Arbuscular mycorrhizal protein mRNA over-expression in bread wheat seedlings by Trichoderma harzianum Raifi (KRL-AG2) elicitation

Association between arbuscular mycorrhizal fungi (AMF) and majority of terrestrial plant species provides many benefits to plants that range from stress alleviation and bioremediation in soils polluted with heavy metals to plant growth promotion and yield quantity. Some non-arbuscular mycorrhizal fungi such as, Trichoderma harzianum, are known to enhance the AMF symbiosis with vascular plants. However, information about their role in AMF symbiosis is still limited. Shoots of (Avocet S) wheat seedlings were sprayed with the fungal culture filtrate and gene expression patterns were analyzed in the treated tissues. An increase in the level of mRNA of arbuscular mycorrhizal protein comparing with control was found. The over-expression of this protein in wheat tissues might contribute in initiation of AMF colonization in wheat tissues. The result of this study can spark future researches to elucidate possible role of this protein in the symbiotic interaction mechanisms between soil AMF and various plant roots.     

丛枝菌根真菌数个种的基因组和转录组研究概况

丛枝菌根真菌（AMF）在自然界分布广泛，能与大部分维管植物的根系形成菌根共生体。它们在调节植物群落结构和全球的碳、氮、磷循环等方面发挥着重要的生态功能，也是农林、环境领域最具应用前景的微生物类群。受限于培养方法、研究手段等，长期以来对AMF基因组、转录组特征的认识非常有限。最近10年，AMF基因组和转录组的相关研究在高通量测序技术的推动下取得了较快发展；研究结果也显著提高了对AMF遗传发育、代谢生理、共生机制等的认识。本文综述了目前已完成测序的AMF种类的基因组、转录组信息。结果发现，已测序的AMF种类普遍具有基因组大、转座子丰富、AT碱基含量高、含大量未知功能基因与特异性基因、缺少部分共生相关基因等特点。在转录层面，总结了不同AMF种类、AMF不同共生结构、共生阶段以及与不同寄主植物共生时的转录本特征。结果发现，不同种类AMF的转录本大小差异明显。不同共生阶段或不同共生结构中的AMF转录本也具有较大的差异，且差异表达的基因大部分与养分交易、信号转导等密切相关。相比之下，同种AMF与不同寄主植物共生时的转录本表现出较高的保守性。最后，本文提出了本领域需要重点关注的研究方向，包括AMF纯培养技术的革新、AMF基因功能的解析、非模式AMF类群的研究以及对AMF蛋白组的研究。     

Water deficit improved the capacity of arbuscular mycorrhizal fungi (AMF) for inducing the accumulation of antioxidant compounds in lettuce leaves

Lettuce, a major food crop within the European Union and the most used for the so-called 'Fourth Range' of vegetables, can associate with arbuscular mycorrhizal fungi (AMF). Mycorrhizal symbiosis can stimulate the synthesis of secondary metabolites, which may increase plant tolerance to stresses and enhance the accumulation of antioxidant compounds potentially beneficial to human health. Our objectives were to assess (1) if the application of a commercial formulation of AMF benefited growth of lettuce under different types and degrees of water deficits; (2) if water restrictions affected the nutritional quality of lettuce; and (3) if AMF improved the quality of lettuce when plants grew under reduced irrigation. Two cultivars of lettuce consumed as salads, Batavia Rubia Munguía and Maravilla de Verano, were used in the study. Four different water regimes were applied to both non-mycorrhizal and mycorrhizal plants: optimal irrigation (field capacity [FC]), a water regime equivalent to 2/3 of FC, a water regime equivalent to 1/2 of FC and a cyclic drought (CD). Results showed that mycorrhizal symbiosis improved the accumulation of antioxidant compounds, mainly carotenoids and anthocyanins, and to a lesser extent chlorophylls and phenolics, in leaves of lettuce. These enhancements were higher under water deficit than under optimal irrigation. Moreover, shoot biomass in mycorrhizal lettuces subjected to 2/3 of FC were similar to those of non-mycorrhizal plants cultivated under well-watered conditions. In addition, lettuces subjected to 2/3 FC had similar leaf RWC than their respective well-watered controls, regardless of mycorrhizal inoculation. Therefore, results suggest that mycorrhizal symbiosis can improve quality of lettuce and may allow restrict irrigation without reducing production.     

