Diverse ALS mutations and cross-and multiple-resistance to ALS and EPSPS inhibitors in flucarbazone­sodium-resistant Bromus japonicus populations from Hebei province, China.

Japanese brome (Bromus japonicus) has become one of the main weeds in wheat fields in Hebei province of China and causes a large decrease of wheat production. A total of 44 putative resistant and 2 susceptible Japanese brome populations were collected in the 2021/2022 crop season from Hebei province of China to determine resistance levels to flucarbazone­sodium and to investigate the diversity of acetolactate synthase (ALS) mutations, as well as to confirm the cross-and multiple-resistance levels to ALS and EPSPS (5-enolpyruvate shikimate-3-phosphate synthetase) inhibitors. Whole plant bioassay results showed that 15 out of 44 populations tested or 34% were resistant to flucarbazone­sodium. The resistance indices of Japanese brome to flucarbazone­sodium ranged from 43 to 1977. The resistant populations were mainly distributed in Baoding and Shijiazhuang districts, and there was only one resistant population in Langfang district. Resistant Japanese brome had diverse ALS mutations, including Pro-197-Ser, -Thr, -Arg and Asp-376-Glu. The incidence of Pro-197-Ser mutation was the highest at 68%. Application of the CYP450 inhibitor malathion suggested that CYP450 was involved in metabolic resistance in a population without an ALS mutation. The population with Pro-197-Thr mutation evolved weak cross-resistance to mesosulfuron-methyl and pyroxsulam, and it is in the process of evolving multiple-resistance to glyphosate.

Biochemical and genetic polymorphism of Bromopsis inermis populations under chronic radiation exposure.

MAIN CONCLUSION: For the subsequent assessment of the genetic mechanisms responsible for the resistance of plants to chronic irradiation, the analysis of RAPD-cDNA with the subsequent isolation, cloning, and sequencing of expressed polymorphic sequences is a promising technique. A study was conducted on Bromopsis inermis populations that have been growing for a long time in the EURT area. Using RAPD primers, we studied the genetic spectra of plants. In analysing the UPGMA algorithm, we identified two well-distinguishable clusters with a high level of bootstrap support (> 85%): background samples hit the first, and impact samples hit the second. Our data indicate a decrease in diversity in the most polluted population, as well as the appearance of new alleles in chronically irradiated samples of the B. inermis. Smooth brome seedlings were characterised by the content of anthocyanins, comparable with other types of cereals. In the gradient of chronic irradiation, the relative content of anthocyanins was not significantly changed. For the first time, the partial nucleotide sequences of the key genes of anthocyanin biosynthesis (Chi and F3h) in the brome were determined, these sequences were found to be 191 and 356 bp in length, respectively, and were cloned and sequenced. Three copies of the Chi gene were identified in the B. inermis genome. One copy (BiChi-1) clustered with the sequences of the Aegilops tauschii gene (D genome), and the other two copies (BiChi-2 and BiChi-3) formed a separate cluster in the Pooideae subfamily adjacent to Hordeum vulgare. In the copy of BiChi-1, a complete deletion of intron 1 was detected. For the F3h gene, one copy of the B. inermis gene was obtained, which forms a separate branch in the subfamily Pooideae.

Remediation of soil polluted with HMW-PAHs by alfalfa or brome in combination with fungi and starch.

