Long-term tropospheric ozone pollution disrupts plant-microbe-soil interactions in the agroecosystem.

Tropospheric ozone (O3 ) threatens agroecosystems, yet its long-term effects on intricate plant-microbe-soil interactions remain overlooked. This study employed two soybean genotypes of contrasting O3 -sensitivity grown in field plots exposed elevated O3 (eO3 ) and evaluated cause-effect relationships with their associated soil microbiomes and soil quality. Results revealed long-term eO3 effects on belowground soil microbiomes and soil health surpass damage visible on plants. Elevated O3 significantly disrupted belowground bacteria-fungi interactions, reduced fungal diversity, and altered fungal community assembly by impacting soybean physiological properties. Particularly, eO3 impacts on plant performance were significantly associated with arbuscular mycorrhizal fungi, undermining their contribution to plants, whereas eO3 increased fungal saprotroph proliferation, accelerating soil organic matter decomposition and soil carbon pool depletion. Free-living diazotrophs exhibited remarkable acclimation under eO3 , improving plant performance by enhancing nitrogen fixation. However, overarching detrimental consequences of eO3 negated this benefit. Overall, this study demonstrated long-term eO3 profoundly governed negative impacts on plant-soil-microbiota interactions, pointing to a potential crisis for agroecosystems. These findings highlight urgent needs to develop adaptive strategies to navigate future eO3 scenarios.

Shifts in the Composition and Activities of Denitrifiers Dominate CO2 Stimulation of N2O Emissions.

Elevated atmospheric CO2 (eCO2) often increases soil N2O emissions, but the underlying mechanisms remain largely unknown. One hypothesis suggests that high N2O emissions may stem from increased denitrification induced by CO2 enhancement of plant carbon (C) allocation belowground. However, direct evidence illustrating linkages among N2O emissions, plant C allocation, and denitrifying microbes under eCO2 is still lacking. We examined the impact of eCO2 on plant C allocation to roots and their associated arbuscular mycorrhizal fungi and its subsequent effects on N2O emissions and denitrifying microbes in the presence of two distinct N sources, ammonium nitrogen (NH4+-N) and nitrate nitrogen (NO3--N). Our results showed that the form of the N inputs dominated the effects of eCO2 on N2O emissions: eCO2 significantly increased N2O emissions with NO3--N inputs but had no effect with NH4+-N inputs. eCO2 increased plant biomass N more with NH4+-N than with NO3--N inputs, likely reducing microbial access to available N under NH4+-N inputs and/or contributing to higher N2O emissions under NO3--N inputs. eCO2 enhanced root and mycorrhizal N uptake and also increased N2O emissions under NO3--N inputs. Further, eCO2 enhancement of N2O emissions under NO3--N inputs concurred with a shift in the soil denitrifier community composition in favor of N2O-producing (nirK- and nirS-type) over N2O-consuming (nosZ-type) denitrifiers. Together, these results indicate that eCO2 stimulated N2O emissions mainly through altering plant N preference in favor of NH4+ over NO3- and thus stimulating soil denitrifiers and their activities. These findings suggest that effective management of N sources may mitigate N2O emissions by negating the eCO2 stimulation of soil denitrifying microbes and their activities.

Contrasting Warming and Ozone Effects on Denitrifiers Dominate Soil N2O Emissions.

Nitrous oxide (N2O) in the atmosphere is a major greenhouse gas and reacts with volatile organic compounds to create ozone (an air pollutant) in the troposphere. Climate change factors such as warming and elevated ozone (eO3) affect N2O fluxes, but the direction and magnitude of these effects are uncertain and the underlying mechanisms remain unclear. We examined the impact of simulated warming (control + 3.6 °C) and eO3 (control + 45 ppb) on soil N2O fluxes in a soybean agroecosystem. Results obtained showed that warming significantly increased soil labile C, microbial biomass, and soil N mineralization, but eO3 reduced these parameters. Warming enhanced N2O-producing denitrifers ( nirS- and nirK-type), corresponding to increases in both the rate and sum of N2O emissions. In contrast, eO3 significantly reduced both N2O-producing and N2O-consuming ( nosZ-type) denitrifiers but had no impact on N2O emissions. Further, eO3 offsets the effects of warming on soil labile C, microbial biomass, and the population size of denitrifiers but still increased N2O emissions, indicating a direct effect of temperature on N2O emissions. Together, these findings suggest that warming may promote N2O production through increasing both the abundance and activities of N2O-producing microbes, positively feeding back to the ongoing climate change.

