scholarly journals Myristate can be used as a carbon and energy source for the asymbiotic growth of arbuscular mycorrhizal fungi

2020 ◽  
Vol 117 (41) ◽  
pp. 25779-25788
Author(s):  
Yuta Sugiura ◽  
Rei Akiyama ◽  
Sachiko Tanaka ◽  
Koji Yano ◽  
Hiromu Kameoka ◽  
...  

Arbuscular mycorrhizal (AM) fungi, forming symbiotic associations with land plants, are obligate symbionts that cannot complete their natural life cycle without a host. The fatty acid auxotrophy of AM fungi is supported by recent studies showing that lipids synthesized by the host plants are transferred to the fungi, and that the latter lack genes encoding cytosolic fatty acid synthases. Therefore, to establish an asymbiotic cultivation system for AM fungi, we tried to identify the fatty acids that could promote biomass production. To determine whether AM fungi can grow on medium supplied with fatty acids or lipids under asymbiotic conditions, we tested eight saturated or unsaturated fatty acids (C12 to C18) and two β-monoacylglycerols. Only myristate (C14:0) led to an increase in the biomass ofRhizophagus irregularis, inducing extensive hyphal growth and formation of infection-competent secondary spores. However, such spores were smaller than those generated symbiotically. Furthermore, we demonstrated thatR. irregulariscan take up fatty acids in its branched hyphae and use myristate as a carbon and energy source. Myristate also promoted the growth ofRhizophagus clarusandGigaspora margarita. Finally, mixtures of myristate and palmitate accelerated fungal growth and induced a substantial change in fatty acid composition of triacylglycerol compared with single myristate application, although palmitate was not used as a carbon source for cell wall biosynthesis in this culture system. Our findings demonstrate that myristate boosts the asymbiotic growth of AM fungi and can also serve as a carbon and energy source.

2019 ◽  
Author(s):  
Yuta Sugiura ◽  
Rei Akiyama ◽  
Sachiko Tanaka ◽  
Koji Yano ◽  
Hiromu Kameoka ◽  
...  

AbstractArbuscular mycorrhizal (AM) fungi, forming symbiotic associations with land plants, are obligate symbionts that cannot complete their natural life cycle without a host. Recently, fatty acid auxotrophy of AM fungi is supported by studies showing that lipids synthesized by the host plants are transferred to the fungi and that the latter lack genes encoding cytosolic fatty acid synthases (1-7). Therefore, to establish an asymbiotic cultivation system for AM fungi, we tried to identify the fatty acids that could promote biomass production. To determine whether AM fungi can grow on medium supplied with fatty acids or lipids under asymbiotic conditions, we tested eight saturated or unsaturated fatty acids (C12–C18) and two β-monoacylglycerols. Only myristate (C14:0) led to an increase in biomass of Rhizophagus irregularis, inducing extensive hyphal growth and formation of infection-competent secondary spores. However, such spores were smaller than those generated symbiotically. Furthermore, we demonstrated that R. irregularis can take up fatty acids in its branched hyphae and use myristate as a carbon and energy source. Myristate also promoted the growth of Rhizophagus clarus and Gigaspora margarita. Finally, mixtures of myristate and palmitate accelerated fungal growth and induced a substantial change in fatty acid composition of triacylglycerol compared with single myristate application, although palmitate was not used as a carbon source for cell wall biosynthesis in this culture system. In conclusion, here we demonstrate that myristate boosts asymbiotic growth of AM fungi and can also serve as a carbon and energy source.Significance statementThe origins of arbuscular mycorrhizal (AM) fungi, which form symbiotic associations with land plants, date back over 460 million years ago. During evolution, these fungi acquired an obligate symbiotic lifestyle, and thus depend on their host for essential nutrients. In particular, fatty acids are regarded as crucial nutrients for the survival of AM fungi owing to the absence of genes involved in de novo fatty acid biosynthesis in the AM fungal genomes that have been sequenced so far. Here, we show that myristate initiates AM fungal growth under asymbiotic conditions. These findings will advance pure culture of AM fungi.


