Transcriptional cascades in the regulation of 2-AP biosynthesis under Zn supply in fragrant rice.

Transcription factors (TFs) regulate gene expression to control certain genetic programs, such as growth and development, phytohormone regulation, and environmental stresses. 2-acetyl-1-pyrroline (2-AP) is the key element involved in aroma biosynthesis pathway, and the application of micronutrients can increase the 2-AP levels. However, little is known about the micronutrient-induced TFs involved in 2-AP biosynthesis. Here, we identify a number of TF families in two fragrant rice varieties, "Meixiangzhan-2" (M) and "Xiangyaxiangzhan" (X), in response to Zinc (Zn) application through transcriptomic analysis. A total of ~678 TFs were identified and grouped into 26 TF families, each of which was found to be involved in numerous signaling pathways. The WRKY TF family was found to be the most abundant, followed by bHLH and MYB. Furthermore, members of the WRKY, bHLH, MYB, ERF, HSF, MADS-box, NFY, and AP2 TF families were significantly upregulated and may be involved in the transcriptional regulation of aroma biosynthesis. In brief, this study enhances our understanding of the molecular mechanism of 2-AP biosynthesis and highlights the key TFs potentially involved in the production of aroma in fragrant rice.

ZnO nanoparticle-based seed priming modulates early growth and enhances physio-biochemical and metabolic profiles of fragrant rice against cadmium toxicity.

BACKGROUND: Cadmium (Cd) is amongst the most toxic heavy metals that severely affects crop growth, whereas application of nanoparticles (NPs) to negate the toxic effects of heavy metals could be an effective management approach. In the present study, the seeds of two fragrant rice varieties i.e., Yuxiangyouzhan and Xiangyaxiangzhan under normal and Cd stress conditions i.e., 0 and 100 mg L- 1 applied with four levels of ZnO NPs i.e., 0, 25, 50, and 100 mg L- 1. RESULTS: Seed priming with ZnO NPs had no significant effect on the seed germination (p > 0.05) however, it substantially improved the seedling growth and other related physiological attributes under the Cd stress. The mean fresh weight of the shoot, and whole seedling was increased by 16.92-27.88% and by 16.92-27.88% after ZnO NPs application. The root fresh weight, root-shoot length was also substantially improved under ZnO NPs treatment. Moreover, application of ZnO NPs induced modulations in physiological and biochemical attributes e.g., the superoxide dismutase (SOD) activity in root and shoot, the peroxidase (POD) activity and metallothionein contents in root were increased under low levels of ZnO NPs. The α-amylase and total amylase activity were improved by ZnO NPs application under Cd Stress. Besides, modulation in Zn concentration and ZnO NPs uptake in the seedling were detected. The metabolomic analysis indicated that various pathways such as alanine, aspartate and glutamate metabolism, phenylpropanoid biosynthesis, and taurine and hypotaurine metabolism were possibly important for rice response to ZnO NPs and Cd. CONCLUSION: Overall, application of ZnO NPs substantially improved the early growth and related physio-biochemical attributes in rice. Our findings provide new insights regarding the effects of ZnO NPs on seed germination, and early growth of rice, and its potential applications in developing crop resilience against Cd contaminated soils.

Ultrasonic seed treatment improved morpho-physiological and yield traits and reduced grain Cd concentrations in rice.

Rice cultivation under cadmium (Cd) contaminated soil often results in reduced growth with excess grain Cd concentrations. A pot experiment was conducted to assess the potential of ultrasonic seed treatment to alleviate Cd stress in rice. Seeds of two aromatic rice cultivars i.e., Xiangyaxiangzhan and Meixiangzhan 2 and two non-aromatic rice cultivars i.e., Huahang 31 and Guangyan 1 were exposed to ultrasonic waves for 1.5 min in 20-40 KHz mixing frequency. The experimental treatments were comprised of untreated seeds (U0) and ultrasonic treated seeds (U1) transplanted in un-contaminated soil (H0) and Cd-contaminated soil (H1). Results revealed that Cd contents and Cd accumulation in grain in U1 were 33.33-42.31% and 12.86-57.58% lower than U0 for fragrant rice cultivars under H1. Meanwhile, biomass production was higher in U1 than U0 under H0 and better yield was assessed in U1 for all cultivars under H1. The activity of peroxidase (POD) in flag leaves was increased by 8.28-115.65% for all cultivars while malondialdehyde (MDA) contents were significantly decreased in U1 compared with U0 under H0. Conclusively, ultrasonic treatment modulated Cd distribution and accumulation in different parts while improved physiological performance as well as yield and grain quality of rice under Cd contaminated conditions.

