The combination of DNA methylation and positive regulation of anthocyanin biosynthesis by MYB and bHLH transcription factors contributes to the petal blotch formation in Xibei tree peony.

Xibei tree peony is a distinctive cultivar group that features red-purple blotches in petals. Interestingly, the pigmentations of blotches and non-blotches are largely independent of one another. The underlying molecular mechanism had attracted lots of attention from investigators, but was still uncertain. Our present work demonstrates the factors that are closely related to blotch formation in Paeonia rockii 'Shu Sheng Peng Mo'. Non-blotch pigmentation is prevented by the silencing of anthocyanin structural genes, among which PrF3H, PrDFR, and PrANS are the three major genes. We characterized two R2R3-MYBs as the key transcription factors that control the early and late anthocyanin biosynthetic pathways. PrMYBa1, which belongs to MYB subgroup 7 (SG7) was found to activate the early biosynthetic gene (EBG) PrF3H by interacting with SG5 member PrMYBa2 to form an 'MM' complex. The SG6 member PrMYBa3 interacts with two SG5 (IIIf) bHLHs to synergistically activate the late biosynthetic genes (LBGs) PrDFR and PrANS, which is essential for anthocyanin accumulation in petal blotches. The comparison of methylation levels of the PrANS and PrF3H promoters between blotch and non-blotch indicated a correlation between hypermethylation and gene silencing. The methylation dynamics of PrANS promoter during flower development revealed a potential early demethylating reaction, which may have contributed to the particular expression of PrANS solely in the blotch area. We suggest that the formation of petal blotch may be highly associated with the cooperation of transcriptional activation and DNA methylation of structural gene promoters.

Ultra-fast 3D imaging by a big codewords space division multiplexing binary coding.

Patternless binary coding strategies have been a challenge for ultra-fast 3D imaging with structured light. This Letter proposes a big codewords space division multiplexing binary coding method. From the third to the multiple order, a special spatial binary coding instead of the Gray code is created for the first time, to the best of our knowledge, to achieve an ultra-wide unambiguous range with only one auxiliary pattern. Advantageously, a connection domain segmentation technique with anomaly detection is proposed to achieve decoding of the fringe order, which cleverly avoids the misalignment problem. Additionally, a center of gravity method is applied to compensate for the codewords of the residual connected domain. The robustness and effectiveness of the proposed method for complex, isolated, and non-uniform reflectivity objects, as well as the ultra-fast 3D imaging of dynamic measurements, are experimentally verified.

Temporal phase unwrapping based on unequal phase-shifting code.

In fringe projection profilometry (FPP) based on temporal phase unwrapping (TPU), reducing the number of projecting patterns has become one of the most important works in recent years. To remove the 2π ambiguity independently, this paper proposes a TPU method based on unequal phase-shifting code. Wrapped phase is still calculated from N-step conventional phase-shifting patterns with equal phase-shifting amount to guarantee the measuring accuracy. Particularly, a series of different phase-shifting amounts relative to the first phase-shifting pattern are set as codewords, and encoded to different periods to generate one coded pattern. When decoding, Fringe order with a large number can be determined from the conventional and coded wrapped phases. In addition, we develop a self-correction method to eliminate the deviation between the edge of fringe order and the 2π discontinuity. Thus, the proposed method can achieve TPU but need to only project one additional coded pattern (e. g. 3+1), which can significantly benefit dynamic 3D shape reconstruction. The theoretical and experimental analysis verify that the proposed method performs high robustness on the reflectivity of the isolated object while ensuring the measuring speed.

Comparative analysis of cold-responsive genes under short-term cold stimulation and cold-adaptive genes under long-term heterogeneous environments reveals a cold adaptation mechanism in weeping forsythia.

