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。     

Cadmium toxicity in crop plants and its alleviation by arbuscular mycorrhizal (AM) fungi: An overview

Cadmium (Cd), a toxic metal released into agricultural settings induces numerous changes in plant growth and physiology. The main known mechanisms of Cd toxicity include its affinity for sulfhydryl groups in proteins and its ability to replace some essential metals in active sites of enzymes, thus causing inhibition of enzyme activities and protein denaturation. This article reviews detrimental effects of Cd toxicity on the functional biology of plants and summarizes the mechanisms that are activated by plants to prevent the absorption or to detoxify Cd ions such as synthesis of antioxidants, osmolytes, phytochelatins, metallothioneins, etc. Arbuscular mycorrhizal (AM) fungi are reported to be present on the roots of plants growing in metal-contaminated soils and play an important role in metal tolerance. Through mycorrhizal symbiosis, heavy metals are immobilized in the rhizosphere through precipitation in the soil matrix, adsorption onto the root surface or accumulation within roots, and compartmentalized in aboveground parts of the plant. This article unfolds the potential role of AM fungi in enhancing Cd tolerance of plants.     

Proteomic analysis as a tool for investigating arsenic stress in Pteris vittata roots colonized or not by arbuscular mycorrhizal symbiosis

Pteris vittata can tolerate very high soil arsenic concentration and rapidly accumulates the metalloid in its fronds. However, its tolerance to arsenic has not been completely explored. Arbuscular mycorrhizal (AM) fungi colonize the root of most terrestrial plants, including ferns. Mycorrhizae are known to affect plant responses in many ways: improving plant nutrition, promoting plant tolerance or resistance to pathogens, drought, salinity and heavy metal stresses. It has been observed that plants growing on arsenic polluted soils are usually mycorrhizal and that AM fungi enhance arsenic tolerance in a number of plant species. The aim of the present work was to study the effects of the AM fungus Glomus mosseae on P. vittata plants treated with arsenic using a proteomic approach. Image analysis showed that 37 spots were differently affected (21 identified). Arsenic treatment affected the expression of 14 spots (12 up-regulated and 2 down-regulated), while in presence of G. mosseae modulated 3 spots (1 up-regulated and 2 down-regulated). G. mosseae, in absence of arsenic, modulated 17 spots (13 up-regulated and 4 down-regulated). Arsenic stress was observed even in an arsenic tolerant plant as P. vittata and a protective effect of AM symbiosis toward arsenic stress was observed.     

Meeting a non-host: the behaviour of AM fungi

 Arbuscular mycorrhizal (AM) fungi are obligately biotrophic organisms that live symbiotically with the roots of most plants. The establishment of a functional symbiosis between AM fungi and host plants involves a sequence of recognition events leading to the morphological and physiological integration of the two symbionts. The developmental switches in the fungi are triggered by host signals which induce changes in gene expression and a process leading to unequivocal recognition between the two partners of the symbiosis. It has been calculated that about 80% of plant families from all phyla of land plants are hosts of AM fungi. The remaining plant species are either non-mycorrhizal or hosts of mycorrhizas other than the arbuscular type. Non-host plants have been used to obtain information on the factors regulating the development of a functional symbiosis. The aim of this present review is to highlight present-day knowledge of the fungal developmental switches involved in the process of host/non-host discrimination. The following stages of the life cycle of AM fungi are analysed in detail: spore germination, presymbiotic mycelial growth, differential branching pattern and chemotropism, appressorium formation, root colonization. Accepted: 17 June 1998     

Proteomic analysis of Pteris vittata fronds: two arbuscular mycorrhizal fungi differentially modulate protein expression under arsenic contamination

Arbuscular mycorrhizae (AM) are the most widespread mutualistic symbioses between the roots of most land plants and a phylum of soil fungi. AM are known to influence plant performance by improving mineral nutrition, protecting against pathogens and enhancing resistance or tolerance to biotic and abiotic stresses. The aim of this study was to investigate the frond proteome of the arsenic hyperaccumulator fern Pteris vittata in plants that had been inoculated with one of the two AM fungi (Glomus mosseae or Gigaspora margarita) with and without arsenic treatment. A protective role for AM fungi colonisation in the absence of arsenic was indicated by the down-regulation of oxidative damage-related proteins. Arsenic treatment of mycorrhizal ferns induced the differential expression of 130 leaf proteins with specific responses in G. mosseae- and Gi. margarita-colonised plants. Up-regulation of multiple forms of glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and enolase, primarily in G. mosseae-inoculated plants, suggests a central role for glycolytic enzymes in arsenic metabolism. Moreover, a putative arsenic transporter, PgPOR29, has been identified as an up-regulated protein by arsenic treatment.     

