Transcriptomic and Proteomic Analyses Unveil the Role of Nitrogen Metabolism in the Formation of Chinese Cabbage Petiole Spot.

Chinese cabbage is the most widely consumed vegetable crop due to its high nutritional value and rock-bottom price. Notably, the presence of the physiological disease petiole spot significantly impacts the appearance quality and marketability of Chinese cabbage. It is well known that excessive nitrogen fertilizer is a crucial factor in the occurrence of petiole spots; however, the mechanism by which excessive nitrogen triggers the formation of petiole spots is not yet clear. In this study, we found that petiole spots initially gather in the intercellular or extracellular regions, then gradually extend into intracellular regions, and finally affect adjacent cells, accompanied by cell death. Transcriptomic and proteomic as well as physiology analyses revealed that the genes/proteins involved in nitrogen metabolism exhibited different expression patterns in resistant and susceptible Chinese cabbage lines. The resistant Chinese cabbage line has high assimilation ability of NH4+, whereas the susceptible one accumulates excessive NH4+, thus inducing a burst of reactive oxygen species (ROS). These results introduce a novel perspective to the investigation of petiole spot induced by the nitrogen metabolism pathway, offering a theoretical foundation for the development of resistant strains in the control of petiole spot.

A BrLINE1-RUP insertion in BrCER2 alters cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis).

Glossiness is an important quality-related trait of Chinese cabbage, which is a leafy vegetable crop in the family Brassicaceae. The glossy trait is caused by abnormal cuticular wax accumulation. In this study, on the basis of a bulked segregant analysis coupled with next-generation sequencing (BSA-seq) and fine-mapping, the most likely candidate gene responsible for the glossy phenotype of Chinese cabbage was identified. It was subsequently named Brcer2 because it is homologous to AtCER2 (At4g24510). A bioinformatics analysis indicated a long interspersed nuclear element 1 (LINE-1) transposable element (named BrLINE1-RUP) was inserted into the first exon of Brcer2 in HN19-G via an insertion-mediated deletion mechanism, which introduced a premature termination codon. Gene expression analysis showed that the InDel mutation of BrCER2 reduced the transcriptional expression levels of Brcer2 in HN19-G. An analysis of cuticular waxes suggested that a loss-of-function mutation to BrCER2 in Chinese cabbage leads to a severe decrease in the abundance of very-long-chain-fatty-acids (> C28), resulting in the production of a cauline leaf, inflorescence stem, flower, and pistil with a glossy phenotype. These findings imply the insertion of the LINE-1 transposable element BrLINE1-RUP into BrCER2 can modulate the waxy traits of Chinese cabbage plants.

Genome-Wide Identification and Expression Analysis of ESPs and NSPs Involved in Glucosinolate Hydrolysis and Insect Attack Defense in Chinese Cabbage (Brassica rapa subsp. pekinensis).

Glucosinolates are secondary plant metabolites that are part of the plant's defense system against pathogens and pests and are activated via enzymatic degradation by thioglucoside glucohydrolases (myrosinases). Epithiospecifier proteins (ESPs) and nitrile-specifier proteins (NSPs) divert the myrosinase-catalyzed hydrolysis of a given glucosinolate to form epithionitrile and nitrile rather than isothiocyanate. However, the associated gene families have not been explored in Chinese cabbage. We identified three ESP and fifteen NSP genes randomly distributed on six chromosomes in Chinese cabbage. Based on a phylogenetic tree, the ESP and NSP gene family members were divided into four clades and had similar gene structure and motif composition of Brassica rapa epithiospecifier proteins (BrESPs) and B. rapa nitrile-specifier proteins (BrNSPs) in the same clade. We identified seven tandem duplicated events and eight pairs of segmentally duplicated genes. Synteny analysis showed that Chinese cabbage and Arabidopsis thaliana are closely related. We detected the proportion of various glucosinolate hydrolysates in Chinese cabbage and verified the function of BrESPs and BrNSPs in glucosinolate hydrolysis. Furthermore, we used quantitative RT-PCR to analyze the expression of BrESPs and BrNSPs and demonstrated that these genes responded to insect attack. Our findings provide novel insights into BrESPs and BrNSPs that can help further promote the regulation of glucosinolate hydrolysates by ESP and NSP to resist insect attack in Chinese cabbage.