Identifying cold-related genes can provide insights into the cold adaptation mechanism of weeping forsythia. In this study, we compared the changes in gene expressions and physiological and biochemical indices under short-term cold stimulation with the changes in gene sequences under a long-term heterogeneous environment to investigate the cold adaptation mechanism in weeping forsythia. The data of adaptive gene sequence changes, e.g., single nucleotide polymorphisms, were obtained from previous landscape genomics studies. The physiological and biochemical indicators and transcriptome results showed that weeping forsythia initiated a series of programs, including increasing cell osmotic pressures, scavenging ROS, activating the defense mechanism that crosses with pathogen infection, and upregulating CBF/DREB1 transcription factor 1, to cope with short-term cold stress. A reanalysis of landscape genomic data suggested that weeping forsythia responded to long-term heterogeneous cold stress by the differentiation of genes related to synthesis of aromatic substances and adenosine triphosphate. Our results supported the hypothesis that the adaptation mechanisms of species to short-term environmental stimulation and long-term stress in heterogeneous environments are different. The differences in cold tolerance among populations are not necessarily obtained by changing cold-responsive gene sequences. This study provides new insights into the cold adaptation mechanisms of plants.

Herbaceous peony PlACLB2 positively regulates red petal formation by promoting anthocyanin accumulation.

ATP-citrate lyase (ACL) gene catalyzes the formation of acetyl-CoA to provide intermediate precursors for many secondary metabolites, and also plays an important role in anthocyanin biosynthesis of plants. Herbaceous peony (Paeonia lactiflora Pall.) is an international cut flower known for its rich flower colors, however, the function of the ACL gene in flower color regulation is still unclear. Here, double-colored P. lactiflora 'Hebao Jinlian' were used to study the molecular mechanism of red petal, and acetyl-CoA and anthocyanin biosynthesis related PlACLB2, PlCHS, PlDFR, PlANS, and PlbHLH1 genes were initially found to highly expressed in the red outer-petals. The expression pattern of PlACLB2 was consistent with the spatial accumulation of anthocyanins. The correlation analysis of PlACLB2 expression pattern, acetyl-CoA content, and anthocyanin accumulation revealed that PlACLB2 was positively correlated with the acetyl-CoA and anthocyanin contents with correlation coefficients of 0.82 and 0.80. Moreover, multiple sequence alignment identified two typical conserved domains in PlACLB2, and phylogenetic analysis clustered PlACLB2 into the ACLB clade. PlACLB2 was localized in the nucleus and cytoplasm. On the one hand, silencing PlACLB2 in P. lactiflora red outer-petal resulted in lighter petal color and decreased acetyl-CoA accumulation, and quantitative analysis detected that PlACLB2-silenced petals lost more anthocyanins than the control groups with a decrease of 31.0%, and the main pigment component cyanidin-3,5-O-diglucoside was reduced by 31.9%. On the other hand, overexpression of PlACLB2 significantly promoted red coloration, acetyl-CoA content, and anthocyanin accumulation in tobacco flowers. These results demonstrated that PlACLB2 promoted anthocyanin accumulation by increasing the abundance of its precursor substrate acetyl-CoA, thereby regulating the formation of the red petals in P. lactiflora.

Nonlinear error full-field compensation method for phase measuring profilometry.

Phase measuring profilometry (PMP) has the highest measuring accuracy among structured light projection-based three-dimensional (3D) sensing methods. Due to their low-cost and high-resolution features, commercial projectors are extensively used in PMP, but they are all designed with a gamma effect purpose that considers the characteristics of human vision. Affected by the gamma effect, a set of phase-shifting sinusoidal deformed patterns captured in PMP may contain high-order harmonics which lead to nonlinear phase errors. Then, a novel nonlinear error full-field compensation method is proposed. First, the unwrapped phases modulated by the reference plane are measured several times, and their average phase is taken as the measured phase modulated by the reference plane to eliminate random errors as much as possible. Second, an expected phase plane is fitted from this average phase with the least-squares method. Third, the nonlinear phase error can be detected by subtracting the fitted expected phase from this average phase. Finally, the full-field look-up table (LUT) can be established between the nonlinear phase error and the measured phase. When an object is measured, the unwrapped phase modulated by the object is taken as the measured phase of the LUT, so the corresponding nonlinear phase error can be directly searched in the LUT. In this way, the full-field nonlinear phase error can be efficiently compensated. Experimental results show the feasibility and validity of the proposed method. The mean absolute error (MAE) can be improved from 0.48 mm to 0.06 mm, and the root mean square error (RMSE) can be improved from 0.55 mm to 0.07 mm.