Application of γ-aminobutyric acid under low light conditions: Effects on yield, aroma, element status, and physiological attributes of fragrant rice.

Cloudy weather with low light is more common during rice growing season of South China which often leads reduced yield and aroma formations in fragrant rice. However, exogenous γ-aminobutyric acid (GABA) application could enhance the 2-acetyl-1-pyrroline (2AP) accumulations and yield of fragrant rice under low light conditions. Field and pot experiments were conducted with three fragrant rice cultivars i.e., Basmati and Yuxiangyouzhan (indica), and Yungengyou 14 (japonica) that were grown under three different treatments i.e., normal light + GABA 0 mg L-1 (CK), low light + GABA 0 mg L-1 (T1), and low light + GABA 250 mg L-1 (T2). The results revealed that the grain 2AP contents were increased by 14.67-34.83% and up to 29.34% under T1 and T2 treatments in pot and field experiments, respectively, as compared with CK. The T1 and T2 treatments improved aroma owing to regulation in the accumulation of micronutrients i.e., Na, Mn, and Fe and enzyme activities involved in 2AP biosynthesis. The grain yield was substantially reduced in T1 as compared with T2 treatment for all rice cultivars. On the other hand, GABA application improved the grain yield under low light conditions by regulating the plant growth, and related physiological and biochemical attributes in all rice cultivars. Thus, GABA could balance low light-induced 2AP content and grain yield by modulating morphological and yield related attributes as well as physio-biochemical responses of fragrant rice.

Molybdenum improves 2-acetyl-1-pyrroline, grain quality traits and yield attributes in fragrant rice through efficient nitrogen assimilation under cadmium toxicity.

Cadmium (Cd) toxicity causes severe perturbations in nitrogen (N) uptake and assimilation, and thereby interrupts normal plant growth. Molybdenum (Mo), a necessary trace element, plays important roles in N metabolism through regulating N assimilatory enzymes activities and expressions in higher plants. Taking this into account, a pot experiment was performed to explore the role of Mo in alleviating Cd-induced inhibitory effects on physio-biochemical processes, N metabolism, yield attributes and grain quality characters of two fragrant rice cultivars; Guixiangzhan and Meixiangzhan-2. Both the fragrant rice cultivars were treated with two levels of each Cd concentrations (0 and 100 mg/kg) and Mo treatments (0 and 0.15 mg/kg). The results revealed that Cd toxicity significantly reduced (p < 0.05) plant dry biomass, gaseous exchange attributes, chlorophyll contents, N utilizing and assimilatory enzymes activities, 2-acetyl-1-pyrroline (2AP) contents and grain yield in both cultivars; however, more severe inhibitions were observed in Meixiangzhan-2 than Guixiangzhan. Nevertheless, Mo application alleviated Cd stress and enhanced 2AP content and grain yield by 75.05% and 67.94% in Guixiangzhan and 87.71% and 83.51% in Meixiangzhan-2, respectively compared with no Mo application. Moreover, Mo application improved photosynthesis, chloroplast configuration, soluble protein and proline contents and also strengthened the N assimilatory pathway through efficient NO3- utilization, higher nitrate reductase, nitrite reductase, glutamine synthetase and glutamate synthase activities and transcript levels under Cd stress. Collectively, our results imply that Mo-induced enhancement in N utilization and assimilation improved yield and grain quality characters of fragrant rice cultivars under Cd stress.

Selenium improved antioxidant response and photosynthesis in fragrant rice (Oryza sativa L.) seedlings during drought stress.

