丛枝菌根利用氮素研究进展

氮素是植物需求量最大的元素,丛枝菌根真菌与植物形成共生体后能从土壤中获取无机氮、简单的氨基酸,还能利用一些复杂的有机态氮.考虑到NH+4在土壤中的移动性低及丛枝菌根真菌的专性共生菌的特点,丛枝菌根真菌吸收NH+4对植物的贡献较大.近年来的研究发现丛枝菌根真菌内存在与氮素代谢有关的鸟氨酸循环,而精氨酸则是菌丝内氮素转移的主要形式.综述最近的AMF对氮素的吸收、转运、同化、交换等方面的文献,旨在揭示丛枝菌根真菌氮素利用特点,阐明丛枝菌根真菌在氮循环系统中的重要作用.     

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。     

丛枝菌根共生体中碳、氮代谢及其相互关系

丛枝菌根共生体(arbuscular mycorrhiza, AM)是丛枝菌根真菌(arbuscular mycorrhizal fungi, AMF)与宿主植物之间形成的互惠共生形式.共生体中的碳、氮交换和代谢影响着宿主植物和共生真菌之间的营养平衡和资源重新分配,在物质和能量循环中发挥着重要作用.宿主植物光合固定的碳输送到真菌内,并且分解和释放真菌所需的生命物质和能量,包括促进孢子萌发、菌丝生长和提高氮等营养元素的吸收;而菌根真菌利用宿主植物提供的碳骨架和能量,发生氮的转化和运输,最终传递给宿主植物供其利用.本文综述了丛枝菌根共生体中碳、氮传输和代谢的主要模式,碳、氮的交互影响和调控机制,以促进丛枝菌根在可持续农业和生态系统中的应用.     

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。     

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。     

丛枝菌根真菌在土壤氮素循环中的作用

作为植物需求量最大的营养元素,氮素是陆地生态系统初级生产力的主要限制因子。丛枝菌根真菌能与地球上80%以上的陆生植物形成菌根共生体,帮助宿主植物吸收土壤中的P、N等矿质养分。目前,丛枝菌根真菌与氮素循环相关研究侧重于真菌对氮素的吸收形态以及共生体中氮的传输代谢机制,却忽略了丛枝菌根真菌在固氮过程、矿化与吸收过程、硝化过程、反硝化过程以及氮素淋洗过程等土壤氮素循环过程中所起到的潜在作用,并且越来越多的证据也表明丛枝菌根真菌是影响土壤氮素循环过程的重要因子。总结了丛枝菌根真菌可利用的氮素形态及真菌的氮代谢转运相关基因的研究现状;重点分析了丛枝菌根真菌在调控土壤氮素循环过程中的潜在作用以及在生态系统中的重要生态学意义,同时提出了丛枝菌根真菌在土壤氮素循环过程中一些需要深入研究的问题。     

脱落酸对丛枝菌根真菌侵染和产孢的调控效应研究

【目的】揭示脱落酸(ABA)对丛枝菌根(AM)真菌侵染和产孢的影响,建立利用外源ABA促进孢子产量的高效菌剂扩繁方法。【方法】利用番茄毛状根和AM真菌Rhizophagus irregularis DAOM 197198建立双重培养体系,通过外源施用ABA、赤霉素(GA)或者使用ABA、GA的缺陷突变体,染色观察菌根侵染,荧光定量PCR测定丛枝发育和脂质合成运输相关基因的表达,统计丛枝和孢子的数量,从而揭示ABA对AM真菌侵染和产孢的影响。【结果】ABA缺陷突变体not中的F%(侵染频率)、a%(丛枝丰度)、丛枝数量,以及丛枝发育特异性相关基因EXO70A1-like (LOC101253481)、脂质合成运输相关基因RAM2和STR2的表达均显著低于其野生型MT;外源施用ABA显著促进了F%、M%(侵染强度)、丛枝数量、孢子产量,以及脂质合成运输相关基因RAM2和STR2的表达,外源添加ABA处理的孢子产量约为不添加处理的4.5倍;外源GA处理极显著抑制了菌根侵染的所有指标和孢子产量;GA缺陷突变体gib3与其野生型MM的AM真菌侵染之间没有显著差异,但gib3的孢子产量显著高于MM。【结论】ABA通过促进脂质的合成和运输,提高AM真菌的侵染和丛枝形成,进而增加AM真菌的孢子产量。     