Identification of DELLA gene family in head cabbage and analysis of mRNA transport in the heterograft.

DELLA gene family is involved in the regulation of signal transduction of plant hormones. mRNAs of GA insensitive (GAI), the member of DELLA gene family, are also signaling molecules of long-distance transport in plants. Genome-wide identification and mRNA transport analysis of the members of DELLA gene family in head cabbage (Brassica oleracea var. capitata) can provide basic data for their application in head cabbage. In this study, five members of DELLA gene family (BoRGA1, BoRGA2, BoRGL1, BoRGL2, and BoRGL3) were identified in head cabbage using genome and transcriptome data. However, head cabbage lacked a GAI gene in its genome. The scion (head cabbage, inbred line G27) and the rootstock Chinese flowering cabbage (Brassica campestris L. ssp. chinensis var. utilis Tsen et Lee) (sijiucaixin) were cleft-grafted together to produce the heterograft. Inflorescence stem of the rootstock and the corresponding inflorescence stem in Chinese flowering cabbage seedlings (as controls) were purified and analyzed with transcriptome sequencing. The total of 8, 9, 3, 5, and 1 exogenous read(s), derived respectively from BoRGA1, BoRGA2, BoRGL1, BoRGL2, and BoRGL3, were identified in the transcriptomes of the rootstocks. Nevertheless, mRNA transport of DELLA family genes from scion to rootstock did not increase the transcriptional level of the members of DELLA gene family in the rootstocks. Correlation analysis suggested that mRNA transport efficiency of the DELLA family genes was correlated with the sequence and the transcriptional level of the respective DELLA gene in the scion (head cabbage). This study lays the foundation for further investigation on the molecular mechanism of mRNA transport of the members of DELLA gene family in head cabbage.

Channel transformer U-Net: an automatic and effective skeleton extraction network for electronic speckle pattern interferometry.

The fringe skeleton extraction method may be the most straightforward method for electronic speckle pattern interferometry (ESPI) phase extraction. Due to ESPI fringe patterns having the characteristics of high noise, low contrast, and different fringe shapes, it is very difficult to extract skeletons from ESPI fringe patterns with high accuracy. To deal with this problem, we propose a skeleton extraction method based on deep learning, called channel transformer U-Net, for directly extracting skeletons from noisy ESPI fringe patterns. In the proposed method, the advanced channel-wise cross fusion transformer module is integrated into the design of deep U-Net architecture, and a loss function by combining binary cross entropy loss and poly focal loss is proposed. In addition, a marking algorithm is proposed for phase extraction, which can realize automatic identification of a skeleton line. The effectiveness of the above proposed algorithms has been verified by computer-simulated and real-dynamic ESPI measurements. The experimental results demonstrate that the proposed channel transformer U-Net can obtain accurate, complete, and smooth skeletons in all cases. The accuracy of the skeleton extraction obtained by our proposed network can reach 0.9878, and the correlation coefficient value can reach 0.9905. The skeleton line automatic marking algorithm has strong universality.

Total Synthesis of Dysifragilones A and B and Dysidavarone C.

The concise synthesis of dysifragilones A and B and dysidavarones has been accomplished for the first time in a divergent way from a common intermediate. The synthetic route features an intramolecular reductive Heck reaction to construct the 6/5/6/6/-tetracycle of dysifragilones A and B and an intramolecular palladium-catalyzed α-arylation of a sterically hindered ketone to forge the tetracyclo[7.7.1.02,7.010,15]heptadecane core structure of dysidavarone C. The late-stage introduction of amino and ethoxy groups is effective.

Distribution of primary and secondary metabolites among the leaf layers of headed cabbage (Brassica oleracea var. capitata).

The present study investigated the distribution of several primary metabolites (soluble sugar, protein, and mineral) and secondary metabolites (carotenoids, vitamin C, anthocyanin, flavonoids, and total phenolic compounds) among the leaf layers of headed cabbage. The leaf layers of two cultivars were separated and numbered sequentially from the outer to the inner leaves. The fructose and glucose content of the inner leaf layers was significantly greater than that of the outer layers. Similarly, the level of glucosinolates increased gradually from the outer leaves to the umbilicus of the leaf head. However, the content of antioxidants decreased from the outer leaves to the core of the leaf head, in line with the antioxidant capacity. The levels of soluble protein and mineral shared the similar decreasing trend. These results provide a reference for consumers to choose optimal fractions of whole cabbage heads in order to cater to their particular dietary needs.

