Photoperiod and gravistimulation-associated Tiller Angle Control 1 modulates dynamic changes in rice plant architecture.

KEY MESSAGE: TAC1 is involved in photoperiodic and gravitropic responses to modulate rice dynamic plant architecture likely by affecting endogenous auxin distribution, which could explain TAC1 widespread distribution in indica rice. Plants experience a changing environment throughout their growth, which requires dynamic adjustments of plant architecture in response to these environmental cues. Our previous study demonstrated that Tiller Angle Control 1 (TAC1) modulates dynamic changes in plant architecture in rice; however, the underlying regulatory mechanisms remain largely unknown. In this study, we show that TAC1 regulates plant architecture in an expression dose-dependent manner, is highly expressed in stems, and exhibits dynamic expression in tiller bases during the growth period. Photoperiodic treatments revealed that TAC1 expression shows circadian rhythm and is more abundant during the dark period than during the light period and under short-day conditions than under long-day conditions. Therefore, it contributes to dynamic plant architecture under long-day conditions and loose plant architecture under short-day conditions. Gravity treatments showed that TAC1 is induced by gravistimulation and negatively regulates shoot gravitropism, likely by affecting auxin distribution. Notably, the tested indica rice containing TAC1 displayed dynamic plant architecture under natural long-day conditions, likely explaining the widespread distribution of TAC1 in indica rice. Our results provide new insights into TAC1-mediated regulatory mechanisms for dynamic changes in rice plant architecture.

STRIPE3, encoding a human dNTPase SAMHD1 homolog, regulates chloroplast development in rice.

Chloroplast is an important organelle for photosynthesis and numerous essential metabolic processes, thus ensuring plant fitness or survival. Although many genes involved in chloroplast development have been identified, mechanisms underlying such development are not fully understood. Here, we isolated and characterized the stripe3 (st3) mutant which exhibited white-striped leaves with reduced chlorophyll content and abnormal chloroplast development during the seedling stage, but gradually produced nearly normal green leaves as it developed. Map-based cloning and transgenic tests demonstrated that a splicing mutation in ST3, encoding a human deoxynucleoside triphosphate triphosphohydrolase (dNTPase) SAMHD1 homolog, was responsible for st3 phenotypes. ST3 is highly expressed in the third leaf at three-leaf stage and expressed constitutively in root, stem, leaf, sheath, and panicle, and the encoded protein, OsSAMHD1, is localized to the cytoplasm. The st3 mutant showed more severe albino leaf phenotype under exogenous 1-mM dATP/dA, dCTP/dC, and dGTP/dG treatments compared with the control conditions, indicating that ST3 is involved in dNTP metabolism. This study reveals a gene associated with dNTP catabolism, and propose a model in which chloroplast development in rice is regulated by the dNTP pool, providing a potential application of these results to hybrid rice breeding.

Tiller Angle Control 1 Is Essential for the Dynamic Changes in Plant Architecture in Rice.

Plant architecture is dynamic as plants develop. Although many genes associated with specific plant architecture components have been identified in rice, genes related to underlying dynamic changes in plant architecture remain largely unknown. Here, we identified two highly similar recombinant inbred lines (RILs) with different plant architecture: RIL-Dynamic (D) and RIL-Compact (C). The dynamic plant architecture of RIL-D is characterized by 'loosetiller angle (tillering stage)-compact (heading stage)-loosecurved stem (maturing stage)' under natural long-day (NLD) conditions, and 'loosetiller angle (tillering and heading stages)-loosetiller angle and curved stem (maturing stage)' under natural short-day (NSD) conditions, while RIL-C exhibits a compact plant architecture both under NLD and NSD conditions throughout growth. The candidate locus was mapped to the chromosome 9 tail via the rice 8K chip assay and map-based cloning. Sequencing, complementary tests, and gene knockout tests demonstrated that Tiller Angle Control 1 (TAC1) is responsible for dynamic plant architecture in RIL-D. Moreover, TAC1 positively regulates loose plant architecture, and high TAC1 expression cannot influence the expression of tested tiller-angle-related genes. Our results reveal that TAC1 is necessary for the dynamic changes in plant architecture, which can guide improvements in plant architecture during the modern super rice breeding.

Reduction of OsMPK6 activity by a R89K mutation induces cell death and bacterial blight resistance in rice.

