Biological culture module for plant research from seed-to-seed on the Chinese Space Station.

The long-term cultivation of higher plants in space plays a substantial role in investigating the effects of microgravity on plant growth and development, acquiring valuable insights for developing a self-sustaining space life supporting system. The completion of the Chinese Space Station (CSS) provides us with a new permanent space experimental platform for long-term plant research in space. Biological Culture Module (GBCM), which was installed in the Wentian experimental Module of the CSS, was constructed with the objective of growing Arabidopsis thaliana and rice plants a full life cycle in space. The techniques of LED light control, gas regulation and water recovery have been developed for GBCM in which dry seeds of Arabidopsis and rice were set in root module of four culture chambers (CCs) and launched with Wentian module on July 24, 2022. These seeds were watered and germinated from July 28 and grew new seeds until November 26 within a duration of 120 days. To this end, both Arabidopsis and rice plants completed a full life cycle in microgravity on the CSS. As we know, this is the first space experiment achieving rice complete life cycle from seed-to-seed in space. This result demonstrates the possibility to cultivate the important food crop rice throughout its entire life cycle under the spaceflight environment and the technologies of GBCM have effectively supported the success of long-term plant culture experiments in space. These results can serve as invaluable references for constructing more expansive and intricate space plant cultivation systems in the future.

How dyadic appraisal moderate the association between dyadic coping and diabetes management efficacy.

To explore the moderating role of dyadic appraisal in the association between dyadic coping and diabetes management efficacy. Two hundred seventy six middle-aged and older couple pairs with one spouse who had diabetes were recruited from 14 community healthcare centers across Guangzhou. The moderating role of dyadic appraisal was investigated using the actor-partner interdependence moderation model. When both couples considered diabetes to be a shared condition, statistically-significant associations were found between patients' negative (ß = -22.7, p = 0.008) and neutral behaviors (ß = 13.6, p = 0.017), plus spouses' positive behaviors (ß = 22.8, p = 0.009) on their own diabetes management efficacy, respectively (i.e. actor effects); as well as between spouses' positive (ß = 16.8, p = 0.028), negative (ß = -28.5, p < 0.001), and neutral behaviors (ß = 16.9, p = 0.006) on patient's diabetes management efficacy (i.e. partner effects). Dyadic appraisal moderates the association between dyadic coping and diabetes management efficacy.

Conditions for successful implementation of couple-based collaborative management model of diabetes among community-dwelling older Chinese: a qualitative comparative analysis.

BACKGROUND: Diabetes mellitus is a prevalent and potentially devastating chronic illness affecting many older adults. Given spousal involvement in many aspects of diabetes management, coping with their partners is increasingly seen as a potential solution to make up for limited resources. This study aimed to identify the key conditions for optimal implementation of couple-based collaborative management model (CCMM) among Chinese older couples with type 2 diabetes mellitus. METHODS: Older couples and community healthcare practitioners were selected according to couples' joint intervention attendance rate and community's average attendance rate. This mixed methods research consisted of a qualitative phase and a quantitative phase. In the qualitative phase, in-depth interviews were conducted among 12 pairs of couples in the intervention group and 4 corresponding practitioners, in the follow-up period of the multicentered RCT from January to April 2022. Qualitative comparative analysis (QCA) in the quantitative phase to identify conditions influencing CCMM's implementation and to explore necessary and sufficient combinations of conditions (i.e., solutions) for improving patients' glycated hemoglobin (HbA1c) control (outcome). RESULTS: Key conditions included implementation process, couple's role in diabetes management, their belief and perception of CCMM, as well as objective obstacles and subjective initiative for behavior change. Accordingly, major barriers in CCMM's implementation were patients' strong autonomy (particularly among husbands), misbelief and misperception about diabetes management as a result of low literacy, and mistrust of the practitioners. QCA further revealed that no single condition was necessary for effective HbA1c control, while three types of their combinations would be sufficient. Solution 1 and 2 both comprised the presence of spousal willingness to help, plus correct belief and perception of diabetes management, well embodying the utility of couple collaborative management in supporting patients' HbA1c control. On the other hand, solution 3 indicated that high-quality implementation even without spousal support, can promote the patient's subjective initiative to overcome objective obstacles, suggesting enhanced self-management for HbA1c control. CONCLUSIONS: Tailored CCMM should be implemented in reference to older couple's preferences and literacy levels, to ensure intervention fidelity, and establish correct understanding of collaborative management among them.

A protocol for measuring the response of Arabidopsis roots to gravity and treatment for simulated microgravity.

We present a protocol to quantify the response of both normal and mutant Arabidopsis seedlings to gravity and simulated microgravity under earth-normal gravity conditions. We describe the steps to simulate microgravity using a three-dimensional (3D) clinostat, which changes the rate and direction at random and consistently rotates the axis horizontally and vertically to counteract the standard gravity at the Earth's surface. We then detail the gravity stimulation experiment, followed by the assessment of root responses using ImageJ-based analysis. For complete details on the use and execution of this protocol, please refer to Xu et al. (2022).1.

Implementation and evaluation of crowdsourcing in global health education.

