The CO2 fertilization effect on leaf photosynthesis of maize (Zea mays L.) depends on growth temperatures with changes in leaf anatomy and soluble sugars.

Understanding the potential mechanisms and processes of leaf photosynthesis in response to elevated CO2 concentration ([CO2]) and temperature is critical for estimating the impacts of climatic change on the growth and yield in crops such as maize (Zea mays L.), which is a widely cultivated C4 crop all over the world. We examined the combined effect of elevated [CO2] and temperature on plant growth, leaf photosynthesis, stomatal traits, and biochemical compositions of maize with six environmental growth chambers controlling two CO2 levels (400 and 800 µmol mol-1) and three temperature regimes (25/19°C, 31/25°C, and 37/31°C). We found that leaf photosynthesis was significantly enhanced by increasing growth temperature from 25/19°C to 31/25°C independent of [CO2]. However, leaf photosynthesis drastically declined when the growth temperature was continually increased to 37/31°C at both ambient CO2 concentration (400 µmol mol-1, a[CO2]) and elevated CO2 concentration (800 µmol mol-1, e[CO2]). Meanwhile, we also found strong CO2 fertilization effect on maize plants grown at the highest temperature (37/31°C), as evidenced by the higher leaf photosynthesis at e[CO2] than that at a[CO2], although leaf photosynthesis was similar between a[CO2] and e[CO2] under the other two temperature regimes of 25/19°C and 31/25°C. Furthermore, we also found that e[CO2] resulted in an increase in leaf soluble sugar, which was positively related with leaf photosynthesis under the high temperature regime of 37/31°C (R 2 = 0.77). In addition, our results showed that e[CO2] substantially decreased leaf transpiration rates of maize plants, which might be partially attributed to the reduced stomatal openness as demonstrated by the declined stomatal width and stomatal area. These results suggest that the CO2 fertilization effect on plant growth and leaf photosynthesis of maize depends on growth temperatures through changing stomatal traits, leaf anatomy, and soluble sugar contents.

Response of soil N2O emission and nitrogen utilization to organic matter in the wheat and maize rotation system.

The appropriate nitrogen (N) fertilizer regulator could increase N utilization of crops and reduce N losses in the North China Plain. We investigated the effects of reduced inorganic-N rate combined with an organic fertilizer on nitrous oxide (N2O) emissions in winter wheat and summer maize rotation system. Simultaneously studied the effect of different treatments on N use efficiency (NUE), N balance and net income. After reducing the amount of nitrogen fertilizer in the wheat-corn rotation system, the results showed that the cumulative emission of soil N2O from the RN40% + HOM [40% of RN (recommended inorganic-N rate) with homemade organic matter] treatment was 41.0% lower than that of the RN treatment. In addition, the N production efficiency, agronomic efficiency, and apparent utilization were significantly increased by 50.2%, 72.4% and 19.5% than RN, respectively. The use of RN40% + HOM resulted in 22.0 and 30.1% lower soil N residual and N losses as compared with RN. After adding organic substances, soil N2O cumulative emission of RN40% + HOM treatment decreased by 20.9% than that of the HAN (zinc and humic acid urea at the same inorganic-N rate of RN) treatment. The N production efficiency, N agronomic efficiency and NUE of RN40% + HOM treatment were 36.6%, 40.9% and 15.3% higher than HAN's. Moreover, soil residual and apparent loss N were 23.3% and 18.0% less than HAN's. The RN40% + HOM treatment appears to be the most effective as a fertilizer control method where it reduced N fertilizer input and its loss to the environment and provided the highest grain yield.

Design of high-oleic tobacco (Nicotiana tabacum L.) seed oil by CRISPR-Cas9-mediated knockout of NtFAD2-2.

