Yasuhiro Usui

Increasing CO2 threatens human nutrition

Dietary deficiencies of zinc and iron are a substantial global public health problem. An estimated two billion people suffer these deficiencies, causing a loss of 63 million life-years annually. Most of these people depend on C3 grains and legumes as their primary dietary source of zinc and iron. Here we report that C3 grains and legumes have lower concentrations of zinc and iron when grown under field conditions at the elevated atmospheric CO2 concentration predicted for the middle of this century. C3 crops other than legumes also have lower concentrations of protein, whereas C4 crops seem to be less affected. Differences between cultivars of a single crop suggest that breeding for decreased sensitivity to atmospheric CO2 concentration could partly address these new challenges to global health.

Rice cultivar responses to elevated CO2 at two free-air CO2 enrichment (FACE) sites in Japan

There is some evidence that rice cultivars respond differently to elevated CO2 concentrations ([CO2]), but [CO2]×cultivar interaction has never been tested under open-field conditions across different sites. Here, we report on trials conducted at free-air CO2 enrichment (FACE) facilities at two sites in Japan, Shizukuishi (2007 and 2008) and Tsukuba (2010). The average growing-season air temperature was more than 5°C warmer at Tsukuba than at Shizukuishi. For four cultivars tested at both sites, the [CO2]×cultivar interaction was significant for brown rice yield, but there was no significant interaction with site-year. Higher-yielding cultivars with a large sink size showed a greater [CO2] response. The Tsukuba FACE experiment, which included eight cultivars, revealed a wider range of yield enhancement (3-36%) than the multi-site experiment. All of the tested yield components contributed to this enhancement, but there was a highly significant [CO2]×cultivar interaction for percentage of ripened spikelets. These results suggest that a large sink is a prerequisite for higher productivity under elevated [CO2], but that improving carbon allocation by increasing grain setting may also be a practical way of increasing the yield response to elevated [CO2].

Do the rich always become richer? Characterizing the leaf physiological response of the high-yielding rice cultivar Takanari to free-air CO2 enrichment.

The development of crops which are well suited to growth under future environmental conditions such as higher atmospheric CO2 concentrations ([CO2]) is essential to meeting the challenge of ensuring food security in the face of the growing human population and changing climate. A high-yielding indica rice variety (Oryza sativa L. cv. Takanari) has been recently identified as a potential candidate for such breeding, due to its high productivity in present [CO2]. To test if it could further increase its productivity under elevated [CO2] (eCO2), Takanari was grown in the paddy field under season-long free-air CO2 enrichment (FACE, approximately 200 µmol mol−1 above ambient [CO2]) and its leaf physiology was compared with the representative japonica variety ‘Koshihikari’. Takanari showed consistently higher midday photosynthesis and stomatal conductance than Koshihikari under both ambient and FACE growth conditions over 2 years. Maximum ribulose-1,5-bisphosphate carboxylation and electron transport rates were higher for Takanari at the mid-grain filling stage in both years. Mesophyll conductance was higher in Takanari than in Koshihikari at the late grain-filling stage. In contrast to Koshihikari, Takanari grown under FACE conditions showed no decrease in total leaf nitrogen on an area basis relative to ambient-grown plants. Chl content was higher in Takanari than in Koshihikari at the same leaf nitrogen level. These results indicate that Takanari maintains its superiority over Koshihikari in regards to its leaf-level productivity when grown in elevated [CO2] and it may be a valuable resource for rice breeding programs which seek to increase crop productivity under current and future [CO2].

Heat-tolerant rice cultivars retain grain appearance quality under free-air CO2 enrichment

BackgroundHeat-tolerant rice cultivars have been developed as a countermeasure to poor grain appearance quality under high temperatures. Recent studies showed that elevated CO2 concentrations (E-[CO2]) also reduce grain quality. To determine whether heat-tolerant cultivars also tolerate E-[CO2], we conducted a free-air CO2 enrichment (FACE) experiment with 12 rice cultivars differing in heat tolerance.ResultsThe percentage of undamaged grains of five standard cultivars (Akitakomachi, Kinuhikari, Koshihikari, Matsuribare, Nipponbare) averaged 61.7% in the ambient [CO2] (AMB) plot and 51.7% in the FACE plot, whereas that of heat-tolerant cultivars (Eminokizuna, Wa2398, Kanto 257, Toyama 80, Mineharuka, Kanto 259, Saikai 290) averaged 73.5% in AMB and 71.3% in FACE. This resulted in a significant [CO2] by cultivar interaction. The percentage of white-base or white-back grains increased from 8.4% in AMB to 17.1% in FACE in the sensitive cultivars, but from only 2.1% in AMB to only 4.4% in FACE in the heat-tolerant cultivars.ConclusionHeat-tolerant cultivars retained their grain appearance quality at E-[CO2] under present air temperatures. Further improvements in appearance quality under present conditions will be needed to achieve improvements under E-[CO2], because E-[CO2] will likely lower the threshold temperature for heat stress.