Atmospheric CO2 Enrichment and Reactive Nitrogen Inputs Interactively Stimulate Soil Cation Losses and Acidification.

Reactive N inputs (Nr) may alleviate N-limitation of plant growth and are assumed to help sustain plant responses to the rising atmospheric CO2 (eCO2). However, Nr and eCO2 may elicit a cascade reaction that alters soil chemistry and nutrient availability, shifting the limiting factors of plant growth, particularly in acidic tropical and subtropical croplands with low organic matter and low nutrient cations. Yet, few have so far examined the interactive effects of Nr and eCO2 on the dynamics of soil cation nutrients and soil acidity. We investigated the cation dynamics in the plant-soil system with exposure to eCO2 and different N sources in a subtropical, acidic agricultural soil. eCO2 and Nr, alone and interactively, increased Ca2+ and Mg2+ in soil solutions or leachates in aerobic agroecosystems. eCO2 significantly reduced soil pH, and NH4+-N inputs amplified this effect, suggesting that eCO2-induced plant preference of NH4+-N and plant growth may facilitate soil acidification. This is, to our knowledge, the first direct demonstration of eCO2 enhancement of soil acidity, although other studies have previously shown that eCO2 can increase cation release into soil solutions. Together, these findings provide new insights into the dynamics of cation nutrients and soil acidity under future climatic scenarios, highlighting the urgency for more studies on plant-soil responses to climate change in acidic tropical and subtropical ecosystems.

Quantitative trait loci associated with soybean seed weight and composition under different phosphorus levels.

Seed size and composition are important traits in food crops and can be affected by nutrient availability in the soil. Phosphorus (P) is a non-renewable, essential macronutrient, and P deficiency limits soybean (Glycine max) yield and quality. To investigate the associations of seed traits in low- and high-P environments, soybean recombinant inbred lines (RILs) from a cross of cultivars Fiskeby III and Mandarin (Ottawa) were grown under contrasting P availability environments. Traits including individual seed weight, seed number, and intact mature pod weight were significantly affected by soil P levels and showed transgressive segregation among the RILs. Surprisingly, P treatments did not affect seed composition or weight, suggesting that soybean maintains sufficient P in seeds even in low-P soil. Quantitative trait loci (QTLs) were detected for seed weight, intact pods, seed volume, and seed protein, with five significant QTLs identified in low-P environments and one significant QTL found in the optimal-P environment. Broad-sense heritability estimates were 0.78 (individual seed weight), 0.90 (seed protein), 0.34 (seed oil), and 0.98 (seed number). The QTLs identified under low P point to genetic regions that may be useful to improve soybean performance under limiting P conditions.

Protecting the photosynthetic performance of snap bean under free air ozone exposure.

Tropospheric ozone (O3) is a major air pollutant and causes serious injury to vegetation. To protect sensitive plants from O3 damage, several agrochemicals have been assessed, including cytokinin (e.g., kinetin, KIN) and ethylenediurea (EDU) with cytokinin-like activity. In higher plant, leaves are primarily injured by O3 and protective agrochemicals are often applied by leaf spraying. To our knowledge, the mitigating abilities of EDU and KIN have not been compared directly in a realistic setup. In the present research, impacts of elevated O3 (2× ambient O3, 24hr per day, for 8days) on an O3 sensitive line (S156) of snap bean (Phaseolus vulgaris), which is often used for biomonitoring O3 pollution, were studied in a free air controlled exposure system. The day before starting the O3 exposure, plants were sprayed with a solution of EDU (300ppm), KIN (1mmol/L) or distilled water, to compare their protective abilities. The results demonstrated that 2× ambient O3 inhibited net photosynthetic rate and stomatal conductance, increased the minimal fluorescence yield of the dark-adapted state, decreased the maximal quantum yield of PSII photochemistry, and led to visible injury. KIN and EDU alleviated the reduction of the photosynthetic performance, and visible injury under O3 fumigation. The plants sprayed with EDU showed greater ability to mitigate the O3 damage than those sprayed with KIN. Chlorophyll fluorescence imaging may have detected more precisely the differences in O3 response across the leaf than the conventional fluorometer.

