Continuing Assessment of the 5-Day Sodium Carbonate-Ammonium Nitrate Extraction Assay as an Indicator Test for Silicon Fertilizers.

The 5-day sodium carbonate-ammonium nitrate extraction assay (5-day method) has been recognized by the American Association of Plant Food Control Officials as a validated test method to identify fertilizers or beneficial substances that provide plant-available silicon (Si). The test method used the molybdenum blue colorimetric assay to quantify percentage Si; however, laboratories may use inductively coupled plasma optical emission spectroscopy (ICP-OES) for elemental analysis. To examine the use of either colorimetric or ICP-OES methods for Si determination, the 5-day method was performed on the following Si-containing compounds; wollastonite, sand, biochar, and a basic oven furnace (BOF) slag. Grow-out studies using Zinnia elegans were also performed using varying rates of the wollastonite, biochar, and BOF slag. Our results show using the 5-day method, wollastonite had the highest extracted amounts of silicic acid (H4SiO4) at 4% followed by biochar (2%), BOF slag (1%), and sand (0%). Extraction values calculated using either the molybdenum blue colorimetric assay or ICP-OES for detection of the H4SiO4 had a significant correlation, supporting the application of either detection method for this type of analysis. However, when extracted values were compared to amounts of Si taken up by the plants, the 5-day method overestimated both wollastonite and biochar. While this method is a valid indicator test for determining a soluble Si source, other plant species and methods should be perused to potentially provide more quantitative analyses for plant-available Si content of all materials.

Impact of a short-term heat event on C and N relations in shoots vs. roots of the stress-tolerant C4 grass, Andropogon gerardii.

Global warming will increase heat waves, but effects of abrupt heat stress on shoot-root interactions have rarely been studied in heat-tolerant species, and abrupt heat-stress effects on root N uptake and shoot C flux to roots and soil remains uncertain. We investigated effects of a high-temperature event on shoot vs. root growth and function, including transfer of shoot C to roots and soil and uptake and translocation of soil N by roots in the warm-season drought-tolerant C4 prairie grass, Andropogon gerardii. We heated plants in the lab and field (lab=5.5days at daytime of 30+5 or 10°C; field=5days at ambient (up to 32°C daytime) vs. ambient +10°C). Heating had small or no effects on photosynthesis, stomatal conductance, leaf water potential, and shoot mass, but increased root mass and decreased root respiration and exudation per g. (13)C-labeling indicated that heating increased transfer of recently-fixed C from shoot to roots and soil (the latter likely via increased fine-root turnover). Heating decreased efficiency of N uptake by roots (uptake/g root), but did not affect total N uptake or the transfer of labeled soil (15)N to shoots. Though heating increased soil temperature in the lab, it did not do so in the field (10cm depth); yet results were similar for lab and field. Hence, acute heating affected roots more than shoots in this stress-tolerant species, increasing root mass and C loss to soil, but decreasing function per g root, and some of these effects were likely independent of direct effects from soil heating.

Proteomic analysis of Arabidopsis thaliana leaves in response to acute boron deficiency and toxicity reveals effects on photosynthesis, carbohydrate metabolism, and protein synthesis.

Boron (B) stress (deficiency and toxicity) is common in plants, but as the functions of this essential micronutrient are incompletely understood, so too are the effects of B stress. To investigate mechanisms underlying B stress, we examined protein profiles in leaves of Arabidopsis thaliana plants grown under normal B (30 µM), compared to plants transferred for 60 and 84 h (i.e., before and after initial visible symptoms) in deficient (0 µM) or toxic (3 mM) levels of B. B-responsive polypeptides were sequenced by mass spectrometry, following 2D gel electrophoresis, and 1D gels and immunoblotting were used to confirm the B-responsiveness of some of these proteins. Fourteen B-responsive proteins were identified, including: 9 chloroplast proteins, 6 proteins of photosynthetic/carbohydrate metabolism (rubisco activase, OEC23, photosystem I reaction center subunit II-1, ATPase δ-subunit, glycolate oxidase, fructose bisphosphate aldolase), 6 stress proteins, and 3 proteins involved in protein synthesis (note that the 14 proteins may fall into multiple categories). Most (8) of the B-responsive proteins decreased under both B deficiency and toxicity; only 3 increased with B stress. Boron stress decreased, or had no effect on, 3 of 4 oxidative stress proteins examined, and did not affect total protein. Hence, our results indicate relatively early specific effects of B stress on chloroplasts and protein synthesis.