Drought stress substantially influences the growth and development of many crops. The present study was conducted to investigate the effects of exogenous selenium on the growth, photosynthesis and antioxidant response of fragrant rice seedlings under drought stress. In a pot experiment, fragrant rice seedlings were subjected to drought stress (soil water potential was controlled at - 0.025 ± 5 MPa) and foliar application of selenium (Se) at 0, 10, 30, and 50 µmol L-1. Rice seedlings not exposed to drought stress and Se were used as control. Exposure of fragrant rice seedlings to drought stress resulted in significant (P < 0.05) decrease in fresh weight, dry weight, plant height and stem diameter relative to the control. Total chlorophyll, chlorophyll a, chlorophyll b and carotenoid were 20.54-27.24%, 20.82-26.83%, 19.45-29.07% and 21.49-29.17% lower with drought stress treatment compared to CK. Drought stress also significantly (P < 0.05) decreased net photosynthetic rate and soluble protein content. However, Se treatments (30 and 50 µmol L-1) substantially improved fresh weight and dry weight of fragrant rice seedlings under drought stress. Net photosynthetic rate, activities of antioxidant enzymes (GPX, SOD and CAT) and soluble protein content in rice seedlings under drought stress improved due to Se treatment. Higher transcript levels of antioxidant-related genes (GPX1, GPX4, CATA and CATC) were also observed with Se treatment.

Foliar application of sodium selenate induces regulation in yield formation, grain quality characters and 2-acetyl-1-pyrroline biosynthesis in fragrant rice.

BACKGROUND: Selenium (Se) is a beneficial element for higher plants and essential for mammals. To study the effect of the foliar application of sodium selenate on fragrant rice performance, a pot experiment was conducted in Guangdong, China. At the initial heading stage, one-time foliar application of sodium selenate with concentrations of 0, 10, 20, 30, 40 and 50 µmol·L- 1 (named CK, Se1, Se2, Se3, Se4 and Se5, respectively) were foliar applied on two fragrant rice varieties, 'Meixiangzhan-2' and 'Xiangyaxiangzhan'. RESULTS: Selenate application at the initial heading stage not only improved the grain yield of fragrant rice by increasing the seed-setting rate and grain weight, but also promoted the grain quality by increasing crude protein contents and lowering the chalky rice rate. Furthermore, Se applications enhanced the biosynthesis of 2-acetyl-1- pyrroline (2-AP), the main aromatic compound, by increasing the contents of precursors (△1- pyrroline, proline and pyrroline-5-carboxylic acid (P5C)) and the activities of enzymes (proline dehydrogenase (PRODH), △1-pyrroline-5-carboxylic acid synthetase (P5CS), and ornithine aminotransferase (OAT)) in fragrant rice. The results also showed that foliar application of sodium selenate enhanced the antioxidant system of both varieties by promoting the activities of peroxidase (POD), superoxide dismutase (SOD), catalase (CAT) and reducing the contents of malondialdehyde (MDA). Furthermore, the real-time PCR analyses depicted that foliar application of selenate up-regulated the GPX1, GPX4 and CATC transcripts. The higher antioxidative enzymatic activities might strength the stress resistant to ensure the stability of yield in fragrant rice form abiotic stress. CONCLUSIONS: Foliar applications of sodium selenate at the initial heading stage increased the grain 2-AP content by enhancing the biosynthesis-related enzymes and precursors. The grain yield and quality of fragrant rice also increased due to selenate application. Furthermore, foliar application of selenate promoted the activities of enzymes such as POD, SOD and CAT and up-regulated the expression of gene GPX4, GPX1 and CATC.

Ultrasonic seed treatment improved cadmium (Cd) tolerance in Brassica napus L.

Cadmium (Cd) affects crop growth and productivity by disrupting normal plant metabolism. To determinate whether ultrasonic (US) seed treatment can alleviate Cd stress in rape (Brassica napus L.), the seeds of two oilseed rape cultivars i.e., 'Youyanzao18' and 'Zaoshu104' were exposed to ultrasonic waves for 1 min at 20 KHz frequency. Seeds without US treatment were taken as control (CK). Results revealed that the germination rate of both cultivars was significantly (P < 0.05) higher in US treatment than CK only at 0 and 10 mg Cd L-1. The shoot and root length of both cultivars were significantly higher in US treatment than CK at all Cd treatments except the root length of Youyanzao18 at 50 mg Cd L-1. The fresh weight Youyanzao18 was significantly (P < 0.05) higher in US than CK except for Youyanzao18 at 25 mg Cd L-1. Moreover, the superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) activities and the proline, glutathione (GSH), and soluble protein contents in Youyanzao18 were relatively higher in the US treatment than CK. The malondialdehyde (MDA) contents were prominently reduced in US treatment than CK. The pods per plant, seeds per pod and rapeseed yield were increased by 15.9, 11.4, and 16.4% in Youyanzao18 and 10.3, 9.5, and 11.5% in Zaoshu104, respectively in US treatment, compared to CK. Moreover, the contents of Cd in root, stem, leaf, rape pod shell, and rapeseeds were comparatively less in US treatment than CK whereas the Cd concentrations in different plant parts of both rape cultivars were recorded as: leaf ˃ root ˃ stem ˃ rape pod shell ˃ rapeseed. In sum, the US treatment improved the morphological growth and rapeseed yield whereas reduced the Cd accumulation in different plant parts of rapeseed under Cd contaminated soil.

