萝卜根培养在两室（菌根室和真菌室）培养皿中。[^15N／^13C]Arg施加在菌根室或真菌室或非侵染根组织中。采用色质联用和液相色谱测定菌根和根外菌丝（extraradical mycelium,ERM）内自由氨基酸的含量和同位素标记。研究发现与外源性Arg相比,AM真菌ERM内Arg的坫N标记在ERM培养于外源性氮源NH^＋4和尿素时是最高的。但是,外源性C源甘油施加于ERM对Arg的^15N标记没有较大的影响。在此期间,ERM在培养于4mmol／LNH^4 6周后,其中Arg从ERM运转至菌根组织,并使菌根组织自由氨基酸中Arg的水平增加至20％左右。同时通过[U-^13C]Arg标记在真菌室或菌根室发现Arg可以沿AM真菌菌丝双向运转。被运转的Arg会进一步分解为Orn和尿素,因为[U-^13C]或[U-^15N／U-^13C]Arg标记于真菌室后完整的[U-^13C]或[U-^15N／U-^13C]Orn在菌根组织中产生。所以,Orn的生成标志着AM真菌氮代谢中发生了Arg的运转和分解代谢。     

不同水肥管理对红花生长的影响及红花根际解磷菌的筛选和鉴定

【目的】分析不同水肥条件对红花生物量、根际土壤磷素及微生物的影响,并从红花根际土壤样品中分离具有高效解磷能力的菌株,为红花科学水肥管理提供理论依据,并为红花的生长发育和根际微环境研究提供优良菌株。【方法】采用不同磷肥梯度处理红花,在红花的莲座期、伸长期、盛花期和种子成熟期检测植株生物量,同时测定植株根际土壤微生物、全磷和速效磷以及土壤磷酸酶活性,并进行差异性和相关性分析。采用抖土法和稀释涂布法分离筛选具有高效解磷能力的菌株。通过16S rRNA基因序列比较分析,对其进行鉴定。通过钼锑抗比色法测定菌株在不同培养基中的溶磷能力。利用灌根法和稀释涂布法接种优势菌株,分析菌株在红花根际定殖能力和促生能力。【结果】W3-P2的水肥处理有利于红花生物量的积累,速效磷含量和磷酸酶活性随施加磷肥浓度的增加呈先增大后减小趋势,水分对土壤全磷、速效磷和磷酸酶的影响与红花发育时期相关。细菌是红花根际土壤的优势菌群,在种子成熟期W4-P2处理组细菌数目最多,分别为3.017×10⁷ CFU/g和3.021×10⁷ CFU/g,远高于相同处理组的真菌和放线菌。从红花根际土壤筛选出5株高效解磷菌株（登录号C1：OR493125;C2：OR493126;C5：OR493127;C6：OR493128;C7：OR493129）,均对以无机磷和有机磷为唯一磷源的培养基具有溶磷能力和降低pH的功能,其中C6的溶磷能力最强,在磷酸三钙、磷酸铝、磷酸铁和植酸钙无机磷培养基中解磷量分别为380.00、269.32、7.15、48.16 mg/L,在有机磷（卵磷脂）培养基中解磷量为18.19 mg/L。通过16S rRNA基因序列分析,C6为假单胞菌属,C1、C2、C5、C7为中华根瘤菌属。在红花植株周围接种2%优势解磷菌C1、C5和C6菌体悬液（10⁸ CFU/mL）,在21 d时仍然保持在10⁵ CFU/g,其中C6定殖能力最强。同时检测盛花期生物量（叶片数、株高、茎粗、茎秆重和根长）,结果显示均能显著促进红花生长,其中C6菌株促生能力最强,分别为122片、115.96 cm、12.49 mm、43.36 g、21.17 cm。【结论】水肥影响红花根际微环境的速效磷含量和微生物数目的变化水平,促进红花根系的生长发育,从而直接或间接影响红花生物量,W3-P2的水肥量相对适合红花的生长。菌株C6是一株高效解磷菌株,能够分解难溶性有机磷和无机磷,盆栽实验表明C6可以在红花根际定殖并显著促进红花生长。     

AM真菌对青枯菌和根际细菌群落结构的影响

利用传统的平板培养与DGGE相结合的技术手段,研究了接种AM真菌对番茄根际土壤中的青枯菌和细菌群落结构的影响。结果表明,菌根根际土壤中的细菌总量和总DNA量都高于非菌根根际土壤,其中前者的青枯菌种群数量比后者低60倍;DGGE图谱也证实了AM真菌对青枯菌的抑制效应,还揭示出接种AM真菌对根际土壤中细菌群落结构所产生的复杂的影响。文章对AM真菌抑制青枯菌的机制进行了探讨。     

Effects of arbuscular mycorrhiza and phosphorus application on arsenic toxicity in sunflower (Helianthus annuus L.) and on the transformation of arsenic in the rhizosphere