Spatial soil heterogeneity has a greater effect on symbiotic arbuscular mycorrhizal fungal communities and plant growth than genetic modification with Bacillus thuringiensis toxin genes

Maize, genetically modified with the insect toxin genes of Bacillus thuringiensis (Bt), is widely cultivated, yet its impacts on soil organisms are poorly understood. Arbuscular mycorrhizal fungi (AMF) form symbiotic associations with plant roots and may be uniquely sensitive to genetic changes within a plant host. In this field study, the effects of nine different lines of Bt maize and their corresponding non‐Bt parental isolines were evaluated on AMF colonization and community diversity in plant roots. Plants were harvested 60 days after sowing, and data were collected on plant growth and per cent AMF colonization of roots. AMF community composition in roots was assessed using 454 pyrosequencing of the 28S rRNA genes, and spatial variation in mycorrhizal communities within replicated experimental field plots was examined. Growth responses, per cent AMF colonization of roots and AMF community diversity in roots did not differ between Bt and non‐Bt maize, but root and shoot biomass and per cent colonization by arbuscules varied by maize cultivar. Plot identity had the most significant effect on plant growth, AMF colonization and AMF community composition in roots, indicating spatial heterogeneity in the field. Mycorrhizal fungal communities in maize roots were autocorrelated within approximately 1 m, but at greater distances, AMF community composition of roots differed between plants. Our findings indicate that spatial variation and heterogeneity in the field has a greater effect on the structure of AMF communities than host plant cultivar or modification by Bt toxin genes.     

Localized versus systemic effect of arbuscular mycorrhizal fungi on defence responses to Phytophthora infection in tomato plants

Development of biological control for plant diseases is accepted as a durable and environmentally friendly alternative for agrochemicals. Arbuscular mycorrhizal fungi (AMF), which form symbiotic associations with root systems of most agricultural, horticultural and hardwood crop species, have been suggested as widespread potential bioprotective agents. In the present study the ability of two AMF (Glomus mosseae and Glomus intraradices) to induce local or systemic resistance to Phytophthora parasitica in tomato roots have been compared using a split root experimental system. Glomus mosseae was effective in reducing disease symptoms produced by P. parasitica infection, and evidence points to a combination of local and systemic mechanisms being responsible for this bioprotector effect. The biochemical analysis of different plant defence-related enzymes showed a local induction of mycorrhiza-related new isoforms of the hydrolytic enzymes chitinase, chitosanase and beta-1,3-glucanase, as well as superoxide dismutase, an enzyme which is involved in cell protection against oxidative stress. Systemic alterations of the activity of some of the constitutive isoforms were also observed in non-mycorrhizal roots of mycorrhizal plants. Studies on the lytic activity against Phytophthora cell wall of root protein extracts also corroborated a systemic effect of mycorrhizal symbiosis on tomato resistance to Phytophthora.     

丛枝菌根对植物根际逆境的生态学意义

近年来,我国在菌根分子生物学、菌根营养学、菌根分类学和菌根生态学等方面取得了令人瞩目的研究成果,其中对丛枝菌根真菌(AMF)的研究居多。AMF能与大部分陆地植物根系形成共生关系,促进植物生长发育,提高植物抗逆性,在保持生态平衡、保护生态环境等方面发挥重要作用。本文主要从非生物胁迫(干旱胁迫、重金属污染、盐碱胁迫)和生物胁迫(致病菌和线虫侵染)方面介绍了AMF在植物根际逆境中发挥的生态功能及作用机制,提出了该研究领域尚存的不足之处和研究前景,为AMF后续研究提供参考。     

