低磷胁迫下AM真菌对枳实生苗吸磷效应及根系分泌有机酸的影响

在温室沙培灭菌条件下,以Al-P为磷源、枳为试材、Glomus mosseae (G.m)和G.versiforme (G.v)为菌剂,研究低磷胁迫下AM真菌对枳实生苗干物重、吸磷效应及根系分泌有机酸的影响。结果表明,接种AM真菌显著增加枳地上部、地下部干物重,增幅16.79%~135.25%;同时显著增加其吸磷量,菌丝对植株的吸磷贡献率为17.04%~71.95%(G.m>G.v),施Al-P显著提高菌丝吸磷贡献率。接种AM真菌的根系分泌的有机酸种类与对照有所不同,未接种处理枳分泌的有机酸有草酸、苹果酸、乳酸、乙酸、顺丁烯二酸和柠檬酸等6种,而接种G.m的则检测到草酸、酒石酸、苹果酸、乳酸、乙酸、柠檬酸、丁二酸等7种,G.v处理的检测到酒石酸,接种处理均未检测到顺丁烯二酸;接种丛枝菌根真菌增加了枳根系分泌有机酸的量(比未接种处理增加19.80~56.87 mg/kg,且施用AlPO₄后有机酸含量显著增加(增加20.06~21.84 mg/kg);未接种植株根系仅分泌少量有机酸;接种植株根系分泌的有机酸以苹果酸(42.87%)、柠檬酸(39.22%)和草酸(12.06%)为主。     

4种植物幼苗根茎叶及根系分泌物中低分子量有机酸的特征

该研究建立了植物根茎叶及根系分泌物中有机酸的离子色谱分析测定方法,并测定了4种不同植物幼苗根茎叶及根系分泌物中低分子量有机酸的组成,为揭示逆境胁迫下植物体内有机酸的作用提供依据。结果表明:离子色谱分析法对植物有机酸的加标回收率为91.10%~105.42%,检测限为0.12~0.36mg/L,方法线性关系良好(R2=0.965 3~0.998 8);4种植物根茎叶及根系分泌物中都可以检测出草酸、柠檬酸、苹果酸、丁二酸和酒石酸,其中草酸、柠檬酸和苹果酸为优势酸;有机酸的组成和含量具有物种以及器官的差异性;根系分泌物中的有机酸与根茎叶中有机酸的相关性也因种属差异而不同。这为研究逆境胁迫下植物器官及根系分泌物提供了可靠方法。     

一种根系分泌物中有机酸的前处理和高效液相色谱检测方法

改进了根系分泌有机酸的前处理和高效液相色谱检测方法.将DEAE纤维素层析法直接用于根系分泌物中有机酸的富集浓缩,并与旋转蒸发浓缩的回收率进行了比较.结果表明,DEAE纤维素柱层析有较高的回收率,可用于根系分泌物中有机酸的富集浓缩,特别适用于大批量样品的前处理.确定了反向高效液相色谱同时检测草酸、酒石酸、马来酸、苹果酸、乙酸、柠檬酸、琥珀酸等7种有机酸的条件.玉米根系分泌物样品中可检出苹果酸等5种有机酸.     

产朊假丝酵母利用不同有机酸作碳源的研究

分别通过不同有机酸为惟一碳源,研究了产朊假丝酵母（Candida utilis）能够利用的有机酸的种类,旨在为微生物的基础研究和应用基础研究提供部分依据。分别通过对培养时间和菌体在不同有机酸合成培养基中OD值进行统计分析,发现培养时间和柠檬酸、乳酸、琥珀酸、L-苹果酸培养基的OD值极显著相关（P〈0.01）;培养时间和乙酸、酒石酸、富马酸和草酸培养基的OD值无相关性。通过对比菌体培养前后有机酸合成培养基pH值的变化发现乙酸、酒石酸、富马酸和草酸合成培养基的pH值没有明显变化,而苹果酸、乳酸、琥珀酸和柠檬酸为碳源的合成培养基的pH值均明显增大。从而说明产朊假丝酵母能利用L-苹果酸、乳酸、琥珀酸、柠檬酸为碳源,而不能利用乙酸、酒石酸、草酸、富马酸为碳源。     