High-molecular weight polycyclic aromatic hydrocarbons (HMW-PAHs) are common pollutants in soil of coal mining areas that affect the safety of crops and the environment. In a pot experiment, we compared the remediation potential of alfalfa (Medicago sativa Linn) and brome (Bromus inermis Leyss.) either alone or in combination with starch or Fusarium sp. strain ZH-H2 for a farmland soil contaminated with 4-6-ring PAHs from a coal mine area. The alfalfa and brome alone treatments reduced the concentrations of most HMW-PAHs. However, when starch was added, the removal rates of indeno(1,2,3-cd)pyrene and benzo(ghi)perylene were significantly higher for brome than for alfalfa. When ZH-H2 was combined with brome, benzo(k)fluoranthene, benzo(a)pyrene, indeno(1,2,3-cd)pyrene, and benzo(ghi)perylene degradation rates were significantly enhanced compared with brome alone. In contrast, an antagonistic effect was observed between alfalfa and Fusarium. The brome, starch and ZH-H2 combination resulted in far better removal rates than the alfalfa combination. Maximum removal rates were obtained with the brome + starch + ZH-H2 combination for benzo(k)fluoranthene (42.64%), benzo(a)pyrene (51.01%), indeno(1,2,3-cd)pyrene (62.29%), and benzo(ghi)perylene (74.85%). These removal rates were 829.78%, 182.34%, 46.13%, and 70.94% higher than the equivalent alfalfa combination treatments. The lignin peroxidase activity was significantly increased in the presence of starch, ZH-H2 and brome, consistent with the increased removal rates of HMW-PAHs.

Effects of carbohydrate and nitrogen supplementation on fermentation of cheatgrass () in a dual-flow continuous culture system.

Cheatgrass (CG; ), an introduced winter annual grass, is an aggressive invader of the sagebrush community in the Western United States. Because of its greater flammability, mature CG constitutes a fire hazard leading to repeated wildfires. One fuel-reduction strategy is livestock grazing. The objective of this study was to evaluate the effects of urea, molasses, or a combination of urea and molasses supplementation of a CG-based diet on digestibility, microbial fermentation, bacterial protein synthesis, and nutrient flow using a dual-flow continuous culture system. Eight fermenters were used in a replicate 4 × 4 Latin square design with four 10-d experimental periods. Experimental treatments (DM basis) were 1) forage only (CON), 2) CG plus urea alone (URE; 1.36% urea), 3) CG plus molasses alone (MOL; 15.9% molasses), and 4) CG plus urea and molasses combined (URE+MOL; 1.28% urea plus 19.3% molasses). Each fermenter was fed 72 g/d of DM, and data were analyzed using the GLIMMIX procedure of SAS (SAS Inst. Inc., Cary, NC). The true digestibilities of NDF and ADF were not affected by diets ( > 0.05). Molasses-containing diets had greater true digestibility of OM ( = 0.02). However, true digestibility of CP was increased when molasses was fed alone ( < 0.01). Molasses-containing diets had lower pH ( < 0.01) and greater VFA concentrations ( < 0.01) compared to those of the other diets. The URE+MOL diet resulted in a greater VFA concentration ( < 0.01). Propionate concentration increased ( < 0.01), whereas acetate concentration decreased ( < 0.01) when molasses alone or in combination with urea was added to the diets. Supplying molasses alone resulted in greater ( = 0.03) total branched-chain VFA compared to the other diets. The concentration of NH-N and total N flow increased ( < 0.01) in response to urea supplementation and was greater ( < 0.01) when urea alone was supplemented in the diet. On the other hand, molasses-supplemented diets yielded more non-ammonia N ( < 0.01) and bacterial N ( = 0.04). Supplementation had no effect ( = 0.83) on bacterial efficiency. Results from this study indicate that the addition of urea and molasses in a CG-based diet could improve nutrient supply to animals, notably VFA supply and microbial N supply; however, in the levels tested in this study, it did not improve CG utilization as assessed by NDF digestion.

Relative abundance of and composition within fungal orders differ between cheatgrass (Bromus tectorum) and sagebrush (Artemisia tridentata)-associated soils.