干旱胁迫条件下AMF促进小马鞍羊蹄甲幼苗生长的机理研究

采用温室水分控制试验,在干旱胁迫条件下,定量化研究优势丛枝菌根真菌(AMF)影响优势乡土植物小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗生长的机理,主要通过研究干旱胁迫条件下摩西球囊霉菌(Funneliformis mosseae)与小马鞍羊蹄甲的共生关系,阐明AMF在植物生长初期的作用。结果表明,干旱胁迫条件下,摩西球囊霉菌能够很好地侵染幼苗,侵染率高达89%—97%,并且不受水分条件影响。接种的幼苗最大光合速率、水分利用效率随着干旱胁迫程度从重度到轻度(水分从低到高)逐渐增大,相反地,叶片脯氨酸含量逐渐减小。接种显著地促进幼苗株高、叶片数、叶面积、根长、根面积等生长指标,提高幼苗各部分生物量、地上地下磷(P)含量。当含水量为60%田间持水量时,AMF促进小马鞍羊蹄甲幼苗吸收P的效果最好。接种还显著影响幼苗的生物量分配,在重度干旱胁迫时影响P分配,水分条件也显著影响幼苗的生物量分配。此外,接种和水分的交互作用对叶生物量、总生物量、生长指标以及地上部氮(N)总量影响显著。结果表明干旱胁迫条件下菌根效应显著,并在干旱条件下显著促进了小马鞍羊蹄甲幼苗的生长,这为进一步干旱河谷植被恢复提供了理论依据。     

Responsiveness of Durum Wheat to Mycorrhizal Inoculation Under Different Environmental Scenarios

A greater understanding of how climate change will affect crop photosynthetic performance has been described as a target goal to improve yield potential. Other concomitant stressors can reduce the positive effect of elevated atmospheric CO₂ on wheat yield. Arbuscular mycorrhizal fungi (AMF) are symbiotic fungi predicted to be important in defining plant responses to rising atmospheric CO₂, but their role in response to global climatic change is still poorly understood. This study aimed to assess if increased atmospheric CO₂ interacting with drought can modify the effects of mycorrhizal symbiosis on flag leaf physiology in winter wheat. The study was performed in climate-controlled greenhouses with ambient (400 ppm, ACO₂) or elevated (700 ppm, ECO₂) CO₂ concentrations in the air. Within each greenhouse half of the plants were inoculated with Rhizophagus intraradices. When ear emergence began, half of the plants from each mycorrhizal and CO₂ treatment were subjected to terminal drought. At ACO₂ AMF improved the photochemistry efficiency of PSII compared with non-mycorrhizal plants, irrespective of irrigation regime. Mycorrhizal wheat accumulated more fructan than non-mycorrhizal plants under optimal irrigation. The level of proline in the flag leaf increased only in mycorrhizal wheat after applying drought. Mycorrhizal association avoided photosynthetic acclimation under ECO₂. However, nitrogen availability to flag leaves in mycorrhizal plants was lower under ECO₂ than at ACO₂. Results suggest that the mechanisms underlying the interactions between mycorrhizal association and atmospheric CO₂ concentration can be crucial for the benefits that this symbiosis can provide to wheat plants undergoing water deficit.     

丛枝菌根真菌与氮添加对不同根形态基因型水稻氮吸收的影响

植物主要依赖自身根系从土壤中获取矿质养分; 具有不同根形态的植物对于养分的吸收能力存在差异。丛枝菌根真菌(AMF)能与陆地植物根系形成共生关系, 帮助植物吸收矿质养分。但是, AMF对于植物根系养分吸收的促进效应是否会受根形态的影响还鲜有研究。该研究选取4种不同根形态基因型水稻(根毛缺陷突变体rhl1、侧根缺陷突变体iaa11、不定根缺失突变体arl1和野生型Kas)为研究对象, 设置2种施氮水平处理(低氮: 20 mg·kg^-1氨氮; 高氮: 100 mg·kg^-1氨氮), 利用稳定同位素¹⁵N示踪标记技术, 探究AMF和氮添加对不同根形态植物氮吸收的影响。研究结果发现, 相比低氮处理, 高氮处理下, rhl1、Kas、iaa11与arl1的茎叶¹⁵N浓度分别提高了60%、72%、128%与118%, 说明氮添加显著促进了水稻氮吸收, 且iaa11与arl1对氮添加的响应更强烈。在低氮水平下, AMF对rhl1、Kas、iaa11与arl1氮吸收的平均效应值分别为17%、31%、42%、51%, 表明AMF对于植物氮吸收的促进效应受根形态影响, iaa11与arl1对AMF的响应明显高于Kas与rhl1; 相较于低氮水平, 高氮水平下AMF对于不同根形态水稻氮吸收的促进效应都会显著降低, 表明氮添加削弱了AMF对植物氮吸收的促进效应。该研究阐明了4种不同根形态基因型水稻氮养分吸收存在显著差异, 其中氮吸收能力较弱的基因型水稻对AMF的响应更强, 该结果补充了植物与AMF在养分吸收上存在功能互补的控制实验证据。     