2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Christopher Ngosong ◽  
Elke Gabriel ◽  
Liliane Ruess

Biomass estimation of arbuscular mycorrhiza (AM) fungi, widespread plant root symbionts, commonly employs lipid biomarkers, predominantly the fatty acid 16:1ω5. We briefly reviewed the application of this signature fatty acid, followed by a case study comparing biochemical markers with microscopic techniques in an arable soil following a change to AM non-host plants after 27 years of continuous host crops, that is, two successive cropping seasons with wheat followed by amaranth. After switching to the non-host amaranth, spore biomass estimated by the neutral lipid fatty acid (NLFA) 16:1ω5 decreased to almost nil, whereas microscopic spore counts decreased by about 50% only. In contrast, AM hyphal biomass assessed by the phospholipid (PLFA) 16:1ω5 was greater under amaranth than wheat. The application of PLFA 16:1ω5 as biomarker was hampered by background level derived from bacteria, and further enhanced by its incorporation from degrading spores used as microbial resource. Meanwhile, biochemical and morphological assessments showed negative correlation for spores and none for hyphal biomass. In conclusion, the NLFA 16:1ω5 appears to be a feasible indicator for AM fungi of the Glomales group in the complex field soils, whereas the use of PLFA 16:1ω5 for hyphae is unsuitable and should be restricted to controlled laboratory studies.


Botany ◽  
2009 ◽  
Vol 87 (4) ◽  
pp. 387-400 ◽  
Author(s):  
Christine Juge ◽  
Annie Champagne ◽  
Andrew P. Coughlan ◽  
Nicolas Juge ◽  
Lael Parrott ◽  
...  

The present study is, to the best of our knowledge, the first to investigate the use of the fractal dimension (FD) to quantify the growth and development of undisturbed, fully functional arbuscular mycorrhizal (AM) hyphae developing in vitro. The majority of the work focused on the model AM fungus Glomus intraradices DAOM 181602. The time course study and final measurements of an intact mature extraradical mycelium allowed us to compare the development of the mycelium and the FD value. The final FD value of 1.62 for the mature mycelium is similar to that obtained for highly branched root systems and tree crowns. The FD method was used to characterize the morphology of germinative and presymbiotic hyphae in the presence of stimulatory (strigolactone GR-24, 0.1 µmol·L–1 and bisphenol A, 10 µmol·L–1) and inhibitory (NaCl, 80 mmol·L–1) molecules, and the extraradical phase in the presence of an inhibitory molecule (NaCl, 80 mmol·L–1). Where possible, results were compared with those obtained using the traditional grid-line (GL) technique. The FD approach allowed treatment effects to be accurately quantified, both in germinative and extraradical phases. In the second case, this technique provided a single quantitative value of extraradical hyphal growth that included runner hyphae (RH) networks, and fine-branching (FB) ramifications. This is in contrast to the GL technique, which provides a value for the estimation of RH, but which is not suitable for accurately measuring FB hyphae. Given the ease with which the FD values can be calculated, and the fact that this method can provide a single value for the quantification of extraradical hyphal growth and development, we suggest that this method is useful for in vitro studies. Furthermore under certain situations of germinative or presymbiotic growth, it may be used in concert with the GL method to provide a greater degree of information about hyphal morphology. The usefulness and limits of the FD method at different stages of the AM fungal growth cycle are discussed.


2007 ◽  
Vol 85 (7) ◽  
pp. 644-651 ◽  
Author(s):  
Donna Glassop ◽  
Rosamond M. Godwin ◽  
Sally E. Smith ◽  
Frank W. Smith

The completed rice-genome sequence was screened with a known inorganic phosphate (Pi) transporter sequence to reveal a family of 13 Pi transporters. This family can be used for studies into Pi acquisition and translocation throughout the plant. Plants that form symbiotic associations with arbuscular mycorrhizal (AM) fungi are of particular interest with respect to Pi acquisition because of their ability to utilize both direct and fungal pathways of uptake. Localization of transcripts of two Pi transporters by real-time RT-PCR and in situ hybridization were conducted in rice subjected to low Pi, high Pi, and AM colonization. One Pi transporter, ORYsa;Pht1;13, was detected in rice roots under all growth conditions. ORYsa;Pht1;11 was only expressed in roots colonized by AM fungi. Antisense RNA probes of ORYsa;Pht1;11 localized to cortical cells containing arbuscules and hyphal coils, formed by Glomus intraradices Schenck and Smith and Scutellospora calospora (Nicolson and Gerdemann) Walker and Sanders, respectively. Localization of the ORYsa;Pht1;13 probes was similar to that observed for ORYsa;Pht1;11 in colonized rice roots. This research proposes that at least two rice Pi transporters are involved in acquiring Pi via AM fungi, emphasising the complexity of Pi acquisition in plants with access to two Pi uptake pathways.