Nonnative Bromus tectorum (cheatgrass) is decimating sagebrush steppe, one of the largest ecosystems in the Western United States, and is causing regional-scale shifts in the predominant plant-fungal interactions. Sagebrush, a native perennial, hosts arbuscular mycorrhizal fungi (AMF), whereas cheatgrass, a winter annual, is a relatively poor host of AMF. This shift is likely intertwined with decreased carbon (C)-sequestration in cheatgrass-invaded soils and alterations in overall soil fungal community composition and structure, but the latter remain unresolved. We examined soil fungal communities using high throughput amplicon sequencing (ribosomal large subunit gene) in the 0-4 cm and 4-8 cm depth intervals of six cores from cheatgrass- and six cores from sagebrush-dominated soils. Sagebrush core surfaces (0-4 cm) contained higher nitrogen and total C than cheatgrass core surfaces; these differences mirrored the presence of glomalin related soil proteins (GRSP), which has been associated with AMF activity and increased C-sequestration. Fungal richness was not significantly affected by vegetation type, depth or an interaction of the two factors. However, the relative abundance of seven taxonomic orders was significantly affected by vegetation type or the interaction between vegetation type and depth. Teloschistales, Spizellomycetales, Pezizales and Cantharellales were more abundant in sagebrush libraries and contain mycorrhizal, lichenized and basal lineages of fungi. Only two orders (Coniochaetales and Sordariales), which contain numerous economically important pathogens and opportunistic saprotrophs, were more abundant in cheatgrass libraries. Pleosporales, Agaricales, Helotiales and Hypocreales were most abundant across all libraries, but the number of genera detected within these orders was as much as 29 times lower in cheatgrass relative to sagebrush libraries. These compositional differences between fungal communities associated with cheatgrass- and sagebrush-dominated soils warrant future research to examine soil fungal community composition across more sites and time points as well as in association with native grass species that also occupy cheatgrass-invaded ecosystems.

The invasive annual cheatgrass increases nitrogen availability in 24-year-old replicated field plots.

Previous studies comparing invaded and non-invaded sites suggest that cheatgrass (Bromus tectorum L.) causes soil N cycling to increase. Unfortunately, these correlative studies fail to distinguish whether cheatgrass caused the differences in N cycling, or if cheatgrass simply invaded sites where N availability was greater. We measured soil C and N concentrations and net and gross N-cycling rates on 24-year-old replicated field plots in a sagebrush-steppe ecosystem that had been plowed, fumigated, and seeded to different plant communities in 1984. Laboratory assays of soil collected throughout the soil profiles (0-60 cm) showed that soil NO3 (-), organic C and N, and net N mineralization, net nitrification, and soil respiration rates were all greater beneath cheatgrass than in sagebrush-perennial grass plots. In surface soils (0-10 cm), field and lab assays on five sampling dates during 2 years showed gross N mineralization, net N mineralization, and net nitrification rates were all faster beneath cheatgrass than in sagebrush-perennial grass plots. Modeling analyses based on soil respiration and gross N-cycling rates suggest that cheatgrass provides soil microbes with lower C:N substrates and that this could explain the faster N-cycling rates beneath cheatgrass. This is the first long-term replicated field study to conclusively show that cheatgrass created greater soil organic N pool sizes and stimulated N-cycling rates compared to similar-aged stands of sagebrush and native perennial grasses. Increased N-cycling rates may represent a positive plant-soil feedback that promotes continued dominance by cheatgrass, even in the absence of soil disturbance or fire.

Contemporary evolution of an invasive grass in response to elevated atmospheric CO(2) at a Mojave Desert FACE site.

Elevated atmospheric CO2 has been shown to rapidly alter plant physiology and ecosystem productivity, but contemporary evolutionary responses to increased CO2 have yet to be demonstrated in the field. At a Mojave Desert FACE (free-air CO2 enrichment) facility, we tested whether an annual grass weed (Bromus madritensis ssp. rubens) has evolved in response to elevated atmospheric CO2 . Within 7 years, field populations exposed to elevated CO2 evolved lower rates of leaf stomatal conductance; a physiological adaptation known to conserve water in other desert or water-limited ecosystems. Evolution of lower conductance was accompanied by reduced plasticity in upregulating conductance when CO2 was more limiting; this reduction in conductance plasticity suggests that genetic assimilation may be ongoing. Reproductive fitness costs associated with this reduction in phenotypic plasticity were demonstrated under ambient levels of CO2 . Our findings suggest that contemporary evolution may facilitate this invasive species' spread in this desert ecosystem.

