葡萄糖、根浸出液对丛枝菌根真菌吸收不同外源氮产生精氨酸的影响

以大葱(Allium fistulosum)为宿主植物, 接种丛枝菌根(Arbuscular mycorrhizal, AM)真菌Glomus intraradices, 采用三室隔离盆栽培养系统, 在菌丝室施加浓度为4 mmol/L的不同形态外源氮、1%葡萄糖及根浸出液, 通过测定根外菌丝(Extraradical mycelium, ERM)和菌根中精氨酸的含量, 探究葡萄糖、根浸出液对AM真菌吸收不同形式外源氮产生精氨酸的影响。结果表明, 不同外源氮对ERM中精氨酸含量的影响为尿素>Gln>NH₄NO₃>Arg/Gly>NH₄Cl>KNO₃, 对菌根中精氨酸含量的影响为Arg>Gln>尿素>NH₄NO₃>Gly>NH₄Cl>KNO₃; 施加葡萄糖和根浸出液在不同程度上提高ERM干重和菌丝室孢子数量, 但使ERM和菌根中的精氨酸含量降低。说明AM真菌吸收同化不同外源氮产生精氨酸的能力不同, 葡萄糖和根浸出液降低AM真菌吸收同化外源氮产精氨酸的能力。     

不同外源氮对丛枝菌根真菌Rhizophagus irregularis侵染棉花植株和氮磷转运的影响

探究供应外源氮对接种AM真菌的棉花植株的侵染率和氮磷转运的影响。本文以棉花为研究对象,接种丛枝菌根真菌(Rhizophagus irregularis),向根外菌丝额外供应不同外源氮,测AM真菌的侵染率、棉花植株株高、地上部和地下部鲜重、叶绿素含量、菌根精氨酸含量、地上部的氮磷含量。试验结果显示:不同外源氮条件下,AM真菌对棉花植株的生物量无显著性影响;外源氮的供应均提高了AM真菌的侵染率和棉花植株地上部的氮含量,但硫酸铵和硝酸钾更能促进AM真菌侵染宿主植物,提高宿主植物菌根精氨酸水平和地上部氮含量;除了尿素,其他氮源处理均能明显提高棉花植株地上部磷含量,其中,精氨酸最为显著。说明在AM共生系统中外源氮的供应对宿主植物生长无显著作用,但促进AM真菌侵染宿主植物,并能提高宿主植物氮磷含量。     

磷水平对接种丛枝菌根真菌甜玉米苗期生长的影响

研究了不同外源磷水平条件下,接种丛枝菌根真菌根内球囊霉(Glomus intraradices)对寄主植物甜玉米菌根侵染率、地上部和地下部鲜重、氮磷含量、精氨酸含量影响。结果表明:丛枝菌根真菌能够很好的侵染于玉米植株根系。且不同磷水平条件下,菌根侵染率差异较显著。在低磷水平下,菌根侵染率较高。孢子数量随着磷水平提高而增加。菌丝室根外菌丝鲜重在P40时最高。菌根化的甜玉米生物量及氮磷含量显著高于对照组。此外,低磷水平促使甜玉米地上部和地下部鲜重显著提高。甜玉米地上部总氮和地下部总氮含量分别在P40、P80和P20、P40时最高。地上部总磷和地下部总磷含量分别在P80和P160时最高。菌根精氨酸含量在低磷(P20)时最高。研究表明接种丛枝菌根真菌可促进甜玉米幼苗生长并与外源磷水平有关。     

