TIME FOR COFFEE regulates phytochrome A-mediated hypocotyl growth through dawn-phased signaling.

To enhance plant fitness under natural conditions, the circadian clock is synchronized and entrained by light via photoreceptors. In turn, the circadian clock exquisitely regulates the abundance and activity of photoreceptors via largely uncharacterized mechanisms. Here we show that the clock regulator TIME FOR COFFEE (TIC) controls the activity of the far-red light photoreceptor phytochrome A (phyA) at multiple levels in Arabidopsis thaliana. Null mutants of TIC displayed dramatically increased sensitivity to light irradiation with respect to hypocotyl growth, especially to far-red light. RNA-sequencing demonstrated that TIC and phyA play largely opposing roles in controlling light-regulated gene expression at dawn. Additionally, TIC physically interacts with the transcriptional repressor TOPLESS (TPL), which was associated with the significantly increased PHYA transcript levels in the tic-2 and tpl-1 mutants. Moreover, TIC interacts with phyA in the nucleus, thereby affecting phyA protein turnover and the formation of phyA nuclear speckles following light irradiation. Genetically, phyA was found to act downstream of TIC in regulating far red light-inhibited growth. Taken together, these findings indicate that TIC acts as a major negative regulator of phyA by integrating transcriptional and post-translational mechanisms at multiple levels.

BBX19 fine-tunes the circadian rhythm by interacting with PSEUDO-RESPONSE REGULATOR proteins to facilitate their repressive effect on morning-phased clock genes.

The core plant circadian oscillator is composed of multiple interlocked transcriptional-translational feedback loops, which synchronize endogenous diel physiological rhythms to the cyclic changes of environmental cues. PSEUDO-RESPONSE REGULATORS (PRRs) have been identified as negative components in the circadian clock, though their underlying molecular mechanisms remain largely unknown. Here, we found that a subfamily of zinc finger transcription factors, B-box (BBX)-containing proteins, have a critical role in fine-tuning circadian rhythm. We demonstrated that overexpressing Arabidopsis thaliana BBX19 and BBX18 significantly lengthened the circadian period, while the null mutation of BBX19 accelerated the circadian speed. Moreover, BBX19 and BBX18, which are expressed during the day, physically interacted with PRR9, PRR7, and PRR5 in the nucleus in precise temporal ordering from dawn to dusk, consistent with the respective protein accumulation pattern of PRRs. Our transcriptomic and genetic analysis indicated that BBX19 and PRR9, PRR7, and PRR5 cooperatively inhibited the expression of morning-phased clock genes. PRR proteins affected BBX19 recruitment to the CCA1, LHY, and RVE8 promoters. Collectively, our findings show that BBX19 interacts with PRRs to orchestrate circadian rhythms, and suggest the indispensable role of transcriptional regulators in fine-tuning the circadian clock.

B-Box proteins BBX28 and BBX29 interplay with PSEUDO-RESPONSE REGULATORS to fine-tune circadian clock in Arabidopsis.

As an endogenous time-keeping mechanism, the circadian clock benefits plant fitness and adaptation to the rhythmically changed diel environments. The key components within the core oscillator of plant circadian clock have been extensively characterised, however, the fine-tuning circadian regulators are still less identified. Here, we demonstrated that BBX28 and BBX29, the two B-Box V subfamily members lacking DNA-binding motifs, are involved in the regulation of Arabidopsis circadian clock. Over-expressing either BBX28 or BBX29 significantly lengthened circadian period, whereas loss-of-function of BBX28 rather than BBX29 displayed a modestly long period in free-running condition. Mechanistically, BBX28 and BBX29 interacted with core clock components PRR5, PRR7 and PRR9 in nucleus to augment their transcriptional repressive activities. RNA-sequencing analysis further revealed that BBX28 and BBX29 shared 686 common differentially expressed genes (DEGs) including a subset of known direct transcriptional targets of PRR proteins within core oscillator, including CCA1, LHY, LNKs and RVE8 etc. Intriguingly, PRR proteins can feedback repress BBX28 and BBX29 transcription by associating with their promoters. Together, our findings unmasked an exquisite mechanism in which BBX28 and BBX29 interplay with PRR proteins to fine-tune the circadian pace.

SDC mediates DNA methylation-controlled clock pace by interacting with ZTL in Arabidopsis.