Nitrogen limitation, 15N tracer retention, and growth response in intact and Bromus tectorum-invaded Artemisia tridentata ssp. wyomingensis communities.

Annual grass invasion into shrub-dominated ecosystems is associated with changes in nutrient cycling that may alter nitrogen (N) limitation and retention. Carbon (C) applications that reduce plant-available N have been suggested to give native perennial vegetation a competitive advantage over exotic annual grasses, but plant community and N retention responses to C addition remain poorly understood in these ecosystems. The main objectives of this study were to (1) evaluate the degree of N limitation of plant biomass in intact versus B. tectorum-invaded sagebrush communities, (2) determine if plant N limitation patterns are reflected in the strength of tracer (15)N retention over two growing seasons, and (3) assess if the strength of plant N limitation predicts the efficacy of carbon additions intended to reduce soil N availability and plant growth. Labile C additions reduced biomass of exotic annual species; however, growth of native A. tridentata shrubs also declined. Exotic annual and native perennial plant communities had divergent responses to added N, with B. tectorum displaying greater ability to use added N to rapidly increase aboveground biomass, and native perennials increasing their tissue N concentration but showing little growth response. Few differences in N pools between the annual and native communities were detected. In contrast to expectations, however, more (15)N was retained over two growing seasons in the invaded annual grass than in the native shrub community. Our data suggest that N cycling in converted exotic annual grasslands of the northern Intermountain West, USA, may retain N more strongly than previously thought.

Variability in C(3)-plant cell-wall biosynthesis in a high-CO(2) atmosphere by solid-state NMR spectroscopy.

We have used a frequency-selective rotational-echo double-resonance (REDOR) solid-state NMR experiment to measure the concentrations of glycine-glycine pairs in proteins (and protein precursors) of intact leaves of plants exposed to both high- and low-CO(2) atomospheres. The results are interpreted in terms of differences in cell-wall biosynthesis between plant species. We illustrate this variability by comparing the assimilation of label in cheatgrass and soybean leaves labeled using (15)N-fertilizer and (13)CO(2) atmospheres. Cheatgrass and soybean are both C(3) plants but differ in their response to a high-CO(2) environment. Based on REDOR results, we determined that cheatgrass (a plant that seems likely to flourish in future low-water, high-CO(2) environments) routes 2% of the assimilated carbon label that remains in the leaf after 1 h in a 600-ppm (13)CO(2) atmosphere to glycine-rich protein (or its precursors), a structural component of cell walls cross-linked to lignins. In contrast, soybean under the same conditions routes none of its assimilated carbon to glycine-rich protein.

Feeding distillers grains supplements to improve amino acid nutriture of lambs consuming moderate-quality forages.

We hypothesized that providing dried distillers grains with solubles (DDGS) would improve the N retention and use of nutrients by wethers fed a moderate-quality bromegrass hay. Additionally, we hypothesized that treatment effects on nutrient fluxes would be similar after 3, 6, or 9 wk on treatment. Chronic indwelling catheters were surgically implanted in a mesenteric artery, mesenteric vein, hepatic vein, and portal vein of 9 Suffolk x Dorset wethers (initial BW +/- SD = 57.4 +/- 6.1 kg). Wethers had ad libitum access to moderate-quality bromegrass hay (8.44% CP, DM basis) and received 100 g/d of either a corn-based (Corn, n = 4) or a DDGS-based (n = 5) supplement. There was no difference in DMI (P = 0.85) or DM digestibility (P = 0.46) between the 2 groups. There was a numerically greater N intake (21.5 vs. 18.4 g/d; P = 0.14) and N retention (4.4 vs. 2.5 g/d; P = 0.15) when wethers were supplemented with DDGS instead of Corn. Wethers fed DDGS had a greater (P = 0.008) release of alpha-amino N from the portal-drained viscera (PDV, 37.9 mmol/h) than those fed Corn (14.1 mmol/h). Similarly, there was a shift (P = 0.004) from a net splanchnic uptake to a net release of alpha-amino N in wethers fed DDGS (9.1 mmol/h) compared with those fed Corn (-9.6 mmol/h). However, there was no difference in ammonia release from the PDV (P = 0.49) or hepatic release of urea-N (P = 0.19) between the 2 treatments. There were very limited interactions between nutrient fluxes and the length of time after the initiation of treatments. However, there was a tendency (interaction, P = 0.07) for the PDV release of alpha-amino N to be greater at 6 and 9 wk after the initiation of the treatments than after 3 wk on treatment for wethers fed DDGS, although there was no difference over time for wethers fed the Corn supplement. Additionally, there were changes in numerous nutrient fluxes between 3 and 6 wk after the initiation of treatments regardless of treatment. These data indicate that DDGS is a viable supplement to enhance the nutriture of ruminants consuming moderate-quality forages. Additionally, these data indicate that the effects are discernible after 3 wk on treatment, with modest alterations in nutrient flux after additional time on treatment.

