Variation in aluminum resistance among arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal (AM) fungi mediate interactions between plants and soils, and are important where nutrient or metal concentrations limit plant growth. Variation in fungal response to edaphic conditions may influence the effectiveness of the plant-mycorrhizal association in some soil environments. Andropogon virginicus (broomsedge) colonizes disturbed sites in the eastern United States, including acidic mine soils where aluminum (Al) is phytotoxic, and Al resistance in broomsedge has been associated with colonization by the AM fungus Glomus clarum. In the present study, inter- and intra-specific variation to confer Al resistance to broomsedge was assessed among selected species of AM fungi. Broomsedge seeds were grown in sand culture inoculated with one of five isolates of three species of fungi (G. clarum, Acaulospora morrowiae, and Scutellospora heterogama). Plants were exposed to 0 or 400 microM Al in nutrient solution and harvested after 4 or 9 weeks of growth. Mean infection percentage, plant biomass, and plant tissue Al and phosphorus (P) concentrations were measured. G. clarum conferred the greatest Al resistance to broomsedge, with the lowest variability among isolates for colonization and growth inhibition by Al [tolerance indices (TI) between 22.4 and 92.7%]. Broomsedge plants colonized by A. morrowiae were consistently the most sensitive to Al, with little variation among isolates (TI between 1.6 and 12.1%). Al resistance by S. heterogama isolates was intermediate and wide-ranging (TI between 3.9 and 40.0%). Across all AM fungal isolates, resistance was associated with high rates of colonization and low tissue Al concentrations of broomsedge plants. The functional diversity in Al resistance displayed by these AM fungi reflect variation in acclimation mechanisms operating in the mycorrhizal symbiosis under environmental stress.

Opposing effects of aluminum on inward-rectifier potassium currents in bean root-tip protoplasts.

Inward currents in root cap protoplasts of the aluminum-tolerant cultivar, Dade, of Phaseolus vulgaris L. were investigated using the whole-cell patch-clamp technique. The properties of these currents were similar to those seen in inward rectifying K+ channels in other plant tissues. Replacing bath K+ with Na+ nearly abolished the observed currents. Higher bath K+ concentrations increased inward currents. AlCl3 in pH 4.7 bath solutions caused inward K+ currents to activate more rapidly and at more positive voltages when compared with AlCl3 free solutions. In 10 microM AlCl3 the activated inward K+ currents were significantly larger than in the AlCl3-free solution at all voltages except at the most negative voltage of -174 mV and the least negative of -74 mV. In contrast, in 80 microM Al3+, when hyperpolarizing voltages were most negative, the inward K+ currents were inhibited relative to the currents in 10 microM AlCl3. Enhancement of inward K+ currents by AlCl3 is consistent with Al3+ binding to the external surface of the root cap protoplast, decreasing the surface charge, thus causing the channels to sense a more negative membrane potential. Inhibition of inward K+ currents with higher AlCl3 concentrations and more negative voltages is consistent with Al3+ block of K+ channels.

Arbuscular mycorrhizal fungi alter phosphorus relations of broomsedge (Andropogon virginicus L.) plants.

Broomsedge (Andropogon virginicus L.) is a dominant grass revegetating many abandoned coal-mined lands in West Virginia, USA. Residual soils on such sites are often characterized by low pH, low nutrients, and high aluminium. Experiments were conducted to assess the resistance of broomsedge to limited phosphorus (Pi) availability and to investigate the role that arbuscular mycorrhizal (AM) fungi play in aiding plant growth under low Pi conditions. Pregerminated mycorrhizal and non-mycorrhizal seedlings were grown in a sand-culture system with nutrient solutions containing Pi concentrations ranging from 10 to 100 microM for 8 weeks. Non-mycorrhizal plants exhibited severe inhibition of growth under Pi limitation (<60 microM). Colonization by AM fungi (combined Glomus clarum Nicolson & Schenck and Gigaspora gigantea (Nicol. & Gerd.) Gerd. & Trappe) greatly enhanced host plant growth at low Pi concentrations, but did not benefit growth when Pi was readily available (100 microM). In comparison to non-mycorrhizal plants, mycorrhizal plants had higher phosphorus use efficiency at low Pi concentrations and maintained nearly constant tissue nutrient concentrations across the gradient of Pi concentrations investigated. Manganese (Mn) and sodium (Na) accumulated in shoots of non-mycorrhizal plants under Pi limitation. Mycorrhizal plants exhibited lower instantaneous Pi uptake rates and significantly lower C(min) values compared to non-mycorrhizal plants. These patterns suggest that the symbiotic association between broomsedge roots and AM fungi effectively maintains nutrient homeostasis through changes in physiological properties, including nutrient uptake, allocation and use. The mycorrhizal association is thus a major adaptation that allows broomsedge to become established on infertile mined lands.

Acid mist and soil Ca and Al alter the mineral nutrition and physiology of red spruce.