Multi-walled carbon nanotubes affect yield, antioxidant response, and rhizosphere microbial community of scented rice under combined cadmium-lead (Cd-Pb) stress.

Rice production is threatened by heavy metal stress. The use of multi-walled carbon nanotubes (MWCNTs) in agriculture has been reported in previous studies. We aimed to quantify the impact of MWCNTs on the growth and physiological characteristics of scented rice under cadmium (Cd) and lead (Pb) stresses. Therefore, a pot experiment was conducted, two scented rice varieties Yuxiangyouzhan and Xiangyaxiangzhan were used as materials grown under different concentrations of MWCNTs (0, 100, and 300 mg kg-1 recorded as CK, CNPs100, and CNPs300, respectively). The yield, antioxidant response, and rhizosphere microbial community of scented rice were studied. The results showed that compared with the CK treatment, the CNPs100 and CNPs300 treatments increased leaf dry weight by 17.95%-56.22% at the heading stage, and the H2O2 content in leaves decreased significantly by 36.64%-42.27% at the maturity stage. Under CNPs100 treatment, the grain yield of two scented rice varieties increased significantly by 17.54% and 27.40%, respectively. The MWCNTs regulated the distribution of the Cd and Pb in different plant tissues. The content of Cd (0.11-0.20 mg kg-1) and Pb (0.01-0.04 mg kg-1) in grain were at a safety level (<0.2 mg kg-1). Moreover, MWCNTs increased soil microbial community abundance and altered community composition structure under Cd-Pb stress, which in turn improved agronomic traits and quality of scented rice. Overall, this study suggested that the application of MWCNTs regulates the growth, yield, physiological response, and soil microbial community, the genotypes response effect of scented rice to MWCNTs is needed further studied.

Substation of vermicompost mitigates Cd toxicity, improves rice yields and restores bacterial community in a Cd-contaminated soil in Southern China.

Cadmium (Cd) contamination in agricultural soil is a global concern for soil health and food sustainability because it can cause Cd accumulation in cereal grains. An in-situ stabilizing technology (using organic amendments) has been widely used for Cd remediation in arable lands. Therefore, the current study examined the influence of vermicompost (VC) on soil biochemical traits, bacterial community diversity and composition, Cd uptake and accumulation in rice plants and grain yield in a Cd-contaminated soil during the late growing season in 2022. Different doses of VC (i.e., V1 = 0 t ha-1, V2 = 3 t ha-1 and V3 = 6 t ha-1) and two concentrations of Cd (i.e., Cd1 = 0 and Cd2 = 50 mg Cd Kg-1 were used. We performed high-throughput sequencing of 16S ribosomal RNA gene amplicons to characterize soil bacterial communities. The addition of VC considerably affected the diversity and composition of the soil bacterial community; and increased the relative abundance of phyla Chloroflexi, Proteobacteria, Acidobacteriota, Plantomycetota, Gemmatimonadota, Patescibacteria and Firmicute. In addition, VC application, particularly High VC treatment, exhibited the highest bacterial diversity and richness (i.e., Simpson, Shannon, ACE, and Chao 1 indexes) of all treatments. Similarly, the VC application increased the soil chemical traits, including soil pH, soil organic carbon (SOC), available nitrogen (AN), total nitrogen (TN), total potassium (TK), total phosphorous (TP) and enzyme activities (i.e., acid phosphatase, catalase, urease and invertase) compared to non-VC treated soil under Cd stress. The average increase in SOC, TN, AN, TK and TP were 5.75%, 41.15%, 18.51%, 12.31%, 25.45% and 29.67%, respectively, in the High VC treatment (Pos-Cd + VC3) compared with Cd stressed soil. Redundancy analysis revealed that the leading bacterial phyla were associated with SOC, AN, TN, TP and pH, although the relative abundance of Firmicutes, Proteobacteria, Bacteroidata, and Acidobacteria on a phylum basis and Actinobacteria, Gammaproteobacteria and Myxococcia on a class basis, were highly correlated with soil environmental factors. Moreover, the VC application counteracted the adverse effects of Cd on plants and significantly reduced the Cd uptake and accumulation in rice organs, such as roots, stem + leaves and grain under Cd stress conditions. Similarly, applying VC significantly increased the fragrant rice grain yield and yield traits under Cd toxicity. The correlation analysis showed that the increased soil quantities traits were crucial in obtaining high rice grain yield. Generally, the findings of this research demonstrate that the application of VC in paddy fields could be useful for growers in Southern China by sustainably enhancing soil functionality and crop production.