Transcriptome-Wide Identification and Characterization of Circular RNAs in Leaves of Chinese Cabbage (Brassica rapa L. ssp. pekinensis) in Response to Calcium Deficiency-Induced Tip-burn.

Circular RNA (circRNA) is a newly discovered non-coding RNA, which play significant roles in the function and transcriptional regulation of microRNA. To date, in Chinese cabbage, the functional characteristic of circRNAs in response to calcium deficiency-induced tip-burn have not been reported. In this study, 730 circRNAs were isolated from Chinese cabbage leaves, of which 23 and 22 were differentially expressed in different calcium deficiency stages compared with the control. Forty-six host genes of the differentially expressed circRNAs were identified, and one circRNA was found to act as miRNAs sponges. Based on the functional analysis of host genes and target mRNAs of the corresponding miRNAs, the identified circRNAs might participated in response to stimulus, electron carrier activity, ATPase activity, cell wall metabolism, transcription factors and plant hormone signal transduction. ABF2, a positive regulator of the abiotic stress response in the abscisic acid (ABA) pathway, may play a role in calcium deficiency tolerance through a circRNA regulatory pathway. Correspondingly, the concentration of ABA is also increased during the Ca2+ deficiency stress. Our results suggest that circRNAs participate in a broad range of biological processes and physiological functions in the response to calcium deficiency-induced tip-burn and provide a basis for further studies of the biological roles that circRNAs play in the plant stress response.

Weighted least-squares phase-unwrapping algorithm based on the orientation coherence for discontinuous optical phase patterns.

Phase unwrapping is one of the key steps of optical interferogram analysis, among which phase discontinuity is still a challenge. In this paper, we propose a new weighted least-squares phase-unwrapping algorithm for discontinuous optical phase patterns. In the proposed algorithm, the orientation coherence is introduced to define the new weighting coefficient, which can accurately show the wrapped phase quality. According to our proposed algorithm, the new weighting coefficient has a good performance on distinguishing the continuous regions and the discontinuous regions in wrapped phase patterns. This advantage of our algorithm can ensure a more reliable unwrapped result for discontinuous optical phase patterns. We test the proposed algorithm on the computer-simulated speckle phase images and two experimentally obtained phase images, respectively, and compare them with the other five widely used methods. The experimental results demonstrate the performance of our new weighted least-squares phase-unwrapping algorithm.

Image decomposition model Shearlet-Hilbert-L2 with better performance for denoising in ESPI fringe patterns.

In this paper, we propose an image decomposition model Shearlet-Hilbert-L2 with better performance for denoising in electronic speckle pattern interferometry (ESPI) fringe patterns. In our model, the low-density fringes, high-density fringes, and noise are, respectively, described by shearlet smoothness spaces, adaptive Hilbert space, and L2 space and processed individually. Because the shearlet transform has superior directional sensitivity, our proposed Shearlet-Hilbert-L2 model achieves commendable filtering results for various types of ESPI fringe patterns, including uniform density fringe patterns, moderately variable density fringe patterns, and greatly variable density fringe patterns. We evaluate the performance of our proposed Shearlet-Hilbert-L2 model via application to two computer-simulated and nine experimentally obtained ESPI fringe patterns with various densities and poor quality. Furthermore, we compare our proposed model with windowed Fourier filtering and coherence-enhancing diffusion, both of which are the state-of-the-art methods for ESPI fringe patterns denoising in transform domain and spatial domain, respectively. We also compare our proposed model with the previous image decomposition model BL-Hilbert-L2.

General filtering method for electronic speckle pattern interferometry fringe images with various densities based on variational image decomposition.