Bromus tectorum invasion alters nitrogen dynamics in an undisturbed arid grassland ecosystem.

The nonnative annual grass Bromus tectorum has successfully replaced native vegetation in many arid and semiarid ecosystems. Initial introductions accompanied grazing and agriculture, making it difficult to separate the effects of invasion from physical disturbance. This study examined N dynamics in two recently invaded, undisturbed vegetation associations (C3 and C4). The response of these communities was compared to an invaded/ disturbed grassland. The invaded/disturbed communities had higher surface NH4+ input in spring, whereas there were no differences for surface input of NO3-. Soil inorganic N was dominated by NH4+, but invaded sites had greater subsurface soil NO3-. Invaded sites had greater total soil N at the surface four years post-invasion in undisturbed communities, but total N was lower in the invaded/disturbed communities. Soil delta15N increased with depth in the noninvaded and recently invaded communities, whereas the invaded/disturbed communities exhibited the opposite pattern. Enriched foliar delta15N values suggest that Bromus assimilated subsurface NO3-, whereas the native grasses were restricted to surface N. A Rayleigh distillation model accurately described decomposition patterns in the noninvaded communities where soil N loss is accompanied by increasing soil delta15N; however, the invaded/ disturbed communities exhibited the opposite pattern, suggesting redistribution of N within the soil profile. This study suggests that invasion has altered the mechanisms driving nitrogen dynamics. Bromus litter decomposition and soil NO3- concentrations were greater in the invaded communities during periods of ample precipitation, and NO3- leached from the surface litter, where it was assimilated by Bromus. The primary source of N input in these communities is a biological soil crust that is removed with disturbance, and the lack of N input by the biological soil crust did not balance N loss, resulting in reduced total N in the invaded/disturbed communities. Bromus produced a positive feedback loop by leaching NO3- from decomposing Bromus litter to subsurface soil layers, accessing that deepsoil N pool with deep roots and returning that N to the surface as biomass and subsequent litter. Lack of new inputs combined with continued loss will result in lower total soil N, evidenced by the lower total soil N in the invaded/disturbed communities.

Identification and immunogold localization of a novel bromegrass (Bromus inermis Leyss) peroxisome channel protein induced by ABA, cold and drought stresses, and late embryogenesis.

A cDNA (BG-15) was isolated through differential screening of a cDNA library made from an ABA-treated bromegrass (Bromus inermis Leyss) suspension cell culture. The 819 bp pair cDNA encoded a 174 amino acid polypeptide with a calculated molecular mass of 18.08 kD and isolectric point of 7.50. The deduced amino acid sequences for the cDNA were 29.5% and 32.6% homologous to the known amino acid-selective channel proteins of the chloroplastic outer membrane in pea and barley, but were highly homologous (55.6% to 83.2%) to the putative membrane channel proteins from rice and Arabidopsis. Immunogold localization demonstrated that the channel protein encoded by this cDNA was present on the peroxisome membrane. High stringency southern analysis revealed that 1 to 2 copies of the peroxisomal channel protein (PCP) genes were present in the bromegrass genome. Northern and Western blots revealed that the PCP gene was responsive to both cold and drought stresses, and was rapidly induced by ABA (75 microM). The transcript of the PCP gene also accumulated during late embryogenesis, but declined rapidly during germination. Data taken together, responsiveness of the PCP to cold and drought stresses, and accumulation during late embryogenesis suggest this novel peroxisomal channel protein is associated with sugar and fatty acid metabolism through fatty acid import or succinate export from peroxisome during desiccation tolerance and energy metabolism.