Effects of arbuscular mycorrhiza (AM) and phosphorus (P) application on arsenic (As) toxicity were studied in a rhizobox system with As-contaminated soil collected from Shimane Prefecture, Japan. The treatments consisted of a combination of two levels of AM (Glomus aggregatum) inoculation (−AM and +AM) and two levels of P application (−P and +P at 30 mg P kg⁻¹). Sunflower (Helianthus annuus L.) seedlings were cultured in rhizoboxes for 6 weeks. Rates of root AM infection in +AM treatments were about 40% regardless of P application. AM inoculation as well as P application reduced As toxicity symptoms, most clearly so in the +AM−P treatment. Plant growth was highest in the +AM + P treatment. Shoot As concentrations were slightly reduced by AM inoculation but enhanced by P application. Shoot P concentration in the +AM−P treatment was similar to that of +P treatments and was higher than in −AM−P. Analyses of rhizosphere soils at the end of the cultivation period indicated that P application increased water-soluble As (WS−As) in all compartments while AM inoculation increased WS−As in the central compartment only. Both the WS−arsenite [WS−As^(III)] and the dominant form, arsenate [WS−As^(V)], showed gradients toward the root surface. Dimethylarsine (DMAA) was detected in the +AM treatments only. To our knowledge, this is the first report of the occurrence of DMAA in the mycorrhizosphere. AM inoculation increased WS−P similarly as +P treatments did and promoted acid phosphatase activity in the soil. In conclusion, AM inoculation alleviated the effects of As toxicity by improving P nutrition without increasing As concentrations in the shoots. Moreover, AM appeared to be involved in the transformation of soil inorganic As into less toxic organic forms.     

Contribution by two arbuscular mycorrhizal fungi to P uptake by cucumber (Cucumis sativus L.) from32P-labelled organic matter during mineralization in soil

An experiment was set up to investigate the role of arbuscular mycorrhiza (AM) in utilization of P from organic matter during mineralization in soil. Cucumber (Cucumis sativus L.) inoculated with one of two AM fungi or left uninoculated were grown for 30 days in cross-shaped PVC pots. One of two horizontal compartments contained 100 g soil (quartz sand: clay loam, 1:1) with 0.5 g ground clover leaves labelled with³²P. The labelled soil received microbial inoculum without AM fungi to ensure mineralization of the added organic matter. The labelling compartment was separated from a central root compartment by either 37 m or 700 m nylon mesh giving only hyphae or both roots and hyphae, respectively, access to the labelled soil. The recovery of³²P from the hyphal compartment was 5.5 and 8.6% for plants colonized withGlomus sp. andG. caledonium, respectively, but only 0.6 % for the non-mycorrhizal controls. Interfungal differences were not related to root colonization or hyphal length densities, which were lowest forG. caledonium. Both fungi depleted the labelled soil of NaHCO₃-extractable P and³²P compared to controls. A 15–25% recovery of³²P by roots was not enhanced in the presence of mycorrhizas, probably due to high root densities in the labelled soil. The experiment confirms that AM fungi differ in P uptake characteristics, and that mycorrhizal hyphae can intercept some P immobilization by other microorganisms and P-sorbing clay minerals.     

Studies on the attractional effect of root exudates on hyphal growth of an arbuscular mycorrhizal fungus in a soil compartment-membrane system

The effects of tomato and bean rhizospheres on hyphal spreading of the arbuscular mycorrhizal (AM) fungus Glomus mosseae were studied using a soil compartment system in combination with hydrophobic polytetrafluorethylene (PTFE) membranes. Both the nylon screen and the PTFE membrane were freely permeable to hyphae but not to roots. Furthermore, the hydrophobic PTFE membrane seemed to be a barrier to the flux of soil solutions containing root exudates. The results show that water soluble exudates of tomato and bean roots greatly stimulate hyphal growth in the soil compartment system used. Moreover, water soluble root exudates of bean exert a clear attractional effect on AM hyphal growth.     

Acquisition of N by external hyphae of an arbuscular mycorrhizal fungus and its impact on physiological responses in maize under drought-stressed and well-watered conditions

This study examined the uptake of nitrogen by external hyphae of an arbuscular mycorrhizal (AM) fungus (Glomus intraradices Schenck &; Smith) and its impact on physiological responses in maize plants subjected to well-watered or drought-stressed conditions. Plants were grown in compartmented boxes divided by a nylon mesh (40?μm) into a root compartment and a hyphal compartment. Maize plants (Zea mays cv. 'Tuxpeño sequia' selection cycle C0) were exposed to 2 weeks of drought 56 days after sowing. A ^[15]N tracer was applied as K^[15]NO_[3] to the hyphal compartment at a distance of 5?cm from the root compartment. Root and shoot samples were then analyzed for ^[15]N atom % excess (APE), glutamine synthetase (GS) activity, protein concentration and nutritional status. Evapotranspiration rate and stomatal resistance were monitored daily to determine the degree of drought stress. The APE values for AM shoots and roots were 32% and 33% higher than non-AM shoots and roots, respectively, under drought conditions. This provides clear evidence that the external mycelium of AM fungus transports considerable amounts of ^[15]NO_[3]^[– ]to the host plant under drought conditions. Drought-stressed AM roots had 28% higher GS activity, possibly as a consequence of higher hyphal acquisition of NO_[3]^[–] ions. Mycorrhizal colonization significantly increased the host plant P status regardless of soil moisture regime. In addition, the N status of drought-stressed AM shoots and roots was slightly higher than stressed non-AM shoots and roots. The improved nutritional status may assist AM plants to exploit available soil moisture more efficiently and to maintain higher leaf relative water content under moderate drought conditions.     