We examined the effects and potential interactions of acid mist and soil solution Ca and Al treatments on foliar cation concentrations, membrane-associated Ca (mCa), ion leaching, growth, carbon exchange, and cold tolerance of red spruce (Picea rubens Sarg.) saplings. Soil solution Ca additions increased foliar Ca and Zn concentrations, and increased rates of respiration early in the growing season (July). Soil Al treatment had a broad impact, reducing foliar concentrations of Ca, Mg, Mn, P and Zn, and resulting in smaller stem diameters, sapling heights and shoot lengths compared with soil treatments with no added Al. Aluminum treatment also reduced respiration when shoots were elongating in July and decreased net photosynthesis at the end of the growing season (September). Three lines of evidence suggest that Al-induced alterations in growth and physiology were independent of foliar Ca status: (1) Ca concentrations in foliage of Al-treated saplings were within the range of sufficiency established for red spruce; (2) mCa concentrations were unaffected by Al treatment; and (3) no Al x Ca interactions were detected. Acid mist treatment increased foliar Fe and K concentrations and increased leaching of Ca, Mg, Mn, Zn, Fe, and Al from foliage. Leaching losses of Ca were more than twice those of the element with the next highest amount of leaching (Zn), and probably led to the reductions in mCa concentration and membrane stability of acid-treated saplings. Acidic mist resulted in enhanced shoot growth, and consistent reductions in foliar cold tolerance in the fall and winter. Of the few significant interactions among treatments, most involved the influence of mist pH and Al treatment on foliar nutrition. In general, reductions in cation concentration associated with Al addition were greater for pH 5.0-treated saplings than for pH 3.0-treated saplings. We propose that H(+)-induced leaching of mCa from mesophyll cells is the mechanism underlying acid-induced reductions in foliar cold tolerance of red spruce.

Effects of serpentine soil factors on Virginia pine (Pinus virginiana) seedlings.

Effects of simulated serpentine soil conditions (elevated Mg:Ca ratio and Ni concentration) on seedlings from populations of Virginia pine (Pinus virginiana Mill.) from serpentine and non-serpentine sites were evaluated in sand culture. We determined (1) how seedlings are affected by elevated Mg:Ca ratio and Ni concentrations, (2) if there are interactive effects between Mg:Ca ratio and Ni concentrations on seedling growth, needle pigment concentrations, and nutrition, and (3) if Virginia pine populations from serpentine areas are edaphic ecotypes. A Mg:Ca ratio of 5 and 50 microM Ni both reduced seedling growth compared with control seedlings grown in the presence of the standard Mg:Ca ratio of 0.5 and no Ni. Interactive effects between Mg:Ca ratio and Ni concentrations were highly significant for growth, foliar pigments, and needle and root elemental concentrations. Nickel-mediated reductions in growth and foliar pigment concentrations were less at the serpentine Mg:Ca ratio of 5 than at the standard (non-serpentine) Mg:Ca ratio of 0.5. Foliar N was reduced by Ni concentrations as low as 10 microM, and foliar and root K, Ca and P concentrations were significantly reduced by Ni concentrations above 25 microM, with greater reductions at a Mg:Ca ratio of 0.5 than at a Mg:Ca ratio of 5. There were no population x serpentine soil factor interactions for seedling growth, foliar pigment concentrations, or nutrition, suggesting that seedlings from trees growing on serpentine soils are not edaphic ecotypes. We conclude that serpentine conditions present at the site of seed collection have not resulted in the selection of edaphic ecotypes of Virginia pine with respect to Mg:Ca ratio and Ni concentration.

Physiological implications of seasonal variation in membrane-associated calcium in red spruce mesophyll cells.

We examined the pattern of seasonal variation in total foliar calcium (Ca) pools and plasma membrane-associated Ca (mCa) in mesophyll cells of current-year and 1-year-old needles of red spruce (Picea rubens Sarg.) and the relationship between mCa and total foliar Ca on an individual plant and seasonal basis. Foliar samples were collected from seedlings and analyzed on 16 dated at 2- to 3-week intervals between June 1994 and March 1995. Concentrations of mCa in current-year needles were more seasonally dynamic and responsive to temporal environmental changes than either mCa concentrations of 1-year-old needles, which were largely stable, or total foliar Ca concentrations in both tissues. In current-year needles, mCa was barely evident in early summer, increased steadily through summer, and then increased dramatically in early fall and surpassed the concentration in 1-year-old needles. Coincident with the first severe frost, mCa concentrations in current-year needles declined significantly and subsequently maintained concentrations comparable to those of 1-year-old needles. Following an extended January thaw, which included 5 days of minimum temperatures > 5 degrees C, mCa concentrations of current-year needles temporarily, but significantly, declined. However, there was no change in mCa concentrations of 1-year-old needles or total Ca concentrations of either tissue. Total Ca concentrations were stable through midsummer in both tissues, doubled in late summer, and then were stable in both tissues throughout fall and winter. Total Ca concentrations were consistently higher in 1-year-old than in current-year needles. Correlations between concentrations of mCa and total foliar Ca were consistently low and mostly nonsignificant. Thus, the dominant, but insoluble, extracellular Ca pool reflected in commonly measured total foliar Ca concentrations is not a meaningful surrogate for the physiologically important and labile pool associated with the plasma membrane-cell wall compartment of red spruce mesophyll cells. It is likely that shifts in the critical mCa compartment would not be detected by analysis of total foliar Ca pools. Seasonal changes in mCa concentration seemed to parallel seasonal changes in membrane structure, and possibly the important role of extracellular Ca in transducing messages associated with environmental signals.