Effects of CO2 on the tolerance of photosynthesis to heat stress can be affected by photosynthetic pathway and nitrogen.

PREMISE OF THE STUDY: Determining effects of elevated CO2 and N on photosynthetic thermotolerance is critical for predicting plant responses to global warming. METHODS: We grew Hordeum vulgare (barley, C3) and Zea mays (corn, C4) at current or elevated CO2 (370, 700 ppm) and limiting or optimal soil N (0.5, 7.5 mmol/L). We assessed thermotolerance of net photosynthesis (Pn), photosystem II efficiency in the light (Fv'/Fm'), photochemical quenching (qp), carboxylation efficiency (CE), and content of rubisco activase and major heat-shock proteins (HSPs). KEY RESULTS: For barley, elevated CO2 had no effect on Pn, qp, and CE at both high and low N and only a positive effect on Fv'/Fm' at high N. However, for corn, Pn, Fv'/Fm', qp, and CE were decreased substantially by elevated CO2 under high and low N, with greater decreases at high N for all but qp. The negative effects of high CO2 during heat stress on photosynthesis were correlated with rubisco activase and HSPs content, which decreased with heat stress, especially for low-N corn. CONCLUSION: These results indicate that stimulatory effects of elevated CO2 at normal temperatures on photosynthesis and growth (only found for high-N barley) may be partly offset by neutral or negative effects during heat stress, especially for C4 species. Thus, CO2 and N effects on photosynthetic thermotolerance may contribute to changes in plant productivity, distribution, and diversity in future.

Effect of volumetric water content and clover (Trifolium incarnatum) on the survival of Escherichia coli O157:H7 in a soil matrix.

Studies aimed at understanding Escherichia coli O157:H7 soil survival dynamics are paramount due to their inevitable introduction into the organic vegetable production systems via animal manure-based fertilizer. Therefore, a greenhouse study was conducted to determine the survival of E. coli O157:H7 in highly controlled soil matrices subjected to two variable environmental stressors: (1) soil volumetric water content (25 or 45 % VWC), and (2) the growth of clover (planted or unplanted). During the 7-week study, molecular-based qPCR analyses revealed that E. coli O157:H7 survival was significantly lower in soils maintained at either near water-holding capacity (45 % VWC) or under clover growth. The significant reduction under clover growth was only observed when E. coli populations were determined relative to all bacteria, indicating the need to further study the competition between E. coli O157:H7 and the total bacterial community in organic soils. Given the significant effect of clover on E. coli O157:H7 survival under different moisture conditions in this greenhouse-based study, this work highlights the antimicrobial potential of clover exudates in arable soils, and future work should concentrate on their specific mechanisms of inhibition; ultimately leading to the development of crop rotations/production systems to improve pre-harvest food safety and security in minimally processed, ready-to-eat and organic production systems.

Silicon delays Tobacco ringspot virus systemic symptoms in Nicotiana tabacum.

Soluble silicon (Si) provides protection to plants against a variety of abiotic and biotic stress. However, the effects of Si on viral infections are largely unknown. To investigate the role of Si in viral infections, hydroponic studies were conducted in Nicotiana tabacum with two pathogens: Tobacco ringspot virus (TRSV) and Tobacco mosaic virus (TMV). Plants grown in elevated Si showed a delay in TRSV systemic symptom formation and a reduction in symptomatic leaf area, compared to the non-supplemented controls. TRSV-infected plants showed significantly higher levels of foliar Si compared to mock-inoculated plants. However, the Si effect appeared to be virus-specific, since the element did not alter TMV symptoms nor did infection by this virus alter foliar Si levels. Hence, increased foliar Si levels appear to correlate with Si-modulated protection against viral infection. This is all the more intriguing since N. tabacum is classified as a low Si accumulator.

Rare excitatory amino acid from flowers of zonal geranium responsible for paralyzing the Japanese beetle.

