Multi-Omics Analysis Reveals the Distinct Features of Metabolism Pathways Supporting the Fruit Size and Color Variation of Giant Pumpkin.

Pumpkin (Cucurbita maxima) is an important vegetable crop of the Cucurbitaceae plant family. The fruits of pumpkin are often used as directly edible food or raw material for a number of processed foods. In nature, mature pumpkin fruits differ in size, shape, and color. The Atlantic Giant (AG) cultivar has the world's largest fruits and is described as the giant pumpkin. AG is well-known for its large and bright-colored fruits with high ornamental and economic value. At present, there are insufficient studies that have focused on the formation factors of the AG cultivar. To address these knowledge gaps, we performed comparative transcriptome, proteome, and metabolome analysis of fruits from the AG cultivar and a pumpkin with relatively small fruit (Hubbard). The results indicate that up-regulation of gene-encoded expansins contributed to fruit cell expansion, and the increased presence of photoassimilates (stachyose and D-glucose) and jasmonic acid (JA) accumulation worked together in terms of the formation of large fruit in the AG cultivar. Notably, perhaps due to the rapid transport of photoassimilates, abundant stachyose that was not converted into glucose in time was detected in giant pumpkin fruits, implying that a unique mode of assimilate unloading is in existence in the AG cultivar. The potential molecular regulatory network of photoassimilate metabolism closely related to pumpkin fruit expansion was also investigated, finding that three MYB transcription factors, namely CmaCh02G015900, CmaCh01G018100, and CmaCh06G011110, may be involved in metabolic regulation. In addition, neoxanthin (a type of carotenoid) exhibited decreased accumulation that was attributed to the down-regulation of carotenoid biosynthesis genes in AG fruits, which may lead to pigmentation differences between the two pumpkin cultivars. Our current work will provide new insights into the potential formation factors of giant pumpkins for further systematic elucidation.

Decoding Natural Grasping Behaviors: Insights Into MRCP Source Features and Coupling Dynamics.

The effective decoding of natural grasping behaviors is crucial for the natural control of neural prosthetics. This study aims to investigate the decoding performance of movement-related cortical potential (MRCP) source features between complex grasping actions and explore the temporal and frequency differences in inter-muscular and cortical-muscular coupling strength during movement. Based on the human grasping taxonomy and their frequency, five natural grasping motions-medium wrap, adducted thumb, adduction grip, tip pinch, and writing tripod-were chosen. We collected 64-channel electroencephalogram (EEG) and 5-channel surface electromyogram (sEMG) data from 17 healthy participants, and projected six EEG frequency bands into source space for further analysis. Results from multi-classification and binary classification demonstrated that MRCP source features could not only distinguish between power grasp and precision grasp, but also detect subtle action differences such as thumb adduction and abduction during the execution phase. Besides, we found that during natural reach-and-grasp movement, the coupling strength from cortical to muscle is lower than that from muscle to cortical, except in the hold phase of γ frequency band. Furthermore, a 12-Hz peak of inter-muscular coupling strength was found in movement execution, which might be related to movement planning and execution. We believe that this research will enhance our comprehension of the control and feedback mechanisms of human hand grasping and contributes to a natural and intuitive control for brain-computer interface.

Explainable cross-task adaptive transfer learning for motor imagery EEG classification.

Objective. In the field of motor imagery (MI) electroencephalography (EEG)-based brain-computer interfaces, deep transfer learning (TL) has proven to be an effective tool for solving the problem of limited availability in subject-specific data for the training of robust deep learning (DL) models. Although considerable progress has been made in the cross-subject/session and cross-device scenarios, the more challenging problem of cross-task deep TL remains largely unexplored.Approach. We propose a novel explainable cross-task adaptive TL method for MI EEG decoding. Firstly, similarity analysis and data alignment are performed for EEG data of motor execution (ME) and MI tasks. Afterwards, the MI EEG decoding model is obtained via pre-training with extensive ME EEG data and fine-tuning with partial MI EEG data. Finally, expected gradient-based post-hoc explainability analysis is conducted for the visualization of important temporal-spatial features.Main results. Extensive experiments are conducted on one large ME EEG High-Gamma dataset and two large MI EEG datasets (openBMI and GIST). The best average classification accuracy of our method reaches 80.00% and 72.73% for OpenBMI and GIST respectively, which outperforms several state-of-the-art algorithms. In addition, the results of the explainability analysis further validate the correlation between ME and MI EEG data and the effectiveness of ME/MI cross-task adaptation.Significance. This paper confirms that the decoding of MI EEG can be well facilitated by pre-existing ME EEG data, which largely relaxes the constraint of training samples for MI EEG decoding and is important in a practical sense.