Molecular bases of eukaryotic circadian clocks mainly rely on transcriptional-translational feedback loops (TTFLs), while epigenetic codes also play critical roles in fine-tuning circadian rhythms. However, unlike histone modification codes that play extensive and well-known roles in the regulation of circadian clocks, whether DNA methylation (5mC) can affect the circadian clock, and the associated underlying molecular mechanisms, remains largely unexplored in many organisms. Here we demonstrate that global genome DNA hypomethylation can significantly lengthen the circadian period of Arabidopsis. Transcriptomic and genetic evidence demonstrate that SUPPRESSOR OF drm1 drm2 cmt3 (SDC), encoding an F-box containing protein, is required for the DNA hypomethylation-tuned circadian clock. Moreover, SDC can physically interact with another F-box containing protein ZEITLUPE (ZTL) to diminish its accumulation. Genetic analysis further revealed that ZTL and its substrate TIMING OF CAB EXPRESSION 1 (TOC1) likely act downstream of DNA methyltransferases to control circadian rhythm. Together, our findings support the notion that DNA methylation is important to maintain proper circadian pace in Arabidopsis, and further established that SDC links DNA hypomethylation with a proteolytic cascade to assist in tuning the circadian clock.

Microbial contamination in distributed drinking water purifiers induced by water stagnation.

Small-scale distributed water purifiers (SSDWPs), providing better quality drinking water, are popularly used both in homes and in the public domain. Non-continuous operation leads to water stagnation and ultimately induces microbial contamination. However, information related to such contamination in these purifiers is reported scarcely. In the present study, an SSDWP, consisting of sand filtration (SF), granular activated carbon (GAC), and ultrafiltration (UF) processes, was established to explore microbial changes induced by water stagnation, based on the aspects of bacterial count, microbial size, microbiome and pathogenic communities. Our results primary showed that: first, compared with drinking water distribution system (DWDS), bacterial counts increased more rapidly in SSDWPs, growing to > 500 cfu/mL after 2.5 h stagnation. The proportion of intact cells also increased with stagnation time. Conversely, microbial size decreased with stagnation time according to changes in forward scatter detected using flow cytometry. Second, microbiome evolution followed the isolated island model, while in stagnated DWDS, microbiome evolved according to the continent island model, and the former had higher abundance of biodiversity. Furthermore, stagnation evidently caused microbiome changes in each unit, and spatial differences contributed to microbiome dissimilarity more significantly than temporal differences. Third, Mycobacterium was the dominant pathogenic genus in the SF and GAC units while Acinetobacter was the most abundant in the UF unit. Pathogenic risks increased with water stagnation time and lower nutrients level contributed to pathogenic community richness. Therefore, terminal disinfection of SSDWPs is strongly advised.

Comparison of real-world effectiveness between valsartan and non-RAS inhibitor monotherapy on the incidence of new diabetes in Chinese hypertensive patients: An electronic health recording system based study.

This study aimed to compare the real-world effectiveness of valsartan and non renin-angiotensin system (non-RAS) agent monotherapy on the incidence of new on-set diabetes (NOD) in Chinese hypertensive patients. It was based on an electronic Health Recording System database from Minhang District of Shanghai. Hypertensive patients aged ≥18 years continuously taking either valsartan or non-RAS agent monotherapy for >12 months were included. Hazard ratios (HR) of NOD events were estimated using propensity score matching method and multivariate regression. Of 29295 patients, there were 2107 in valsartan group, 21397 in CCB group, 4094 in ß-blockers group and 1697 in diuretics group. Two-year follow-up revealed NOD rates of 11.09 and 14.22 per 100 persons per year in valsartan and non-RAS inhibitor groups (HR = 0.77, 95% confidence interval 0.65-0.93, P = 0.006), respectively. Among non-RAS agents, CCB group had the highest incidence of NOD (21.72 per 100 persons per year). Comparisons between CCB sub-groups revealed the highest NOD incidence for nifedipine, followed by amlodipine and felodipine. NOD incidences in ß-blockers and diuretics groups (11.70 and 10.50 per 100 persons per year, respectively) were not significantly different from valsartan group. Compared with non-RAS inhibitors, particularly CCBs, valsartan could significantly reduce the incidence of NOD.

Aschoff's rule on circadian rhythms orchestrated by blue light sensor CRY2 and clock component PRR9.