丛枝菌根真菌萌发孢子利用不同氮素及外源葡萄糖对其代谢的影响

对各种含氮基质、葡萄糖和(或)根浸出液中培养的丛枝菌根真菌Glomus intraradices孢子,在萌发过程中对不同氮素的利用及其氨基酸的生物合成进行了研究.用稳定同位素标记及质谱仪来分析不同氮素的利用和氨基酸的生物合成.以高效液相色谱测量氨基酸的浓度.在缺少外源氮素的情况下,丛枝菌根真菌孢子萌发时可以利用内部储存的含氮化合物生物合成游离氨基酸.其中,丝氨酸和甘氨酸是大量合成的氨基酸.合成的氨基酸浓度在2周内随着萌发时间的增加而增加.在有可利用的外源无机氮(铵盐、硝酸盐和尿素)和有机氮(氨基酸)时,铵盐和尿素比硝酸盐更容易被AM真菌萌发孢子利用,而其利用氨基酸中的氮比无机氮源慢的多.孢子吸收同化外源无机氮,且将其整合到游离氨基酸中,这些新生氨基酸浓度比无外源氮添加时要高得多.在无葡萄糖添加的硝酸盐培养液中,AM真菌孢子中积累大量天冬酰胺.然而,在含有葡萄糖的培养液中,萌发孢子因葡萄糖的吸收促进了对外源氮的吸收,产生的游离氨基酸是无葡萄糖时的5倍,并且发现精氨酸转为含量最多的游离氨基酸.并且,从外源氮吸收同化的氮可以储存于精氨酸中,随之,精氨酸被整合到AM真菌孢子储存的蛋白质中.此外,根浸出原液在AM真菌孢子萌发2周后对氮的吸收作用不明显.     

宿主植物栽培密度对AM真菌生长发育的影响

温室盆栽条件下宿主植物高粱(SorghumvulgarePers.)的栽培密度对丛枝菌根(Arbuscularmycorrhizae,AM)真菌Glomusmosseae(Nicol.&Gerd.)Gerdemann&Trappe生长发育的影响试验结果表明:60株/盆密度处理的根外菌丝量及孢子数均高于其它处理。在一定栽培密度下(20~60株/盆),植株根系可溶性糖浓度与根外菌丝量呈显著负相关,与菌根侵染率呈显著正相关。植株根中磷浓度与根外菌丝量、根外菌丝量与孢子数均呈显著正相关。植株根中磷浓度与菌根侵染率呈显著负相关。结果说明:适当密植虽对植株生长有一定影响,但却促进了真菌的生长,此时菌根共生体有可能由互惠共生开始向偏利共生或弱寄生转化。密植作为一种调控手段,在菌剂生产中能获得较大数量的侵染根段、菌丝及孢子等繁殖体。     

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．     

宿主植物栽培密度对AM真菌生长发育的影响*

温室盆栽条件下宿主植物高粱(SorghumvulgarePers.)的栽培密度对丛枝菌根(Arbuscularmycorrhizae,AM)真菌Glomusmosseae(Nicol.&Gerd.)Gerdemann&Trappe生长发育的影响试验结果表明:60株/盆密度处理的根外菌丝量及孢子数均高于其它处理。在一定栽培密度下(20~60株/盆),植株根系可溶性糖浓度与根外菌丝量呈显著负相关,与菌根侵染率呈显著正相关。植株根中磷浓度与根外菌丝量、根外菌丝量与孢子数均呈显著正相关。植株根中磷浓度与菌根侵染率呈显著负相关。结果说明:适当密植虽对植株生长有一定影响,但却促进了真菌的生长,此时菌根共生体有可能由互惠共生开始向偏利共生或弱寄生转化。密植作为一种调控手段,在菌剂生产中能获得较大数量的侵染根段、菌丝及孢子等繁殖体。     

菌丝室接种解磷细菌Bacillus megaterium C4对土壤有机磷矿化和植物吸收的影响

通过30μm尼龙网将根盒分成根室和菌丝室,菌丝室中的低磷土壤施加75 mg P/kg土壤的植酸钙,研究了菌丝室土壤中丛枝菌根(AM)真菌Glomus intraradices和解磷细菌Bacillus megaterium C4对有机磷的矿化和吸收.结果表明,在试验条件下,植酸钙的溶解性很低,对土壤溶液有机磷的贡献不大.接种解磷细菌C4提高了土壤中磷酸酶的活性,减少了土壤中有机磷的含量.但是,由于存在解磷细菌与AM真菌对磷的竞争,解磷细菌矿化出的磷大部分被自身利用,AM真菌的生长受到抑制,解磷细菌对植物磷营养的改善没有表现出显著的贡献.     