University MOOC should be added with farmer interested sections and provide individualized service to adapt to farmer training.

Vegetables represent an important agricultural industry in China. New farmers and new technologies for vegetable production have emerged in recent years, which makes farmer training very necessary. On the other hand, massive open online courses (MOOCs) are currently widely used in universities. The purpose of this study is to investigate the importance of different sections of a university MOOC focused on olericulture to farmers with different demographic characteristics and provide a basis to improve university MOOCs for farmer training. The survey results suggest that the age, education level, gender, farmer scale, facility type and profit of farmer learners are important factors determining evaluations of the importance of different MOOC sections, indicating that services customized to different farmer populations are necessary. Among different sections of MOOC "Olericulture", farmers with younger age, higher education, larger farm, more advanced facility and more profit were more interesting in sections include cultural, social and theoretical knowledge, and less interesting in practical skill sections. Based on the survey, eight new sections including one marketing subsection (new agricultural supplies and market news), one social subsection (laws and regulations), two practical subsections (practice videos, photos and videos from other farms), and three comprehensive subsections (discussion of practical issues, mechanization, and smart olericulture) were added to the original MOOC, and the results indicate that this improvement is efficient in enhancing the importance evaluations and profits of all farmer learners, especially among those with high education levels.

Genome-Wide Identification and Expression Patterns of Cucumber Invertases and Their Inhibitor Genes.

Invertases and their inhibitors play important roles in sucrose metabolism, growth and development, signal transduction, and biotic and abiotic stress tolerance in many plant species. However, in cucumber, both the gene members and functions of invertase and its inhibitor families remain largely unclear. In this study, in comparison with the orthologues of Citrullus lanatus (watermelon), Cucumis melo (melon), and Arabidopsis thaliana (Arabidopsis), 12 invertase genes and 12 invertase inhibitor genes were identified from the genome of Cucumis sativus (cucumber). Among them, the 12 invertase genes were classified as 4 cell wall invertases, 6 cytoplasmic invertases, and 2 vacuolar invertases. Most invertase genes were conserved in cucumber, melon, and watermelon, with several duplicate genes in melon and watermelon. Transcriptome analysis distinguished these genes into various expression patterns, which included genes CsaV3_2G025540 and CsaV3_2G007220, which were significantly expressed in different tissues, organs, and development stages, and genes CsaV3_7G034730 and CsaV3_5G005910, which might be involved in biotic and abiotic stress. Six genes were further validated in cucumber based on quantitative real-time PCR (qRT-PCR), and three of them showed consistent expression patterns as revealed in the transcriptome. These results provide important information for further studies on the physiological functions of cucumber invertases (CSINVs) and their inhibitors (CSINHs).

The Potential Role of bZIP55/65 in Nitrogen Uptake and Utilization in Cucumber Is Revealed via bZIP Gene Family Characterization.

The bZIP (basic leucine zipper) proteins play crucial roles in various biological functions. Nitrogen (N) is an essential element for plant growth, especially in cucumber (Cucumis sativus) due to its shallow roots. However, the regulation of bZIP genes in cucumber nitrogen metabolism has not been studied yet. In this study, we identified a total of 72 bZIP genes (CsbZIPs) in the cucumber genome that could be classified into 13 groups. These genes were unevenly distributed on seven chromosomes, and synteny analysis showed that the CsbZIP genes were expanded in a segmentally duplicating manner. Furthermore, our genome-wide expression analysis suggested that CsbZIP genes had different patterns and that five CsbZIP genes were regulated by nitrogen treatment in both leaves and roots. Consistent with CsNPF, CsbZIP55 and CsbZIP65 were regulated by nitrogen treatment in leaves and roots. Moreover, the subcellular localization showed that CsbZIP55 and CsbZIP65 were specifically located in the nucleus, and the transcriptional activation assay showed that CsbZIP55 and CsbZIP65 have transcriptional activation activity. Additionally, in the CsbZIP55 and CsbZIP65 overexpression plants, most nitrogen-regulated CsNPF genes were downregulated. Taken together, our comprehensive analysis of the bZIP gene family lays a foundation for understanding the molecular and physiological functions of CsbZIPs.