Phenotypic variation and identification of quantitative trait loci for ozone tolerance in a Fiskeby III × Mandarin (Ottawa) soybean population.

KEY MESSAGE: Soybean quantitative trait loci for ozone response. Ground-level ozone reduces yield in crops such as soybean (Glycine max (L.) Merr.). Phenotypic variation has been observed for this trait in multiple species; however, breeding for ozone tolerance has been limited. A recombinant inbred population was developed from soybean genotypes differing in tolerance to ozone: tolerant Fiskeby III and sensitive Mandarin (Ottawa). Plants were exposed to ozone treatment for 5 days in greenhouse chambers followed by visual scoring for foliar injury. Mean injury score in the mid-canopy was 16 % for Fiskeby III, and 81 % for Mandarin (Ottawa). Injury scores were lower in younger leaves for both parents and progeny, compared to scores in the older leaves. Segregation was consistent with multigenic inheritance. Correlation coefficients for injury between leaf positions ranged from 0.34 to 0.81, with the closer leaf positions showing the greater correlation. Narrow sense heritability within an ozone treatment chamber was 0.59, 0.40, 0.29, 0.30, 0.19, and 0.35 for the 2nd, 3rd, 4th, 5th, 6th, and combined 3rd-5th main stem leaf positions (numbered acropetally), respectively, based on genotypic means over three independent replications. Quantitative trait loci (QTL) analysis showed that loci were associated with distinct leaf developmental stages. QTL were identified on Chromosome 17 for the 2nd and 3rd leaf positions, and on Chromosome 4 for the 5th and 6th leaf positions. Additional loci were identified on Chromosomes 6, 18, 19, and 20. Interacting loci were identified on Chromosomes 5 and 15 for injury on trifoliate 4. The ozone sensitive parent contributed one favorable allele for ozone response.

Alteration of the alkaloid profile in genetically modified tobacco reveals a role of methylenetetrahydrofolate reductase in nicotine N-demethylation.

Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme of the tetrahydrofolate (THF)-mediated one-carbon (C1) metabolic network. This enzyme catalyzes the reduction of 5,10-methylene-THF to 5-methyl-THF. The latter donates its methyl group to homocysteine, forming methionine, which is then used for the synthesis of S-adenosyl-methionine, a universal methyl donor for numerous methylation reactions, to produce primary and secondary metabolites. Here, we demonstrate that manipulating tobacco (Nicotiana tabacum) MTHFR gene (NtMTHFR1) expression dramatically alters the alkaloid profile in transgenic tobacco plants by negatively regulating the expression of a secondary metabolic pathway nicotine N-demethylase gene, CYP82E4. Quantitative real-time polymerase chain reaction and alkaloid analyses revealed that reducing NtMTHFR expression by RNA interference dramatically induced CYP82E4 expression, resulting in higher nicotine-to-nornicotine conversion rates. Conversely, overexpressing NtMTHFR1 suppressed CYP82E4 expression, leading to lower nicotine-to-nornicotine conversion rates. However, the reduced expression of NtMTHFR did not affect the methionine and S-adenosyl-methionine levels in the knockdown lines. Our finding reveals a new regulatory role of NtMTHFR1 in nicotine N-demethylation and suggests that the negative regulation of CYP82E4 expression may serve to recruit methyl groups from nicotine into the C1 pool under C1-deficient conditions.

Differential expression of a novel gene EaF82a in green and yellow sectors of variegated Epipremnum aureum leaves is related to uneven distribution of auxin.

EaF82, a gene identified in previous studies of the variegated plant Epipremnum aureum, exhibited a unique expression pattern with greater transcript abundance in yellow sectors than green sectors of variegated leaves, but lower abundance in regenerated pale yellow plants than in green plants derived from leaf tissue culture. Studies of its full-length cDNA and promoter region revealed two members with only the EaF82a expressed. Immunoblotting confirmed that EaF82a encodes a 12 kDa protein and its accumulation consistent with its gene expression patterns in different color tissues. Transient expression of EaF82a-sGFP fusion proteins in protoplasts showed that EaF82a seems to be present in the cytosol as unidentified spots. Sequence motif search reveals a potential auxin responsive element in promoter region. Using transgenic Arabidopsis seedlings carrying EaF82a promoter driving the bacterial uidA (GUS) gene, an increased GUS activity was observed when IAA (indole-3-acetic acid) concentration was elevated. In E. aureum, EaF82a is more abundant at the site where axillary buds emerge and at the lower side of bending nodes where more IAA accumulates relative to the upper side. The measurement of endogenous IAA levels in different color tissues revealed the same pattern of IAA distribution as that of EaF82a expression, further supporting that EaF82a is an IAA responsive gene. EaF82a expression in etiolated transgenic Arabidopsis seedlings responded to IAA under the influence of light suggesting a microenvironment of uneven light condition affects the EaF82a transcript levels and protein accumulation in variegated leaves.

