Thomas W. Rufty

Arbuscular Mycorrhizal Fungi Increase Organic Carbon Decomposition Under Elevated CO2

A Fungal Culprit to Carbon Loss In some ecosystems, such as in the layer of soil containing plant roots, fungi, and bacteria, increased levels of CO2 should stimulate more efficient aboveground photosynthesis, which in turn should promote increased sequestration of organic carbon in soil through the protective action of arbuscular mycorrhizal fungi. However, in a series of field and microcosm experiments performed under elevated levels of CO2 thought to be consistent with future emissions scenarios, Cheng et al. (p. 1084; see the Perspective by Kowalchuk) observed that these fungi actually promote degradation of soil organic carbon, releasing more CO2 in the process. Counter to expectations, fungi associated with plant roots diminish the carbon pool in soil ecosystems under elevated levels of carbon dioxide. The extent to which terrestrial ecosystems can sequester carbon to mitigate climate change is a matter of debate. The stimulation of arbuscular mycorrhizal fungi (AMF) by elevated atmospheric carbon dioxide (CO2) has been assumed to be a major mechanism facilitating soil carbon sequestration by increasing carbon inputs to soil and by protecting organic carbon from decomposition via aggregation. We present evidence from four independent microcosm and field experiments demonstrating that CO2 enhancement of AMF results in considerable soil carbon losses. Our findings challenge the assumption that AMF protect against degradation of organic carbon in soil and raise questions about the current prediction of terrestrial ecosystem carbon balance under future climate-change scenarios.

Rapid Uptake of Aluminum into Cells of Intact Soybean Root Tips (A Microanalytical Study Using Secondary Ion Mass Spectrometry).

A wide range of physiological disorders has been reported within the first few hours of exposing intact plant roots to moderate levels of Al3+. Past microanalytic studies, largely limited to electron probe x-ray microanalysis, have been unable to detect intracellular Al in this time frame. This has led to the suggestion that Al exerts its effect solely from extracellular or remote tissue sites. Here, freeze-dried cryosections (10 [mu]m thick) collected from the soybean (Glycine max) primary root tip (0.3–0.8 mm from the apex) were analyzed using secondary ion mass spectrometry (SIMS). The high sensitivity of SIMS for Al permitted the first direct evidence of early entry of Al into root cells. Al was found in cells of the root tip after a 30-min exposure of intact roots to 38 [mu]M Al3+. The accumulation of Al was greatest in the first 30 [mu]m, i.e. two to three cell layers, but elevated Al levels extended at least 150 [mu]m inward from the root edge. Intracellular Al concentrations at the root periphery were estimated to be about 70 nmol g-1 fresh weight. After 18 h of exposure, Al was evident throughout the root cross-section, although the rate of accumulation had slowed considerably from that during the initial 30 min. These results are consistent with the hypothesis that early effects of Al toxicity at the root apex, such as those on cell division, cell extension, or nutrient transport, involve the direct intervention of Al on cell function.

Growth, productivity, and competitiveness of introgressed weedy Brassica rapa hybrids selected for the presence of Bt cry1Ac and gfp transgenes

Concerns exist that transgenic crop × weed hybrid populations will be more vigorous and competitive with crops compared with the parental weed species. Hydroponic, glasshouse, and field experiments were performed to evaluate the effects of introgression of Bacillus thuringiensis (Bt) cry1Ac and green fluorescent protein (GFP) transgenes on hybrid productivity and competitiveness in four experimental Brassica rapa × transgenic Brassica napus hybrid generations (F1, BC1F1, BC2F1 and BC2F2). The average vegetative growth and nitrogen (N) use efficiency of transgenic hybrid generations grown under high N hydroponic conditions were lower than that of the weed parent (Brassica rapa, AA, 2n = 20), but similar to the transgenic crop parent, oilseed rape (Brassica napus, AACC, 2n = 38). No generational differences were detected under low N conditions. In two noncompetitive glasshouse experiments, both transgenic and nontransgenic BC2F2 hybrids had on average less vegetative growth and seed production than B. rapa. In two high intraspecific competition field experiments with varied herbivore pressure, BC2F2 hybrids produced less vegetative dry weight than B. rapa. The competitive ability of transgenic and nontransgenic BC2F2 hybrids against a neighbouring crop species were quantified in competition experiments that assayed wheat (Triticum aestivum) yield reductions under agronomic field conditions. The hybrids were the least competitive with wheat compared with parental Brassica competitors, although differences between transgenic and nontransgenic hybrids varied with location. Hybridization, with or without transgene introgression, resulted in less productive and competitive populations.