Fresh perspectives on the roles of arbuscular mycorrhizal fungi in plant nutrition and growth

Recent research on arbuscular mycorrhizas has demonstrated that AM fungi play a significant role in plant phosphorus (P) uptake, regardless of whether the plant responds positively to colonization in terms of growth or P content. Here we focus particularly on implications of this finding for consideration of the balance between organic carbon (C) use by the fungi and P delivery (i.e. the C-P trade between the symbionts). Positive growth responses to arbuscular mycorrhizal (AM) colonization are attributed frequently to increased P uptake via the fungus, which results in relief of P deficiency and increased growth. Zero AM responses, compared with non-mycorrhizal (NM) plants, have conventionally been attributed to failure of the fungi to deliver P to the plants. Negative responses, combined with excessive C use, have been attributed to this failure. The fungi were viewed as parasites. Demonstration that the AM pathway of P uptake operates in such plants indicates that direct P uptake by the roots is reduced and that the fungi are not parasites but mutualists because they deliver P as well as using C. We suggest that poor plant growth is the result of P deficiency because AM fungi lower the amount of P taken up directly by roots but the AM uptake of P does compensate for the reduction. The implications of interplay between direct root uptake and AM fungal uptake of P also include increased tolerance of AM plants to toxins such as arsenate and increased success when competing with NM plants. Finally we discuss the new information on C-P trade in the context of control of the symbiosis by the fungus or the plant, including new information (from NM plants) on sugar transport and on the role of sucrose in the signaling network involved in responses of plants to P deprivation.     

Effect of arbuscular mycorrhizal (AM) colonization on terpene emission and content of Artemisia annua L

Plant roots interact with a wide variety of rhizospheric microorganisms, including bacteria and the symbiontic arbuscular mycorrhizal (AM) fungi. The mycorrhizal symbiosis represents a series of complex feedbacks between plant and fungus regulated by their physiology and nutrition. Despite the widespread distribution and ecological significance of AM symbiosis, little is known about the potential of AM fungi to affect plant VOC metabolism. The purpose of this study was to investigate whether colonization of plant roots by AM fungi and associated soil microorganisms affects VOC emission and content of Artemisia annua L. plants (Asteraceae). Two inoculum types were evaluated: one consisted of only an arbuscular mycorrhizal (AM) fungus species (Glomus spp.), and the other was a mixture of different Glomus species and associated soil bacteria. Inoculated plants were compared with non-inoculated plants and with plants supplemented with extra phosphorus (P) to obtain plants of the same size as mycorrhizal plants, thus excluding potentially-confounding mycorrhizal effects on shoot growth. VOC emissions of Artemisia annua plants were analyzed by leaf cuvette sampling followed by off-line measurements with pre-concentration and gas chromatography mass spectrometry (GC-MS). Measurements of CO(2) and H(2)O exchanges were conducted simultaneously. Several volatile monoterpenes were identified and characterized from leaf emissions of Artemisia annua L. by GC-MS analysis. The main components identified belong to different monoterpene structures: alpha-pinene, beta-pinene, camphor, 1,8-cineole, limonene, and artemisia ketone. A good correlation between monoterpene leaf concentration and leaf emission was found. Leaf extracts included also several sesquiterpenes. Total terpene content and emission was not affected by AM inoculation with or without bacteria, while emission of limonene and artemisia ketone was stimulated by this treatment. No differences were found among treatments for single monoterpene content, while accumulation of specific sesquiterpenes in leaves was altered in mycorrhizal plants compared to control plants. Growth conditions seemed to have mainly contributed to the outcome of the symbiosis and influenced the magnitude of the plant response. These results highlight the importance of considering the below-ground interaction between plant and soil for estimating VOC emission rates and their ecological role at multitrophic levels.     