Exogenous TiO2 Nanoparticles Alleviate Cd Toxicity by Reducing Cd Uptake and Regulating Plant Physiological Activity and Antioxidant Defense Systems in Rice (Oryza sativa L.).

Cadmium (Cd) is a potentially hazardous element with significant biological toxicity, negatively affecting plant growth and physio-biochemical metabolism. Thus, it is necessary to examine practical and eco-friendly approaches to reduce Cd toxicity. Titanium dioxide nanoparticles (TiO2-NPs) are growth regulators that help in nutrient uptake and improve plant defense systems against abiotic and biological stress. A pot experiment was performed in the late rice-growing season (July-November) 2022 to explore the role of TiO2-NPs in relieving Cd toxicity on leaf physiological activity, biochemical attributes, and plant antioxidant defense systems of two different fragrant rice cultivars, i.e., Xiangyaxiangzhan (XGZ) and Meixiangzhan-2 (MXZ-2). Both cultivars were cultivated under normal and Cd-stress conditions. Different doses of TiO2-NPs with and without Cd-stress conditions were studied. The treatment combinations were: Cd-, 0 mg/kg CdCl2·2.5 H2O; Cd+, 50 mg/kg CdCl2·2.5 H2O; Cd + NP1, 50 mg/kg Cd + 50 TiO2-NPs mg/L; Cd + NP2, 50 mg/kg Cd + 100 TiO2-NPs mg/L; Cd + NP3, 50 mg/kg Cd + 200 TiO2-NPs mg/L; Cd + NP4, 50 mg/kg Cd + 400 TiO2-NPs mg/L. Our results showed that the Cd stress significantly (p < 0.05) decreased leaf photosynthetic efficiency, stomatal traits, antioxidant enzyme activities, and the expression of their encoding genes and protein content. Moreover, Cd toxicity destabilized plant metabolism owing to greater accretion of hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels at vegetative and reproductive stages. However, TiO2-NPs application improved leaf photosynthetic efficacy, stomatal traits, and protein and antioxidant enzyme activities under Cd toxicity. Application of TiO2-NPs decreased the uptake and accumulation of Cd in plants and levels of H2O2 and MDA, thereby helping to relieve Cd-induced peroxidation damage of leaf membrane lipids by enhancing the activities of different enzymes like ascorbate peroxidase (APX), catalase (CAT), peroxidase (POS), and superoxide dismutase (SOD). Average increases in SOD, APX, CAT, and POS activities of 120.5 and 110.4%, 116.2 and 123.4%, 41.4 and 43.8%, and 36.6 and 34.2% in MXZ-2 and XGZ, respectively, were noted in Cd + NP3 treatment across the growth stages as compared with Cd-stressed plants without NPs. Moreover, the correlation analysis revealed that the leaf net photosynthetic rate is strongly associated with leaf proline and soluble protein content, suggesting that a higher net photosynthetic rate results in higher leaf proline and soluble protein content. Of the treatments, the Cd + NP3 (50 mg/kg Cd + 200 mg/L TiO2-NPs) performed the best for both fragrant rice cultivars under Cd toxicity. Our results showed that TiO2-NPs strengthened rice metabolism through an enhanced antioxidant defense system across the growth stages, thereby improving plant physiological activity and biochemical characteristics under Cd toxicity.

A 40 % paddy surface soil organic carbon increase after 5-year no-tillage is linked with shifts in soil bacterial composition and functions.