Continuous shared control of a mobile robot with brain-computer interface and autonomous navigation for daily assistance.

Although the electroencephalography (EEG) based brain-computer interface (BCI) has been successfully developed for rehabilitation and assistance, it is still challenging to achieve continuous control of a brain-actuated mobile robot system. In this study, we propose a continuous shared control strategy combining continuous BCI and autonomous navigation for a mobile robot system. The weight of shared control is designed to dynamically adjust the fusion of continuous BCI control and autonomous navigation. During this process, the system uses the visual-based simultaneous localization and mapping (SLAM) method to construct environmental maps. After obtaining the global optimal path, the system utilizes the brain-based shared control dynamic window approach (BSC-DWA) to evaluate safe and reachable trajectories while considering shared control velocity. Eight subjects participated in two-stage training, and six of these eight subjects participated in online shared control experiments. The training results demonstrated that naïve subjects could achieve continuous control performance with an average percent valid correct rate of approximately 97 % and an average total correct rate of over 80 %. The results of online shared control experiments showed that all of the subjects could complete navigation tasks in an unknown corridor with continuous shared control. Therefore, our experiments verified the feasibility and effectiveness of the proposed system combining continuous BCI, shared control, autonomous navigation, and visual SLAM. The proposed continuous shared control framework shows great promise in BCI-driven tasks, especially navigation tasks for brain-driven assistive mobile robots and wheelchairs in daily applications.

The Roles of Hormone Signals Involved in Rhizosphere Pressure Response Induce Corm Expansion in Sagittaria trifolia.

Soil is the base for conventional plant growth. The rhizosphere pressure generated from soil compaction shows a dual effect on plant growth in agricultural production. Compacted soil leads to root growth stagnation and causes bending or thickening, thus affecting the growth of aboveground parts of plants. In arrowhead (Sagittaria trifolia L.), the corms derived from the expanded tips of underground stolons are its storage organ. We found that the formation of corms was significantly delayed under hydroponic conditions without rhizosphere pressure originating from soil/sand. In the initial stage of corm expansion, the anatomic structure of arrowhead corm-forming parts harvested from hydroponics and sand culture was observed, and we found that the corm expansion was derived from cell enlargement and starch accumulation. Comparative transcriptome analysis indicated that the corm expansion was closely related to the change in endogenous hormone levels. Endogenous abscisic acid and salicylic acid concentrations were significantly increased in sand-cultured corms. Higher ethylene and jasmonic acid contents were also detected in all arrowhead samples, demonstrating that these hormones may play potential roles in the rhizosphere pressure response and corm expansion. The expression of genes participating in hormone signaling could explain the rising accumulation of certain hormones. Our current results draw an extensive model to reveal the potential regulation mechanism of arrowhead corm expansion promoted by rhizosphere pressure, which will provide important references for further studying the molecular mechanism of rhizosphere pressure modulating the development of underground storage organs in other plants.

Metabolomics and Transcription Profiling of Pumpkin Fruit Reveals Enhanced Bioactive Flavonoids and Coumarins in Giant Pumpkin (Cucurbita maxima).