小麦-蚕豆间作对根系分泌低分子量有机酸的影响

通过盆栽试验收集了不同生育期单作和间作小麦、蚕豆的根系分泌物,用HPLC分析了根系分泌物中低分子量有机酸的含量和种类.结果表明: 小麦-蚕豆间作显著提高了有机酸的分泌量,在小麦分蘖期(57 d)、孕穗期(120 d)和灌浆期(142 d),间作使小麦根系有机酸分泌量分别提高155%、35.6%和92.6%;在蚕豆分枝期(57 d)和籽粒膨大期(142 d),间作使蚕豆根系有机酸分泌量提高87.4%和38.7%.小麦-蚕豆间作改变了根系分泌物中有机酸的种类,与单作小麦相比,在分蘖期,间作小麦根系分泌物中增加了乳酸;在拔节期(98 d),间作小麦根系分泌物中增加了柠檬酸,但未检测到乙酸;在蚕豆分枝期,间作蚕豆根系分泌物中增加了乙酸,但未检测到乳酸;在蚕豆籽粒膨大期,间作蚕豆根系分泌物中增加了乳酸.小麦-蚕豆间作提高了小麦根系有机酸的分泌速率,在小麦孕穗期,间作小麦分泌柠檬酸、富马酸的速率是单作小麦的179和184倍;在小麦灌浆期,间作小麦分泌乳酸的速率是单作的2.53倍.总之,小麦-蚕豆间作增加了有机酸的分泌量,改变了根系分泌物中有机酸的种类,提高了小麦根系有机酸的分泌速率.     

秋茄(Kandelia candel (L))根系分泌低分子量有机酸及其对重金属生物有效性的影响

应用电感耦合等离子体质谱(ICP-Ms)与高效液相色谱(HPLC)分别对福建漳江口红树林湿地不同土壤结构(砂质与泥质)根际与非根际沉积物中重金属(Cu,Pb,Cd,Zn)含量,以及生长于砂质与泥质滩涂上的红树植物秋茄(Kandelia candel(L)Druce))幼苗根系分泌物中的低分子量有机酸进行测定。在室内模拟秋茄根系分泌的低分子量有机酸,作为重金属提取剂提取沉积物中可溶解态与碳酸盐结合态重金属,并与欧盟标准物质局(BCR)连续提取法相比较,探讨红树根系分泌的低分子量有机酸对红树林沉积物重金属生物有效性的影响。研究结果表明:漳江口红树林泥质沉积物中重金属含量大于砂质沉积物,根际沉积物大于非根际沉积物。两样地沉积物中重金属的赋存形态主要以铁锰氧化物结合态为主,根际沉积物中可交换态与碳酸盐结合态重金属含量均大于非根际沉积物。秋茄根系分泌低分子量有机酸为甲酸,丁酸,苹果酸,柠檬酸,乳酸。不同土壤结构对秋茄根系分泌的苹果酸,柠檬酸,乳酸有显著影响(P<0.05)。以低分子量有机酸作为提取剂对沉积物中可溶解态与碳酸盐结合态重金属的提取率表现为:柠檬酸>混合酸>苹果酸>乳酸>乙酸,低分子量有机酸对红树林沉积物重金属的生物有效性有促进作用。     

棉花根际解磷菌的解磷能力和分泌有机酸的初步测定

利用特殊培养基对盐碱地棉花根际解磷菌进行了分离以及pH值和分泌有机酸能力的初步测定。利用溶磷圈法筛选出10个解磷能力较高的菌株进行深入研究,其中液体培养条件下测定了菌株的溶磷能力,有效磷在4.04~185.63 mg/L,其中wpL2溶磷量达到185.63 mg/L;测定了培养液pH值,下降到5.12~6.67,但是pH与溶磷量之间没有线性关系,测定了培养液的有机酸含量,菌株溶磷量与有机酸总量没有线性相关性,其中所分离到的解无机磷菌株均可以分泌酒石酸,除此之外,wpc1还分泌乙酸,wpc2和wpL2还分泌柠檬酸;分离到的解有机磷菌株均可分泌乙酸,除此之外,ypL1和ypc2分泌酒石酸,ypL3分泌柠檬酸,ypL2和ypc3分泌柠檬酸和丁二酸,均不能产生苹果酸。     