Response of snapdragon (<Emphasis Type="Italic">Antirrhinum majus</Emphasis> L.) to blended water irrigation and arbuscular mycorrhizal fungi inoculation: uptake of minerals and leaf water relations

A greenhouse study was performed in order to investigate the effects of three arbuscular mycorrhizal fungi (AMF) species on vegetative growth, water relations, and mineral composition parameters of snapdragon (Antirrhinum majus cv. Bells white) under irrigation from different water sources. Five irrigation treatments included using purely desalinized (fresh) water (DW), as a control, three different blends of DW with saline ground water from a well with increasing salinity, and one with 100% of saline well water. Inoculation with AMF enhanced growth rates and a relative water content of snapdragon plants grown under well-water irrigation. AMF also improved the leaf water potential and increased water-use efficiency of the plants. Shoot and root dry masses were higher in the AMF-treated plants than those in AMF-free plants. In both shoots and roots, concentrations of total P, Ca²⁺, N, Mg²⁺, and K⁺ were higher in the AMF-treated plants compared with AMF-free plants under salt-stress conditions. Shoot Cl^- and Na⁺ concentrations were lower in the AMF-treated plants than those in the AMF-free plants grown under well-water irrigation. Snapdragon plants exhibited a high degree of dependency on AMF; it improved plant growth rates and leaf water relations, particularly, with increasing salinity of irrigation water.     

丛枝菌根真菌对小麦生长的影响

为了促进经济作物小麦的生长,提高土壤氮磷循环与转化效率,选择两种优良丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)——摩西球囊霉(GM,Glomus mosseae)、根内球囊霉(GI,Glomus intraradices),研究AMF在小麦整个营养生长阶段中对其生长以及对土壤中植物生长需求的大量元素——氮、磷的作用及影响。结果表明:人工施加菌剂可显著提高AMF对小麦的侵染率,施加GM菌剂时,小麦侵染率提高24.54%,同时,株高提高14.08%,小麦地上生物量提高24.05%。GM效果优于GI。施加菌剂后,小麦侵染率与土壤中水解性氮呈显著正相关;植物地上生物量与土壤中总氮,水解性氮呈显著负相关。表明AMF可活化土壤中的氮元素,同时促进作物生长,强化对土壤中氮元素的利用。     

丛枝菌根真菌提高盐胁迫植物抗氧化机制的研究进展

土地盐渍化是在自然环境和人为活动的双重作用下形成的全球性的重要生态问题,其会对植物造成渗透失衡、离子胁迫、氧化损伤等危害,导致植物生长缓慢、生物量减少甚至是绝产。丛枝菌根真菌(AMF)是一种普遍存在于土壤中的有益微生物,能够与大多数植物根系形成共生关系,其共生关系在多种逆境生态系统中均具有重要生态意义。AMF-植物共生体具有高效抗氧化系统,能够提高植物在盐胁迫下的抗氧化反应进而增强耐盐性。本文从氧化损伤、渗透调节、抗氧化机制和生物活性分子等角度,系统地阐述了丛枝菌根真菌提高植物抗氧化机制的研究进展,并提出了研究展望,以期为利用菌根生物技术提高植物耐盐性提供理论参考。     

Transmembrane H + and Ca 2+ fluxes through extraradical hyphae of arbuscular mycorrhizal fungi in response to drought stress