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。     

丛枝菌根真菌和施氮量对茶树生长、矿质元素吸收与茶叶品质的影响

采用盆栽法研究了不同施氮水平下接种丛枝菌根（arbuscular mycorrhiza,AM）真菌Glomus mosseae对茶树生长、矿质元素吸收及茶叶品质的影响。结果表明,适量的施氮利于AM真菌的侵染和菌根发育,当施氮过量时则会抑制菌根发育。在不同施氮水平下接种AM真菌均提高了茶树地上部、地下部和总干物质量,其中又以接种AM真菌同时施氮量为0.53g kg-1的茶树总干物质量最大,为对照的1.63倍。不同矿质元素受AM真菌和氮肥的影响不一致,在一定施氮水平下接种AM真菌可提高茶树叶片中N、P、K、Ca、Zn和Fe含量,降低Mn和Cu含量;显著增加根中N、P、K、Mg和Zn含量,降低Mn含量,施高浓度的氮（1.06 g kg-1）显著降低了根系Ca和Fe含量。不同施氮水平下AM真菌处理可增加茶叶中可溶性糖和可溶性蛋白含量,提高了茶叶中茶多酚、咖啡碱、氨基酸和水浸出物含量,降低酚氨比,显著改善茶叶品质。本实验条件下,茶树施氮量为0.53 g kg-1时,接种AM真菌改善茶叶品质的效果最佳。     

Nitrogen transfer and assimilation between the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith and Ri T-DNA roots of Daucus carota L. in an in vitro compartmented system

Nitrogen metabolism was examined in monoxenic cultures of carrot roots (Daucus carota L.) colonized with the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith. Glutamine synthetase and glutamate dehydrogenase activities were significantly increased in mycorrhizal roots for which only the extraradical mycelium had exclusive access to NH4NO3 in a distinct hyphal compartment inaccessible to the roots. This was in comparison with the water controls but was similar to the enzyme activities of non-arbuscular-mycorrhizal (non-AM) roots that had direct access to NH4NO3. In addition, glutamate dehydrogenase activity was significantly enhanced in AM roots compared with non-AM roots. Carrot roots took up 15NH4+ more efficiently than 15NO3-, and the extraradical hyphae transfered 15NH4+ to host roots from the hyphal compartment but did not transfer 15NO3-. The extraradical mycelium was shown, for the first time, to have a different glutamine synthetase monomer than roots. Our overall results highlight the active role of AM fungi in nitrogen uptake, transfer, and assimilation in their symbiotic root association.     

Studies of iron transport by arbuscular mycorrhizal hyphae from soil to peanut and sorghum plants

 The influence of an arbuscular mycorrhizal (AM) fungus on phosphorus (P) and iron (Fe) uptake of peanut (Arachis hypogea L.) and sorghum (Sorghum bicolor L.) plants was studied in a pot experiment under controlled environmental conditions. The plants were grown for 10 weeks in pots containing sterilised calcareous soil with two levels of Fe supply. The soil was inoculated with rhizosphere microorganisms only or with rhizosphere microorganisms together with an AM fungus (Glomus mosseae [Nicol. & Gerd.] Gerdemann & Trappe). An additional small soil compartment accessible to hyphae but not roots was added to each pot after 6 weeks of plant growth. Radiolabelled P and Fe were supplied to the hyphae compartment 2 weeks after addition of this compartment. After a further 2 weeks, plants were harvested and shoots were analysed for radiolabelled elements. In both plant species, P uptake from the labelled soil increased significantly more in shoots of mycorrhizal plants than non-mycorrhizal plants, thus confirming the well-known activity of the fungus in P uptake. Mycorrhizal inoculation had no significant influence on the concentration of labelled Fe in shoots of peanut plants. In contrast, ⁵⁹Fe increased in shoots of mycorrhizal sorghum plants. The uptake of Fe from labelled soil by sorghum was particularly high under conditions producing a low Fe nutritional status of the plants. These results are preliminary evidence that hyphae of an arbuscular mycorrhizal fungus can mobilise and/or take up Fe from soil and translocate it to the plant. Accepted: 6 March 1998     