硅和稻瘟病菌接种对水稻植株有机酸含量的影响

以抗病性不同的一对水稻近等基因系——感病CO39和抗病C101LAC(Pi-1)为实验材料,在水培条件下研究了施硅和稻瘟病接菌对水稻根系和叶片抗性物质有机酸含量的影响,揭示硅提高水稻对稻瘟病抗性的机理。结果表明:接菌条件下硅处理显著降低了2个基因型材料的叶片反丁烯二酸、柠檬酸的含量,而增加了叶片中草酸、顺丁烯二酸的含量;加硅显著降低接菌后第3天的CO39叶片酒石酸含量,但增加了C101LAC(Pi-1)接菌后第7天的叶片酒石酸含量;硅处理还显著增加了根系中苹果酸和草酸的含量;各种有机酸在水稻植株体内的分布也不尽相同,如柠檬酸主要分布在叶片中,苹果酸主要分布在根系中,顺丁烯二酸、反丁烯二酸、酒石酸和草酸在叶片和根系中都有分布。研究表明,硅可能通过影响植株体内的有机酸代谢而增强稻瘟病的抗性。     

缺磷白羽扇豆排根与非排根区根尖分泌有机酸的比较

采用根系分泌有机酸原位收集方法及高效液相色谱技术分析了供磷及缺磷后不同时间白羽扇豆(LupinusalbusL .)非排根区根尖和排根分泌有机酸的种类和数量 ,以及相应的根尖、排根组织 ,茎木质部、韧皮部汁液中有机酸含量的变化。结果表明 :(1)缺磷能够诱导白羽扇豆根系产生大量排根 ,根系的有机酸分泌量也明显增加。 (2 )无论在供磷或缺磷条件下 ,排根与非排根区根尖组织中的有机酸种类相同 ,但排根主要分泌柠檬酸和苹果酸 ,而非排根区根尖主要分泌苹果酸和乙酸。 (3)缺磷后非排根区根尖分泌苹果酸的量增加 ,至第 17天达到高峰 ;排根开始分泌柠檬酸的时间相对较晚。缺磷后排根分泌柠檬酸的量随缺磷时间的延长不断增加。 (4 )在缺磷的排根与非排根区根尖组织和茎木质部伤流液中含有大量柠檬酸和苹果酸 ,但在茎韧皮部汁液中则几乎检测不到这两种有机酸。上述结果表明 ,尽管排根和非排根区根尖组织中的有机酸种类相同 ,但它们向外分泌的有机酸种类不同。缺磷后排根及非排根区根尖增加向外分泌的有机酸主要在根中合成     

外源有机酸对镉胁迫下秋华柳镉积累特征的影响

为探究外源有机酸在加强秋华柳(Salix variegata Franch.)镉积累效率中的应用潜力,采用营养液培养方法,研究了5种100μmol/L外源有机酸(草酸、柠檬酸、酒石酸、苹果酸和琥珀酸)对50μmol/L Cd胁迫下秋华柳生长适应性及Cd积累特征的影响,并通过化学平衡程序VISUAL MINTEQ v3.0计算溶液中不同化学形态Cd(游离态和螯合态Cd)的含量。结果表明:柠檬酸、酒石酸、苹果酸和琥珀酸的添加有效缓解了Cd对秋华柳的毒害,明显促进了秋华柳的生长。除草酸处理组外,其余4个有机酸处理组培养液中游离Cd~(2+)含量均得到较大提升,显著促进了秋华柳对Cd的吸收。柠檬酸、酒石酸、琥珀酸和苹果酸的添加显著提高了秋华柳植株的Cd积累量,分别是未加入有机酸Cd处理组的210%、190%、190%和178%。外源有机酸的施加提高了介质中游离Cd~(2+)的含量,并通过与重金属的络合等作用提升了植株对Cd的吸收和积累能力。但不同有机酸对秋华柳Cd积累特征的影响差异明显,柠檬酸加入后通过提高秋华柳根生物量和根中Cd含量,显著增加了根系Cd积累量;酒石酸、苹果酸和琥珀酸的应用则明显提高了秋华柳地上部分Cd积累量。因此,外源酒石酸、琥珀酸和苹果酸的应用可提高秋华柳地上部分的Cd积累效率,更有利于对Cd污染土壤的修复。     