Atlantic giant (AG, Cucurbita maxima) is a type of giant pumpkin in the Cucurbitaceae family and has the world's largest fruit. AG possesses excellent ornamental and economic value due to its well-known large fruit. However, giant pumpkins are usually thrown away after viewing, thus generating a waste of resources. To explore the additional value of giant pumpkins, a metabolome assay was performed between AG and Hubbard (a small fruit pumpkin) fruits. We found that bioactive compounds, especially flavonoids (including 8-prenylnaringenin, tetrahydrocurcumin, galangin, and acacetin) and coumarins (including coumarin, umbelliferone, 4-coumaryl alcohol, and coumaryl acetate), with extensive antioxidant and pharmacological functions, showed higher accumulation in AG fruit than in Hubbard fruits. Comparative transcriptomics of the two pumpkin fruits indicated that the differentially expressed genes (DEGs) encoding PAL, C4H, 4CL, CSE, HCT, CAD, and CCoAOMT were relatively highly expressed, which promoted an increased accumulation of the identified flavonoids and coumarins in giant pumpkins. In addition, the construction of a co-expression network and cis-element analysis of the promoter demonstrated that differentially expressed MYB, bHLH, AP2, and WRKY transcription factors might play vital roles in regulating the expression of DEGs involved in the biosynthesis of several flavonoids and coumarins. Our current results provide new insights into the accumulation of active compounds in giant pumpkins.

Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia.

Real-time quantitative PCR (RT-qPCR) is a method with high sensitivity and convenience that has been extensively used to analyze the expression level of target genes. A reference gene with a highly stable expression is required to ensure the accuracy of experimental results. However, the report on appropriate reference genes in arrowheads (Sagittaria trifolia) is still limited. In this study, eight candidate reference genes (ACT5, UBQ, GAPDH, CYP, NAC, IDH, SLEEPER and PLA) were selected. The candidate genes were employed in a RT-qPCR assay in different tissues at different developmental stages of the same tissue (including corm, leaf and leafstalk) in arrowheads. Five statistical algorithms, GeNorm, NormFinder, BestKeeper, delta cycle threshold (ΔCt) and RefFinder, were used to evaluate the stability of these genes' expressions in order to identify the appropriate reference genes. The results showed that UBQ was the optimum reference gene in leaf, leafstalk, root, stolon and corm, IDH exhibited the most stable expression during the expansion of corm, UBQ and PLA were the most stable reference genes in developmental stages of leaf and leafstalk, respectively. Finally, the reliability of reference genes was further confirmed by the normalization of PDS and EXP1 genes under different arrowhead tissues and developmental stages of corm, respectively. This study constitutes important guidance for the selection of reliable reference genes for analyzing the tissue- and developmental-stage-specific expression of genes in arrowheads.

Genome-Wide Identification and Expression Analysis of NPF Genes in Cucumber (Cucumis sativus L.).

The NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER family (NPF) proteins perform an essential role in regulating plant nitrate absorption and distribution and in improving plant nitrogen use efficiency. In this study, cucumber (Cucumis sativus L.) NPF genes were comprehensively analyzed at the whole genome level, and 54 NPF genes were found to be unevenly distributed on seven chromosomes in the cucumber genome. The phylogenetic analysis showed that these genes could be divided into eight subfamilies. We renamed all CsNPF genes according to the international nomenclature, based on their homology with AtNPF genes. By surveying the expression profiles of CsNPF genes in various tissues, we found that CsNPF6.4 was specifically expressed in roots, indicating that CsNPF6.4 may play a role in N absorption; CsNPF6.3 was highly expressed in petioles, which may be related to NO3- storage in petioles; and CsNPF2.8 was highly expressed in fruits, which may promote NO3- transport to the embryos. We further examined their expression patterns under different abiotic stress and nitrogen conditions, and found that CsNPF7.2 and CsNPF7.3 responded to salt, cold, and low nitrogen stress. Taken together, our study lays a foundation for further exploration of the molecular and physiological functions of cucumber nitrate transporters.

Genome-Wide Identification of WD40 Proteins in Cucurbita maxima Reveals Its Potential Functions in Fruit Development.

WD40 proteins, a super gene family in eukaryotes, are involved in multiple biological processes. Members of this family have been identified in several plants and shown to play key roles in various development processes, including acting as scaffolding molecules with other proteins. However, WD40 proteins have not yet been systematically analyzed and identified in Cucurbita maxima. In this study, 231 WD40 proteins (CmWD40s) were identified in C. maxima and classified into five clusters. Eleven subfamilies were identified based on different conserved motifs and gene structures. The CmWD40 genes were distributed in 20 chromosomes; 5 and 33 pairs of CmWD40s were distinguished as tandem and segmental duplications, respectively. Overall, 58 pairs of orthologous WD40 genes in C. maxima and Arabidopsis thaliana, and 56 pairs of orthologous WD40 genes in C. maxima and Cucumis sativus were matched. Numerous CmWD40s had diverse expression patterns in fruits, leaf, stem, and root. Several genes were involved in responses to NaCl. The expression pattern of CmWD40s suggested their key role in fruit development and abiotic stress response. Finally, we identified 14 genes which might be involved in fruit development. Our results provide valuable basis for further functional verification of CmWD40s in C. maxima.