植物中丛枝菌根形成的信号途径研究进展

丛枝菌根(arbuscular mycorrhizal,AM)共生是丛枝菌根真菌与大多数陆地植物的根系之间形成的一种互利共生关系。植物给菌根真菌提供碳水化合物;作为回报,菌根真菌能够增强植物对矿质营养元素(尤其是磷)的吸收。菌根的形成过程是一系列信号交换和转导的结果,具有严格并且一致的顺序。本文以植物中菌根形成的信号途径为主线,对菌根真菌的形成过程和信号转导途径及其方式进行了分析和讨论。高等植物中菌根形成的信号途径与豆科植物的结瘤信号途径部分共享,并且与钙离子信号途径相关,但前者更为广泛。尽管该途径中很多过程目前还不十分清楚,但是相信在不久的将来就可以揭开菌根形成过程中的众多谜团。     

植物中丛枝菌根形成的信号途径研究进展

丛枝菌根(arbuscular mycorrhizal, AM)共生是丛枝菌根真菌与大多数陆地植物的根系之间形成的一种互利共生关系。植物给菌根真菌提供碳水化合物; 作为回报, 菌根真菌能够增强植物对矿质营养元素(尤其是磷)的吸收。菌根的形成过程是一系列信号交换和转导的结果, 具有严格并且一致的顺序。本文以植物中菌根形成的信号途径为主线, 对菌根真菌的形成过程和信号转导途径及其方式进行了分析和讨论。高等植物中菌根形成的信号途径与豆科植物的结瘤信号途径部分共享, 并且与钙离子信号途径相关, 但前者更为广泛。尽管该途径中很多过程目前还不十分清楚, 但是相信在不久的将来就可以揭开菌根形成过程中的众多谜团。     

Aquaporin genes GintAQPF1 and GintAQPF2 from Glomus intraradices contribute to plant drought tolerance

Arbuscular mycorrhizal (AM) symbiosis, established between AM fungi (AMF) and roots of higher plants, occurs in most terrestrial ecosystems. It has been well demonstrated that AM symbiosis can improve plant performance under various environmental stresses, including drought stress. However, the molecular basis for the direct involvement of AMF in plant drought tolerance has not yet been established. Most recently, we cloned two functional aquaporin genes, GintAQPF1 and GintAQPF2, from AM fungus Glomus intraradices. By heterologous gene expression in yeast, aquaporin localization, activities and water permeability were examined. Gene expressions during symbiosis in expose to drought stress were also analyzed. Our data strongly supported potential water transport via AMF to host plants. As a complement, here we adopted the monoxenic culture system for AMF, in which carrot roots transformed by Ri-T DNA were cultured with Glomus intraradices in two-compartment Petri dishes, to verify the aquaporin gene functions in assisting AMF survival under polyethylene glycol (PEG) treatment. Our results showed that 25% PEG significantly upregulated the expression of two aquaporin genes, which was in line with the gene functions examined in yeast. We therefore concluded that the aquaporins function similarly in AMF as in yeast subjected to osmotic stress. The study provided further evidence to the direct involvement of AMF in improving plant water relations under drought stresses.     

Phosphate transporters of Medicago truncatula and arbuscular mycorrhizal fungi

Most vascular plants acquire phosphate from their environment either directly, via the roots, or indirectly, via a symbiotic interaction with arbuscular mycorrhizal (AM) fungi. The symbiosis develops in the plant roots where the fungi colonize the cortex of the root to obtain carbon from the plant host, while assisting the plant with acquisition of phosphate and other mineral nutrients from the soil solution. As a first step toward understanding the molecular basis of the symbiosis and phosphate utilization, we have cloned and characterized phosphate transporter genes from the AM fungi Glomus versiforme and Glomus intraradices, and from the roots of a host plant, Medicago truncatula. Expression analyses and localization studies indicate that each of these transporters has a role in phosphate uptake from the soil solution.     

Arbuscular mycorrhizal influence on leaf water potential, solute accumulation, and oxidative stress in soybean plants subjected to drought stress

This study investigated several aspects related to drought tolerance in arbuscular mycorrhizal (AM) soybean plants. The investigation included both shoot and root tissues in order to reveal the preferred target tissue for AM effects against drought stress. Non-AM and AM soybean plants were grown under well-watered or drought-stressed conditions, and leaf water status, solute accumulation, oxidative damage to lipids, and other parameters were determined. Results showed that AM plants were protected against drought, as shown by their significantly higher shoot-biomass production. The leaf water potential was also higher in stressed AM plants (-1.9 MPa) than in non-AM plants (-2.5 MPa). The AM roots had accumulated more proline than non-AM roots, while the opposite was observed in shoots. Lipid peroxides were 55% lower in shoots of droughted AM plants than in droughted non-AM plants. Since there was no correlation between the lower oxidative damage to lipids in AM plants and the activity of antioxidant enzymes, it seems that first the AM symbiosis enhanced osmotic adjustment in roots, which could contribute to maintaining a water potential gradient favourable to the water entrance from soil into the roots. This enabled higher leaf water potential in AM plants during drought and kept the plants protected against oxidative stress, and these cumulative effects increased the plant tolerance to drought.     