Filtering off speckle noise from a fringe image is one of the key tasks in electronic speckle pattern interferometry (ESPI). In general, ESPI fringe images can be divided into three categories: low-density fringe images, high-density fringe images, and variable-density fringe images. In this paper, we first present a general filtering method based on variational image decomposition that can filter speckle noise for ESPI fringe images with various densities. In our method, a variable-density ESPI fringe image is decomposed into low-density fringes, high-density fringes, and noise. A low-density fringe image is decomposed into low-density fringes and noise. A high-density fringe image is decomposed into high-density fringes and noise. We give some suitable function spaces to describe low-density fringes, high-density fringes, and noise, respectively. Then we construct several models and numerical algorithms for ESPI fringe images with various densities. And we investigate the performance of these models via our extensive experiments. Finally, we compare our proposed models with the windowed Fourier transform method and coherence enhancing diffusion partial differential equation filter. These two methods may be the most effective filtering methods at present. Furthermore, we use the proposed method to filter a collection of the experimentally obtained ESPI fringe images with poor quality. The experimental results demonstrate the performance of our proposed method.

Single-lens Fourier-transform-based optical color image encryption using dual two-dimensional chaotic maps and the Fresnel transform.

We propose an optical color image encryption system based on the single-lens Fourier transform, the Fresnel transform, and the chaotic random phase masks (CRPMs). The proposed encryption system contains only one optical lens, which makes it more efficient and concise to implement. The introduction of the Fresnel transform makes the first phase mask of the proposed system also act as the main secret key when the input image is a non-negative amplitude-only map. The two CRPMs generated by dual two-dimensional chaotic maps can provide more security to the proposed system. In the proposed system, the key management is more convenient and the transmission volume is reduced greatly. In addition, the secret keys can be updated conveniently in each encryption process to invalidate the chosen plaintext attack and the known plaintext attack. Numerical simulation results have demonstrated the feasibility and security of the proposed encryption system.

Gradient vector fields based on variational image decomposition for skeletonization of electronic speckle pattern interferometry fringe patterns with variable density and their applications.

The skeletonization methods based on gradient vector fields (GVFs) have been a powerful tool for electronic speckle pattern interferometry (ESPI) fringe patterns. However, the skeletonization of ESPI fringe patterns with variable density has been an open problem in this area. In this paper, we propose a novel method for calculating GVFs based on the variational image decomposition of ESPI fringe patterns with variable density. In the proposed method, the GVFs of low-density regions are described in Beppo-Levi space, the high-density regions in Hilbert space and the noise regions in curvelet space. The GVFs of a whole image are the sum of the decomposed GVFs of low-density regions and high-density regions. The skeletons of ESPI fringe patterns with variable density can be obtained based on the topological analysis of the GVFs of a whole image. We apply the proposed method to a computer-simulated and two experimentally obtained ESPI fringe patterns with variable density and compare them with the related skeleton methods based on GVFs. The experimental results have demonstrated that the proposed method outperforms the other methods, even when the quality of the ESPI fringe patterns is considerably low.

Genome-wide identification, classification, and analysis of NADP-ME family members from 12 crucifer species.

NADP-dependent malic enzymes (NADP-MEs) play essential roles in both normal development and stress responses in plants. Here, genome-wide analysis was performed to identify 65 putative NADP-ME genes from 12 crucifer species. These NADP-ME genes were grouped into five categories of syntenic orthologous genes and were divided into three clades of a phylogenic tree. Promoter motif analysis showed that NADP-ME1 genes in Group IV were more conserved with each other than the other NADP-ME genes in Groups I and II. A nucleotide motif involved in ABA responses, desiccation and seed development was found in the promoters of most NADP-ME1 genes. Generally, the NADP-ME genes of Brassica rapa, B. oleracea and B. napus had less introns than their corresponding Arabidopsis orthologs. In these three Brassica species, the NADP-ME genes derived from the least fractionated subgenome have lost less introns than those from the medium fractionated and most fractionated subgenomes. BrNADP-ME1 showed the highest expression in petals and mature embryos. Two paralogous NADP-ME2 genes (BrNADP-ME2a and BrNADP-ME2b) shared similar expression profiles and differential expression levels. BrNADP-ME3 showed down-regulation during embryogenesis and reached its lowest expression in early cotyledonary embryos. BrNADP-ME4 was expressed widely in multiple organs and showed high expression during the whole embryogenesis process. Different NADP-ME genes of B. rapa showed differential gene expression profiles in young leaves after ABA treatment or cold stress. Our genome-wide identification and characterization of NADP-ME genes extend our understanding of the evolution or function of this family in Brassicaceae.