KEY MESSAGE: The R89 is essential for the kinase activity of OsMPK6 which negatively regulates cell death and defense response in rice. Mitogen-activated protein kinase cascade plays critical roles in various vital activities, including the plant immune response, but the mechanisms remain elusive. Here, we identified and characterized a rice lesion mimic mutant osmpk6 which displayed hypersensitive response-like lesions in company with cell death and hydrogen peroxide hyperaccumulation. Map-based cloning and complementation demonstrated that a G702A single-base substitution in the second exon of OsMPK6 led to the lesion mimic phenotype of the osmpk6 mutant. OsMPK6 encodes a cytoplasm and nucleus-targeted mitogen-activated protein kinase and is expressed in the various organs. Compared with wild type, the osmpk6 mutant exhibited high resistance to the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo), likely due to the increased ROS production induced by flg22 and chitin and up-regulated expression of genes involved in pathogenesis, as well as activation of SA and JA signaling pathways after inoculation. By contrast, the OsMPK6-overexpression line (OE-1) was found to be susceptible to the bacterial pathogens, indicating that OsMPK6 negatively regulated Xoo resistance. Furthermore, the G702A single-base substitution caused a R89K mutation at both polypeptide substrate-binding site and active site of OsMPK6, and kinase activity assay revealed that the R89K mutation led to reduction of OsMPK6 activity, suggesting that the R89 is essential for the function of OsMPK6. Our findings provide insight into a vital role of the R89 of OsMPK6 in regulating cell death and defense response in rice.

Characterization and genetic analysis of the oshpl3 rice lesion mimic mutant showing spontaneous cell death and enhanced bacterial blight resistance.

Plant lesion mimic mutants have been used as ideal materials for studying pathogen defense mechanisms due to their spontaneous activation of defense responses in plants. Here, we report the identification and characterization of a rice lesion mimic mutant, oshpl3. The oshpl3 mutant initially displayed white spots on leaves of 7-day-old seedlings, and the white spots gradually turned into large brown spots during plant development, accompanied by poor metrics of major agronomic traits. Histochemical analysis showed that spontaneous cell death and H2O2 hyperaccumulation occurred in oshpl3. Defense responses were induced in the oshpl3 mutant, such as enhanced ROS signaling activated by recognition of pathogen-associated molecular patterns, and also upregulated expression of genes involved in pathogenesis and JA metabolism. These defense responses enhanced resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae. The mutated gene was identified as OsHPL3 (LOC_Os02g02000) by map-based cloning. A G1006A mutation occurred in OsHPL3, causing a G-to-D mutation of the 295th amino acid in the transmembrane region of OsHPL3. OsHPL3 localized to the chloroplast, cytoplasm, and another unknown organelle, while the mutated protein OsHPL3G295D was not obviously observed in the chloroplast, suggesting that the G295D mutation affected its chloroplast localization. Based on our findings, the G295D mutation in OsHPL3 is most likely responsible for the phenotypes of the oshpl3 mutant. Our results provide new clues for studying the function of the OsHPL3 protein.

Evaluation of a novel automated water analyzer for continuous monitoring of toxicity and chemical parameters in municipal water supply.

A novel tool, the DAMTA analyzer (Device for Analytical Monitoring and Toxicity Assessment), designed for fully automated toxicity measurements based on luminescent bacteria as well as for concomitant determination of chemical parameters, was developed and field-tested. The instrument is a robotic water analyzer equipped with a luminometer and a spectrophotometer, integrated on a thermostated reaction plate which contains a movable carousel with 80 cuvettes. Acute toxicity is measured on-line using a wild type Photobacterium phosphoreum strain with measurable bioluminescence and unaltered sensitivity to toxicants lasting up to ten days. The EC50 values of reference compounds tested were consistent with A. fischeri and P. phosphoreum international standards and comparable to previously published data. Concurrently, a laboratory trial demonstrated the feasibility of use of the analyzer for the determination of nutrients and metals in parallel to the toxicity measurements. In a prolonged test, the system was installed only in toxicity mode at the premises of the World Fair "Expo Milano-2015″, a high security site to ensure the quality of the supplied drinking water. The monitoring program lasted for six months during which ca. 2400 toxicity tests were carried out; the results indicated a mean non-toxic outcome of -5.5 ±â€¯6.2%. In order to warrant the system's robustness in detecting toxic substances, Zn was measured daily with highly reproducible inhibition results, 70.8 ±â€¯13.6%. These results assure that this novel toxicity monitor can be used as an early warning system for protection of drinking water sources from emergencies involving low probability/high impact contamination events in source water or treated water.

Warming increases nutrient mobilization and gaseous nitrogen removal from sediments across cascade reservoirs.