Effects of Elevated Carbon Dioxide, Elevated Temperature, and Rice Growth Stage on the Community Structure of Rice Root–Associated Bacteria

The effects of free-air carbon dioxide enrichment (FACE) and elevated soil and water temperature (warming) on the rice root–associated bacterial community were evaluated by clone library analysis of the 16S ribosomal RNA gene. Roots were sampled at the panicle initiation and ripening stages 41 and 92 days after transplanting (DAT), respectively. The relative abundances of the methanotrophs Methylosinus and Methylocystis were increased by warming and decreased by FACE at 92 DAT, which indicated that microbial methane (CH4) oxidation in rice roots may have been influenced by global warming. The relative abundance of Burkholderia kururiensis was increased by warming at 41 DAT and by FACE or warming at 92 DAT. The abundances of methanotrophs increased during rice growth, which was likely induced by an enhancement in the emission of CH4 from the paddy fields, suggesting that CH4 is one of the predominant factors affecting the structure of the microbial community in rice roots. Marked variations in the community structure were also observed during rice growth in other genera: Bradyrhizobium, Clostridium, and an unknown genus close to Epsilonproteobacteria were abundant at 92 DAT, whereas Achromobacter was abundant at 41 DAT. These results demonstrated that the community structures of rice root-associated bacteria were markedly affected by FACE, temperature, and the rice growth stage.

The effects of free-air CO2 enrichment (FACE) on carbon and nitrogen accumulation in grains of rice (Oryza sativa L.)

Rising atmospheric CO2 concentrations will probably increase rice (Oryza sativa L.) yield but decrease grain nitrogen (GN) concentration. Grains attached to different positions in the panicles differ greatly in weight and quality, but their responses to elevated CO2 (e[CO2]) are poorly understood, which limits our understanding of the mechanisms of yield enhancement and quality degradation. Thus a free-air CO2 enrichment experiment was conducted to examine the effects of e[CO2] on grain mass (GM), grain carbon (GC), and GN accumulation in the spikelets attached to the upper primary rachis branch (superior spikelets; SS) and those attached to the lower secondary rachis (inferior spikelets; IS). e[CO2] stimulated the rice yield by 13% but decreased the N concentration in the panicle by 7% when averaged over two levels of N fertilizations (P < 0.01). The responses of SS and IS to e[CO2] were different particularly under higher N supply. For SS, e[CO2] decreased GN by 24% (P < 0.01) but did not affect GM. For IS, e[CO2] increased GM by 13% (P < 0.05) but GN was not affected. The reduction of GN due to e[CO2] started to appear at the beginning of grain filling. These results suggest that future [CO2] levels probably stimulate the grain growth of IS, most of which are not marketable due to limited size, at the expense of GN reduction in SS. Translocation of N from SS to IS may be a possible mechanism for reduction in GN of SS. This may degrade the grain quality of marketable rice under e[CO2].

Impacts of elevated atmospheric CO2 on nutrient content of important food crops

One of the many ways that climate change may affect human health is by altering the nutrient content of food crops. However, previous attempts to study the effects of increased atmospheric CO2 on crop nutrition have been limited by small sample sizes and/or artificial growing conditions. Here we present data from a meta-analysis of the nutritional contents of the edible portions of 41 cultivars of six major crop species grown using free-air CO2 enrichment (FACE) technology to expose crops to ambient and elevated CO2 concentrations in otherwise normal field cultivation conditions. This data, collected across three continents, represents over ten times more data on the nutrient content of crops grown in FACE experiments than was previously available. We expect it to be deeply useful to future studies, such as efforts to understand the impacts of elevated atmospheric CO2 on crop macro- and micronutrient concentrations, or attempts to alleviate harmful effects of these changes for the billions of people who depend on these crops for essential nutrients.