A super-grayscale and real-time computer-generated Moiré profilometry using video grating projection.

By using the time-division multiplexing characteristics of the projector and the integral exposure characteristics of the charge coupled device (CCD) camera, a super-grayscale and real-time computer-generated Moiré profilometry based on video grating projection is proposed. The traditional digital static grating is of 256-grayscale at most. If an expected super-grayscale grating with a maximum grayscale of 766 is designed and divided into three 256-grayscale fringe patterns with balanced grayscale as far as possible, they can be synthesized into a repeated playing video grating instead of the traditional static grating. When the video grating is projected onto the measured object, as long as the exposure time is set to three times the refresh cycle of the video grating, the super-grayscale deformed patterns in the 766-grayscale can be captured with a 10-bit CCD camera, so that the deformed patterns are realistic. The digital error in computer-generated Moiré profilometry is effectively reduced. In addition, this method can expand the linear range of the deformed pattern by 20% in computer Moiré profilometry. Therefore, the proposed method has the perspectives of high accuracy and real-time measurement. Theoretical analysis and experimental results demonstrate the validity and capability of the proposed method.

Color-encoded single-shot computer-generated Moiré profilometry.

A color-encoded single-shot computer-generated Moiré profilometry (CSCGMP) is proposed. Two sinusoidal gratings with a π phase difference are encoded in red and blue channels respectively to combine a composite color grating. While this composite color grating is projected onto the measured object, the corresponding color deformed pattern can be captured. So two deformed patterns with a π phase difference are separated from its red and blue components respectively. After normalization and subtraction, the AC component of both separated deformed patterns can be extracted. If this AC component respectively multiplied by the two AC components of fringe patterns of reference plane with a π/2 phase difference prepared and saved on the computer in advance, two computer-generated Moiré fringes just respectively standing for sine and cosine of phase which is modulated by the height of the object relative to the reference plane are figured out. So the 3D shape of the measured object can be reconstructed with normal computer-generated Moiré profilometry. Both simulation and experimental results show the feasibility and validity of the proposed method. It has potential in real-time 3D measurement due to its single-shot feature.

Improved computer-generated moiré profilometry with flat image calibration.

An improved computer-generated moiré profilometry (CGMP) with flat image calibration is proposed. In CGMP, the purification of the AC component plays a decisive role. While a composite grating modulated with both the sinusoidal grating and its background light substitutes for the sinusoidal grating itself, the sinusoidal deformed pattern and flat image can be demodulated from the captured pattern. It is found that the sinusoidal deformed pattern and flat image may deviate, which is caused by ambient light. So flat image calibration is conducted to obtain a purer AC component that can effectively suppress the influence of ambient light and ensure the measurement accuracy, even if spectrum aliasing exists. Experimental results show the feasibility and validity of the proposed method.

Desktop lighting for comfortable use of a computer screen.

BACKGROUND: The contrast between a bright computer screen and a dark ambient environment may influence comfort of the users, especially on their eyes. OBJECTIVE: The objective of this research is to identify the optimal desktop lighting for the comfortable use of the computer screen in a dark environment. METHODS: An experiment was designed where seven illumination setups were introduced for the users to perform their leisure tasks on a computer screen. Fifteen healthy subjects participated in the experiments. During each session, durations of the eye blinks, fixations and saccades of the user were recorded by an eye tracker. His/her neck and trunk movements were recorded by a motion tracking system as well. The comfort/discomfort questionnaire, localized postural discomfort questionnaire, NASA task load index and computer user questionnaire were used to record the overall comfort/discomfort, the local perceived physical discomfort, the cognitive workload, and general/eye health problems, respectively. RESULTS: Subjective and objective measurement results indicated that users felt more comfortable with high intensity warm lights using a computer screen. We also identified that the eye fixation durations, as well as the scores of two questions in the computer user questionnaire, have significant negative correlations with comfort. On the other side, the durations of blinks and the scores of three questions in the computer user questionnaire, were significantly correlated with discomfort. CONCLUSION: The warm (3000K) and high intensity (1500 lux) light reduced the visual and cognitive fatigue of the user and therefore improve the comfort of the user during the use of a computer screen.