Sucrose synthase levels do not limit or regulate carbon transfer in the arbuscular mycorrhizal symbiosis

Abstract

Sucrose synthase (SuSy) is the main sucrose breakdown enzyme in plant sink tissues, including nodules, and is a possible candidate for the diversion of plant carbon to arbuscular mycorrhizal (AM) fungi in roots. We tested the involvement of SuSy in AM symbiosis of Glomus intraradices and Pisum sativum (pea). We observed that peas deficient in the predominant root isoform of SuSy were colonized successfully by AM fungi similar to wild-type roots. SuSy protein levels did not increase in roots as AM symbiosis developed, although SuSy protein levels did increase in nodules as the rhizobium symbiosis developed. Our results lead us to conclude that, unlike nodule symbiosis, SuSy protein does not limit or regulate carbon transfer in the AM symbiosis.     

Cloning of cDNAs encoding SODs from lettuce plants which show differential regulation by arbuscular mycorrhizal symbiosis and by drought stress

In the present study three cDNA fragments were cloned using degenerate primers for Mn-sod genes and PCR: two showed a high degree of identity with Mn-sods from plants and the third with Fe-sod. Arbuscular mycorrhizal (AM) symbiosis down-regulated their expression pattern under well-watered conditions. In contrast, AM symbiosis in combination with drought stress considerably increased the expression of the Mn-sod II gene and this correlated well with plant tolerance to drought. These results would suggest that mycorrhizal protection against oxidative stress caused by drought may be an important mechanism by which AM fungi protect the host plant against drought.     

MycoRed: Betalain pigments enable in vivo real-time visualisation of arbuscular mycorrhizal colonisation

Arbuscular mycorrhiza (AM) are mutualistic interactions formed between soil fungi and plant roots. AM symbiosis is a fundamental and widespread trait in plants with the potential to sustainably enhance future crop yields. However, improving AM fungal association in crop species requires a fundamental understanding of host colonisation dynamics across varying agronomic and ecological contexts. To this end, we demonstrate the use of betalain pigments as in vivo visual markers for the occurrence and distribution of AM fungal colonisation by Rhizophagus irregularis in Medicago truncatula and Nicotiana benthamiana roots. Using established and novel AM-responsive promoters, we assembled multigene reporter constructs that enable the AM-controlled expression of the core betalain synthesis genes. We show that betalain colouration is specifically induced in root tissues and cells where fungal colonisation has occurred. In a rhizotron setup, we also demonstrate that betalain staining allows for the noninvasive tracing of fungal colonisation along the root system over time. We present MycoRed, a useful innovative method that will expand and complement currently used fungal visualisation techniques to advance knowledge in the field of AM symbiosis.

Arbuscular mycorrhiza are mutualistic interactions formed between soil fungi and plant roots. This study presents the MycoRed system, which uses red plant pigments derived from beetroot to reveal how fungi establish symbiosis with living legume and wild tobacco roots.     

Regulatory mechanisms of host plant defense responses to arbuscular mycorrhiza

Arbuscular mycorrhizal (AM) fungi colonize the roots of over 80% of terrestrial plant species, forming mutually beneficial symbioses. During the colonization process, symbiotic partners recognize each other, and undergo observable morphological and physiological changes; indicating that symbiosis formation involves multiple factors that are finely regulated. Sometimes host plants generate a transient, weak, defense response. This response and its down-regulation play a very important role in the development of AM symbioses. Although AM fungi can infect a wide range of host root tissues, which host defense may play a crucial role is hypothesized from the fact that hyphal expansion is only observed in the root cortex.
We discuss five defense mechanisms. (1) The degradation of exogenous elicitors. The host’s weak defense response may be due to the degradation of the exogenous elicitor chitin, or the prevention of release of an endogenous inductor from the plant cell wall. (2) The inactivation of defense signal molecules. Some defense signal molecules such as hydrogen peroxidase, salicylic acid (SA), and jasmonic acid (JA), are inactivated in host plants. This helps to avoid the turn-on of defense-related genes and facilitate mycorrhizal formation. (3) The regulation of plant hormones and plant photosynthates. Plant hormone levels and plant photosynthate metabolism both change during AM colonization. These mechanisms need further exploration. (4) Changes in levels of phosphorous (P), and (iso)flavonoids. High P levels can induce some defense genes to express hydrogen peroxidase, chitinase, and glucanase. These gene products can repress colonization by AM fungi. The plant defense response regulatory effect for different (iso)flavonoids varies, and their levels are regulated by P. (5) The suppressed expression of symbiotic genes. Some symbiosis-related genes inhibit plant defense responses, but it is still unclear which mechanisms underlie gene regulation. We provide here a theoretical basis for research into AM symbiosis that may promote study of host plant resistance and the mechanisms of symbiosis formation.
We provide a deeper insight into the signal transduction pathways of mycorrhization that will aid understanding and analysis of plant defense mechanisms in the AM context. The on-going development of genome sequencing technology will contribute greatly to the detailed study of symbiosis-related genes, and pathogenesis-related protein genes. These related genes may be induced to express corresponding proteins, be repressed, postpone expression or even shutdown, or both may work together to form symbioses. Elucidation of these features will help us understand the roles that plant defenses play in mycorrhizal formation; providing an unprecedented opportunity for research into mycorrhizal molecular biology and the interaction of symbiotic partners, and allowing the underlying mechanisms to be gradually uncovered.     