Cucumber STACHYOSE SYNTHASE is regulated by its cis-antisense RNA asCsSTS to balance source-sink carbon partitioning.

Photosynthate partitioning between source and sink is a key determinant of crop yield. In contrast to sucrose-transporting plants, cucumber (Cucumis sativus) plants mainly transport stachyose and stachyose synthase (CsSTS) synthesizes stachyose in the vasculature for loading. Therefore, CsSTS is considered a key regulator of carbon partitioning. We found that CsSTS expression and CsSTS enzyme activity were upregulated in the vasculature and downregulated in mesophyll tissues at fruiting. In situ hybridization and tissue enrichment experiments revealed that a cis-natural antisense noncoding transcript of CsSTS, named asCsSTS, is mainly expressed in mesophyll tissues. In vitro overexpression (OE), RNA interference (RNAi), and dual luciferase reporter experiments indicated that CsSTSs are negatively regulated by asCsSTS. Fluorescence in situ hybridization revealed that asCsSTS transcript localized in leaf cytoplasm, indicating that the regulation of CsSTS by asCsSTS is a posttranscriptional process. Further investigation revealed that this regulation occurred by reducing CsSTS transcript stability through a DICER-like protein-mediated pathway. Chemically induced OE and RNAi of asCsSTS led to promotion or inhibition, respectively, of assimilate export from leaves and altered fruit growth rates. Our results suggest that the regulation of CsSTSs between the mesophyll and vasculature reduces sugar storage in mesophyll tissue and promotes assimilate export from the leaf when the plant carries fruit.

The Sink-Source Relationship in Cucumber (Cucumis sativus L.) Is Modulated by DNA Methylation.

The optimization of the sink-source relationship is of great importance for crop yield regulation. Cucumber is a typical raffinose family oligosaccharide (RFO)-transporting crop. DNA methylation is a common epigenetic modification in plants, but its role in sink-source regulation has not been demonstrated in RFO-translocating species. Here, whole-genome bisulfite sequencing (WGBS-seq) was conducted to compare the nonfruiting-node leaves (NFNLs) and leaves of fruit setting (FNLs) at the 12th node by removing all female flowers in other nodes of the two treatments. We found considerable differentially methylated genes enriched in photosynthesis and carbohydrate metabolic processes. Comparative transcriptome analysis between FNLs and NFNLs indicated that many differentially expressed genes (DEGs) with differentially methylated regions were involved in auxin, ethylene and brassinolide metabolism; sucrose metabolism; and RFO synthesis pathways related to sink-source regulation. Moreover, DNA methylation levels of six sink-source-related genes in the pathways mentioned above decreased in leaves after 5-aza-dC-2'-deoxycytidine (5-Aza-dC, a DNA methyltransferase inhibitor) treatment on FNLs, and stachyose synthase (CsSTS) gene expression, enzyme activity and stachyose content in RFO synthesis pathway were upregulated, thereby increasing fruit length and dry weight. Taken together, our findings proposed an up-to-date inference for the potential role of DNA methylation in the sink-source relationship, which will provide important references for further exploring the molecular mechanism of DNA methylation in improving the yield of RFO transport plants.

Effects of Exogenous α-Naphthaleneacetic Acid and 24-Epibrassinolide on Fruit Size and Assimilate Metabolism-Related Sugars and Enzyme Activities in Giant Pumpkin.