Circadian pace is modulated by light intensity, known as the Aschoff's rule, with largely unrevealed mechanisms. Here we report that photoreceptor CRY2 mediates blue light input to the circadian clock by directly interacting with clock core component PRR9 in blue light dependent manner. This physical interaction dually blocks the accessibility of PRR9 protein to its co-repressor TPL/TPRs and the resulting kinase PPKs. Notably, phosphorylation of PRR9 by PPKs is critical for its DNA binding and repressive activity, hence to ensure proper circadian speed. Given the labile nature of CRY2 in strong blue light, our findings provide a mechanistic explanation for Aschoff's rule in plants, i.e., blue light triggers CRY2 turnover in proportional to its intensity, which accordingly releasing PRR9 to fine tune circadian speed. Our findings not only reveal a network mediating light input into the circadian clock, but also unmask a mechanism by which the Arabidopsis circadian clock senses light intensity.

Coordinative regulation of plants growth and development by light and circadian clock.

The circadian clock, known as an endogenous timekeeping system, can integrate various cues to regulate plant physiological functions for adapting to the changing environment and thus ensure optimal plant growth. The synchronization of internal clock with external environmental information needs a process termed entrainment, and light is one of the predominant entraining signals for the plant circadian clock. Photoreceptors can detect and transmit light information to the clock core oscillator through transcriptional or post-transcriptional interactions with core-clock components to sustain circadian rhythms and regulate a myriad of downstream responses, including photomorphogenesis and photoperiodic flowering which are key links in the process of growth and development. Here we summarize the current understanding of the molecular network of the circadian clock and how light information is integrated into the circadian system, especially focus on how the circadian clock and light signals coordinately regulate the common downstream outputs. We discuss the functions of the clock and light signals in regulating photoperiodic flowering among various crop species.

Producing and storing self-sustaining drinking water from rainwater for emergency response on isolated island.

Drinking water on isolated islands includes treated rainwater, water shipped from the mainland, and desalinated seawater. However, marine transportation and desalination plants are vulnerable to emergencies, such as extreme weather, making self-sustaining stand-by water for emergency response essential. Rainwater is ideal for producing the stand-by water, and rainwater harvesting is sustainable and clean, and prolonged biostability can be ensured by managing biological and chemical parameters. The present study applied a stand-by drinking water purification system (primarily including nanofiltration and low-dose chlorination) to explore the feasibility of producing and storing cleaner drinking water from rainwater and the following conclusions were drawn. First, treatment of rainwaters ensures biosafety for seven days, which is longer than that for untreated rainwater; the proportion of opportunistic pathogens decreased from 23.40-7.77% after nanofiltration, and it was proposed that the microbial community converges after advanced water treatment. Second, chemical qualities were improved. Local resource coral sand prevents pH in rainwater from decreasing below 6.5, and treated rainwater had lower disinfection by-product potential and higher disinfection efficiency, allowing periodical rainwater recycling. Third, harvesting rainwater was extremely cost-effective, with an operation cost of 1.5-2.5 RMB/m3. From biosafety, chemical safety, and economic cost perspectives, self-sustaining water from rainwater can contributes to the development of sustainable and cost-effective water supply systems on isolated islands. Mixing treated rainwater and desalinated seawater reasonably guarantees sufficiency and safety.

Microplastics as carbon-nutrient sources and shaper for microbial communities in stagnant water.

Microplastics (MPs) are emerging pollutants as vectors for microbial colonization, but their role as nutrients sources for microbial communities has rarely been reported. This study explored the impact of six types of MPs on assimilable organic carbon (AOC) and microbial communities over eight weeks. The following were the primary conclusions: (1) MPs contributed to AOC increment and subsequently increased bacterial regrowth potential. The maximum AOC reached 722.03 µg/L. The increase in AOC formation corresponded to AOC NOX, except in PVC samples where AOC P17 primarily increased. (2) The MPs accelerated bacterial growth and changed the bacterial distribution between the biofilm and water phases. A high MP surface-area-to-volume ratio or low MPs density contributed to bacterial accumulation and biofilm formation around the plastisphere, thereby decreasing the relative microbial proportion in the water phase. (3) High-throughput sequencing and scanning electron microscope revealed that different MPs shaped various microbial communities temporally and spatially. (4) Biofilm formatting and formatted models were established and simulated to explain the kinetic interaction between the AOC and bacteria inhabiting the plastisphere. Finally, the challenges that plastic-deprived AOC represent in terms of anti-bacterial measures and chemical safety are discussed.