感染羽茅的香柱菌属内生真菌对丛枝菌根真菌孢子萌发的影响

内生真菌与丛枝菌根(AM)真菌是构成草原生态系统的重要组成部分.内生真菌会抑制其宿主植物的AM真菌侵染率.本研究以感染2种香柱菌属内生真菌[Epichloё gansuensis(Eg)和E. sibirica(Es)]的天然禾草羽茅为供试材料,进行体外纯培养的内生真菌培养滤液、感染内生真菌的羽茅叶片(包括鲜叶和枯叶)浸提液,以及根系分泌物对摩西球囊霉(Gm)和幼套球囊霉(Ge)2种AM真菌孢子萌发影响试验.结果表明: 香柱菌属内生真菌的培养滤液会显著抑制2种AM真菌孢子的萌发,而感染香柱菌属内生真菌的羽茅根系分泌物只对Ge孢子萌发有显著抑制作用,且上述抑制作用与内生真菌种类无关;鲜叶浸提液对Gm和Ge的孢子萌发率均无显著影响,而枯叶浸提液对Ge的孢子萌发有显著抑制作用.在自然生态系统中,香柱菌属内生真菌通常存在于宿主植物体内,可能通过影响宿主植物的根系分泌物来影响AM真菌孢子的萌发.     

多胺对离体培养条件下丛枝菌根真菌生长发育的影响*

研究离体培养条件下多胺 (PUT,SPD,SPM) 及多胺生物合成抑制剂 (MGBG) 对丛枝菌根真菌 (Glomus mosseae, Gigaspora margarita) 孢子萌发特性及菌丝生长发育的影响。试验结果表明,3种多胺类物质在50 ~200g/ml浓度范围内,对丛枝菌根真菌生长发育具显著促进作用,而500礸/ml浓度处理对丛枝菌根真菌生长发育表现强烈的抑制效应。MGBG (50 ~500g/ml) 对丛枝菌根真菌生长发育有较强的抑制作用,且可被外源多胺部分解除,但随浓度升高外源多胺的恢复作用降低,500礸/ml时无效。多胺对丛枝菌根真菌生长发育的促进作用因多胺类型及真菌菌种的变化而有不同的最适浓度范围。作者认为丛枝菌根真菌体内内源多胺的含量也许是其生长发育的限制因子。     

菌根真菌促进植物吸收利用氮素机制的研究进展

作为自然界最为普遍的一种植物共生体,菌根能够极大地促进植物对氮素的吸收和利用,其中菌根真菌在共生结构功能中发挥了重要作用。本文分别从菌根解剖构造、生理生化和分子生物学方面系统总结了菌根真菌促进植物吸收和利用氮素的研究现状。重点介绍了菌根真菌可利用的氮素形态及影响其利用的主要因素、菌根真菌的氮代谢途径GS-GOGAT以及菌根真菌中存在的鸟氨酸循环途径,指出精氨酸是菌丝内氮转运的主要形式,NH3可能为菌根真菌和植物界面质外体的主要转运形式。     

The initial metabolism of germinating fern spores

Label from tritiated water is distributed in a manner that suggests that germinating fern spores initially convert amino acids and hydroxy acids into oxo acids. Enzymes capable of catalysing these processes occur in resting spores.     