Metabolic profiling and gene expression analysis provides insights into flavonoid and anthocyanin metabolism in poplar.

Poplar, a woody perennial model, is a common and widespread tree genus. We cultivated two red leaf poplar varieties from bud mutation of Populus sp. Linn. '2025' (also known as Zhonglin 2025, L2025 for shot): Populus deltoides varieties with bright red leaves (LHY) and completely red leaves (QHY). After measuring total contents of flavonoid, anthocyanin, chlorophyll and carotenoid metabolites, a liquid chromatography-electrospray ionization-tandem mass spectrometry system was used for the relative quantification of widely targeted metabolites in leaves of three poplar varieties. A total of 210 flavonoid metabolites (89 flavones, 40 flavonols, 25 flavanones, 18 anthocyanins, 16 isoflavones, 7 dihydroflavonols, 7 chalcones, 5 proanthocyanidins and 3 other flavonoid metabolites) were identified. Compared with L2025, 48 and 8 flavonoids were more and less abundant, respectively, in LHY, whereas 51 and 9 flavonoids were more and less abundant in QHY, respectively. On the basis of a comprehensive analysis of the metabolic network, gene expression levels were analyzed by deep sequencing to screen for potential reference genes for the red leaves. Most phenylpropanoid biosynthesis pathway-involved genes were differentially expressed among the examined varieties. Gene expression analysis also revealed several potential anthocyanin biosynthesis regulators including three MYB genes. The study results provide new insights into poplar flavonoid metabolites and represent the theoretical basis for future studies on leaf coloration in this model tree species.

Computer-generated moiré profilometry based on fringe-superposition.

A computer-generated moiré profilometry based on algebraic addition instead of algebraic multiplication is proposed. Firstly, the two AC components of the captured fringe patterns on the reference plane with [Formula: see text] phase difference are retrieved and saved in advance. While measuring, two sinusoidal gratings with [Formula: see text] phase difference are projected onto the measured object alternatively, and the corresponding deformed patterns are captured. Then the AC component of the captured deformed pattern can be separated exactly. When the positive and negative AC component of the captured deformed pattern are added to the two prestored AC components respectively, two moiré fringes only reflect sine and cosine of the object's phase information can be successfully generated via a series of data processing procedures. Finally, the phase distribution of the measured object can be extracted by arctangent of the ratio of these two moiré fringes. Compared with computer-generated moiré profilometry based on algebraic multiplication, this proposed method can reduce the effect of high frequency noise and residual DC component on measurement and improve the measurement accuracy. While compared with [Formula: see text] phase shifting FTP, this method can measure more complex objects with better measurement capability. Experimental results verify the feasibility and validity of the proposed method.

Identification and functional analysis of three new anthocyanin R2R3-MYB genes in Petunia.