Functional aspects of root architecture and mycorrhizal inoculation with respect to nutrient uptake capacity

The aim of this research was to investigate the effect of arbuscular mycorrhizal (AM) colonisation on root morphology and nitrogen uptake capacity of carob ( Ceratonia siliqua L.) under high and low nutrient conditions. The experimental design was a factorial arrangement of presence/absence of mycorrhizal fungus inoculation ( Glomus intraradices) and high/low nutrient status. Percent AM colonisation, nitrate and ammonium uptake capacity, and nitrogen and phosphorus contents were determined in 3-month-old seedlings. Grayscale and colour images were used to study root morphology and topology, and to assess the relation between root pigmentation and physiological activities. AM colonisation lead to a higher allocation of biomass to white and yellow parts of the root. Inorganic nitrogen uptake capacity per unit root length and nitrogen content were greatest in AM colonised plants grown under low nutrient conditions. A better match was found between plant nitrogen content and biomass accumulation, than between plant phosphorus content and biomass accumulation. It is suggested that the increase in nutrient uptake capacity of AM colonised roots is dependent both on changes in root morphology and physiological uptake potential. This study contributes to an understanding of the role of AM fungi and root morphology in plant nutrient uptake and shows that AM colonisation improves the nitrogen nutrition of plants, mainly when growing at low levels of nutrients.     

不同基因型水生植物对铵态氮和硝态氮吸收动力学特性研究

针对不同营养状况的富营养化水体修复而选择吸收养分效率较强的水生植物,采用改进常规耗竭法比较研究了6种不同基因型水生植物凤眼莲(Eichhornia crassipes Solms)、黄花水龙(Jussiaea stipulacea Ohwi)、再力花(Thalia dealbata Fraser)、美人蕉(Canna glauca L.)、水芹[Oenanthe javanica(Bl).DC]和豆瓣菜(Nasturtium officinaleR.Br.)对铵态氮和硝态氮吸收动力学特性。结果表明,不同基因型水生植物吸收铵态氮和硝态氮的动力学特性可用Michaelis-Menten方程来描述。在低浓度培养下,不同基因型水生植物对NH4+-N和NO3--N吸收的动力学参数Imax和Km差异较大,其吸收NH4+-N和NO3--N的Imax最大是水芹,其次是豆瓣菜;Km值最小的是水芹,其次是豆瓣菜;且水芹对NH4+-N和NO3--N不仅具有较强的亲和力,还具有较高的离子吸收速率。结果还表明,当介质中氮浓度较低时,水芹有优先吸收硝态氮的趋势,而豆瓣菜和再力花有优先吸收铵态氮的趋势。     

Resource allocation to growth or luxury consumption drives mycorrhizal responses

Highly variable phenotypic responses in mycorrhizal plants challenge our functional understanding of plant‐fungal mutualisms. Using non‐invasive high‐throughput phenotyping, we observed that arbuscular mycorrhizal (AM) fungi relieved phosphorus (P) limitation and enhanced growth of Brachypodium distachyon under P‐limited conditions, while photosynthetic limitation under low nitrogen (N) was exacerbated by the fungus. However, these responses were strongly dependent on host genotype: only the faster growing genotype (Bd3‐1) utilised P transferred from the fungus to achieve improved growth under P‐limited conditions. Under low N, the slower growing genotype (Bd21) had a carbon and N surplus that was linked to a less negative growth response compared with the faster growing genotype. These responses were linked to the regulation of N : P stoichiometry, couples resource allocation to growth or luxury consumption in diverse plant lineages. Our results attest strongly to a mechanism in plants by which plant genotype‐specific resource economics drive phenotypic outcomes during AM symbioses.     