Elevated CO2 spurs reciprocal positive effects between a plant virus and an arbuscular mycorrhizal fungus.

Plants form ubiquitous associations with diverse microbes. These interactions range from parasitism to mutualism, depending partly on resource supplies that are being altered by global change. While many studies have considered the separate effects of pathogens and mutualists on their hosts, few studies have investigated interactions among microbial mutualists and pathogens in the context of global change. Using two wild grass species as model hosts, we grew individual plants under ambient or elevated CO(2), and ambient or increased soil phosphorus (P) supply. Additionally, individuals were grown with or without arbuscular mycorrhizal inoculum, and after 2 wk, plants were inoculated or mock-inoculated with a phloem-restricted virus. Under elevated CO(2), mycorrhizal association increased the titer of virus infections, and virus infection reciprocally increased the colonization of roots by mycorrhizal fungi. Additionally, virus infection decreased plant allocation to root biomass, increased leaf P, and modulated effects of CO(2) and P addition on mycorrhizal root colonization. These results indicate that plant mutualists and pathogens can alter each other's success, and predict that these interactions will respond to increased resource availability and elevated CO(2). Together, our findings highlight the importance of interactions among multiple microorganisms for plant performance under global change.

The interaction of O3 and CO2 concentration, exposure timing and duration on stem rust severity on winter wheat variety 'Coker 9553'.

Wheat rusts, elevated ozone (O3), and carbon dioxide (CO2) are simultaneously impacting wheat production worldwide, but their interactions are not well understood. This study investigated whether near-ambient O3 is suppressive or conducive to stem rust (Sr) of wheat, considering the interactions with ambient and elevated CO2. Winter wheat variety 'Coker 9553' (Sr-susceptible; O3 sensitive) was inoculated with Sr (race QFCSC) following pre-treatment with four different concentrations of O3 (CF, 50, 70, and 90 ppbv) at ambient CO2 levels. Gas treatments were continued during the development of disease symptoms. Disease severity, measured as percent sporulation area (PSA), significantly increased relative to the CF control only under near-ambient O3 conditions (50 ppbv) in the absence of O3-induced foliar injury. Disease symptoms at higher O3 exposures (70 and 90 ppbv) were similar to or less than the CF control. When Coker 9553 was inoculated with Sr while exposed to CO2 (400; 570 ppmv) and O3 (CF; 50 ppbv) in four different combinations, and seven combinations of exposure timing and duration, PSA significantly increased only under continuous treatment with O3 for six weeks or pre-inoculation treatment for three weeks, suggesting that O3-predisposes wheat to the disease rather than enhancing disease post-inoculation. O3 singly and in combination with CO2 increased PSA on flag leaves of adult Coker 9553 plants while elevated CO2 alone had little effect on PSA. These findings show that sub-symptomatic O3 conditions are conducive to stem rust, contradicting the current consensus that biotrophic pathogens are suppressed by elevated O3. This suggests that sub-symptomatic O3 stress may enhance rust diseases in wheat-growing regions.

Microbial community dynamics responding to nutrient allocation associated with soybean cultivar 'Jake' ozone adaptation.