Forms of nitrogen uptake, translocation, and transfer via arbuscular mycorrhizal fungi: A review

Arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the roots of more than 80% of land plants. Experiments on the relationship between the host plant and AM in soil or in sterile root-organ culture have provided clear evidence that the extraradical mycelia of AM fungi uptake various forms of nitrogen (N) and transport the assimilated N to the roots of the host plant. However, the uptake mechanisms of various forms of N and its translocation and transfer from the fungus to the host are virtually unknown. Therefore, there is a dearth of integrated models describing the movement of N through the AM fungal hyphae. Recent studies examined Ri T-DNA-transformed carrot roots colonized with AM fungi in ¹⁵N tracer experiments. In these experiments, the activities of key enzymes were determined, and expressions of genes related to N assimilation and translocation pathways were quantified. This review summarizes and discusses the results of recent research on the forms of N uptake, transport, degradation, and transfer to the roots of the host plant and the underlying mechanisms, as well as research on the forms of N and carbon used by germinating spores and their effects on amino acid metabolism. Finally, a pathway model summarizing the entire mechanism of N metabolism in AM fungi is outlined.     

Role of amino acids and endogenous protein in the germination of Mucor racemosus sporangiospores

The role of amino acids as triggers of Mucor racemosus sporangiospore germination was investigated. No single amino acid was effective as glucose or peptone at triggering germination. Germination induced by glucose or peptone was pH-independent, whereas germination induced by glutamate was pH-dependent. The composition of the free amino acid pools of M-spores (those unable to germinate on glutamate) was qualitatively and quantitatively similar to that of C-spores (those capable of germinating on glutamate) with the exceptions of hydroxyproline and methionine and methionine whose concentrations were several-fold higher in C-spores. Glutamate and leucine were taken up by germinating and nongerminating spores; however, significant protein synthesis occurred only in germinating spores. Spores not triggered by 3-O-methyl-D-glucose (M-spores) contained about one-half the protein of those triggered by 3-O-methyl-D-glucose (C-spores). C-spores initiated to germinate by 3-O-methyl-D-glucose decreased in total organic carbon and protein over a 6 h period; removal of the 3-O-methyl-D-glucose resulted in an immediate halt of protein degradation and spore swelling. These results suggest that protein is a major endogenous reserve in M. racemosus sporangiospores and that its turnover is a necessary event for glucose-triggered germination.     

ORIGIN OF AMINO ACIDS CONSTITUTING CELLULAR PROTEIN IN GERMINATING CONIDIOSPORES OF ASPERGILLUS NIGER

Formation of pool amino acids in germinating spores of Aspergillusniger strain 1617 was investigated. The pool amino acids comprisedmainly glutamic acid and alanine. Small amounts of pyruvateand -ketoglutarate were found to increase almost in parallelwith the course of increase in the amount of free amino acidsup to the stage of onset of active protein synthesis. Asparticglutamictransaminase activity was exhibited even in dormant spores andit developed in response to the increase in cellular protein.Alanine-glutamic transaminase activity, on the other hand, waslacking in dormant spores and appeared at the stage of accumulationof amino acids preceding protein synthesis. It was revealed from the experiments with ³⁵S-labeled sporesthat the dormant spores of this fungus contain two unidentifiedsulfur substances, and the sulfur of these substances is incorporatedinto the sulfur amino acids of the protein synthesized in germinatingspores. ¹Present address: Institute of Applied Microbiology, Universityof Tokyo, Tokyo (Received September 11, 1959; )     

Endobacteria affect the metabolic profile of their host Gigaspora margarita, an arbuscular mycorrhizal fungus