接种AMF对菌根植物和非菌根植物竞争的影响

为了研究丛枝菌根真菌(arbuscular mycorrhizal fungus, AMF)对菌根植物与非菌根植物种间竞争的影响,以玉米(菌根植物)和油菜(非菌根植物)作为供试植物,分别进行间作、尼龙网分隔和单作,模拟这两种植物之间不同的竞争状态,接种丛枝菌根真菌Glomus intraradices和Glomus mosseae,比较菌根植物和非菌根植物的生长和磷营养状况,分析AMF侵染对植物种间竞争作用的影响。结果显示,与单作相比,间作模式下玉米的生物量及磷营养状况有所降低,但其菌根依赖性却有所提高。与不接种相比,接种处理显著降低了间作体系油菜根系的磷含量和磷吸收量,但趋于改善菌根植物玉米的磷营养状况。因此,接种AMF可以降低非菌根植物的磷营养状况及生物量,使得菌根植物的相对竞争能力明显提高,说明AMF在维持物种多样性方面有着重要的作用。     

Development of the arbuscular mycorrhizal symbiosis

The arbuscular mycorrhizal (AM) symbiosis formed between plant roots and fungi is one of the most widespread symbiotic associations found in plants, yet our understanding of events underlying its development are limited. The recent integration of biochemical, molecular and genetic approaches into analyses of the symbiosis is providing new insights into various aspects of its development. In the past year there have been advances in our understanding of the signals required for the formation of appressoria, the molecular changes in the root in response to colonisation, and components of the signal transduction pathways common to both the AM and Rhizobium symbioses.     

The mycorrhizal dependence of subordinates determines the effect of arbuscular mycorrhizal fungi on plant diversity

It is widely recognized that arbuscular mycorrhizal fungi (AMF) influence plant community structure. Several studies in the past few years have focused on the effect of AMF on plant diversity, showing contradictory results. On the basis of studies carried out in grasslands, it has been proposed that these results may not be in conflict but rather depend on mycorrhizal dependence of dominant and subordinate species in the plant community. Here, we propose a simple conceptual model to explain the effect of AMF on plant diversity and dominance hierarchy. Our model suggests that the effect of AMF on plant diversity can be explained mainly by mycorrhizal dependence of subordinates. It also gives predictions for a wider range of community types, and helps in identifying the systems in which empirical data are most critically needed.     

Molecular genetics of the arbuscular mycorrhizal symbiosis

During arbuscular mycorrhiza (AM) development, fungal hyphae grow throughout root epidermal, exodermal and cortical cell layers to reach the inner cortex where the symbiosis' functional units, the arbuscles, develop. Three essential components of a plant signalling network, a receptor-like kinase, a predicted ion-channel and a calmodulin-dependent protein kinase have been identified. A detailed morphological study of symbiotic plant mutants revealed that different subsets of plant genes support the progress of fungal infection in successive root cell layers. Moreover, evidence of a diffusible fungal signalling factor that triggers gene activation in the root has recently been obtained.     

