Mycorrhizal impact on drought stress tolerance of rose plants probed by chlorophyll a fluorescence, proline content and visual scoring

Micropropagated rose plants (Rosa hybrida L., cv. New Dawn) were inoculated with the arbuscular mycorrhizal (AM) fungus Glomus intraradices (Schenk and Smith) and subjected to different drought regimens. The dual objectives of these experiments were to investigate the mechanism and the extent to which AM can prevent drought damages and whether physiological analyses reveal enhanced drought tolerance of an economically important plant such as the rose. In a long-term drought experiment with four different water regimens, visual scoring of wilt symptoms affirmed that AM in a selected host–symbiont combination increased plant performance. This effect was mostly expressed if moderate drought stress was constantly applied over a long period. In a short-term experiment in which severe drought stress was implemented and plants were allowed to recover after 4 or 9 days, no visual differences between mycorrhizal and non-mycorrhizal roses were observed. Therefore, the early physiological steps conferring drought tolerance were prone to investigation. Proline content in leaves proved to be an unsuitable marker for AM-induced drought tolerance, whereas analysis of chlorophyll a fluorescence using the JIP test (collecting stress-induced changes of the polyphasic O-J-I-P fluorescence kinetics in a non-destructive tissue screening) was more explanatory. Parameters derived from this test could describe the extent of foliar stress response and help to differentiate physiological mechanisms of stress tolerance. AM led to a more intense electron flow and a higher productive photosynthetic activity at several sites of the photosynthetic electron transport chain. A K step, known as a stress indicator of general character, appeared in the fluorescence transient only in drought-stressed non-mycorrhizal plants; conversely, the data elucidate a stabilising effect of AM on the oxygen-evolving complex at the donor site of photosystem (PS) II and at the electron-transport chain between PS II and PS I. If drought stress intensity was reduced by a prolonged and milder drying phase, these significant tolerance features were less pronounced or missing, indicating a possible threshold level for mycorrhizal tolerance induction.     

Mycorrhiza and root hairs in barley enhance acquisition of phosphorus and uranium from phosphate rock but mycorrhiza decreases root to shoot uranium transfer

Some phosphate rocks (PR) contain high concentrations of uranium (U), which are potentially toxic via accumulation in soils and food chains, and plant uptake of U is likely to be influenced by characteristics of roots and associated microorganisms. The relative importance of root hairs and mycorrhiza in U uptake from PR was studied using a root hairless barley (Hordeum vulgare) mutant (Brb) and its wild type (WT). Both plant genotypes were grown in pots with Glomus intraradices BEG 87, or in the absence of mycorrhiza, and three P treatments were included: nil P, 2% (w/w) PR and 50 mg KH(2)PO(4)-P kg(-1) soil. Mycorrhiza markedly increased d. wts and P contents of Brb amended with nil P or PR, but generally depressed d. wts of WT plants, irrespective of P amendments. Mycorrhiza had contrasting effects on U contents in roots and shoots, in particular in Brb where mycorrhiza increased root U concentrations but decreased U translocation from roots to shoots. The experiment supports our understanding of arbuscular mycorrhiza as being multifunctional by not only improving the utilization of PR by the host plant but also by contributing to the phytostabilization of uranium.     

VA-mycorrhiza mediated P effect on growth and yield of sunflower (Helianthus annuus L.) at different P levels

A field trial was conducted to study the response of sunflower (Helianthus annuus L.) to different phosphorus levels (16, 24 or 32 kg P ha^–1) and inoculation with vesicular-arbuscular mycorrhizal fungus, Glomus fasciculatum on vertisol during summer 1993. At the vegetative stage of sunflower, percent mycorrhizal root colonization, spore count, dry biomass and P uptake did not differ significantly between inoculated and uninoculated control plants. However, at later stages (flowering and maturity) percent root colonization, spore count, total dry biomass and total P uptake were significantly higher in inoculated plants than in uninoculated control plants. The total dry biomass, P content and seed yield increased with increasing P level in uninoculated plants, whereas no significant difference was observed between 16 and 32 kg P ha^–1 in inoculated plants. The positive effect of mycorrhizal inoculation decreased with increasing P level above 16 kg P ha^–1, due to decreased percent root colonization and spore count at higher P levels.     

Functional diversity of arbuscular mycorrhizal fungal isolates in relation to extraradical mycelial networks

We investigated the functional significance of extraradical mycorrhizal networks produced by geographically different isolates of the arbuscular mycorrhizal fungal (AMF) species Glomus mosseae and Glomus intraradices. A two-dimensional experimental system was used to visualize and quantify intact extraradical mycelium (ERM) spreading from Medicago sativa roots. Growth, phosphorus (P) and nitrogen (N) nutrition were assessed in M. sativa plants grown in microcosms. The AMF isolates were characterized by differences in extent and interconnectedness of ERM. Phenotypic fungal variables, such as total hyphal length, hyphal density, hyphal length per mm of total or colonized root length, were positively correlated with M. sativa growth response variables, such as total shoot biomass and plant P content. The utilization of an experimental system in which size, growth rate, viability and interconnectedness of ERM extending from mycorrhizal roots are easily quantified under realistic conditions allows the simultaneous evaluation of different isolates and provides data with a predictive value for selection of efficient AMF.     