The aim of this paper was to understand whether the endobacterium identified as Candidatus Glomeribacter gigasporarum has an effect on the biology of its host, the arbuscular mycorrhizal fungus Gigaspora margarita, through the study of the modifications induced on the fungal proteome and lipid profile. The availability of G. margarita cured spores (i.e. spores that do not contain bacteria), represented a crucial tool to enable the comparison between two fungal homogeneous populations in the presence and the absence of the bacterial components. Our results demonstrate that the endobacterial presence leads to a modulation of fungal protein expression in all the different conditions we tested (quiescent, germinating and strigolactone-elicited germinating spores), and in particular after treatment with a strigolactone analogue. The fungal fatty acid profile resulted to be modified both quantitatively and qualitatively in the absence of endobacteria, being fatty acids less abundant in the cured spores. The results offer one of the first comparative metabolic studies of an AM fungus investigated under different physiological conditions, reveal that endobacteria have an important impact on the host fungal activity, influencing both protein expression and lipid profile, and suggest that the bacterial absence is perceived by G. margarita as a stimulus which activates stress-responsive proteins.     

Cell cycle progression and cell polarity require sphingolipid biosynthesis in Aspergillus nidulans

Sphingolipids are major components of the plasma membrane of eukaryotic cells and were once thought of merely as structural components of the membrane. We have investigated effects of inhibiting sphingolipid biosynthesis, both in germinating spores and growing hyphae of Aspergillus nidulans. In germinating spores, genetic or pharmacological inactivation of inositol phosphorylceramide (IPC) synthase arrests the cell cycle in G(1) and also prevents polarized growth during spore germination. However, inactivation of IPC synthase not only eliminates sphingolipid biosynthesis but also leads to a marked accumulation of ceramide, its upstream intermediate. We therefore inactivated serine palmitoyltransferase, the first enzyme in the sphingolipid biosynthesis pathway, to determine effects of inhibiting sphingolipid biosynthesis without an accumulation of ceramide. This inactivation also prevented polarized growth but did not affect nuclear division of germinating spores. To see if sphingolipid biosynthesis is required to maintain polarized growth, and not just to establish polarity, we inhibited sphingolipid biosynthesis in cells in which polarity was already established. This inhibition rapidly abolished normal cell polarity and promoted cell tip branching, which normally never occurs. Cell tip branching was closely associated with dramatic changes in the normally highly polarized actin cytoskeleton and found to be dependent on actin function. The results indicate that sphingolipids are essential for the establishment and maintenance of cell polarity via control of the actin cytoskeleton and that accumulation of ceramide is likely responsible for arresting the cell cycle in G(1).     

Protein synthesis in germinating Saccharomyces cerevisiae ascospores

The uptake and incorporation of macromolecular precursors in germinating Saccharomyces cerevisiae ascospores were investigated. Addition of cycloheximide at various times during germination revealed that protein synthesis can occur within 20 min after the spores are shifted to glucose-containing media. The time of initiation of uptake and incorporation of several amino acids differed; this can be attributed to differing amino acid pool levels in the spores, as well as differing transport activities. Two-dimensional gel electrophoresis of proteins labeled with [35S]methionine for various 20-min periods after germination began showed at least one protein whose synthesis begins well after the bulk of the proteins.     

Activation and expression of proteins during synchronous germination of aerial spores of Streptomyces granaticolor

Synchronously germinating aerial spores of Streptomyces granaticolor were used to study protein activation and expression during the transition from dormant to metabolically active vegetative forms. The first phase of protein activation is associated with the solubility of proteins. Three major chaperones, DnaK, Trigger factor, and GroEL, were identified in spores. Enhancement in rate of protein synthesis during germination was accompanied by the association of TF and DnaK with ribosomes. During germination, the chaperones TF, GroEL, and DnaK undergo reversible phosphorylation. GroEL was phosphorylated on both Ser and Thr, whereas phosphorylation of DnaK and TF was detected on Thr only. A proteomic approach was used to gain more information on protein expression during germination on two types of media differing in the ability of cells to produce antibiotic granaticin. To obtain an overview of the metabolic activity of germinating spores, glycolytic enzymes, enzymes of citric acid cycle, metabolism of amino acids and nucleic acids, and components of the protein synthesis system were identified and analyzed using the proteomic database. The results were deposited on the SWICZ proteomic server and are accessible on http://proteom.biomed.cas.cz.