Dual mycorrhizal colonization of forest-dominating tropical trees and the mycorrhizal status of non-dominant tree and liana species

The contribution of mycorrhizal associations to maintaining tree diversity patterns in tropical rain forests is poorly known. Many tropical monodominant trees form ectomycorrhizal (EM) associations, and there is evidence that the EM mutualism contributes to the maintenance of monodominance. It is assumed that most other tropical tree species form arbuscular mycorrhizal (AM) associations, and while many mycorrhizal surveys have been done, the mycorrhizal status of numerous tropical tree taxa remains undocumented. In this study, we tested the assumption that most tropical trees form AM associations by sampling root vouchers from tree and liana species in monodominant Dicymbe corymbosa forest and an adjacent mixed rain forest in Guyana. Roots were assessed for the presence/absence of AM and EM structures. Of the 142 species of trees and lianas surveyed, three tree species (the monodominant D. corymbosa, the grove-forming D. altsonii, and the non-dominant Aldina insignis) were EM, 137 were exclusively AM, and two were non-mycorrhizal. Both EM and AM structures were observed in D. corymbosa and D. altsonii. These results provide empirical data supporting the assumption that most tropical trees form AM associations for this region in the Guiana Shield and provide the first report of dual EM/AM colonization in Dicymbe species. Dual colonization of the Dicymbe species should be further explored to determine if this ability contributes to the establishment and maintenance of site dominance. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Albert Howard and the mycorrhizal association

Albert Howard worked as an imperial agronomist for the British Government in India. Following his retirement in 1931, he returned to England and embarked on a passionate global campaign to reform agricultural practices. Central to Howard's project was the mycorrhizal association, a symbiotic relationship between plant roots and subterranean fungi, believed to play an important part in plant nutrition. I show that there are a number of close parallels between Howard's work in India and his portrayal of the mycorrhizal association, and argue that Howard used these fungi to naturalise his imperial project. Understood in this way, these mycorrhizal and imperial associations reveal ways that Howard was able to negotiate the boundaries between the local and global, England and India, science and agriculture, institute and village, and soil and plant. In contrast to Thomas Gieryn's work on hybridisation at the cultural boundaries between science and non-science, I concentrate on Howard's use of intermediaries to negotiate and articulate specific boundaries within his imperial project. Arguing that this approach reveals limitations in Gieryn's hybrid framework, I suggest that a focus on Howard's dependence on intermediaries draws attention to the discontinuities between entities, besides the dynamic ways that they might be coupled.     

Polyphosphate dynamics in mycorrhizal roots during colonization of an arbuscular mycorrhizal fungus

Inorganic polyphosphate (poly P) has been considered to be a translocatable form of phosphate (Pi) in arbuscular mycorrhizal fungi (AMF). Here we examined time-course changes in poly P content during the AMF colonization process. Onion (Allium cepa) plants were cultured with or without inoculation with Gigaspora margarita for 2-8 wk with periodic sampling. Poly P in the extracts, purified through gel filtration, was quantified by the reverse reaction of polyphosphate kinase. The length of poly P in mycorrhizal roots appeared to be shorter than in extraradical hyphae or in spores of the AMF, indicating that AMF depolymerize poly P before providing Pi to the host. The poly P content increased as colonization proceeded, and was highly correlated with the weight of the colonized roots. These results support the model that AMF supply Pi to the host through the poly P pool, and that the poly P content of a mycorrhizal root can be a good indicator of the Pi-supplying activity of AMF.     

Different levels of hyphal self-incompatibility modulate interconnectedness of mycorrhizal networks in three arbuscular mycorrhizal fungi within the Glomeraceae

Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21 % of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9 %. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2–7.7, while it ranged from 25.8–48 to 35.6–53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2 %). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots.     

Functional complementarity in the arbuscular mycorrhizal symbiosis

The causes and consequences of biodiversity are central themes in ecology. Perhaps one reason for much of the current interest in biodiversity is the belief that the loss of species (by extinction) or their gain (by invasion) will significantly influence ecosystem function. Arbuscular mycorrhizal (AM) fungi are components of most terrestrial ecosystems and, while many research programs have shown that variability among species or isolates of AM fungi does occur (Giovannetti & Gianinazzi-Pearson, 1994), the basis for this variability and its consequences to the function of communities and ecosystems remains largely unexplored. Smith et al . (pp. 357–366 in this issue) now show clearly that ecologically significant functional diversity exists among AM fungal species in the regions of the soil from which they absorb phosphate, and their results suggest that such diversity may have significant ecological consequences.