Tropospheric ozone (O3), a major air pollutant, leads to significant global yield loss in soybean [Glycine max (L.) Merr.]. Soybean cultivar 'Jake' shows O3 resilient traits in above-ground organs, but the root system remains sensitive to elevated O3 (eO3). Changing carbon (C) and nitrogen (N) resource composition during eO3 stress suggests that eO3 presumably alters belowground soil microbial communities and their driven nutrient transformation. Yet, the responses of belowground microbes to eO3 and their feedback on nutrient cycling in 'Jake' are unknown. In this study, we holistically investigated soil microbial communities associated with C and N dynamics and bacterial-fungal inter-kingdom networks in the rhizosphere and bulk soil at different developmental stages of 'Jake' grown under sub-ambient O3 [charcoal-filtered (CF) air, 12 h mean: 20 ppb] or eO3 (12 h mean: 87 ppb). The results demonstrated eO3 significantly decreased fungal diversity and complexity of microbial networks at different 'Jake' developmental stages, whereas bacterial diversity was more tolerant to eO3 in both bulk soil and rhizosphere. In the bulk soil, no O3-responsive microbial biomarkers were found to be associated with C and N content, implying eO3 may stimulate niche-based processes during 'Jake' growth. In contrast, this study identified O3-responsive microbial biomarkers that may contribute to the N acquisition (Chloroflexales) and C dynamics (Caldilineales, Thermomicrobiales, and Hypocreales) in the rhizosphere, which may support the O3 resilience of the 'Jake' cultivar. However, further investigation is required to confirm their specific contributions by determining changes in microbial gene expression. Overall, these findings conduce to an expanding knowledge base that O3 induces temporal and spatial changes in the effects of microbial and nutrient networks in the O3-tolerant agriculture ecosystems.

A Rapid Alkalinization Factor-like Peptide EaF82 Impairs Tapetum Degeneration during Pollen Development through Induced ATP Deficiency.

In plants, the timely degeneration of tapetal cells is essential for providing nutrients and other substances to support pollen development. Rapid alkalinization factors (RALFs) are small, cysteine-rich peptides known to be involved in various aspects of plant development and growth, as well as defense against biotic and abiotic stresses. However, the functions of most of them remain unknown, while no RALF has been reported to involve tapetum degeneration. In this study, we demonstrated that a novel cysteine-rich peptide, EaF82, isolated from shy-flowering 'Golden Pothos' (Epipremnum aureum) plants, is a RALF-like peptide and displays alkalinizing activity. Its heterologous expression in Arabidopsis delayed tapetum degeneration and reduced pollen production and seed yields. RNAseq, RT-qPCR, and biochemical analyses showed that overexpression of EaF82 downregulated a group of genes involved in pH changes, cell wall modifications, tapetum degeneration, and pollen maturation, as well as seven endogenous Arabidopsis RALF genes, and decreased proteasome activity and ATP levels. Yeast two-hybrid screening identified AKIN10, a subunit of energy-sensing SnRK1 kinase, as its interacting partner. Our study reveals a possible regulatory role for RALF peptide in tapetum degeneration and suggests that EaF82 action may be mediated through AKIN10 leading to the alteration of transcriptome and energy metabolism, thereby causing ATP deficiency and impairing pollen development.

Re-evaluating the role of ascorbic acid and phenolic glycosides in ozone scavenging in the leaf apoplast of Arabidopsis thaliana L.

Phenolic glycosides are effective reactive oxygen scavengers and peroxidase substrates, suggesting that compounds in addition to ascorbate may have functional importance in defence responses against ozone (O(3)), especially in the leaf apoplast. The apoplastic concentrations of ascorbic acid (AA) and phenolic glycosides in Arabidopsis thaliana L. Col-0 wild-type plants were determined following exposure to a range of O(3) concentrations (5, 125 or 175 nL L(-1)) in controlled environment chambers. AA in leaf apoplast extracts was almost entirely oxidized in all treatments, suggesting that O(3) scavenging by direct reactions with reduced AA was very limited. In regard to phenolics, O(3) stimulated transcription of numerous phenylpropanoid pathway genes and increased the apoplastic concentration of sinapoyl malate. However, modelling of O(3) scavenging in the apoplast indicated that sinapoyl malate concentrations were too low to be effective protectants. Furthermore, null mutants for sinapoyl esters (fah1-7), kaempferol glycosides (tt4-1) and the double mutant (tt4-1/fah1-7) were equally sensitive to chronic O(3) as Ler-0 wild-type plants. These results indicate that current understanding of O(3) defence schemes deserves reassessment as mechanisms other than direct scavenging of O(3) by extracellular AA and antioxidant activity of some phenolics may predominate in some plant species.