Effects of elevated nitrate and aluminum on the growth and nutrition of red spruce (Picea rubens) seedlings.

Acidic deposition in high-elevation forests in the Appalachian Mountains of the eastern United States has been implicated in the decline of red spruce (Picea rubens Sarg.). Elevated soil acidity may increase soil Al availability and toxicity to roots. Enhanced soil solution NO(3) (-) concentrations, resulting from precipitation inputs and enhanced soil organic matter mineralization, may exacerbate Al toxicity by increasing root Al uptake. We exposed red spruce seedlings to 350, 500, 800 or 1400 micro M NO(3) (-) and 0 or 200 micro M Al in a factorial design in sand-nutrient solution culture to test if increased NO(3) (-) concentrations enhance Al uptake and toxicity. In addition to significant reductions in seedling growth parameters resulting from Al exposure, we found significant interactions between NO(3) (-) and Al for seedling height growth rate, needle weight, shoot weight and root weight. Differences in these parameters between Al treatments became more pronounced as solution NO(3) (-) concentration increased and reflected an Al-mediated inhibition of seedling response to increasing NO(3) (-) concentration. Solution NO(3) (-) concentrations above 500 micro M induced root nitrate reductase (NR) activity, whereas shoot NR activity increased in response to NO(3) (-) up to 500 micro M and declined above that concentration. In contrast, exposure to Al depressed NR activity of roots but tended to stimulate needle NR activity. Foliar N concentrations increased in seedlings grown in cultures containing between 350 and 500 micro M NO(3) (-), with no change above 500 micro M. Increasing concentrations of NO(3) (-) depressed foliar P concentrations, with reductions being greatest in seedlings exposed to 1400 micro M NO(3) (-). Exposure to Al increased foliar Ca, K and Al concentrations, decreased foliar P concentrations, and inhibited increases in foliar Mg concentration in response to increasing NO(3) (-). The consistent interactions between NO(3) (-) and Al for growth, root NR activity and foliar Mg concentration were the result of an inhibition of seedling response to NO(3) (-) mediated by Al in solution, rather than enhanced Al toxicity resulting from growth in the presence of elevated NO(3) (-) concentrations.

Growth and nutrition of nonmycorrhizal and mycorrhizal pitch pine (Pinus rigida) seedlings under phosphorus limitation.

The association of ectomycorrhizal fungi with tree roots enhances the acquisition of phosphorus (P) from the soil. In addition to increasing the uptake of H(2)PO(4) (-) (P(i)), mycorrhizal fungi may increase the spectrum of P sources utilized by tree roots by mediating the dissolution of insoluble metallophosphate salts or the hydrolysis of organic P compounds. To investigate the role of ectomycorrhizal fungi in enhancing P acquisition by tree roots, pitch pine (Pinus rigida Mill.) seedlings were grown in sand culture with or without the ectomycorrhizal symbiont Pisolithus tinctorius Coker and Couch under various conditions of P limitation. Compared with nonmycorrhizal seedlings, seedlings inoculated with P. tinctorius exhibited a greater capacity to function under P limitation as evidenced by superior growth and the maintenance of normal foliar ion composition at low P(i) concentrations. Nonmycorrhizal seedlings subjected to P-limiting conditions exhibited depressed K and P and elevated Na concentrations in foliage. The association of P. tinctorius with pitch pine seedling roots maintained foliar K concentrations and prevented the accumulation of Na under P limitation. Nonmycorrhizal seedlings were unable to obtain P from either solid AlPO(4) or inositol hexaphosphate (IHP), whereas seedlings inoculated with P. tinctorius utilized AlPO(4), but not IHP as a P source. Root surface acid phosphatase (APase) activity was depressed in roots infected with the mycorrhizal symbiont and was negatively correlated with seedling growth on all P sources. Root APase activity was negatively correlated with foliar P concentrations in seedlings grown on P(i), but was not correlated with foliar P concentrations in seedlings cultured with AlPO(4) or IHP.

Treatment of acute pancreatitis with fresh frozen plasma.

A consecutive series of 239 patients experiencing 336 separate attacks of acute pancreatitis have been treated with fresh frozen plasma during the first 5 days of the illness. Using established criteria, 96 attacks occurring in 69 patients were judged to be severe at the time of admission. The overall mortality was 3.7 per cent per patient population and 2.7 per cent per total number of separate attacks. Four of 69 patients with severe pancreatitis (5.8 per cent) have died and the mortality rate for the 96 severe attacks was 4.2 per cent. No complications which could be directly attributed to fresh frozen plasma were encountered in the series. The results of this prospective pilot study with naturally occurring human proteinase inhibitors have been encouraging and indicate the need for randomized clinical trials.