接种丛枝菌根真菌对模拟大气氮沉降下灌木铁线莲根系形态及养分承载的影响

为了解菌根化处理的灌木铁线莲（Clematis fruticosa）苗木根系形态及养分承载对氮沉降的应激响应,以1年生盆栽灌木铁线莲为对象,分别采用单接种和混合接种,即：单接种根内根孢囊霉（Rhizophagus intraradices,以下简称+R）,单接种摩西斗管囊霉（Funneliformis mosseae,以下简称+F）;混合接菌（上述2菌种菌剂按体积1∶1混合,以下简称+RF）的菌根苗。以非菌根苗（未接菌,以下简称-M）为对照。氮沉降处理设置4个梯度（不施氮（0N,0 g·m^-2·a^-1）、低氮（LN,3 g·m^-2·a^-1）、中氮（MN,6 g·m^-2·a^-1）、高氮（HN,9 g·m^-2·a^-1））,1年后测定各处理细根形态（直径≤0.5 mm的总根长、总表面积、总体积、根尖数量）、菌根侵染率、土壤孢子密度及根、茎、叶各器官的养分（碳、氮、磷）含量等指标。①在+R和+RF处理下,LN处理的苗木菌根侵染率和孢子密度达到最大,且LN处理的苗木菌根侵染率显著高于HN处理;而+F处理的苗木菌根侵染率随氮沉降递增无显著差异。②0N处理下,+F和+R处理的灌木铁线莲苗木细根（直径≤0.5 mm）的总根长、总表面积、总体积和根尖数量均显著高于-M处理。然而,+F和+R处理的灌木铁线莲苗木上述根系形态指标随着氮沉降量的增加均呈下降的趋势。③+F和+R处理下,苗木养分承载量随氮沉降量增加呈增加的趋势。氮沉降处理下,接菌处理的苗木碳、氮、磷养分含量显著高于-M处理,其中+F处理下苗木碳氮磷养分含量最高。④直径≤0.5 mm细根形态指标与养分含量指标均呈正相关关系。综上,接菌处理可改变灌木铁线莲苗木细根形态对氮沉降的响应规律,接种摩西斗管囊霉有效增强苗木对氮沉降的适应能力,提高了高氮沉降处理下苗木的养分承载量。     

Improved tolerance of maize plants to salt stress by arbuscular mycorrhiza is related to higher accumulation of soluble sugars in roots

The effect of colonization with the arbuscular mycorrhizal (AM) fungus Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe on the growth and physiology of NaCl-stressed maize plants ( Zea mays L. cv. Yedan 13) was examined in the greenhouse. Maize plants were grown in sand with 0 or 100 mM NaCl and at two phosphorus (P) (0.05 and 0.1 mM) levels for 34 days, following 34 days of non-saline pre-treatment. Mycorrhizal plants maintained higher root and shoot dry weights. Concentrations of chlorophyll, P and soluble sugars were higher than in non-mycorrhizal plants under given NaCl and P levels. Sodium concentration in roots or shoots was similar in mycorrhizal and non-mycorrhizal plants. Mycorrhizal plants had higher electrolyte concentrations in roots and lower electrolyte leakage from roots than non-mycorrhizal plants under given NaCl and P levels. Although plants in the low P plus AM fungus treatment and those with high P minus AM fungus had similar P concentrations, the mycorrhizal plants still had higher dry weights, soluble sugars and electrolyte concentrations in roots. Similar relationships were observed regardless of the presence or absence of salt stress. Higher soluble sugars and electrolyte concentrations in mycorrhizal plants suggested a higher osmoregulating capacity of these plants. Alleviation of salt stress of a host plant by AM colonization appears not to be a specific effect. Furthermore, higher requirement for carbohydrates by AM fungi induces higher soluble sugar accumulation in host root tissues, which is independent of improvement in plant P status and enhances resistance to salt-induced osmotic stress in the mycorrhizal plant.