The Japanese beetle (JB), Popillia japonica, exhibits rapid paralysis after consuming flower petals of zonal geranium, Pelargonium x hortorum. Activity-guided fractionations were conducted with polar flower petal extracts from P. x hortorum cv. Nittany Lion Red, which led to the isolation of a paralysis-inducing compound. High-resolution-MS and NMR ((1)H, (13)C, COSY, heteronuclear sequential quantum correlation, heteronuclear multiple bond correlation) analysis identified the paralytic compound as quisqualic acid (C(5)H(7)N(3)O(5)), a known but rare agonist of excitatory amino acid receptors. Optical rotation measurements and chiral HPLC analysis determined an L-configuration. Geranium-derived and synthetic L-quisqualic acid demonstrated the same positive paralytic dose-response. Isolation of a neurotoxic, excitatory amino acid from zonal geranium establishes the phytochemical basis for induced paralysis of the JB, which had remained uncharacterized since the phenomenon was first described in 1920.

A model of canopy photosynthesis incorporating protein distribution through the canopy and its acclimation to light, temperature and CO2.

BACKGROUND AND AIMS: The distribution of photosynthetic enzymes, or nitrogen, through the canopy affects canopy photosynthesis, as well as plant quality and nitrogen demand. Most canopy photosynthesis models assume an exponential distribution of nitrogen, or protein, through the canopy, although this is rarely consistent with experimental observation. Previous optimization schemes to derive the nitrogen distribution through the canopy generally focus on the distribution of a fixed amount of total nitrogen, which fails to account for the variation in both the actual quantity of nitrogen in response to environmental conditions and the interaction of photosynthesis and respiration at similar levels of complexity. MODEL: A model of canopy photosynthesis is presented for C(3) and C(4) canopies that considers a balanced approach between photosynthesis and respiration as well as plant carbon partitioning. Protein distribution is related to irradiance in the canopy by a flexible equation for which the exponential distribution is a special case. The model is designed to be simple to parameterize for crop, pasture and ecosystem studies. The amount and distribution of protein that maximizes canopy net photosynthesis is calculated. KEY RESULTS: The optimum protein distribution is not exponential, but is quite linear near the top of the canopy, which is consistent with experimental observations. The overall concentration within the canopy is dependent on environmental conditions, including the distribution of direct and diffuse components of irradiance. CONCLUSIONS: The widely used exponential distribution of nitrogen or protein through the canopy is generally inappropriate. The model derives the optimum distribution with characteristics that are consistent with observation, so overcoming limitations of using the exponential distribution. Although canopies may not always operate at an optimum, optimization analysis provides valuable insight into plant acclimation to environmental conditions. Protein distribution has implications for the prediction of carbon assimilation, plant quality and nitrogen demand.

Influence of silicon on resistance of Zinnia elegans to Myzus persicae (Hemiptera: Aphididae).

Studies were conducted to examine the effect of treating Zinnia elegans Jacq. with soluble silicon on the performance of the green peach aphid, Myzus persicae (Sulzer). Z. elegans plants were irrigated every 2 d throughout the duration of the experiment with a nutrient solution amended with potassium silicate (K2SiO2), or a nutrient solution without K2SiO2. Length of the prereproductive period and survivorship of M. persicae were not affected by K2SiO2 treatment, but total cumulative fecundity and the intrinsic rate of increase (r(m)) were slightly reduced on Z. elegans plants receiving soluble silicon. Quantification of silicon content in leaf tissues using inductively coupled plasma optical emission spectroscopy (ICP-OES) confirmed significantly higher silicon concentrations in plants treated with K2SiO2 compared with control plants. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis was used to identify and quantify phenolic acids and flavonols in leaf tissue of Z. elegans. Compared with untreated control plants, significant elevations in 5-caffeoylquinic acid, p-coumaroylquinic acid, and rutin were detected in leaves of Z. elegans plants treated with K2SiO2, but none of seven other phenolics were significantly affected. Similarly, a slight elevation in guaiacol peroxidase activity was detected in plants treated with K2SiO2 Overall, these results indicate treatment of Z. elegans with soluble silicon provides a modest increase in resistance levels to M. persicae, which may be caused in part by defense-related compounds.

Interactive effects of elevated CO2 and ozone on leaf thermotolerance in field-grown Glycine max.