2020 ◽  
Vol 57 (1) ◽  
pp. 47-64
Author(s):  
Alberto Andrino ◽  
Georg Guggenberger ◽  
Leopold Sauheitl ◽  
Stefan Burkart ◽  
Jens Boy

AbstractTo overcome phosphorus (P) deficiency, about 80% of plant species establish symbiosis with arbuscular mycorrhizal fungi (AMF), which in return constitute a major sink of photosynthates. Information on whether plant carbon (C) allocation towards AMF increases with declining availability of the P source is limited. We offered orthophosphate (OP), apatite (AP), or phytic acid (PA) as the only P source available to arbuscular mycorrhiza (AM) (Solanum lycopersicum x Rhizophagus irregularis) in a mesocosm experiment, where the fungi had exclusive access to each P source. After exposure, we determined P contents in the plant, related these to the overall C budget of the system, including the organic C (OC) contents, the respired CO2, the phospholipid fatty acid (PLFA) 16:1ω5c (extraradical mycelium), and the neutral fatty acid (NLFA) 16:1ω5c (energy storage) at the fungal compartment. Arbuscular mycorrhizal (AM) plants incorporated P derived from the three P sources through the mycorrhizal pathway, but did this with differing C-P trading costs. The mobilization of PA and AP by the AM plant entailed larger mycelium infrastructure and significantly larger respiratory losses of CO2, in comparison with the utilization of the readily soluble OP. Our study thus suggests that AM plants invest larger C amounts into their fungal partners at lower P availability. This larger C flux to the AM fungi might also lead to larger soil organic C contents, in the course of forming larger AM biomass under P-limiting conditions.


Agriculture ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 520 ◽  
Author(s):  
Thomas I. Wilkes ◽  
Douglas J. Warner ◽  
Keith G. Davies ◽  
Veronica Edmonds-Brown

Zero till cropping systems typically apply broad-spectrum herbicides such as glyphosate as an alternative weed control strategy to the physical inversion of the soil provided by cultivation. Glyphosate targets 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase in plants. There is growing evidence that this may have a detrimental impact on non-target organisms such as those present in the soil microbiome. Species of commercial importance, such as arbuscular mycorrhizal (AM) fungi that form a symbiotic relationship with plant roots are an important example. This study investigates the impact of soil cultivation and glyphosate application associated with conventional tillage (CT) and zero tillage (ZT) respectively on AM fungi populations under field and glasshouse conditions. Topsoil (<10 cm) was extracted from CT and ZT fields cropped with winter wheat, plus non-cropped control plots within the same field boundary, throughout the cropping year. Glyphosate was applied in glasshouse experiments at rates between 0 and 350 g L−1. Ergosterol, an indicator of fungal biomass, was measured using high performance liquid chromatography before and after glyphosate application. Fungal root arbuscules, an indicator of AM fungi–root symbiosis, were quantified from the roots of wheat plants. Under glasshouse conditions root arbuscules were consistently higher in wheat grown in ZT field extracted soils (P = 0.01) compared to CT. Glyphosate application however inhibited fungal biomass in both the ZT (P < 0.00001) and CT (P < 0.001) treatments. In the absence of glyphosate, the number of stained root arbuscules increased significantly. Ergosterol levels, used as a proxy for fungal biomass, remained lower in the soil post glyphosate application. The results suggest that CT has a greater negative impact on AM fungal growth than ZT and glyphosate, but that glyphosate is also detrimental to AM fungal growth and hinders subsequent population recovery.


2003 ◽  
Vol 69 (11) ◽  
pp. 6762-6767 ◽  
Author(s):  
Ingrid M. van Aarle ◽  
Pål Axel Olsson

ABSTRACT We monitored the development of intraradical and extraradical mycelia of the arbuscular mycorrhizal (AM) fungi Scutellospora calospora and Glomus intraradices when colonizing Plantago lanceolata. The occurrence of arbuscules (branched hyphal structures) and vesicles (lipid storage organs) was compared with the amounts of signature fatty acids. The fatty acid 16:1ω5 was used as a signature for both AM fungal phospholipids (membrane constituents) and neutral lipids (energy storage) in roots (intraradical mycelium) and in soil (extraradical mycelium). The formation of arbuscules and the accumulation of AM fungal phospholipids in intraradical mycelium followed each other closely in both fungal species. In contrast, the neutral lipids of G. intraradices increased continuously in the intraradical mycelium, while vesicle occurrence decreased after initial rapid root colonization by the fungus. S. calospora does not form vesicles and accumulated more neutral lipids in extraradical than in intraradical mycelium, while the opposite pattern was found for G. intraradices. G. intraradices allocated more of its lipids to storage than did S. calospora. Thus, within a species, the fatty acid 16:1ω5 is a good indicator for AM fungal development. The phospholipid fatty acid 16:1ω5 is especially suitable for indicating the frequency of arbuscules in the symbiosis. We propose that the ratio of neutral lipids to phospholipids is more important than is the presence of vesicles in determining the storage status of AM fungi.