Soil organic carbon (SOC) is related to soil fertility, crop yield, and climate change mitigation. Paddy soil is a significant carbon (C) sink, but its C sequestration potential has not been realized as the various driving factors are still not fully understood. We performed a 5-year paddy field experiment in southern China to estimate tillage effects on SOC accumulation and its relation with soil bacteria. The C input from rice residue, SOC content, CO2 flux, soil bacterial community composition, and predicted functions were analyzed. No-tillage (NT) increased (p < 0.05) rice residue C inputs (by 12.6 %-15.9 %), SOC (by 40 % at the surface soil layer compared with conventional tillage, CT), and CO2 fluxes compared with reduced tillage (RT) and CT. Also, NT significantly altered the soil bacterial community. The random forest model showed that the predicted bacterial functions of "Degradation/Utilization/Assimilation Other", "C1 Compound Assimilation", and "Amin and Polyamine Degradation" were the most important functions associated with SOC accumulation. Analysis of metabolic pathway differences indicated that NT significantly decreased the BENZCOA-PWY (anaerobic aromatic compound degradation) and the AST-PWY (L-arginine degradation II). Therefore, the rapid paddy SOC increase is associated with both residue C input (from higher rice yields) and the degradation functions regulated by soil bacteria.

Effect of Exogenous Melatonin Application on the Grain Yield and Antioxidant Capacity in Aromatic Rice under Combined Lead-Cadmium Stress.

This study aimed to determine the mechanism of exogenous melatonin application in alleviating the combined Pb and Cd (Pb-Cd) toxicity on aromatic rice (Oryza sativa L.). In this study, a pot experiment was conducted; two aromatic rice varieties, Yuxiangyouzhan and Xiangyaxiangzhan, were selected, and sprays using 50, 100, 200, and 400 µmol L-1 melatonin (denoted as S50, S100, S200, and S400) and irrigation using 100, 300, and 500 µmol L-1 melatonin (denoted as R100, R300, and R500) were also selected. The results showed that, under the S50, S100, and S200 treatments, the Pb content of aromatic rice grain decreased, and the grain yield increased significantly. Moreover, the application of exogenous melatonin significantly reduced the accumulation of H2O2 in rice leaves at maturity under Cd-Pb stress and reduced the MDA content in Xiangyaxiangzhan leaves. In addition, the microbial community structure changed significantly under S50 and R300 treatments. Some pathways, such as the synthesis of various amino acids and alanine, aspartate, and glutamate metabolism, were regulated by S50 treatment. Overall, melatonin application improved aromatic rice grain yield while reducing heavy metal accumulation by regulating the antioxidant capacity and metabolites in aromatic rice plants and altering the physicochemical properties and microbial community structures of the soil.

Post-transcriptional regulation of 2-acetyl-1-pyrroline (2-AP) biosynthesis pathway, silicon, and heavy metal transporters in response to Zn in fragrant rice.

Fragrant rice (Oryza sativa L.) has a high economic and nutritional value, and the application of micronutrients regulates 2-acetyl-1-pyrroline (2-AP) production, which is responsible for aroma in fragrant rice. Alternative splicing (AS) is an important post-transcriptional regulatory mechanism to generate transcript variability and proteome diversity in plants. However, no systematic investigation of AS events in response to micronutrients (Zn) has been performed in fragrant rice. Furthermore, the post-transcriptional regulation of genes involved in 2-AP biosynthesis is also not known. In this study, a comprehensive analysis of AS events under two gradients of Zn treatment in two different fragrant rice cultivars (Meixiangzhan-2 and Xiangyaxiangzhan) was performed based on RNA-seq analysis. A total of 386 and 598 significant AS events were found in Meixiangzhan-2 treated with low and high doses of Zn, respectively. In Xiangyaxiangzhan, a total of 449 and 598 significant AS events were found in low and high doses of Zn, respectively. Go analysis indicated that these genes were highly enriched in physiological processes, metabolism, and cellular processes in both cultivars. However, genotype and dose-dependent AS events were also detected in both cultivars. By comparing differential AS (DAS) events with differentially expressed genes (DEGs), we found a weak overlap among DAS and DEGs in both fragrant rice cultivars indicating that only a few genes are post-transcriptionally regulated in response to Zn treatment. We further report that Zn differentially regulates the expression of 2-AP biosynthesis-related genes in both cultivars and Zn treatment altered the editing frequency of single nucleotide polymorphism (SNPs) in the genes involved in 2-AP biosynthesis. Finally, we showed that epigenetic modifications associated with active gene transcription are generally enriched over 2-AP biosynthesis-related genes. Similar to the 2-AP pathway, we found that heavy metal transporters (genes related to silicon, iron, Zn and other metal transport) are also regulated at transcriptional and post-transcriptional levels in response to Zn in fragrant rice. Taken together, our results provide evidence of the post-transcriptional gene regulation in fragrant rice in response to Zn treatment and highlight that the 2-AP biosynthesis pathway and heavy metal transporters may also be regulated through epigenetic modifications. These findings will serve as a cornerstone for further investigation to understand the molecular mechanisms of 2-AP biosynthesis and regulation of heavy metal transporters in fragrant rice.