Humans are increasing atmospheric CO2, ground-level ozone (O3), and mean and acute high temperatures. Laboratory studies show that elevated CO2 can increase thermotolerance of photosynthesis in C3 plants. O3-related oxidative stress may offset benefits of elevated CO2 during heat-waves. We determined effects of elevated CO2 and O3 on leaf thermotolerance of field-grown Glycine max (soybean, C3). Photosynthetic electron transport (et) was measured in attached leaves heated in situ and detached leaves heated under ambient CO2 and O3. Heating decreased et, which O3 exacerbated. Elevated CO2 prevented O3-related decreases during heating, but only increased et under ambient O3 in the field. Heating decreased chlorophyll and carotenoids, especially under elevated CO2. Neither CO2 nor O3 affected heat-shock proteins. Heating increased catalase (except in high O3) and Cu/Zn-superoxide dismutase (SOD), but not Mn-SOD; CO2 and O3 decreased catalase but neither SOD. Soluble carbohydrates were unaffected by heating, but increased in elevated CO2. Thus, protection of photosynthesis during heat stress by elevated CO2 occurs in field-grown soybean under ambient O3, as in the lab, and high CO2 limits heat damage under elevated O3, but this protection is likely from decreased photorespiration and stomatal conductance rather than production of heat-stress adaptations.

Night temperature has a minimal effect on respiration and growth in rapidly growing plants.

BACKGROUND AND AIMS: Carbon gain depends on efficient photosynthesis and adequate respiration. The effect of temperature on photosynthetic efficiency is well understood. In contrast, the temperature response of respiration is based almost entirely on short-term (hours) measurements in mature organisms to develop Q(10) values for maintenance and whole-plant respiration. These Q(10) values are then used to extrapolate across whole life cycles to predict the influence of temperature on plant growth. METHODS: In this study, night temperature in young, rapidly growing plant communities was altered from 17 to 34 degrees C for up to 20 d. Day temperature was maintained at 25 degrees C. CO(2) gas-exchange was continuously monitored in ten separate chambers to quantify the effect of night-temperature on respiration, photosynthesis and the efficiency of carbon gain (carbon use efficiency). KEY RESULTS: Respiration increased only 20-46 % for each 10 degrees C rise in temperature (total respiratory Q(10) of between 1.2 to about 1.5). This change resulted in only a 2-12 % change in carbon use efficiency, and there was no effect on cumulative carbon gain or dry mass. No acclimation of respiration was observed after 20 d of treatment. CONCLUSIONS: These findings indicate that whole-plant respiration of rapidly growing plants has a small sensitivity to temperature, and that the sensitivity does not change among the species tested, even after 20 d of treatment. Finally, the results support respiration models that separate respiration into growth and maintenance components.

Exploring the limits of crop productivity: beyond the limits of tipburn in lettuce.

The productivity of lettuce in a combination of high light, high temperature, and elevated CO2 has not been commonly studied because rapid growth usually causes a calcium deficiency in meristems called tipburn, which greatly reduces quality and marketability. We eliminated tipburn by blowing air directly onto the meristem, which allowed us to increase the photosynthetic photon flux (PPF) to 1000 micromoles m-2 s-1 (57.6 mol m-2 d-1); two to three times higher than normally used for lettuce. Eliminating tipburn doubled edible yield at the highest PPF level. In addition to high PPF, CO2 was elevated to 1200 micromoles m-2 mol-1, which increased the temperature optimum from 25 to 30 degrees C. The higher temperature increased leaf expansion rate, which improved radiation capture and more than doubled yield. Photosynthetic efficiency, measured as canopy quantum yield in a whole-plant gas exchange system, steadily increased up to the highest temperature of 32 degrees C in high CO2. The highest productivity was 19 g m-2 d-1 of dry biomass (380 g d-1 fresh mass) averaged over the 23 days the plants received light. Without the limitation of tipburn, the combination of high PPF, high temperature, and elevated CO2 resulted in a 4-fold increase in growth rate over productivity in conventional environments.

Anaerobic conditions improve germination of a gibberellic acid deficient rice.

Dwarf plants are useful in research because multiple plants can be grown in a small area. Rice (Oryza sativa L.) is especially important since its relatively simple genome has recently been sequenced. We are characterizing a gibberellic acid (GA) mutant of rice (japonica cv 'Shiokari,' line N-71) that is extremely dwarf (20 cm tall). Unfortunately, this GA mutation is associated with poor germination (70%) under aerobic conditions. Neither exogenous GA nor a dormancy-breaking heat treatment improved germination. However, 95% germination was achieved by germinating the seeds anaerobically, either in a pure N2 environment or submerged in unstirred tap water. The anaerobic conditions appear to break a mild post-harvest dormancy in this rice cultivar.