Influence of atmospheric vapour pressure deficit on ozone responses of snap bean (Phaseolus vulgaris L.) genotypes.

Environmental conditions influence plant responses to ozone (O(3)), but few studies have evaluated individual factors directly. In this study, the effect of O(3) at high and low atmospheric vapour pressure deficit (VPD) was evaluated in two genotypes of snap bean (Phaseolus vulgaris L.) (R123 and S156) used as O(3) bioindicator plants. Plants were grown in outdoor controlled-environment chambers in charcoal-filtered air containing 0 or 60 nl l(-1) O(3) (12 h average) at two VPDs (1.26 and 1.96 kPa) and sampled for biomass, leaf area, daily water loss, and seed yield. VPD clearly influenced O(3) effects. At low VPD, O(3) reduced biomass, leaf area, and seed yield substantially in both genotypes, while at high VPD, O(3) had no significant effect on these components. In clean air, high VPD reduced biomass and yield by similar fractions in both genotypes compared with low VPD. Data suggest that a stomatal response to VPD per se may be lacking in both genotypes and it is hypothesized that the high VPD resulted in unsustainable transpiration and water deficits that resulted in reduced growth and yield. High VPD- and water-stress-induced stomatal responses may have reduced the O(3) flux into the leaves, which contributed to a higher yield compared to the low VPD treatment in both genotypes. At low VPD, transpiration increased in the O(3) treatment relative to the clean air treatment, suggesting that whole-plant conductance was increased by O(3) exposure. Ozone-related biomass reductions at low VPD were proportionally higher in S156 than in R123, indicating that differential O(3) sensitivity of these bioindicator plants remained evident when environmental conditions were conducive for O(3) effects. Assessments of potential O(3) impacts on vegetation should incorporate interacting factors such as VPD.

Xanthophyll cycle pigment and antioxidant profiles of winter-red (anthocyanic) and winter-green (acyanic) angiosperm evergreen species.

Leaves of many angiosperm evergreen species change colour from green to red during winter, corresponding with the synthesis of anthocyanin pigments. The ecophysiological function of winter colour change (if any), and why it occurs in some species and not others, are not yet understood. It was hypothesized that anthocyanins play a compensatory photoprotective role in species with limited capacity for energy dissipation. Seasonal xanthophyll pigment content, chlorophyll fluorescence, leaf nitrogen, and low molecular weight antioxidants (LMWA) of five winter-red and five winter-green angiosperm evergreen species were compared. Our results showed no difference in seasonal xanthophyll pigment content (V+A+Z g(-1) leaf dry mass) or LMWA between winter-red and winter-green species, indicating red-leafed species are not deficient in their capacity for non-photochemical energy dissipation via these mechanisms. Winter-red and winter-green species also did not differ in percentage leaf nitrogen, corroborating previous studies showing no difference in seasonal photosynthesis under saturating irradiance. Consistent with a photoprotective function of anthocyanin, winter-red species had significantly lower xanthophyll content per unit chlorophyll and less sustained photoinhibition than winter-green species (i.e. higher pre-dawn F(v)/F(m) and a lower proportion of de-epoxidized xanthophylls retained overnight). Red-leafed species also maintained a higher maximum quantum yield efficiency of PSII at midday (F'(v)/F'(m)) during winter, and showed characteristics of shade acclimation (positive correlation between anthocyanin and chlorophyll content, and negative correlation with chlorophyll a/b). These results suggest that the capacity for photon energy dissipation (photochemical and non-photochemical) is not limited in red-leafed species, and that anthocyanins more likely function as an alternative photoprotective strategy to increased VAZ/Chl during winter.

Impact of elevated ozone on yield and carbon-nitrogen content in soybean cultivar 'Jake'.

Tropospheric ozone (O3) is a pollutant that leads to significant global yield loss in soybean [Glycine max (L.) Merr.]. To ensure soybean productivity in areas of rising O3, it is important to identify tolerant genotypes. This work describes the response of the high-yielding soybean cultivar 'Jake' to elevated O3 concentrations. 'Jake' was treated with either low O3 [charcoal-filtered (CF) air, 12 h mean: 20 ppb] or with O3-enriched air (12 h mean: 87 ppb) over the course of the entire growing season. In contrast to the absence of O3-induced leaf injury under low O3, elevated O3 caused severe leaf injury and decreased stomatal conductance and photosynthesis. Although elevated O3 reduced total leaf area, leaf number, and plant height at different developmental stages, above-ground and root biomass remained unchanged. Analyzing carbon and nitrogen content, we found that elevated O3 altered allocation of both elements, which ultimately led to a 15 % yield loss by decreasing seed size but not seed number. We concluded that cultivar 'Jake' possesses developmental strength to tolerate chronic O3 conditions, attributes that make it suitable breeding material for the generation of new O3 tolerant lines.