不同丛枝菌根真菌侵染对土壤结构的影响

为了定量化比较研究接种丛枝菌根真菌后,根际、菌根际和菌丝际土壤结构的变化,采用四室分根装置,比较中性紫色土接种不同AM真菌后,菌根际、根际、菌丝际和非根际土壤平均重量直径(MWD)、几何平均直径(GMD)和大于0.25mm团聚体总量(R_0.25)的变化。结果表明:接种3个菌种后菌丝际EEG和有机质含量均呈高于菌根际的趋势。菌丝密度和易提取球囊霉素相关蛋白(EEG)与MWD、GMD和R_0.25呈显著正相关,菌根际和菌丝际土壤水稳性R_0.25与菌丝密度显著正相关,相关系数分别为0.777和0.671。接种G. mosseae的菌根际土壤R_0.25值显著高于其它分室土壤,而接种G.etunicatum的菌丝际土壤R_0.25值显著高于其它分室土壤。试验结果在一定程度上说明不同菌种对土壤结构均有不同程度的影响,反映了丛枝菌根真菌生态功能的多样性。     

Spatial changes of arbuscular mycorrhizal fungi in peach and their correlation with soil properties

Arbuscular mycorrhizal (AM) fungi have beneficial effects on host plants, but their growth is influenced by various factors. This study was carried out to analyze the variation of AM fungi in soils and roots of peach (Prunus persica L. var. Golden Honey 3, a yellow-flesh variety) trees in different soil layers (0–40 cm) and their correlation with soil properties. The peach tree could be colonized by indigenous AM fungi (2.2–8.7 spores/g soil and 1.63–3.57 cm hyphal length/g soil), achieving 79.50–93.55% of root AM fungal colonization degree. The mycorrhizal growth, root sugars, soil three glomalins, NH₄⁺-N, NO₃⁻-N, available P and K, and soil organic matter (SOM) had spatial heterogeneity. Soil spores, but not soil hyphae contributed to soil glomalin, and soil glomalin also contributed to SOM. There was a significant correlation of soil hyphae with spore density, soil NO₃⁻-N, and SOM. Root mycorrhiza was positively correlated with spore density, NH₄⁺-N, NO₃⁻-N, and easily extractable glomalin-related soil protein. Notably, spore density positively correlated with NO₃⁻-N, available K, SOM, and root fructose and glucose, while negatively correlated with available P and root sucrose. These findings concluded that mycorrhiza of peach showed spatial distribution, and soil properties mainly affected/altered based on the soil spore density.     

Contribution of the VA mycorrhizal hyphae in acquisition of phosphorus and zinc by maize grown in a calcareous soil

An investigation was carried out to test whether the mechanism of increased zinc (Zn) uptake by mycorrhizal plants is similar to that of increased phosphorus (P) acquisition. Maize (Zea mays L.) was grown in pots containing sterilised calcareous soil either inoculated with a mycorrhizal fungus Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappe or with a mixture of mycorrhizal fungi, or remaining non-inoculated as non-mycorrhizal control. The pots had three compartments, a central one for root growth and two outer ones for hyphal growth. The compartmentalization was done using a 30-m nylon net. The root compartment received low or high levels of P (50 or 100 mg kg^–1 soil) in combination with low or high levels of P and micronutrients (2 or 10 mg kg^–1 Fe, Zn and Cu) in the hyphal compartments.Mycorrhizal fungus inoculation did not influence shoot dry weight, but reduced root dry weight when low P levels were supplied to the root compartment. Irrespective of the P levels in the root compartment, shoots and roots of mycorrhizal plants had on average 95 and 115% higher P concentrations, and 164 and 22% higher Zn concentrations, respectively, compared to non-mycorrhizal plants. These higher concentrations could be attributed to a substantial translocation of P and Zn from hyphal compartments to the plant via the mycorrhizal hyphae. Mycorrhizal inoculation also enhanced copper concentration in roots (135%) but not in shoots. In contrast, manganese (Mn) concentrations in shoots and roots of mycorrhizal plants were distinctly lower, especially in plants inoculated with the mixture of mycorrhizal fungi.The results demonstrate that VA mycorrhizal hyphae uptake and translocation to the host is an important component of increased acquisition of P and Zn by mycorrhizal plants. The minimal hyphae contribution (delivery by the hyphae from the outer compartments) to the total plant acquisition ranged from 13 to 20% for P and from 16 to 25% for Zn.