Nitrogen Transfer Between Plants: A 15N Natural Abundance Study with Crop and Weed Species

An increasing amount of evidence indicates that N can be transferred between plants. Nonetheless, a number of fundamental questions remain. A series of experiments was initiated in the field to examine N transfer between N2-fixing soybean (Glycine max [L.] Merr.) varieties and a non-nodulating soybean, and between N2-fixing peanut (Arachis hypogaea L.) or soybean and neighboring weed species. The experiments were conducted in soils with low N fertilities and used differences in N accumulation and/or 15N natural abundance to estimate N transfer. Mixtures of N2-fixing and non-nod soybean indicated that substantial inter-plant N transfer occurred. Amounts were variable, ranging from negligible levels to 48% of the N found in the non-nod at maturity. Transfer did not appear to strongly penalize the N2-fixing donor plants. But, in cases where high amounts of N were transferred, N content of donors was noticeably lowered. Differences were evident in the amount of N transferred from different N2-fixing donor genotypes. Results of experiments with N2-fixing crops and the weed species prickly sida (Sida spinosa L.) and sicklepod (Senna obtusifolia [L.] Irwin & Barneby) also indicated substantial N transfer occurred over a 60-day period, with amounts accounting for 30–80% of the N present in the weeds. Transfer of N, however, was generally very low in weed species that are known to be non-hosts for arbuscular mycorrhizae (yellow nutsedge, Cyperus esculentus L. and Palmer amaranth, Amaranthus palmeri [S.] Watson). The results are consistent with the view that N transfer occurs primarily through mycorrhizal hyphal networks, and they reveal that N transfer may be a contributing factor to weed problems in N2-fixing crops in low N fertility conditions.

Atmospheric CO2 enrichment facilitates cation release from soil

Atmospheric CO(2) enrichment generally stimulates plant photosynthesis and nutrient uptake, modifying the local and global cycling of bioactive elements. Although nutrient cations affect the long-term productivity and carbon balance of terrestrial ecosystems, little is known about the effect of CO(2) enrichment on cation availability in soil. In this study, we present evidence for a novel mechanism of CO(2)-enhancement of cation release from soil in rice agricultural systems. Elevated CO(2) increased organic C allocation belowground and net H(+) excretion from roots, and stimulated root and microbial respiration, reducing soil redox potential and increasing Fe(2+) and Mn(2+) in soil solutions. Increased H(+), Fe(2+), and Mn(2+) promoted Ca(2+) and Mg(2+) release from soil cation exchange sites. These results indicate that over the short term, elevated CO(2) may stimulate cation release from soil and enhance plant growth. Over the long-term, however, CO(2)-induced cation release may facilitate cation losses and soil acidification, negatively feeding back to the productivity of terrestrial ecosystems.

The Early Entry of Al into Cells of Intact Soybean Roots (A Comparison of Three Developmental Root Regions Using Secondary Ion Mass Spectrometry Imaging)

Al localization was compared in three developmental regions of primary root of an Al-sensitive soybean (Glycine max) genotype using secondary ion mass spectrometry. In cryosections obtained after a 4-h exposure to 38 [mu]M [Al3+], Al had penetrated across the root and into the stele in all three regions. Although the greatest localized Al concentration was consistently at the root periphery, the majority of the Al in each region had accumulated in cortical cells. It was apparent that the secondary ion mass spectrometry 27Al+ mass signal was spread throughout the intracellular area and was not particularly intense in the cell wall. Inclusion of some cell wall in determinations of the Al levels across the root radius necessitated that these serve as minimal estimates for intracellular Al. Total accumulation of intracellular Al for each region was 60, 73, and 210 nmol g-1 fresh weight after 4 h, increasing with root development. Early metabolic responses to external Al, including those that have been reported deep inside the root and in mature regions, might result directly from intracellular Al. These responses might include ion transport events at the endodermis of mature roots or events associated with lateral root emergence, as well as events within the root tip.

