Ecto- and arbuscular mycorrhizal symbiosis can induce tolerance to toxic pulses of phosphorus in jarrah (Eucalyptus marginata) seedlings

In common with many plants native to low P soils, jarrah (Eucalyptus marginata) develops toxicity symptoms upon exposure to elevated phosphorus (P). Jarrah plants can establish arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) associations, along with a non-colonizing symbiosis described recently. AM colonization is known to influence the pattern of expression of genes required for P uptake of host plants and our aim was to investigate this phenomenon in relation to P sensitivity. Therefore, we examined the effect on hosts of the presence of AM and ECM fungi in combination with toxic pulses of P and assessed possible correlations between the induced tolerance and the shoot P concentration. The P transport dynamics of AM (Rhizophagus irregularis and Scutellospora calospora), ECM (Scleroderma sp.), non-colonizing symbiosis (Austroboletus occidentalis), dual mycorrhizal (R. irregularis and Scleroderma sp.), and non-mycorrhizal (NM) seedlings were monitored following two pulses of P. The ECM and A. occidentalis associations significantly enhanced the shoot P content of jarrah plants growing under P-deficient conditions. In addition, S. calospora, A. occidentalis, and Scleroderma sp. all stimulated plant growth significantly. All inoculated plants had significantly lower phytotoxicity symptoms compared to NM controls 7 days after addition of an elevated P dose (30 mg P kg−1 soil). Following exposure to toxicity-inducing levels of P, the shoot P concentration was significantly lower in R. irregularis-inoculated and dually inoculated plants compared to NM controls. Although all inoculated plants had reduced toxicity symptoms and there was a positive linear relationship between rank and shoot P concentration, the protective effect was not necessarily explained by the type of fungal association or the extent of mycorrhizal colonization.

Moderating mycorrhizas: arbuscular mycorrhizas modify rhizosphere chemistry and maintain plant phosphorus status within narrow boundaries

Pastures often experience a pulse of phosphorus (P) when fertilized. We examined the role of arbuscular mycorrhizal fungi (AMF) in the uptake of P from a pulse. Five legumes (Kennedia prostrata, Cullen australasicum, Bituminaria bituminosa, Medicago sativa and Trifolium subterraneum) were grown in a moderate P, sterilized field soil, either with (+AMF) or without (−AMF) addition of unsterilized field soil. After 9–10 weeks, half the pots received 15 mg P kg−1 of soil. One week later, we measured: shoot and root dry weights; percentage of root length colonized by AMF; plant P, nitrogen and manganese (Mn) concentrations; and rhizosphere carboxylates, pH and plant-available P. The P pulse raised root P concentration by a similar amount in uncolonized and colonized plants, but shoot P concentration increased by 143% in uncolonized plants and 53% in colonized plants. Inoculation with AMF decreased the amount of rhizosphere carboxylates by 52%, raised rhizosphere pH by ∼0.2–0.7 pH units and lowered shoot Mn concentration by 38%. We conclude that AMF are not simply a means for plants to enhance P uptake when P is limiting, but also act to maintain shoot P within narrow boundaries and can affect nutrient uptake through their influence on rhizosphere chemistry.

Do arbuscular mycorrhizas or heterotrophic soil microbes contribute toward plant acquisition of a pulse of mineral phosphate?

Aims: We investigated the role of arbuscular mycorrhizal fungi (AMF) and heterotrophic soil microbes in the uptake of phosphorus (P) by Trifolium subterraneum from a pulse. Methods: Plants were grown in sterilised pasture field soil with a realistic level of available P. There were five treatments, two of which involved AMF: 1) unsterilised field soil containing a community of AMF and heterotrophic organisms; 2) Scutellospora calospora inoculum (AMF); 3) microbes added as filtrate from the field soil; 4) microbes added as filtrate from the S. calospora inoculum; 5) no additions, i.e. sterilised field soil. After 11 weeks, plants were harvested: 1 day before (day 0), 1 day after (day 2) and 7 days after (day 8) the pulse of P (10 mg kg−1). Results: There was no difference among treatments in shoot and root dry weight, which increased from day 0 to day 8. At day 0, shoots and roots of plants in the colonised treatments had higher P and lower Mn concentrations. After the pulse, the rate of increase in P concentration in the shoots was slower for the colonised plants, and the root Mn concentration declined by up to 50 % by day 2. Conclusions: Plants colonised by AMF had a lower rate of increase in shoot P concentration after a pulse, perhaps because intraradical hyphae accumulated P and thus reduced its transport to the shoots.