We identified three novel members of the R2R3-MYB clade of anthocyanin regulators in the genome of the purple flowering Petunia inflata S6 wild accession, and we called them ANTHOCYANIN SYNTHESIS REGULATOR (ASR). Two of these genes, ASR1 and ASR2, are inactivated by two different single base mutations in their coding sequence. All three of these genes are absent in the white flowering species P. axillaris N and P. parodii, in the red flowering P. exserta, and in several Petunia hybrida lines, including R27 and W115. P. violacea and other P. hybrida lines (M1, V30, and W59) instead harbor functional copies of the ASR genes. Comparative, functional and phylogenic analysis of anthocyanin R2R3-MYB genes strongly suggest that the ASR genes cluster is a duplication of the genomic fragment containing the other three R2R3-MYB genes with roles in pigmentation that were previously defined, the ANTHOCYANIN4-DEEP PURPLE-PURPLE HAZE (AN4-DPL-PHZ) cluster. An investigation of the genomic fragments containing anthocyanin MYBs in different Petunia accessions reveals that massive rearrangements have taken place, resulting in large differences in the regions surrounding these genes, even in closely related species. Yeast two-hybrid assays showed that the ASR proteins can participate in the WMBW (WRKY, MYB, B-HLH, and WDR) anthocyanin regulatory complex by interacting with the transcription factors AN1 and AN11. All three ASRs can induce anthocyanin synthesis when ectopically expressed in P. hybrida lines, but ASR1 appeared to be the most effective. The expression patterns of ASR1 and ASR2 cover several different petunia tissues with higher expression at early stages of bud development. In contrast, ASR3 is only weakly expressed in the stigma, ovary, and anther filaments. The characterization of these novel ASR MYB genes completes the picture of the MYB members of the petunia anthocyanin regulatory MBW complex and suggests possible mechanisms of the diversification of pigmentation patterns during plant evolution.

Chalcone synthase is ubiquitinated and degraded via interactions with a RING-H2 protein in petals of Paeonia 'He Xie'.

Flavonoids are secondary metabolites widely distributed among angiosperms, where they play diverse roles in plant growth, development, and evolution. The regulation of flavonoid biosynthesis in plants has been extensively studied at the transcriptional level, but post-transcriptional, translational, and post-translational control of flavonoid biosynthesis remain poorly understood. In this study, we analysed post-translational regulation of flavonoid biosynthesis in the ornamental plant Paeonia, using proteome and ubiquitylome profiling, in conjunction with transcriptome data. Three enzymes involved in flavonoid biosynthesis were identified as being putative targets of ubiquitin-mediated degradation. Among these, chalcone synthase (PhCHS) was shown to have the greatest number of ubiquitination sites. We examined PhCHS abundance in petals using PhCHS-specific antibody and found that its accumulation decreased at later developmental stages, resulting from 26S proteasome-mediated degradation. We further identified a ring domain-containing protein (PhRING-H2) that physically interacts with PhCHS and demonstrated that PhRING-H2 is required for PhCHS ubiquitination. Taken together, our results suggest that PhRING-H2-mediates PhCHS ubiquitination and degradation is an important mechanism of post-translational regulation of flavonoid biosynthesis in Paeonia, providing a theoretical basis for the manipulation of flavonoid biosynthesis in plants.

Expression profiling and functional characterization of a CBL-interacting protein kinase gene from Populus euphratica.

KEY MESSAGE: This is the first report on the function of a member of the CIPK family in Populus euphratica. The Ca(2+)-dependent salt overly sensitive (SOS) pathway has been shown to play an essential role in maintaining ion homeostasis and conferring salt tolerance. One component of the SOS pathway, SOS1, was identified in the salt-resistant tree P. euphratica. In this study, we identified and functionally characterized another component of the SOS pathway in this tree called PeSOS2 or PeCIPK26. On the basis of protein sequence similarity and complementation studies in Arabidopsis, PeCIPK26 was concluded to be the functional homolog of Arabidopsis AtSOS2. Yeast two-hybrid assays revealed that PeCIPK26 can interact with four calcineurin B-like (CBL) genes, i.e., PeCBL1, PeCBL4/PeSOS3, PeCBL9 and PeCBL10. Autophosphorylation assays showed that PeCIPK26 is an active protein kinase. Expression profile analysis demonstrated that PeCIPK26 is expressed in root, stem and leaf, and throughout the cell including cell membrane, cytoplasm and nucleus; in addition, it can be induced under salt-stress treatment. Functions of PeCIPK26 in salt tolerance were evaluated by gene overexpression in Arabidopsis cipk24 mutants. The better salt tolerance of transgenic plants relative to mutants was shown by their higher germination rate, lower Na(+) accumulation and higher capacity to discharge Na(+) when grown with NaCl. These results suggest the involvement of PeCIPK26 in the salt-stress response of P. euphratica.