Size is the most important quality attribute of giant pumpkin fruit. Different concentrations and application frequencies of α-naphthaleneacetic acid (NAA) and 24-epibrassinolide (EBR) were sprayed on the leaves and fruits of giant pumpkin at different growth stages to determine their effects and the mechanism responsible for fruit size increase. NAA+EBR application improved source strength, and further analysis indicated that NAA+EBR markedly boosted net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr) and the expression level and activity of galactitol synthetase (GolS), raffinose synthetase (RS), and stachyose synthetase (STS), resulting in an increase in the synthesis of photoassimilate, especially stachyose. Concomitantly, NAA+EBR spray increased stachyose and sucrose contents throughout pumpkin fruit growth and the concentrations of glucose and fructose at 0 and 20 days post-anthesis (DPA) in peduncle phloem sap, implying that such treatment improved the efficiency of assimilate transport from the peduncle to the fruit. Furthermore, it improved the expression and activity of alkaline α-galactosidase (AGA), facilitating assimilate unloading, providing carbon skeletons and energy for fruit growth, and increasing fruit weight by more than 44.1%. Therefore, exogenous NAA and EBR increased source capacity, transportation efficiency, and sink strength, overall promoting the synthesis and distribution of photoassimilate, ultimately increasing fruit size.

The Effects of Bilateral Phase-Dependent Closed-Loop Vibration Stimulation With Motor Imagery Paradigm.

Vibration stimulation has been shown to have the potential to improve the activation pattern of unilateral motor imagery (MI) and to promote motor recovery. However, in the widely used left and right hand MI brain-computer interface (BCI) paradigm, the vibration stimuli cannot be directly applied to the imaginary side due to the spontaneity of imagery. In this study, we proposed a method of phase-dependent closed-loop vibration stimulation to be applied on both hands, and explored the effects of different vibration stimuli on the left and right hand MI-BCI. Eighteen healthy subjects were recruited and asked to perform, in sequence, MI tasks under three different conditions of vibratory feedback, which were no vibration stimulus (MI), phase-dependent closed-loop vibration stimulus (PDS), and continuous vibration stimulus (CS). Then the performance of the left and right hand MI-BCI and the patterns of brain oscillation were compared and analyzed under these different stimulation conditions. The results showed that vibration stimulation effectively boosted the activation of the sensorimotor cortex and enhanced the functional connectivity among sensorimotor-related brain regions during MI. The closed-loop stimulation evoked stronger event-related desynchronization patterns on the contralateral side of the imagined hand compared to continuous stimulation. There was a more obvious distinction between left hand task and right hand task. In addition, phase-dependent closed-loop vibration stimulation increased classification accuracy by approximately 7% (paired t-test, p=0.004, n=18) compared to MI alone, while continuous vibration stimulation only increased it by 4% (paired t-test, p=0.067, n=18). This result further demonstrated the effectiveness of the phase-dependent closed-loop vibration stimulation method in improving the overall performance of the MI paradigm and is expected to be further applied in areas such as stroke rehabilitation in the future.

Whole-genome resequencing identified QTLs, candidate genes and Kompetitive Allele-Specific PCR markers associated with the large fruit of Atlantic Giant (Cucurbita maxima).

Atlantic Giant (AG) pumpkin (Cucurbita maxima) produces the world's largest fruit. Elucidating the molecular mechanism of AG fruit formation is of scientific and practical importance. In this research, genome-wide resequencing of an F2 population produced by a cross between AG and its small-fruit ancestor Hubbard was used to identify quantitative trait loci (QTLs) and candidate genes. Transgressive segregation of fruit size-related traits was observed in the F2 population, suggesting that fruit size was a quantitative trait controlled by multiple genes. A genetic map with an average physical distance of 154 kb per marker was constructed, and 13 QTLs related to fruit size were identified using bin-map construction. RNA sequencing analysis revealed that pathways associated with assimilate accumulation into the fruit, including carbohydrate metabolism, were significantly enriched in differentially expressed genes. According to the predicted impact of mutation on the biological function of certain proteins, 13 genes were selected as candidate genes associated with fruit size, among which two phytohormone-related genes, CmaCh17G011340 (a flavin-containing monooxygenase) and CmaCh04G029660 (a leucine-rich repeat protein kinase) were chosen for further investigation. Finally, one insertion-deletion (inDel) and three single nucleotide polymorphisms (SNPs) were successfully transformed to Kompetitive Allele-Specific PCR (KASP) markers. The novel QTLs and candidate genes identified provide insights into the genetic mechanism of large fruit formation of AG, and the genetic map and tightly linked KASP markers developed in this study can be employed for marker-assisted breeding to alter fruit size of C. maxima.