Carbon trading for phosphorus gain: the balance between rhizosphere carboxylates and arbuscular mycorrhizal symbiosis in plant phosphorus acquisition

Two key plant adaptations for phosphorus (P) acquisition are carboxylate exudation into the rhizosphere and mycorrhizal symbioses. These target different soil P resources, presumably with different plant carbon costs. We examined the effect of inoculation with arbuscular mycorrhizal fungi (AMF) on amount of rhizosphere carboxylates and plant P uptake for 10 species of low-P adapted Kennedia grown for 23 weeks in low-P sand. Inoculation decreased carboxylates in some species (up to 50%), decreased plant dry weight (21%) and increased plant P content (23%). There was a positive logarithmic relationship between plant P content and the amount of rhizosphere citric acid for inoculated and uninoculated plants. Causality was indicated by experiments using sand where little citric acid was lost from the soil solution over 2 h and citric acid at low concentrations desorbed P into the soil solution. Senesced leaf P concentration was often low and P-resorption efficiencies reached >90%. In conclusion, we propose that mycorrhizally mediated resource partitioning occurred because inoculation reduced rhizosphere carboxylates, but increased plant P uptake. Hence, presumably, the proportion of plant P acquired from strongly sorbed sources decreased with inoculation, while the proportion from labile inorganic P increased. Implications for plant fitness under field conditions now require investigation.

The diversity of arbuscular mycorrhizas of selected Australian Fabaceae

Members of the Australian native perennial Fabaceae have been little explored with regard to their root biology and the role played by arbuscular mycorrhizal (AM) fungi in their establishment, nutrition and long-term health. The ultimate goal of our research is to determine the dependency of native perennial legumes on their co-evolved AM fungi and conversely, the impact of AM fungal species in agricultural fields on the productivity of sown native perennial legume pastures. In this paper we investigate the colonisation morphology in roots and the AMF, identified by spores extracted from rhizosphere soil, from three replicate plots of each of the native legumes, Cullen australasicum, C. tenax and Lotus australis and the exotic legumes L. pedunculatus and Medicago sativa. The plants were grown in an agricultural field. The level and density of colonisation by AM fungi, and the frequency of intraradical and extraradical hyphae, arbuscules, intraradical spores and hyphal coils all differed between host plants and did not consistently differ between native and exotic species. However, there were strong similarities between species in the same genus. The three dominant species of AM fungi in rhizosphere soil also differed with host plant, but one fungus (Glomus mosseae) was always the most dominant. Sub-dominant AM species were the same between species in the same genus. No consistent differences in dominant spores were observed between the exotic and native Fabaceae species. Our results suggest that plant host influences the mycorrhizal community in the rhizosphere soil and that structural and functional differences in the symbiosis may occur at the plant genus level, not the species level or due to provenance.

Seedling response to phosphate addition and inoculation with arbuscular mycorrhizas and the implications for old-field restoration in Western Australia

We predicted that P-fertiliser residues will limit the establishment of native plant species and their mycorrhizas to old-fields in the wheat-growing region (i.e. the wheatbelt) of Western Australia. To test this prediction, we assessed the growth and P uptake of seedlings of three native plant species to phosphate addition and inoculation with arbuscular mycorrhizas (AM) in a pot study. The native plant species were Acacia acuminata Benth. (Mimosaceae), Eucalyptus loxophleba Benth. subsp. loxophleba (Myrtaceae) and Hakea preissii Meisn. (Proteaceae); and each pot contained one seedling. P was added to field soil to mimic pre-agricultural (P0), old-field (P1) and 10 times old-field (P10) soils. AM inoculant, which was a mix of Scutellospora calospora (Nicolson and Gerdemann) Walker and Sanders, Glomus intraradices Schenck and Smith and Glomus mosseae (Nicolson and Gerdemann) Gerdemann and Trappe, was added to half of the pots. After 12 weeks, the biomass and P uptake of the mycorrhizal A. acuminata were greater than those of the non-mycorrhizal plants across all P treatments. Plant biomass decreased significantly with increasing P addition, yet this species was apparently unable to suppress its mycorrhizal colonisation at high P despite this reduction in growth. In contrast, mycorrhizal and non-mycorrhizal E. loxophleba subsp. loxophleba were of a similar biomass after 12 weeks; maximum biomass was attained at intermediate (old-field) levels of P. P uptake increased with increasing P supply, beyond that required to attain maximum biomass. AM did not form on H. preissii. P uptake increased with increasing P supply for this species also. Overall, it is the apparent inability of these species to down-regulate P uptake rather than a lack of mycorrhizal symbiosis that will constrain their establishment on wheatbelt old-fields.