The calcium sensor PeCBL1, interacting with PeCIPK24/25 and PeCIPK26, regulates Na(+)/K (+) homeostasis in Populus euphratica.

KEY MESSAGE: This paper is the first to directly link two types of ion channel regulation pathway into an emerging and complex CBL-CIPK signal system in wooden plant. In Arabidopsis thaliana, the calcineurin b-like (CBL) 1 gene has been shown to be necessary in response to abiotic stresses. In this study, we identified CBL1 in the woody plant Populus euphratica, designated as PeCBL1. Heterologous expression of PeCBL1 could build the resistance of sensitive phenotypes to low K(+) stress in the corresponding Arabidopsis cbl1/cbl9 mutant, and display a salt-sensitive phenotype compared with the mutant. Protein interaction analysis showed that PeCBL1 can interact with PeCIPK24, 25 and 26, and form different complexes of PeCBL-PeCIPK. To further investigate the mechanism of PeCBL1, we analyzed the fluxes of K(+) and Na(+) in roots of the wild-type Arabidopsis, cbl1/9 mutant, and PeCBL1 transgenic plants under low K(+) stress and high Na(+) stress. These analyses revealed that, compared to the cbl1/9 mutant, the PeCBL1 transgenic plant roots exhibited a higher capacity to absorb K(+) after exposure to low K(+) stress, and a lower capacity to discharge Na(+) after exposure to salt stress. The results suggest that CBL1 interacts with CIPK24, CIPK25 and CIPK26 to regulate Na(+)/K(+) homeostasis in Populus euphratica.

Shaker-like potassium channels in Populus, regulated by the CBL-CIPK signal transduction pathway, increase tolerance to low-K+ stress.

Shaker-like potassium channels in plants play an important role in potassium absorption and transport. Here, we characterized 11 genes encoding shaker-like channels from Populus trichocarpa. Furthermore, two homologs from this family were isolated from Populus euphratica and named PeKC1 and PeKC2. Subcellular localization analysis of them in Nicotiana benthamiana revealed that they are located in the cell membrane. Yeast two-hybrid assays showed that they not only interacted strongly with PeCIPK24, a homolog of AtCIPK23, but also interacted with several other CIPK members, including PeCIPK10 and PeCIPK17. To further analyze their function, we over-expressed PeKC1 or PeKC2 in akt1 mutant, the results show that the transgenic plant can recover the mutant phonotype sensitive to low-K(+) stress. This means PeKC1 or PeKC2 can complement the function of AKT1 in akt1 mutant, involved in the CBL1-CIPK23 signal transduction pathway and play an important role under low-K(+) stress.

Molecular characterization of putative vacuolar NHX-type Na(+)/H(+) exchanger genes from the salt-resistant tree Populus euphratica.

The vacuolar NHX-type Na(+)/H(+) exchangers play a key role in salt tolerance in plants. However, little is known about the Na(+)/H(+) exchangers in the salt-resistant tree, Populus euphratica. In this study, we identified six putative vacuolar Na(+)/H(+) exchanger genes from P. euphratica, designated as PeNHX1-6. Real-time polymerase chain reaction indicated that the PeNHX1/3/6 transcripts were abundant compared with the other three PeNHX genes in the three tissues (roots, stems and leaves) examined. After NaCl treatment for 6 h, the transcript levels of PeNHX1-6 were upregulated in the roots. To address the function of PeNHX1-6, complementation studies were performed with the salt-sensitive yeast mutant strain R100, which lacks activity of the endosomal Na(+)/H(+) antiporter NHX1. The results showed that PeNHX1-6 compensates, at least in part, for the function of yeast NHX1. Moreover, PeNHX3 was targeted to the tonoplast when transiently expressed in onion. Together, these results suggest that PeNHX1-6 function as vacuolar Na(+)/H(+) exchangers and that PeNHX products play an important role in the salt resistance of P. euphratica.