Application of inorganic passivators reduced Cd contents in brown rice in oilseed rape-rice rotation under Cd contaminated soil.

Immobilization of heavy metals by the application of chemical amendments is an eco-friendly, economical and effective method to remediate cadmium (Cd) -contaminated soils. Field experiments were conducted during 2016-2018 following oilseed rape-rice rotation with the application of inorganic passivators i.e., slaked lime (CaOH2>95%) and sepiolite (SiO2>50%, MgO>20%). The experimental treatments were comprised of: rice planting in winter fallow and without passivator (F-PA0); fallow with slaked lime (F-PA1); fallow with sepiolite (F-PA2); oilseed rape-rice rotation without passivator (R-PA0); rotation with slaked lime (R-PA1); and rotation with sepiolite (R-PA2). The slaked lime and sepiolite were applied after the harvest of rape at 2000 and 5000 kg ha-1, respectively. Results revealed that the Cd contents were reduced by 47.44-49.03% in brown rice for F-PA2 and 9.54-42.66% in soil for R-PA2. The superoxide dismutase (SOD) activity was decreased by 10.65-17.98%, the malondialdehyde (MDA) contents were reduced by 28.57% whereas the proline content was decreased by 32.61% under R-PA2. In addition, the filled grain percentage was improved by 6.87% (F-PA2) and 3.70% (R-PA1), respectively. Overall, rice sown after oilseed rape gave better yields than sown after fallow fields while application of slaked lime and sepiolite as passivator could be a potential management option to grow crops in metal polluted soils.

Light and water treatment during the early grain filling stage regulates yield and aroma formation in aromatic rice.

The effect of light and water on aromatic rice remain largely unclear. A pot experiment was conducted to investigate the influences of light-water treatments (CK: natural light and well-watered conditions, WS: natural light and water-stressed conditions, LL: low light and well-watered conditions, LL-WS: low light and water-stressed treatment) on yield and 2-acetyl-1-pyrroline (2AP) formation in aromatic rice. Compared with CK, the light-water treatments decreased grain yield (10.32-39.19%) due to reductions in the filled grain percentage and total dry weight, in the regulation of biomass distribution, and in the attributes of gas exchange and antioxidant response parameters. The 2AP content in grains increased in the LL treatment (5.08-16.32%) but decreased in the WS treatment compared with that in CK. The changes in 2AP were associated with changes in 2AP formation-related traits and element content. Low light and water stress led to yield declines in aromatic rice, but low light alleviated the decrease in 2AP content caused by water stress.

Exogenous application of zinc (Zn) at the heading stage regulates 2-acetyl-1-pyrroline (2-AP) biosynthesis in different fragrant rice genotypes.

Zinc (Zn) is an important microelement for rice and plays a key role in many physiological processes. This study assessed the physio-biochemical responses involved in biosynthesis of 2-acety-1-pyrroline (2-AP), which is a key compound in the aroma of fragrant rice, in four different fragrant rice varieties, i.e., Meixiangzhan-2, Xiangyaxiangzhan, Ruanhuayou-134, and Yunjingyou. Four concentrations (0, 0.50, 1.00 and 2.00 g L-1) of zinc chloride were applied to fragrant rice foliage at the heading stage and named CK, Zn1, Zn2 and Zn3, respectively. Our results showed that compared with CK, the Zn1, Zn2 and Zn3 treatments all significantly increased the 2-AP concentration in mature grains of the four fragrant rice genotypes. Furthermore, exogenous application of Zn not only enhanced the activities of enzymes, including proline dehydrogenase (PDH), △1-pyrroline-5-carboxylic acid synthetase (P5CS), and diamine oxidase (DAO), which are involved in 2-AP biosynthesis, but also improved the contents of the related precursors, such as Δ1-pyrroline, proline and pyrroline-5-carboxylic acid (P5C). In addition, compared to the CK treatment, the Zn2 treatment markedly increased the net photosynthetic rate of fragrant rice during the grain filling stage and increased the seed-setting rate, 1000-grain weight and grain yield in all fragrant rice genotypes. Foliar application of Zn also markedly increased the grain Zn content. In general, 1.00 g L-1 seemed to be the most suitable application concentration because the highest 2-AP content and grain weight were recorded with this treatment.