Effect of restricted forage intake on ruminal disappearance of bromegrass hay and a blood meal, feather meal, and fish meal supplement.

Two experiments were conducted to determine in situ disappearance of bromegrass hay and a ruminally undegraded protein (RUP) supplement in beef cattle fed restricted amounts of forage. Six Angus crossbred cattle (BW = 589 +/- 44.4 kg; three steers and three heifers) fitted with ruminal cannulas were fed chopped (2.54 cm) bromegrass hay (8.9% CP) at one of three percentages of maintenance intake (30, 55, or 80%; one steer and one heifer per treatment). In both experiments, the cattle were allowed 7 d for diet adaptation followed by 3 d of sample collection. In Exp 1, in situ bags (50 microm pore size) containing 4.1 g of brome-grass hay (OM basis) were inserted into the rumen and subsequently removed at 3, 6, 9, 12, 15, 18, 24, 36, and 48 h after insertion. Nonlinear regression models were used to determine the rapidly solubilized protein Fraction A, the potentially ruminal degradable protein Fraction B, the ruminally undegraded protein Fraction C, and protein degradation rate. Intake level did not affect (P = 0.15 to 0.95) forage protein remaining after in situ incubation or Fractions A, B, and C; however, effective ruminal degradation of hay protein tended to increase quadratically (P = 0.12) as forage intake increased. In Exp 2, 4.2 g (OM basis) of an RUP supplement (6.8% porcine blood meal, 24.5% hydrolyzed feather meal, and 68.7% menhaden fish meal) formulated to provide equal amounts of metabolizable protein across all levels of hay consumption was evaluated in a similar manner as in Exp 1. The undegraded protein fraction of the supplement did not differ (P = 0.16 to 0.74) across treatments at 3, 6, 9, and 18 h; however, increasing forage intake resulted in a linear increase (P < or = 0.06) in undegraded protein remaining at 12, 15, 24, 36, and 48 h. Dietary treatment had no affect (P = 0.30) on protein Fractions A, B, or C; however, protein degradation rate of the supplement decreased linearly (P = 0.03) as forage intake increased. Therefore, effective ruminal degradation of the supplement decreased linearly (P = 0.01) from 50.8 to 40.9% as forage intake increased from 30 to 80% of maintenance. Corresponding estimates of supplement RUP were 49.2, 56.5, and 59.1% for the 30, 55, and 80% of maintenance intake treatments, respectively. Restricting dietary intake can decrease the quantity of dietary protein that escapes ruminal degradation. Tabular estimates of RUP may not be appropriate for formulating diets to balance metabolizable protein in beef cattle consuming limited quantities of forage.

Supplementing a ruminally undegradable protein supplement to maintain essential amino acid supply to the small intestine when forage intake is restricted in beef cattle.