Galactinol synthase 1 improves cucumber performance under cold stress by enhancing assimilate translocation.

Cucumber (Cucumis sativus L.) predominately translocates raffinose family oligosaccharides (RFOs) in the phloem and accumulates RFOs in leaves. Galactinol synthase (GolS) catalyzes the critical step of RFO biosynthesis, and determining the functional diversity of multiple GolS isoforms in cucumber is of scientific significance. In this study, we found that all four isoforms of CsGolS in the cucumber genome were upregulated by different abiotic stresses. ß-glucuronidase staining and tissue separation experiments suggested that CsGolS1 is expressed in vascular tissues, whereas the other three CsGolSs are located in mesophyll cells. Further investigation indicates that CsGolS1 plays double roles in both assimilate loading and stress response in minor veins, which could increase the RFO concentration in the phloem sap and then improve assimilate transport under adverse conditions. Cold-induced minor vein-specific overexpression of CsGolS1 enhanced the assimilate translocation efficiency and accelerated the growth rates of sink leaves, fruits and whole plants under cold stress. Finally, our results demonstrate a previously unknown response to adverse environments and provide a potential biotechnological strategy to improve the stress resistance of cucumber.

Automated CCA-MWF Algorithm for Unsupervised Identification and Removal of EOG Artifacts From EEG.

Affective brain computer interface (ABCI) enables machines to perceive, understand, express and respond to people's emotions. Therefore, it is expected to play an important role in emotional care and mental disorder detection. EEG signals are most frequently adopted as the physiology measurement in ABCI applications. Eye blinking and movements introduce lots of artifacts into raw EEG data, which seriously affect the quality of EEG signal and the subsequent emotional EEG feature engineering and recognition. In this paper, we propose a fully automatic and unsupervised ocular artifact identification and removal algorithm named automated canonical correlation analysis (CCA)-multi-channel wiener filter (MWF) (ACCAMWF). Firstly, spatial distribution entropy (SDE) and spectral entropy (SE) are computed to automatically annotate artifact segments. Then, CCA algorithm is used to extract neural signal from artifact contaminated data to further supplement the clean EEG data. Finally, MWF is trained to remove ocular artifacts from multiple channel EEG data adaptively. Extensive experiments have been carried out on semi-simulated EEG/EOG dataset and real eye blinking-contaminated EEG dataset to verify the effectiveness of our method when compared to two state-of-the-art algorithms. The results clearly demonstrate that ACCAMWF is a promising solution for removing EOG artifacts from emotional EEG data.

CsAGA1 and CsAGA2 Mediate RFO Hydrolysis in Partially Distinct Manner in Cucumber Fruits.

A Raffinose family oligosaccharides (RFOs) is one of the major translocated sugars in the vascular bundle of cucumber, but little RFOs can be detected in fruits. Alpha-galactosidases (α-Gals) catalyze the first catabolism step of RFOs. Six α-Gal genes exist in a cucumber genome, but their spatial functions in fruits remain unclear. Here, we found that RFOs were highly accumulated in vascular tissues. In phloem sap, the stachyose and raffinose content was gradually decreased, whereas the content of sucrose, glucose and fructose was increased from pedicel to fruit top. Three alkaline forms instead of acid forms of α-Gals were preferentially expressed in fruit vascular tissues and alkaline forms have stronger RFO-hydrolysing activity than acid forms. By inducible gene silencing of three alkaline forms of α-Gals, stachyose was highly accumulated in RNAi-CsAGA2 plants, while raffinose and stachyose were highly accumulated in RNAi-CsAGA1 plants. The content of sucrose, glucose and fructose was decreased in both RNAi-CsAGA1 and RNAi-CsAGA2 plants after ß-estradiol treatment. In addition, the fresh- and dry-weight of fruits were significantly decreased in RNAi-CsAGA1 and RNAi-CsAGA2 plants. In cucurbitaceous plants, the non-sweet motif within the promoter of ClAGA2 is widely distributed in the promoter of its homologous genes. Taken together, we found RFOs hydrolysis occurred in the vascular tissues of fruits. CsAGA1 and CsAGA2 played key but partly distinct roles in the hydrolysis of RFOs.