Dual inoculation with Aspergillus fumigatus and Glomus mosseae enhances biomass production and nutrient uptake in wheat (Triticum aestivum L.) supplied with organic phosphorus as Na-phytate

In a pot experiment, wheat was grown for 50 days in two heat-sterilized low-phosphorus (P) soils supplied with organic P as Na-phytate. Seed inoculation with the phosphatase-producing fungus (PPF) Aspergillus fumigatus or soil inoculation with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae increased shoot and root dry weight and root length, phosphatase activity in the rhizosphere and shoot concentrations of P and to a lesser extent of K and Mg. As a rule, the greatest effects on those parameters were most in the combined inoculation treatment (PPF + VAM). Shoot concentrations of Cu and Zn were only enhanced by VAM, not by PPF. At harvest, depletion of organic P in the rhizosphere soil increased in the order of: sterilized soil < PPF < VAM < PPF + VAM which corresponded with the enhanced P concentrations in the plants. The results demonstrate that organic P in form of Na-Phytate is efficiently used by VAM and that use of organic P can be increased by simultaneous inoculation with phosphatase-producing fungi.     

Endocytotic uptake of cationized ferritin tracer into glomus cells dissociated from the adult rat carotid body

Summary Glomus cells from carotid bodies of adult rats dissociated by means of collagenase or collagenase + trypsin were used to study by electron microscopy the endocytotic uptake of cationized ferritin (CF) tracer into subcellular compartments. The glomus cells were incubated with the tracer (1) in a basic salt medium (BM), or (2) in the BM into which calcium ionophore A23187 had been added, or (3) in a potassium-rich medium.Incubation of the cells in BM containing CF for 30 min resulted in attachment of the tracer to the cell membrane and uptake of a few solitary tracer particles into small vesicles and multivesicular bodies. No uptake into the cisternae of the Golgi apparatus was observed. Further incubation in BM containing CF for another 30 min resulted in increased uptake of the tracer into small vesicles and multivesicular bodies. A similar pattern of uptake was observed when the dissociated glomus cells were first preincubated in BM with CF for 30 min and then incubated for 1 min or 30 min in the BM solution containing both the ionophore and CF. Upon such incubation, CF particles were seen to penetrate into coated pits and sites of exocytosis at the cell surface. When the 30-min preincubation in BM was followed by incubation in a CF-containing potassium-rich medium for 15–30 min, uptake into vesicles, small lysosomes and occasionally also into profiles of the smooth endoplasmic reticulum was seen. Endocytotic mechanisms of the glomus cells are outlined.     

Evidence that differences in phosphate metabolism in mycorrhizas formed by species of Glomus and Gigaspora might be related to their life-cycle strategies

A glasshouse experiment was done to assess the development and phosphate metabolism of mycorrhizas formed by species of arbuscular mycorrhizal fungi (AMF) from two different genera, Gigaspora and Glomus on Desmodium ovalifolium plants at three concentrations of a phosphate source. The addition of phosphate (0–100 mg P kg⁻¹) had no effect on the alkaline phosphatase activity, stained histochemically, in the intra-radical mycelium of Gigaspora rosea (BEG111), but decreased that of Glomus manihotis (BEG112) over a 10-wk period. The alkaline phosphatase activity of the extra-radical mycelium was unaffected by increasing phosphate addition (0–100 mg P kg⁻¹) in both species of AMF over a 10-wk period. The extra-radical mycelium of Gi. rosea (BEG111) accumulated polyphosphate, determined by staining with 4',6-diamidino-2-phenylindole, whereas polyphosphate was not detected in the extra-radical mycelium of G. manihotis (BEG112). This work indicates differences in the mechanisms of phosphate metabolism in the mycelium of AMF from different genera on a tropical host. This might be determined by the life-cycle strategies of these fungi, in particular the formation of auxiliary cells in Gigaspora . The possibility of a negative-feedback mechanism between alkaline phosphatase and polyphosphate in the extra-radical mycelium of Gi. rosea (BEG111) and the role of polyphosphate in the symbiosis are discussed.     

Nuclei of symbiotic arbuscular mycorrhizal fungi as revealed by in vivo two-photon microscopy

Summary The present work reports the results obtained from in vivo studies on the distribution and behavior of nuclei of two arbuscular mycorrhizal (AM) fungi growing in symbiosis with tomato root organ cultures (AM monoxenic cultures). Upon staining with 4′,6-diamidino-2-phenylindole and two-photon microscopy (2PM) observations, symbiotic thick runner hyphae appeared mostly opaque to 2PM and did not reveal nuclei within them; thin runner hyphae showed dimly stained nuclei along them, whereas nuclei were clearly visible within the branches of the so-called branched absorbing structures. When visible, nuclei appeared anchored laterally at regular intervals along the symbiotic AM extraradical hyphae. Other nuclei migrate through the hyphal central core; this migration occurs in pulses. Simultaneous observations on different areas of extraradical AM mycelium revealed the existence of lysed compartments along the fungal hyphae, containing nuclei remnants and/or chromatin masses. All these results give new insights in (i) the differential permeability of AM hyphae in the symbiotic versus the asymbiotic state; (ii) the behavior and distribution of nuclei along the symbiotic extraradical mycelium; (iii) the occurrence of ageing events within the AM fungal colony; and (iv) the existence of “healing” mechanisms aiming to restrict the damage induced by such ageing or lytic events. An AM fungal strategy for hyphal survival under adverse conditions is also suggested.