Twelve Angus crossbred cattle (eight heifers and four steers; average initial BW = 594 +/- 44.4 kg) fitted with ruminal and duodenal cannulas and fed restricted amounts of forage plus a ruminally undegradable protein (RUP) supplement were used in a triplicated 4 x 4 Latin square design experiment to determine intestinal supply of essential AA. Cattle were fed four different levels of chopped (2.54 cm) bromegrass hay (11.4% CP, 57% NDF; OM basis): 30, 55, 80, or 105% of the forage intake required for maintenance. Cattle fed below maintenance were given specified quantities of a RUP supplement (6.8% porcine blood meal, 24.5% hydrolyzed feather meal, and 68.7% menhaden fish meal; DM basis) designed to provide duodenal essential AA flow equal to that of cattle fed forage at 105% of maintenance. Experimental periods lasted 21 d (17 d of adaptation and 4 d of sampling). Total OM intake and duodenal OM flow increased linearly (P < 0.001) as cattle consumed more forage; however, OM truly digested in the rumen (% of intake) did not change (P = 0.43) as intake increased. True ruminal N degradation (% of intake) tended (P = 0.07) to increase linearly, and true ruminal N degradation (g/d) decreased quadratically (P = 0.02) as intake increased from 30 to 105%. Duodenal N flow was equal (P = 0.33) across intake levels, even though microbial N flow increased linearly (P < 0.001) as forage OM intake increased. Total and individual essential AA intake decreased (cubic; P < 0.001) as forage intake increased because the supply of nonammonia, nonmicrobial N flow from RUP was decreased (linear; P < 0.001) by design. Total duodenal flow of essential AA did not differ (P = 0.39) across these levels of forage intake. Although the profile of essential AA reaching the duodenum differed (P < or = 0.02) for all 10 essential AA, the range of each essential AA as a proportion of total essential AA was low (11.1 to 11.2% of total essential AA for phenylalanine to 12.3 to 14.3% of total essential AA for lysine). Duodenal essential AA flow did not differ (P = 0.10 to 0.65) with forage intake level for eight of the 10 essential AA. Duodenal flow of arginine decreased linearly (P = 0.01), whereas duodenal flow of tryptophan increased linearly (P = 0.002) as forage intake increased from 30 to 105% of maintenance. Balancing intestinal essential AA supply in beef cattle can be accomplished by varying intake of a RUP supplement.

Abrupt rise in atmospheric CO2 overestimates community response in a model plant-soil system.

Attempts to understand the ecological effect of increasing atmospheric CO2 concentration, [CO2], usually involve exposing today's ecosystems to expected future [CO2] levels. However, a major assumption of these approaches has not been tested--that exposing ecosystems to a single-step increase in [CO2] will yield similar responses to those of a gradual increase over several decades. We tested this assumption on a mycorrhizal fungal community over a period of six years. [CO2] was either increased abruptly, as is typical of most [CO2] experiments, or more gradually over 21 generations. The two approaches resulted in different structural and functional community responses to increased [CO2]. Some fungi were sensitive to the carbon pulse of the abrupt [CO2] treatment. This resulted in an immediate decline in fungal species richness and a significant change in mycorrhizal functioning. The magnitude of changes in fungal diversity and functioning in response to gradually increasing [CO2] was smaller, and not significantly different to those with ambient [CO2]. Our results suggest that studies may overestimate some community responses to increasing [CO2] because biota may be sensitive to ecosystem changes that occur as a result of abrupt increases.

A lipid transfer protein gene BG-14 is differentially regulated by abiotic stress, ABA, anisomycin, and sphingosine in bromegrass (Bromus inermis).

The objective was to investigate the expression of a lipid transfer protein gene (LTP) both in bromegrass (Bromus inermis) cells and seedlings after exposure to abiotic stresses, abscisic acid (ABA), anisomycin, and sphingosine. A full-length cDNA clone BG-14 isolated from bromegrass suspension cell culture encodes a polypeptide of 124 amino acids with typical LTP characteristics, such as a conserved arrangement of cysteine residues. During active stages of cold acclimation LTP expression was up-regulated, whereas at the final stage of cold acclimation LTP transcript level declined to pre-acclimation level. A severe drought stress induced the LTP gene; yet, LTP expression doubled 3 d after re-hydration. Both temperature and heat shock duration influence LTP induction; however temperature is the primary factor. Treatment with NaCl stimulated accumulation of LTP mRNA within 15 min and the transcripts remained at elevated levels for the duration of the salinity stress. Most interestingly, Northern blots showed LTP was rapidly induced not only by ABA, but also by anisomycin and sphingosine in suspension cell cultures. Of the three chemicals, ABA induced the most rapid and highest response in LTP expression as well as highest freezing tolerance, whereas sphingosine was the least active for both LTP expression and freezing tolerance.