Correction to: Nitrogen application and different water regimes at booting stage improved yield and 2-acetyl-1-pyrroline (2AP) formation in fragrant rice.

It was highlighted that the original article (Mo et al. 2019) contained an error in Fig. 1f which revealed the biosynthesis pathway of 2AP. This Correction article shows the correct Fig. 1 and incorrect Fig. 1. The original article has been updated.

Nitrogen application and different water regimes at booting stage improved yield and 2-acetyl-1-pyrroline (2AP) formation in fragrant rice.

BACKGROUND: Water (W) and nitrogen (N) management generally cause regulations in the 2-acetyl-1-pyrroline (2AP) accumulation in fragrant rice; nevertheless, the feasibility of such management strategies at booting stage in improving 2AP accumulation has not been examined in details. METHODS: Field experiments were conducted in the early season (March-July) and repeated in the late season (July-November) in 2013. The treatments were applied urea (90 kg ha- 1), calcium super phosphate (90 kg ha- 1) and potassium chloride (195 kg ha- 1) as basal fertilizer, and urea (65 kg ha- 1) at tillering stage. Three N levels i.e., 0 kg N ha- 1 (N1), 30 kg N ha- 1 (N2), and 60 kg N ha- 1 (N3) and three water levels i.e., W1 treatment (well-watered treatment with water layer of 2-4 cm), W2 treatment (soil water potential was - 15 ± 5 kPa), and W3 treatment (soil water potential was - 25 ± 5 kPa) at booting stage was set up for three rice varieties i.e., Nongxiang 18, Yungengyou 14 and Basmati. The grain yield, head milled rice yield, 2AP contents and the biochemical parameters related to 2AP formation were investigated. RESULTS: Result indicated that W and N dynamics regulated the grain yield, head milled rice yield, and 2AP contents in brown rice across three varieties. The N2 and N3 treatment significantly increased the 2AP contents in brown rice by 9.54% and 11.95%, and 8.88% and 32.54% in the early and the late season, respectively; improved grain yield and head milled rice yield. The W3 treatment improved grain yield, head milled rice yield and 2AP content. Significant W and N interaction effect on 2AP content in brown rice was detected, where the W3 N3 treatment showed the strongest interaction regarding improvement of 2AP contents in brown rice. The 2AP accumulation and its related biochemical parameters and their relationships in different plant tissues at different growth stages under W and N treatments had also been assessed. The 2AP content, P5C content and DAO activity during grain filling periods was highly related to the 2AP content in brown rice. CONCLUSION: This study revealed that the 60 kg N ha- 1 coupled with - 25 ± 5 kPa treatment showed the best positive effects on yield and aroma in fragrant rice, suggested that water and nitrogen management at booting stage can improve grain yield and fragrance in fragrant rice. However, further study to evaluate the metabolic and molecular basis of 2AP accumulation in fragrant rice is needed.

Responses of plant growth, physiological, gas exchange parameters of super and non-super rice to rhizosphere temperature at the tillering stage.

Temperature is one of the critical factors affecting rice growth and yield production. This study investigated the effects of rhizosphere temperature at the tillering stage on the growth, physiological parameters and gas exchange attributes of two rice cultivars, i.e., the super rice cultivar Yuxiangyouzhan and the non-super rice cultivar Xiangyaxiangzhan, under hydroponic conditions. Three rhizosphere temperature treatments, i.e., a low-rhizosphere-temperature treatment (LT, nutrient solution at 17.5 ± 1.5 °C), an ambient-temperature treatment (AT, nutrient solution at 27.5 ± 1.5 °C), and a high-rhizosphere-temperature treatment (HT, nutrient solution at 37.5 ± 1.5 °C), were applied in this study. The results showed significant differences in most of the rice growth and physiological and gas exchange parameters as a result of cultivar and rhizosphere temperature as well as their interaction. In addition, the marked reduction in total dry weight was positively correlated with a notable reduction in plant morphological parameters, such as the fresh and dry weight of the leaves and stem sheaths, and changes in gas exchange parameters. Moreover, antioxidant reactions were active in response to high and low rhizosphere temperatures, which varied in different plant tissues. These results suggested that the super and non-super rice were sensitive to high and low rhizosphere temperatures, respectively.