丛枝菌根真菌(AMF)能够和大多数陆地植物形成共生体系, 对于植物生长发育和适应各种逆境胁迫具有重要作用。很多研究表明干旱胁迫下AMF能够促进宿主植物对水分的吸收从而增强植物抗旱能力, 但目前针对AMF根外菌丝响应水分胁迫的生理变化以及AMF与宿主植物逆境信号交流的研究并不多。该研究利用AMF Rhizophagus irregularis和胡萝卜(Daucus carota var. sativa)毛状根双重无菌培养体系获得纯净根外菌丝, 向培养基添加聚乙二醇(PEG)模拟干旱胁迫, 运用场发射扫描电子显微镜(FE-SEM-EDS)观察干旱胁迫对AMF根外菌丝形态的影响, 同时采用非损伤微测技术(NMT)观测根外菌丝跨膜H ⁺和Ca ²⁺离子流变化。结果发现, PEG处理1 h后菌丝尖端和侧面发生H ⁺外流和强烈的Ca ²⁺内流, 荧光探针分析也显示菌丝胞内pH值显著上升、Ca ²⁺浓度增加; PEG处理24 h后菌丝形态发生明显变化, 培养基pH值降低, P、Ca、Fe等元素在菌丝际积累。这些试验结果表明, 干旱胁迫下AMF根外菌丝跨膜H ⁺和Ca ²⁺流发生变化, 促进了菌丝与环境之间的物质交换。菌丝酸化生长环境有利于养分吸收, 并促进AMF与宿主植物之间的信号交流以增强植物的耐旱性。     

Common mycorrhizal networks provide a potential pathway for the transfer of hydraulically lifted water between plants

Plant roots may be linked by shared or common mycorrhizal networks (CMNs) that constitute pathways for the transfer of resources among plants. The potential for water transfer by such networks was examined by manipulating CMNs independently of plant roots in order to isolate the role(s) of ectomycorrhizal (EM) and arbuscular mycorrhizal fungal (AMF) networks in the plant water balance during drought (soil water potential -5.9 MPa). Fluorescent tracer dyes and deuterium-enriched water were used to follow the pathways of water transfer from coastal live oak seedlings (Quercus agrifolia Nee; colonized by EM and AMF) conducting hydraulic lift (HL) into the roots of water-stressed seedlings connected only by EM (Q. agrifolia) or AMF networks (Q. agrifolia, Eriogonum fasciculatum Benth., Salvia mellifera Greene, Keckiella antirrhinoides Benth). When connected to donor plants by hyphal linkages, deuterium was detected in the transpiration flux of receiver oak plants, and dye-labelled extraradical hyphae, rhizomorphs, mantles, and Hartig nets were observed in receiver EM oak roots, and in AMF hyphae of Salvia. Hyphal labelling was scarce in Eriogonum and Keckiella since these species are less dependent on AMF. The observed patterns of dye distribution also indicated that only a small percentage of mycorrhizal roots and extraradical hyphae were involved with water transfer among plants. Our results suggest that the movement of water by CMNs is potentially important to plant survival during drought, and that the functional ecophysiological traits of individual mycorrhizal fungi may be a component of this mechanism.     

The impact of arbuscular mycorrhizal fungi on tomato plant resistance against Tuta absoluta (Meyrick) in greenhouse conditions

The symbiotic relationship between plants and arbuscular mycorrhizal fungi (AMF) improves plant growth and increases its resistance to pests and diseases. Mycorrhizal fungi are among the specialized fungi associated with the rhizosphere and are completely dependent on plant organic carbon. In this research tomato, Solanum lycopersicum L. was used as the host plant to evaluate the interaction effects between inoculation of tomato plant with AMF and feeding of tomato leaf miner, Tuta absoluta (Meyrick). In addition, plant growth parameters and growth rate of insect were assessed. The mycorrhizal treatment included a mixture of four fungal species (Funneliformis mosseae, Rhizophagus intraradices, R. irregularis and Glomus iranicus). The results of the experiment showed that tomato plant roots were well colonized (66.29%) by AMF and there was a significant mutual relationship between the insects feeding on the plants and the fungi. Feeding by the insects on plants inoculated with the fungus increased percentage of colonization by AMF in plants infested with the insect as compared to the control plants. The results also indicated that growth parameters and phosphorus content of the plants inoculated with fungi significantly increased compared to the control group. Moreover, significantly lower growth rate and consumption index observed in the T. absoluta larvae were fed on the leaves of plants treated with AMF compared to leaves of plants not inoculated with AMF.