New Forests ◽  
2021 ◽  
Author(s):  
Magdalena Kulczyk-Skrzeszewska ◽  
Barbara Kieliszewska-Rokicka

AbstractPopulus nigra ‘Italica’ (Lombardy poplar) is a breeding cultivar of black poplar, widely used as a street tree or windbreak, often exposed to salinity and limited water availability. Populus roots can develop dual mycorrhizal associations with ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) fungi, and with non-mycorrhizal fungal endophytes (FE). The symbiotic fungi may alleviate the effects of adverse environmental conditions. We investigated the performance (growth and symbiotic associations) of one-year-old Populus nigra ‘Italica’ grown from woody cuttings in soil from natural poplar habitat and subjected to water scarcity and soil salinity (50 mM NaCl, 150 mM NaCl, 250 mM NaCl). With increasing soil salinity, a decrease in the growth parameters of the aboveground parts of the poplar plantlets and their fine roots were found; however, the roots were more resistant to the stress factors analyzed than the shoots. ECMF, AMF, and non-mycorrhizal FE were all tolerant to increased salt levels in the soil, and the ECM abundance was significantly higher under conditions of mild salinity (50 mM NaCl, 150 mM NaCl) compared to the control plants and those treated with 250 mM NaCl. Our results indicated that enhanced soil salinity increased the content of sodium and chlorine in leaves, but did not affect significantly the concentrations potassium, magnesium, calcium, phosphorus, or nitrogen. Significant accumulation of proline in leaves suggest salt stress of P. nigra ‘Italica’ treated with 250 mM NaCl and contribution of proline to the plant defense reactions.


2005 ◽  
Vol 71 (9) ◽  
pp. 5341-5347 ◽  
Author(s):  
Martin Trépanier ◽  
Guillaume Bécard ◽  
Peter Moutoglis ◽  
Claude Willemot ◽  
Serge Gagné ◽  
...  

ABSTRACT Lipids are the major form of carbon storage in arbuscular-mycorrhizal fungi. We studied fatty acid synthesis by Glomus intraradices and Gigaspora rosea. [14C]Acetate and [14C]sucrose were incorporated into a synthetic culture medium to test fatty acid synthetic ability in germinating spores (G. intraradices and G. rosea), mycorrhized carrot roots, and extraradical fungal mycelium (G. intraradices). Germinating spores and extraradical hyphae could not synthesize 16-carbon fatty acids but could elongate and desaturate fatty acids already present. The growth stimulation of germinating spores by root exudates did not stimulate fatty acid synthesis. 16-Carbon fatty acids (16:0 and 16:1) were synthesized only by the fungi in the mycorrhized roots. Our data strongly suggest that the fatty acid synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that fatty acid metabolism may play a major role in the obligate biotrophism of arbuscular-mycorrhizal fungi.


2000 ◽  
Vol 13 (6) ◽  
pp. 693-698 ◽  
Author(s):  
M. Buee ◽  
M. Rossignol ◽  
A. Jauneau ◽  
R. Ranjeva ◽  
G. Bécard

Arbuscular mycorrhizal (AM) symbiosis is an association between obligate biotrophic fungi and more than 80% of land plants. During the pre-symbiotic phase, the host plant releases critical metabolites necessary to trigger fungal growth and root colonization. We describe the isolation of a semipurified fraction from exudates of carrot hairy roots, highly active on germinating spores of Gigaspora gigantea, G. rosea, and G. margarita. This fraction, isolated on the basis of its activity on hyphal branching, contains a root factor (one or several molecules) that stimulates, directly or indirectly, G. gigantea nuclear division. We demonstrate the presence of this active factor in root exudates of all mycotrophic plant species tested (eight species) but not in those of nonhost plant species (four species). We negatively tested the hypothesis that it was a flavonoid or a compound synthesized via the flavonoid pathway. We propose that this root factor, yet to be chemically characterized, is a key plant signal for the development of AM fungi.


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