Navigating the labyrinth: a guide to sequence-based, community ecology of arbuscular mycorrhizal fungi

Data generated from next generation sequencing (NGS) will soon comprise the majority of information about arbuscular mycorrhizal fungal (AMF) communities. Although these approaches give deeper insight, analysing NGS data involves decisions that can significantly affect results and conclusions. This is particularly true for AMF community studies, because much remains to be known about their basic biology and genetics. During a workshop in 2013, representatives from seven research groups using NGS for AMF community ecology gathered to discuss common challenges and directions for future research. Our goal was to improve the quality and accessibility of NGS data for the AMF research community. Discussions spanned sampling design, sample preservation, sequencing, bioinformatics and data archiving. With concrete examples we demonstrated how different approaches can significantly alter analysis outcomes. Failure to consider the consequences of these decisions may compound bias introduced at each step along the workflow. The products of these discussions have been summarized in this paper in order to serve as a guide for any researcher undertaking NGS sequencing of AMF communities.

Cultivo in vitro de raízes de tomateiro, menta e videira para produção de inóculo de fungo micorrízico arbuscular

Os Fungos Micorrízicos Arbusculares (FMAs) desempenham um papel importante na sustentabilidade dos ecossistemas, devido a importância da simbiose que formam com a maioria das plantas. A associação destes fungos com as raízes possibilita uma melhor nutrição das plantas, além de promover o crescimento e a resistência a fatores causadores de estresse. Desta forma, auxiliam no aumento da produtividade das mesmas, e na redução do uso de insumos químicos, como fertilizantes e agrotóxicos, na agricultura. Em função ao caráter de simbiontes obrigatórios, é de grande interesse o desenvolvimento de processos que permitam o isolamento, caracterização, manutenção de isolados e produção de inóculo desses fungos. Assim, com o objetivo de produzir um método de cultivo axênico, e que também possa auxiliar o estudo destes em áreas como a biologia molecular, testou-se protocolos de micropropagação para a produção in vitro de Glomus etunicatum W. N. Becker & Gerd., utilizando-se como hospedeiro raízes de tomateiro, videira e menta. Além destas culturas, que foram propagadas vegetativamente para a manutenção das culturas estoques e enraizadas in vitro com a aplicação exógena do regulador de crescimento ácido indolbutírico (AIB), trabalhou-se também com a cultura da menta, mas devido a problemas que ocorreram durante a etapa de enraizamento, não foi possível efetivar a associação do inóculo Na inoculação de esporos em raízes da videira cultivadas in vitro, não se obteve êxito. Com o cultivo in vitro de raízes de tomateiro foi possível conduzir os trabalhos até a etapa de colonização do FMA. No enraizamento in vitro das três culturas trabalhadas foi avaliado o número e o comprimento médio das raízes cultivadas nos meios de enraizamento nas doses de 0, 1,0 e 2,0 mg.L-1 de AIB. Para o tomateiro e a menta, o número e o comprimento médio de raízes por explante foram significativamente maiores com a dose de 1,0 mg.L-1 de AIB. Entretanto, para a cultura da videira, embora o uso de AIB, em ambas as doses de 1,0 e 2,0 mg.L-1 induziram um maior número de raízes, o comprimento médio de raízes foi maior quando AIB não foi adicionado no meio de enraizamento.

Efeito do fósforo, fumigação do substrato e fungo micorrízico arbuscular sobre o crescimento de plantas de mamoeiro

Estudou-se o efeito da inoculação com o fungo micorrízico arbuscular (FMA), Glomus macrocarpum, da fumigação do substrato e da adição de fósforo solúvel (60, 120, 240 e 480 mg kg-1 de P no solo) sobre as variáveis altura, número de folhas e diâmetro do caule de plantas de mamoeiro cv. Sunrise Solo.O FMA edoses crescentes de fósforo, isoladamente, exerceram efeitos significativos sobre essas variáveis. Não houve efeito significativo do fator fumigação do substrato. O efeito da inoculação foi mais acentuado no tratamento com adição de 60 mg kg-1 de P no solo. A inoculação com G. macrocarpum reduziu a necessidade de fósforo para o mamoeiro, tanto que as variáveis estudadas em plantas inoculadas na ausência de adubação fosfática não diferiram de plantas não inoculadas em substrato adicionado de mais de 240 mg kg-1 de P no solo.

Contribuição de fungos micorrízicos arbusculares autóctones no crescimento de Guazuma ulmifolia em solo de cerrado degradado

Ensaios foram conduzidos, em casa de vegetação, com solos de pastagem degradada reflorestada e cerrado preservado (controle) visando avaliar a contribuição de fungos micorrízicos arbusculares (FMA) autóctones no crescimento de mutambo (Guazuma ulmifolia Lamb.). As mudas foram transplantadas para sacos de plástico (2 kg) com substratos esterilizados na proporção 4:1 (solo:areia), e o tratamento inoculado recebeu 300 esporos de FMA por saco. A inoculação não proporcionou aumento significativo na produção da matéria seca da parte aérea, matéria fresca das raízes e altura da planta, sugerindo que a G. ulmifolia não é responsiva à micorrização.