Genome-wide identification and characterization of aquaporin genes (AQPs) in Chinese cabbage (Brassica rapa ssp. pekinensis).

Aquaporins (AQPs) are members of a superfamily of integral membrane proteins and play a significant role in the transportation of small molecules across membranes. However, currently little is known about the AQP genes in Chinese cabbage (Brassica rapa ssp. pekinensis). In this study, a genome-wide analysis was carried out to identify the AQP genes in Chinese cabbage. In total, 53 non-redundant AQP genes were identified that were located on all of the 10 chromosomes. The number of AQP genes in Chinese cabbage was greater than in Arabidopsis. They were classified into four subfamilies, including PIP, TIP, NIP, and SIP. Thirty-three groups of AQP orthologous genes were identified between Chinese cabbage and Arabidopsis, but orthologs corresponding to AtNIP1;1 and AtPIP2;8 were not detected. Seventeen groups of paralogous genes were identified in Chinese cabbage. Three-dimensional models of the AQPs of Chinese cabbage were constructed using Phyre2, and ar/R selectivity filters were analyzed comparatively between Chinese cabbage and Arabidopsis. Generally, gene structure was conserved within each subfamily, especially in the SIP subfamily. Intron loss events have occurred during the evolution of the PIP, TIP, and NIP subfamilies. The expression of AQP genes in Chinese cabbage was analyzed in different organs. Most AQP genes were downregulated in response to salt stress. This work shows that the AQP genes of Chinese cabbage have undergone triplication and subsequent biased gene loss.

Using microfluidic chip to form brain-derived neurotrophic factor concentration gradient for studying neuron axon guidance.

Molecular gradients play a significant role in regulating biological and pathological processes. Although conventional gradient-generators have been used for studying chemotaxis and axon guidance, there are still many limitations, including the inability to maintain stable tempo-spatial gradients and the lack of the cell monitoring in a real-time manner. To overcome these shortcomings, microfluidic devices have been developed. In this study, we developed a microfluidic gradient device for regulating neuron axon guidance. A microfluidic device enables the generation of Brain-derived neurotrophic factor (BDNF) gradient profiles in a temporal and spatial manner. We test the effect of the gradient profiles on axon guidance, in the BDNF concentration gradient axon towards the high concentration gradient. This microfluidic gradient device could be used as a powerful tool for cell biology research.

Comparative analysis of alternative splicing, alternative polyadenylation and the expression of the two KIN genes from cytoplasmic male sterility cabbage (Brassica oleracea L. var. capitata L.).

The KIN genes are crucial members of the cold-regulated gene family. They play exclusive roles during the developmental processes of many organs and respond to various abiotic stresses in plants. However, little is known about the regulation of KIN gene expression in cytoplasmic male sterility (CMS) cabbages (Brassica oleracea L. var. capitata L.). We carried out a genome-wide analysis to identify the KIN genes in the CMS cabbage. Two non-redundant KIN genes, named BoKIN1 (Bol021262) and BoKIN2 (Bol030498), were identified. Reverse transcriptase PCR detected alternative splicing (AS) products of BoKIN1 (four AS products) and BoKIN2 (three AS products). In addition, alternative polyadenylation (APA) was observed for BoKIN1 and BoKIN2 in the CMS cabbage, resulting in variable 3'UTRs in their transcripts. Furthermore, the transcription levels of BoKIN1-0 and BoKIN2-0, the introns of which were spliced completely, were analyzed in various organs and young leaves treated by abiotic stresses. Our data indicated that BoKIN1-0 is highly expressed in various organs, whereas BoKIN2-0 is expressed exclusively in the stamen. Our study also suggested that BoKIN1-0 was upregulated significantly in young leaves of plants exposed to abscisic acid treatment, and cold and heat stress. BoKIN1 and BoKIN2 had differential AS and APA patterns in pre-mRNA processing, and showed differences in their expression patterns and transcript levels. BoKIN1 participates widely in organ development and responds to diverse abiotic stresses, whereas BoKIN2 plays a main role in stamen development in the CMS cabbage.