Increases in water temperature, as a result of climate change, may influence biogeochemical cycles, sediment-water fluxes and consequently environmental sustainability. Effects of rising temperature on dynamics of nitrate, nitrite, ammonium, dissolved inorganic nitrogen (DIN), dissolved reactive phosphorus (DRP), dissolved organic carbon (DOC) and gaseous nitrogen (N2 and N2O) were examined in a subtropical river (the Jiulong River, southeast China) by microcosm experiments. Slurry sediments and overlying water were collected from three continuous cascade reservoirs, and laboratory incubations were performed at four temperature gradients (5 °C, 15 °C, 25 °C and 35 °C). Results indicated: (1) warming considerably increased sediment ammonium, DIN and DOC fluxes to overlying water; (2) warming increased retention of nitrate, and to a lesser extent, nitrite, corresponding to increases in N2 and N2O emission; (3) DRP was retained but released from Fe/Al-P enriched sediments at high temperature (35 °C) due to enhanced coupled transformation of carbon and nitrogen with oxygen deficiency. Using relationships between sediment fluxes and temperature, a projected 2.3°C-warming in future would increase ammonium flux from sediment by 7.0%-16.8%, while increasing nitrate flux into sediment by 8.9%-28.6%. Moreover, substrates (e.g., grain size, carbon availability) influenced nutrient delivery and cycling across cascade reservoirs. This study highlights that warming would increase bioreactive nutrient (i.e., ammonium and phosphate) mobilization with limited gaseous N removal from sediments, consequently deteriorating water quality and increasing eutrophication with future climate change.

Ammonium and phosphate enrichment across the dry-wet transition and their ecological relevance in a subtropical reservoir, China.

Small river reservoirs are widespread and can be ecologically sensitive across the dry-wet transition under monsoon climate with respect to nutrient loading and phenology. Monthly sampling and high-frequency in situ measurements were conducted for a river reservoir (southeast China) in 2013-2014 to examine the seasonal pattern of nutrients and phytoplankton. We found that nutrient concentrations were runoff-mediated and determined by watershed inputs and, in some cases, by internal cycling depending on hydrology and temperature. Ammonium and phosphate were relatively enriched in February-March (a transitional period from dry/cold to wet/hot climate), which can be ascribed to initial flushing runoff from human/animal waste and spring fertilizer use. A phytoplankton bloom (mainly Chlorophyta) occurred during April after a surge of water temperature, probably due to the higher availability of inorganic nutrients and sunlight and suitable hydraulic residence time (medium flow) in the transitional period. The concentration of phytoplankton was low during May-June (wet-hot climate) when the concentrations of total suspended matter (TSM) were highest, likely owing to the "shading" effect of TSM and turbulence of high flow conditions. Nutrient-algae shifts across the dry-wet season and vertical profiles suggested that algal blooms seem to be fueled primarily by phosphate and ammonium rather than nitrate. Current findings of a strong temporal pattern and the relationship between physical parameters, nutrient and biota would improve our understanding of drivers of change in water quality and ecosystem functions with dam construction.

[Sediment-water flux and processes of nutrients and gaseous nitrogen release in a China River Reservoir].

The key processes and fluxes of nutrients (N and P) and gaseous N (N2 and N2O) across the sediment-water interface in a river reservoir (Xipi) of the Jiulong River watershed in southeast China were studied. Intact core sediment incubation of nutrients exchange, in-situ observation and lab incubation of excess dissolved N2 and N2O (products of nitrification, denitrification and Anammox), and determination of physiochemical and microbe parameters were carried out in 2013 for three representative sites along the lacustrine zone of the reservoir. Results showed that ammonium and phosphate were generally released from sediment to overlying water [with averaged fluxes of N (479.8 ± 675.4) mg. (m2. d)-1 and P (4. 56 ± 0.54) mg. (m2 d) -1] , while nitrate and nitrite diffused into the sediment. Flood events in the wet season could introduce a large amount of particulate organic matter that would be trapped by the dam reservoir, resulting in the high release fluxes of ammonium and phosphate observed in the following low-flow season. No clear spatial variation of sediment nutrient release was found in the lacustrine zone of the reservoir. Gaseous N release was dominated by excess dissolved N2 (98% of total), and the N2 flux from sediment was (15.8 ± 12. 5) mg (m2. d) -1. There was a longitudinal and vertical variation of excess dissolved N2, reflecting the combined results of denitrification and Anammox occurring in anoxic sediment and fluvial transport. Nitrification mainly occurred in the lower lacustrine zone, and the enrichment of N2O was likely regulated by the ratio of ammonium to DIN in water.

Control-oriented modeling and real-time control for the ozone dosing process of drinking water treatment.

Ozonation is one of the most important steps during drinking water treatment. To improve the efficiency of ozonation and to stabilize the quality of the treated water, control-oriented modeling and a real-time control method for the ozone dosing process are developed in this study. Compared with existing ozonation models developed by bench-scale and pilot-scale batch experiments, the model reported herein is control-oriented and based on plant-scale batch experiments. A real-time control strategy for maintaining a constant ozone exposure is attempted to meet primary disinfection requirements. An internal model control scheme is proposed to maintain a constant ozone exposure by adjusting the ozone dosage. The proposed real-time control method can cope with changing water quality, water flow rate, and process operational conditions. Both simulations and experimental studies have been carried out and implemented for the ozone dosing process control system, and the results demonstrate the effectiveness and practicality of this real-time control method.