BACKGROUND: Tobacco seed oil could be used as an appropriate feedstock for biodiesel production. However, the high linoleic acid content of tobacco seed oil makes it susceptible to oxidation. Altering the fatty acid profile by increasing the content of oleic acid could improve the properties of biodiesel produced from tobacco seed oil. RESULTS: Four FAD2 genes, NtFAD2-1a, NtFAD2-1b, NtFAD2-2a, and NtFAD2-2b, were identified in allotetraploid tobacco genome. Phylogenetic analysis of protein sequences showed that NtFAD2-1a and NtFAD2-2a originated from N. tomentosiformis, while NtFAD2-1b and NtFAD2-2b from N. sylvestris. Expression analysis revealed that NtFAD2-2a and NtFAD2-2b transcripts were more abundant in developing seeds than in other tissues, while NtFAD2-1a and NtFAD2-1b showed low transcript levels in developing seed. Phylogenic analysis showed that NtFAD2-2a and NtFAD2-2b were seed-type FAD2 genes. Heterologous expression in yeast cells demonstrated that both NtFAD2-2a and NtFAD2-2b protein could introduce a double bond at the Δ12 position of fatty acid chain. The fatty acid profile analysis of tobacco fad2-2 mutant seeds derived from CRISPR-Cas9 edited plants showed dramatic increase of oleic acid content from 11% to over 79%, whereas linoleic acid decreased from 72 to 7%. In addition, the fatty acid composition of leaf was not affected in fad2-2 mutant plants. CONCLUSION: Our data showed that knockout of seed-type FAD2 genes in tobacco could significantly increase the oleic acid content in seed oil. This research suggests that CRISPR-Cas9 system offers a rapid and highly efficient method in the tobacco seed lipid engineering programs.

Enhancement of Tobacco (Nicotiana tabacum L.) Seed Lipid Content for Biodiesel Production by CRISPR-Cas9-Mediated Knockout of NtAn1.

Tobacco (Nicotiana tabacum L.) seed lipid is a promising non-edible feedstock for biodiesel production. In order to meet the increasing demand, achieving high seed lipid content is one of the major goals in tobacco seed production. The TT8 gene and its homologs negatively regulate seed lipid accumulation in Arabidopsis and Brassica species. We speculated that manipulating the homolog genes of TT8 in tobacco could enhance the accumulation of seed lipid. In this present study, we found that the TT8 homolog genes in tobacco, NtAn1a and NtAn1b, were highly expressed in developing seed. Targeted mutagenesis of NtAn1 genes was created by the CRISPR-Cas9-based gene editing technology. Due to the defect of proanthocyanidin (PA) biosynthesis, mutant seeds showed the phenotype of a yellow seed coat. Seed lipid accumulation was enhanced by about 18 and 15% in two targeted mutant lines. Protein content was also significantly increased in mutant seeds. In addition, the seed yield-related traits were not affected by the targeted mutagenesis of NtAn1 genes. Thus, the overall lipid productivity of the NtAn1 knockout mutants was dramatically enhanced. The results in this present paper indicated that tobacco NtAn1 genes regulate both PAs and lipid accumulation in the process of seed development and that targeted mutagenesis of NtAn1 genes could generate a yellow-seeded tobacco variety with high lipid and protein content. Furthermore, the present results revealed that the CRISPR-Cas9 system could be employed in tobacco seed de novo domestication for biodiesel feedstock production.

Elevated CO2 concentration induces photosynthetic down-regulation with changes in leaf structure, non-structural carbohydrates and nitrogen content of soybean.

BACKGROUND: Understanding the mechanisms of crops in response to elevated CO2 concentrations is pivotal to estimating the impacts of climate change on the global agricultural production. Based on earlier results of the "doubling-CO2 concentration" experiments, many current climate models may overestimate the CO2 fertilization effect on crops, and meanwhile, underestimate the potential impacts of future climate change on global agriculture ecosystem when the atmospheric CO2 concentration goes beyond the optimal levels for crop growth. RESULTS: This study examined the photosynthetic response of soybean (Glycine max (L.) Merr.) to elevated CO2 concentration associated with changes in leaf structure, non-structural carbohydrates and nitrogen content with environmental growth chambers where the CO2 concentration was controlled at 400, 600, 800, 1000, 1200, 1400, 1600 ppm. We found CO2-induced down-regulation of leaf photosynthesis as evidenced by the consistently declined leaf net photosynthetic rate (An) with elevated CO2 concentrations. This down-regulation of leaf photosynthesis was evident in biochemical and photochemical processes since the maximum carboxylation rate (Vcmax) and the maximum electron transport rate (Jmax) were dramatically decreased at higher CO2 concentrations exceeding their optimal values of about 600 ppm and 400 ppm, respectively. Moreover, the down-regulation of leaf photosynthesis at high CO2 concentration was partially attributed to the reduced stomatal conductance (Gs) as demonstrated by the declines in stomatal density and stomatal area as well as the changes in the spatial distribution pattern of stomata. In addition, the smaller total mesophyll size (palisade and spongy tissues) and the lower nitrogen availability may also contribute to the down-regulation of leaf photosynthesis when soybean subjected to high CO2 concentration environment. CONCLUSIONS: Down-regulation of leaf photosynthesis associated with the changes in stomatal traits, mesophyll tissue size, non-structural carbohydrates, and nitrogen availability of soybean in response to future high atmospheric CO2 concentration and climate change.