Characterization of Leaf Blade- and Leaf Sheath-Associated Bacterial Communities and Assessment of Their Responses to Environmental Changes in CO2, Temperature, and Nitrogen Levels under Field Conditions

Rice shoot-associated bacterial communities at the panicle initiation stage were characterized and their responses to elevated surface water-soil temperature (ET), low nitrogen (LN), and free-air CO2 enrichment (FACE) were assessed by clone library analyses of the 16S rRNA gene. Principal coordinate analyses combining all sequence data for leaf blade- and leaf sheath-associated bacteria revealed that each bacterial community had a distinct structure, as supported by PC1 (61.5%), that was mainly attributed to the high abundance of Planctomycetes in leaf sheaths. Our results also indicated that the community structures of leaf blade-associated bacteria were more sensitive than those of leaf sheath-associated bacteria to the environmental factors examined. Among these environmental factors, LN strongly affected the community structures of leaf blade-associated bacteria by increasing the relative abundance of Bacilli. The most significant effect of FACE was also observed on leaf blade-associated bacteria under the LN condition, which was explained by decreases and increases in Agrobacterium and Pantoea, respectively. The community structures of leaf blade-associated bacteria under the combination of FACE and ET were more similar to those of the control than to those under ET or FACE. Thus, the combined effects of environmental factors need to be considered in order to realistically assess the effects of environmental changes on microbial community structures.

Elevated atmospheric CO2 levels affect community structure of rice root-associated bacteria

A number of studies have shown that elevated atmospheric CO2 ([CO2]) affects rice yields and grain quality. However, the responses of root-associated bacteria to [CO2] elevation have not been characterized in a large-scale field study. We conducted a free-air CO2 enrichment (FACE) experiment (ambient + 200 μmol.mol−1) using three rice cultivars (Akita 63, Takanari, and Koshihikari) and two experimental lines of Koshihikari [chromosome segment substitution and near-isogenic lines (NILs)] to determine the effects of [CO2] elevation on the community structure of rice root-associated bacteria. Microbial DNA was extracted from rice roots at the panicle formation stage and analyzed by pyrosequencing the bacterial 16S rRNA gene to characterize the members of the bacterial community. Principal coordinate analysis of a weighted UniFrac distance matrix revealed that the community structure was clearly affected by elevated [CO2]. The predominant community members at class level were Alpha-, Beta-, and Gamma-proteobacteria in the control (ambient) and FACE plots. The relative abundance of Methylocystaceae, the major methane-oxidizing bacteria in rice roots, tended to decrease with increasing [CO2] levels. Quantitative PCR revealed a decreased copy number of the methane monooxygenase (pmoA) gene and increased methyl coenzyme M reductase (mcrA) in elevated [CO2]. These results suggest elevated [CO2] suppresses methane oxidation and promotes methanogenesis in rice roots; this process affects the carbon cycle in rice paddy fields.

Fully automated, high-throughput instrumentation for measuring the δ13C value of methane and application of the instrumentation to rice paddy samples

RATIONALE The stable carbon isotope ratio ((13)C/(12)C or δ(13)C value) of methane (CH4) produced in methanogenic environments contains information about primary source material, CH4 production pathways, degree of oxidation, and transport. However, the availability of δ(13)C-CH4 data is severely limited because isotope analysis methods are low throughput, owing primarily to the need for manual processing steps. High-throughput, fully automated measurement is necessary to facilitate the use of the δ(13)C signature in understanding CH4 biogeochemistry. METHODS We modified a conventional continuous-flow (CF) gas chromatography/combustion/isotope ratio mass spectrometry (IRMS) instrument system by incorporating (i) automated sample injection, (ii) a newly developed temperature-control unit for preconcentration and cryofocus traps, and (iii) an automatic system for liquid-nitrogen refilling. The system, which could run unattended for 1 day, was used to obtain δ(13)C-CH4 data for CH4 samples collected from an irrigated rice paddy with an automated closed-chamber system. RESULTS Using the fully automated CF-IRMS system, we measured δ(13)C-CH4 data for 77 samples during a 21.5-h continuous run (17 min per sample) with high precision (1σ = 0.11‰, reproducibility) and moderate consumption of liquid nitrogen (11 L). Application of the system to CH4 samples obtained from the rice paddy revealed distinct seasonal and diurnal variations in δ(13)C values with the highest temporal resolution ever reported. CONCLUSIONS A fully automated, high-throughput system for the measurement of δ(13)C-CH4 values was developed and used to analyze air samples obtained from a rice paddy. Our results demonstrate the high potential of this system for obtaining δ(13)C data useful for process-level understanding of CH4 biogeochemistry with respect to spatiotemporal variation of CH4 sources and how that variation is affected by environmental and management factors.