Tropospheric ozone rapidly decreases root growth by altering carbon metabolism and detoxification capability in growing soybean roots.

High tropospheric ozone (O3) concentrations lead to significant global soybean (Glycine max) yield reductions. Research concerning O3 impacts on soybean has focused on the contributions of above-ground tissues. In this study, Mandarin (Ottawa) (O3-sensitive) and Fiskeby III (O3-tolerant) soybean genotypes provide contrasting materials to investigate O3 effects on root growth. We compared root morphological and proteomic changes when 16-day-old plants were treated with charcoal-filtered (CF) air or elevated O3 (80 ppb O3 for 7 h/day) in continuously stirred-tank reactors (CSTR) for 7 days. Our results showed that in Mandarin (Ottawa), decreased expression of enzymes involved in the tricarboxylic acid (TCA) cycle contributes to reduction of root biomass and diameter under elevated O3. In contrast, O3 tolerance in Fiskeby III roots was associated with O3-dependent induction of enzymes involved in glycolysis and O3-independent expression of enzymes involved in the ascorbate-glutathione cycle. We conclude that a decreased abundance of key redox enzymes in roots due to limited carbon availability rapidly alters root growth under O3 stress. However, maintaining a high abundance of enzymes associated with redox status and detoxification capability contributes to overall O3 tolerance in roots.

Transformation of Long-Lived Albino Epipremnum aureum 'Golden Pothos' and Restoring Chloroplast Development.

Chloroplasts are organelles responsible for chlorophyll biosynthesis, photosynthesis, and biosynthesis of many metabolites, which are one of key targets for crop improvement. Elucidating and engineering genes involved in chloroplast development are important approaches for studying chloroplast functions as well as developing new crops. In this study, we report a long-lived albino mutant derived from a popular ornamental plant Epipremnum aureum 'Golden Pothos' which could be used as a model for analyzing the function of genes involved in chloroplast development and generating colorful plants. Albino mutant plants were isolated from regenerated populations of variegated 'Golden Pothos' whose albino phenotype was previously found to be due to impaired expression of EaZIP, encoding Mg-protoporphyrin IX monomethyl ester cyclase. Using petioles of the mutant plants as explants with a traceable sGFP gene, an efficient transformation system was developed. Expressing Arabidopsis CHL27 (a homolog of EaZIP) but not EaZIP in albino plants restored green color and chloroplast development. Interestingly, in addition to the occurrence of plants with solid green color, plants with variegated leaves and pale-yellow leaves were also obtained in the regenerated populations. Nevertheless, our study shows that these long-lived albino plants along with the established efficient transformation system could be used for creating colorful ornamental plants. This system could also potentially be used for investigating physiological processes associated with chlorophyll levels and chloroplast development as well as certain biological activities, which are difficult to achieve using green plants.

Warming and elevated ozone induce tradeoffs between fine roots and mycorrhizal fungi and stimulate organic carbon decomposition.

Climate warming and elevated ozone (eO3) are important climate change components that can affect plant growth and plant-microbe interactions. However, the resulting impact on soil carbon (C) dynamics, as well as the underlying mechanisms, remains unclear. Here, we show that warming, eO3, and their combination induce tradeoffs between roots and their symbiotic arbuscular mycorrhizal fungi (AMF) and stimulate organic C decomposition in a nontilled soybean agroecosystem. While warming and eO3 reduced root biomass, tissue density, and AMF colonization, they increased specific root length and promoted decomposition of both native and newly added organic C. Also, they shifted AMF community composition in favor of the genus Paraglomus with high nutrient-absorbing hyphal surface over the genus Glomus prone to protection of soil organic C. Our findings provide deep insights into plant-microbial interactive responses to warming and eO3 and how these responses may modulate soil organic C dynamics under future climate change scenarios.