Dinâmica sazional de fungos micorrízicos arbusculares (Glomeromycota) em cerradão e brejo de altitude na Chapada do Araripe, CE

Arbuscular mycorrhizal fungi (AMF) are obligatory symbiotic organisms that associate with roots of a large number of plant taxa, and are found in all terrestrial ecosystems. These fungi promote greater tolerance to environmental stresses to associated plant, favoring the establishment of plant communities, especially where soil fertility is a limiting factor, as in the Caatinga, an exclusively Brazilian domain that has been focus of research due to its great biodiversity that can help clarify the history of vegetation in South America. Because of the ecological importance of AMF, the limited number of jobs and the potential diversity of the Caatinga, this work aims to inventory the diversity and determine AMF communities in areas with different faces occurrent in FLONA Araripe, Ceará (CE). The sample collection occurred in four periods at the beginning and end of the dry season (August and December 2011, respectively) and rainy (February and June 2012, respectively) in an area of marsh and woodland altitude of the Araripe, Crato, CE. The glomerosporos were extracted by wet sieving and centrifugation in water and sucrose (50%) mounted between slide and coverslip using PVLG and PVLG + Reagent Melzer. In total, we found 46 species of AMF distributed in eight families and 16 genera: Acaulospora (6), Ambispora (1), Cetraspora (2), Dentiscutata (5), Fuscutata (2), Gigaspora (6), Glomus (13) Intraornatospora (1), Kuklospora (1), Orbispora (1), Paradentiscutata (1), Quatunica (1), Racocetra (1), Scutellospora (2), Septoglomus (2) and a new genus. analysis showed that ecological each area of study has its own seasonal dynamics, with an area of woodland with a greater diversity of species throughout the year, while the marsh elevation showed greater variation in species found among the collection periods, showing that vegetation and rainfall has strong influence on the seasonal dynamics of AMF, as well as the availability of nutrients and soil pH so

Nitrogen-tixing and vesicular-arbuscular mycorrhizal symbioses in some tropical legume trees of tribe Mimoseae

Response to mineral fertilization and inoculation with rhizobia and/or arbuscular mycorrhiza fungi (AMF) of the Anadenanthera colubrina, Mimosa bimucronata and Parapiptadenia rigida (Leguminosae-Mimosoideae) native trees from Brazilian riparian forests, were studied in nursery conditions. Each species was submitted to seven treatments, varying nitrogen and phosphorous fertilization and inoculation with rhizobia (r), mycorrhiza (m) or both (rm): NP, P, P + r, P + rm, N, N + m and N + rm. Results showed that AMF inoculations did not enhance the mycorrhizal colonization, and P uptake was not sufficient to sustain good growth of plants. The level of P mineral added affected negatively the AMF colonization in A. colubrina and M. bimucronata, but not in P. rigida. Native fungi infected the three legume hosts. The absence of mineral N limited growth of A. colubrina and P. rigida, but in M. bimucronata the lack of N was corrected by biological nitrogen fixation. N mineral added inhibited the nodulation, although spontaneous nodulation had occurred in A. colubrina and M. bimucronata. Rhizobia inoculation enhanced the number of nodules, nitrogenase activity and leghemoglobin content of these two species. Thus, the extent of rhizobial and mycorrhizal symbiosis in these species under nursery conditions can affect growth and consequently the post-planting success. (C) 2004 Elsevier B.V. All rights reserved.

Occurrence of symbiotic fungi and rhizospheric phosphate solubilization in weeds

Estudos sobre a ecologia de organismos envolvidos no processo de produção são necessários para o desenvolvimento de uma agricultura sustentável, e hoje, a sustentabilidade está intimamente ligada à rentabilidade de produção. Objetivou-se com este trabalho verificar a ocorrência de fungos micorrízicos arbusculares em plantas daninhas de lavouras brasileiras e avaliar o potencial de solubilização de fosfato inorgânico da microbiota associada. 36 espécies de plantas daninhas foram avaliadas quanto à ocorrência de micorrizas, e 11 foram selecionadas para avaliação do potencial de solubilização total e relativa de fosfato. Todas as espécies apresentaram colonização por micorrizas, inclusive um exemplar da família Brassicaceae, geralmente enquadrada como não micorrizada. Na maioria das espécies, os tipos morfológicos de arbúsculos e enovelados de hifas foram observados, sendo os enovelados mais comuns entre as gramíneas. Fungos endofíticos do tipo dark septate foram visualizados na maioria das plantas. As plantas daninhas apresentaram distintos potenciais de solubilização de P na rizosfera. Amaranthus retroflexus, Bidens pilosa e Leonotis nepetaefolia apresentaram elevados valores de solubilização relativa de fosfato. Este é o primeiro relato de micorrizas e da atividade de solubilização de fosfato em plantas daninhas no Brasil.