PIP Aquaporin Gene Expression in Arbuscular Mycorrhizal Glycine max and Lactuca sativa Plants in Relation to Drought Stress Tolerance

Although the discovery of aquaporins in plants has resulted in a paradigm shift in the understanding of plant water relations, the relationship between aquaporins and plant responses to drought still remains elusive. Moreover, the contribution of aquaporin genes to the enhanced tolerance to drought in arbuscular mycorrhisal (AM) plants has never been investigated. Therefore, we studied, at a molecular level, whether the expression of aquaporin-encoding genes in roots is altered by the AM symbiosis as a mechanism to enhance host plant tolerance to water deficit. In this study, genes encoding plasma membrane aquaporins (PIPs) from soybean and lettuce were cloned and their expression pattern studied in AM and nonAM plants cultivated under well-watered or drought stressed conditions. Results showed that AM plants responded to drought stress by down-regulating the expression of the PIP genes studied and anticipating its down-regulation as compared to nonAM plants. The possible physiological implications of this down-regulation of PIP genes as a mechanism to decrease membrane water permeability and to allow cellular water conservation is further discussed.     

Exogenous ABA accentuates the differences in root hydraulic properties between mycorrhizal and non mycorrhizal maize plants through regulation of PIP aquaporins

The arbuscular mycorrhizal (AM) symbiosis has been shown to modulate the same physiological processes as the phytohormone abscisic acid (ABA) and to improve plant tolerance to water deficit. The aim of the present research was to evaluate the combined influence of AM symbiosis and exogenous ABA application on plant root hydraulic properties and on plasma-membrane intrinsic proteins (PIP) aquaporin gene expression and protein accumulation after both a drought and a recovery period. Results obtained showed that the application of exogenous ABA enhanced osmotic root hydraulic conductivity (L) in all plants, regardless of water conditions, and that AM plants showed lower L values than nonAM plants, a difference that was especially accentuated when plants were supplied with exogenous ABA. This effect was clearly correlated with the accumulation pattern of the different PIPs analyzed, since most showed reduced expression and protein levels in AM plants fed with ABA as compared to their nonAM counterparts. The possible involvement of plant PIP aquaporins in the differential regulation of L by ABA in AM and nonAM plants is further discussed.     

纳米氧化锌和接种丛枝菌根真菌对大豆生长及营养吸收的影响

纳米氧化锌是应用最广的人工纳米颗粒(nanoparticles,NPs)之一,具有一定生物毒性。丛枝菌根(arbuscular mycorrhizal,AM)真菌能与陆地上80%以上的高等植物形成丛枝菌根共生体,并能改善宿主植物矿质营养,提高其抗逆性。然而纳米ZnO与丛枝菌根的关系尚不清楚。通过温室沙培盆栽试验,研究了施加不同水平纳米ZnO(0、500、1000、2000、3000 mg/kg)和接种AM真菌Acaulospora mellea对大豆生长及营养状况的影响。结果表明,3000 mg/kg的纳米ZnO显著抑制大豆植株生长,表现出植物毒性,在其他水平时没有显著影响。纳米ZnO在施加水平500、1000 mg/kg时没有抑制AM真菌对大豆根系的侵染,但是高施加水平(2000 mg/kg)时对AM真菌产生毒害,几乎完全抑制大豆根系菌根侵染。接种AM真菌仅在500 mg/kg纳米ZnO时显著促进大豆生长,增加大豆植株对P、K、N的吸收,降低根系Zn含量。纳米ZnO可能会持续释放锌离子,并抑制大豆根系对矿质营养元素的吸收,从而产生生物毒性,而AM真菌与大豆根系的共生可起到有益作用。