Altered aluminum inhibition of soybean root elongation in the presence of magnesium

Variations in genotype rankings among screenings for Al tolerance in hydroponics may be related to differences in the composition of the solutions. In the present study, we investigated the involvement of Mg ions in modifying Al rhizotoxicity in soybeans. Root elongation was strongly inhibited by Al in a simple, 800 μM CaSO4 solution, but elongation increased noticeably when the solutions also contained Mg. Amelioration of Al rhizotoxicity was not associated with an increase in ionic strength of treatment solutions because Al3+ activities were kept constant. Concentration series experiments indicated that the Mg effect occurred in the μM range, while Ca amelioration of Al toxicity occurred at mM concentrations. The positive effect of Mg on root elongation was greatest for Al-sensitive genotypes and minimized genotypic differences for Al-tolerance. The Mg protection against Al rhizotoxicity apparently does not occur with all species, because it was not observed in Atlas and Scout 66 wheat varieties. The ability of Mg to ameliorate Al toxicity in soybean at μM levels suggests the involvement of distinct physiological factors.

Alterations in Soybean Leaf Development and Photosynthesis in a Co2- Enriched Atmosphere

This study was conducted to characterize changes in the canopy photosynthetic leaf area of developing soybean (Glycine max [L.] Merr. cv Lee) exposed to a CO2-enriched atmosphere. Young, vegetative plants were exposed to 350 or 700 μL L-1 CO2 for 15 d. Plant dry mass and total leaf area were greater in the CO2-enriched environment. Emergence and expansion rates of main stem leaves increased at high CO2, but the areas of individual leaves at full expansion were affected very little (5%-10% greater than controls). More rapid leaf expansion rates occurred in the light and dark. Under CO2-enriched conditions, the net CO2 exchange rates of all leaves on the main stem were higher before and after full expansion. Stomatal conductance was lower in high CO2 only after leaves approached full expansion. Leaf development on the lateral branches also was increased at high CO2, accounting for 40% of the total increase in leaf area by the end of the experiment. We conclude that more rapid rates of leaf development under CO2 enrichment likely resulted from increased photosynthesis rates and that both direct and indirect effects were involved.

Spatial and temporal patterns of green fluorescent protein (GFP) fluorescence during leaf canopy development in transgenic oilseed rape, Brassica napus L.

The green fluorescent protein (GFP) holds promise as a field-level transgene marker. One obstacle to the use of GFP is fluorescence variability observed within leaf canopies. In growth chamber and field experiments, GFP fluorescence in transgenic oilseed rape (Brassica napus) was shown to be variable at each leaf position over time and among different leaves on the same plant. A leaf had its highest GFP fluorescence after emergence and, subsequently, its fluorescence intensity decreased. GFP fluorescence intensity was directly correlated with the concentration of soluble protein. The concentration of the genetically linked recombinant Bacillus thuringiensis (Bt) cry1Ac endotoxin protein also was examined, and GFP fluorescence was positively correlated with Bt throughout development. The results show that GFP can be used as an accurate transgene marker but that aspects of plant developmental should be taken into account when interpreting fluorescence measurements.

Growth and Nitrogen Assimilation of Soybeans in Response to Ammonium and Nitrate Nutrition

Plants supplied with moderate concentrations of NH4 + in solution generally grow poorly compared with plants supplied with other sources of nitrogen. Experiments were conducted with a flowing solution culture system to determine whether growth restrictions could be avoided over an extended period in the presence of NH4 + if root-zone pH were strictly controlled and if plants were exposed to NH4 + during exponential growth when carbohydrate fluxes to the root are coordinated with the rate of nitrogen acquisition. Vegetative soybeans (Glycine max [L.] Merrill) initially were exposed to complete nutrient solutions containing NO3 - until the exponential growth stage and then were exposed for 4 wk to solutions in which nitrogen was supplied as either 1.0 mM NH4 +, 1 0 mM NO3 -, or 0.5 mM NH4 + plus 0.5 mM NO3 -. Acidity of the solutions was constantly maintained at pH 5 8 ± 0 1 by automated control In separate experiments, irradiance (photosynthetic photon flux density [PPFD] of 700 and 325 μE m-2 s-1) levels were controlled to produce distinct steady-state rates of leaf, root, and whole-plant growth The source of nitrogen did not alter growth or nitrogen accumulation within either environment Growth of whole plants and plant parts and accumulation of nitrogen remained exponential The results support the conclusion that plants can effectively utilize NH4 + as a nitrogen source as long as root-zone pH is strictly controlled and a balance is maintained between carbohydrate availability and acquisition of NH4 +