The optimal CO2 concentrations for the growth of three perennial grass species.

BACKGROUND: Grasslands are one of the most representative vegetation types accounting for about 20% of the global land area and thus the response of grasslands to climate change plays a pivotal role in terrestrial carbon balance. However, many current climate change models, based on earlier results of the doubling-CO2 experiments, may overestimate the CO2 fertilization effect, and as a result underestimate the potentially effects of future climate change on global grasslands when the atmospheric CO2 concentration goes beyond the optimal level. Here, we examined the optimal atmospheric CO2 concentration effect on CO2 fertilization and further on the growth of three perennial grasses in growth chambers with the CO2 concentration at 400, 600, 800, 1000, and 1200 ppm, respectively. RESULTS: All three perennial grasses featured an apparent optimal CO2 concentration for growth. Initial increases in atmospheric CO2 concentration substantially enhanced the plant biomass of the three perennial grasses through the CO2 fertilization effect, but this CO2 fertilization effect was dramatically compromised with further rising atmospheric CO2 concentration beyond the optimum. The optimal CO2 concentration for the growth of tall fescue was lower than those of perennial ryegrass and Kentucky bluegrass, and thus the CO2 fertilization effect on tall fescue disappeared earlier than the other two species. By contrast, the weaker CO2 fertilization effect on the growth of perennial ryegrass and Kentucky bluegrass was sustained for a longer period due to their higher optimal CO2 concentrations than tall fescue. The limiting effects of excessively high CO2 concentrations may not only associate with changes in the biochemical and photochemical processes of photosynthesis, but also attribute to the declines in stomatal conductance and nitrogen availability. CONCLUSIONS: In this study, we found apparent differences in the optimal CO2 concentrations for the growth of three grasses. These results suggest that the growth of different types of grasses may respond differently to future elevated CO2 concentrations through the CO2 fertilization effect, and thus potentially alter the community composition and structure of grasslands. Meanwhile, our results may also be helpful for improving current process-based ecological models to more accurately predict the structure and function of grassland ecosystems under future rising atmospheric CO2 concentration and climate change scenarios.

[Changes of non-structural carbohydrates and its impact factors in trees: a review].

Non-structural carbohydrates (NSCs) are an important energy source for the metabolism of plants. The size of the NSC pool is likely to mirror the overall carbon supply status and its dynamics strongly influences physiological processes in plants. In order to predict the response and adaptation of trees to climate change, this review summarized the current understanding of NSC pool in trees, and mainly focused on its seasonal and spatial variation for analyzing the relationships between environmental factors and NSC allocation. Moreover, the response and adaptation strategies of NSC pool in trees to climate change were also discussed. Finally, some suggestions were proposed for the potential study orientation of NSC pool in trees in future climate conditions.

Effects of experimental warming on stomatal traits in leaves of maize (Zea may L.).

We examined the warming effects on the stomatal frequency, stomatal aperture size and shape, and their spatial distribution pattern of maize (Zea may L.) leaves using a light microscope, an electron scanning microscope, and geostatistic techniques. A field manipulative experiment was conducted to elevate canopy temperature by 2.08°C, on average. We found that experimental warming had little effect on stomatal density, but significantly increased stomatal index due to the reduction in the number of epidermal cells under the warming treatment. Warming also significantly decreased stomatal aperture length and increased stomatal aperture width. As a result, warming significantly increased the average stomatal aperture area and stomatal aperture circumference. In addition, warming dramatically changed the stomatal spatial distribution pattern with a substantial increase in the average nearest neighbor distance between stomata on both adaxial and abaxial surfaces. The spatial distribution pattern of stomata was scale dependent with regular patterns at small scales and random patterns at larger scales on both leaf surfaces. Warming caused the stomatal distribution to become more regular on both leaf surfaces with smaller L(t) values (Ripley's K-function, L(t) is an expectation of zero for any value of t) in the warming plots than the control plots.