BACKGROUND: Current global health course is most set as elective course taught in traditional teacher-taught model with low credit and short term. Innovate teaching models are required. Crowdsourcing characterized by high flexibility and strong application-orientation holds its potential to enhance global health education. We applied crowdsourcing to global health teaching for undergraduates, aiming to develop and evaluate a new teaching model for global health education. METHODS: Crowdsourcing was implemented into traditional course-based teaching via introducing five COVID-19 related global health debates. Undergraduate students majoring in preventative medicine and nursing grouped in teams of 5-8, were asked to resolve these debates in reference to main content of the course and with manner they thought most effective to deliver the messages. Students' experience and teaching effect, were evaluated by questionnaires and teachers' ratings, respectively. McNemar's test was used to compare the difference in students' experience before and after the course, and regression models were used to explore the influencing factors of the teaching effect. RESULTS: A total of 172 undergraduates were included, of which 122 (71%) were females. Students' evaluation of the new teaching model improved after the course, but were polarized. Students' self-reported teaching effect averaged 67.53 ± 16.8 and the teachers' rating score averaged 90.84 ± 4.9. Students majoring in preventive medicine, participated in student union, spent more time on revision, and had positive feedback on the new teaching model tended to perform better. CONCLUSION: We innovatively implemented crowdsourcing into global health teaching, and found this new teaching model was positively received by undergraduate students with improved teaching effects. More studies are needed to optimize the implementation of crowdsourcing alike new methods into global health education, to enrich global health teaching models.

Gender-Specific Effect of Couple-Based Intervention on Behavioral and Psychological Outcomes of Older Adults with Type 2 Diabetes during the COVID-19 Partial Lockdown in Guangzhou, China.

This study aimed to evaluate the gender-specific effect of a couple-based intervention on the management behaviors and mental well-being of community-dwelling older adults with type 2 diabetes mellitus during the COVID-19 partial lockdown in Guangzhou. Out of 207 participants involved in a prior randomized controlled trial (Trial no. ChiCTR1900027137), 156 (75%) completed the COVID-19 survey. Gendered differences in management behaviors and depressive symptoms between the couple-based intervention group and the patient-only control group were compared by distance to the high-risk areas cross-sectionally and longitudinally using random intercept models. Cross-sectionally, female patients of the intervention group had more positive behavior change scores (ß = 1.53, p = 0.002) and fewer depressive symptoms (ß = −1.34, p = 0.02) than the control group. Over time, female patients lived closer to the high-risk areas (<5 km) and showed decreasing depressive symptoms (ß = −4.48, p = 0.008) in the intervention group vs. the control group. No statistically significant between-group difference was found for males. Females tended to benefit more from the coupled-based intervention than males did, particularly among these closer to the high-risk areas. Chronic disease management can be better sustained with active spousal engagement.

BREVIPEDICELLUS and ERECTA control the expression of AtPRX17 to prevent Arabidopsis callus browning.

Efficient in vitro callus generation is required for tissue culture propagation, a process that allows for plant regeneration and transgenic breeding for desired phenotypes. Identifying genes and regulatory elements that prevent impaired callus growth and callus browning is essential for the development of in vitro callus systems. Here, we show that the BREVIPEDICELLUS and ERECTA pathways in Arabidopsis calli converge to prevent callus browning, and positively regulate the expression of the isoperoxidase gene AtPRX17 in rapidly growing calli. Loss-of-function mutations in both BREVIPEDICELLUS and ERECTA resulted in markedly increased callus browning. Transgenic lines expressing 35S pro::AtPRX17 in the bp-5 er105 double mutant background fully rescued this phenotypic abnormality. Using in vivo (chromatin immunoprecipitation-PCR and transient expression) and in vitro (electrophoretic mobility shift assays) assays, we observed that the BREVIPEDICELLUS protein binds directly to the upstream sequence of AtPRX17 to promote its transcription during callus growth. ERECTA is a ubiquitous factor required for cell proliferation and growth. We show that ERECTA positively regulates the expression of the transcription factor WRKY6, which directly binds to a separate site on the AtPRX17 promoter, further increasing its expression. Our data reveal an important molecular mechanism involved in the regulation of peroxidase isozyme expression to reduce Arabidopsis callus browning.

Transcriptomic Analysis of the Interaction Between FLOWERING LOCUS T Induction and Photoperiodic Signaling in Response to Spaceflight.

Spaceflight has an impact on the growth and development of higher plants at both the vegetative stage and reproductive stage. A great deal of information has been available on the vegetative stage in space, but relatively little is known about the influence of spaceflight on plants at the reproductive stage. In this study, we constructed transgenic Arabidopsis thaliana plants expressing the flowering control gene, FLOWERING LOCUS T (FT), together with the green fluorescent protein gene (GFP) under control of a heat shock-inducible promoter (HSP17.4), by which we induced FT expression inflight through remote controlling heat shock (HS) treatment. Inflight photography data showed that induction of FT expression in transgenic plants in space under non-inductive short-day conditions could promote flowering and reduce the length of the inflorescence stem in comparison with that of wild-type plants under the same conditions. Whole-genome microarray analysis of gene expression changes in leaves of wild-type and these transgenic plants grown under the long-day and short-day photoperiod conditions in space indicated that the function of the photoperiod-related spaceflight responsive genes is mainly involved in protein synthesis and post-translation protein modulation, notably protein phosphorylation. In addition, changes of the circadian component of gene expression in response to spaceflight under different photoperiods indicated that roles of the circadian oscillator could act as integrators of spaceflight response and photoperiodic signals in Arabidopsis plants grown in space.

Molecular Basis to Integrate Microgravity Signals into the Photoperiodic Flowering Pathway in Arabidopsis thaliana under Spaceflight Condition.

Understanding the effects of spaceflight on plant flowering regulation is important to setup a life support system for long-term human space exploration. However, the way in which plant flowering is affected by spaceflight remains unclear. Here, we present results from our latest space experiments on the Chinese spacelab Tiangong-2, in which Arabidopsis wild-type and transgenic plants pFT::GFP germinated and grew as normally as their controls on the ground, but the floral initiation under the long-day condition in space was about 20 days later than their controls on the ground. Time-course series of digital images of pFT::GFP plants showed that the expression rhythm of FT in space did not change, but the peak appeared later in comparison with those of their controls on the ground. Whole-genome microarray analysis revealed that approximately 16% of Arabidopsis genes at the flowering stage changed their transcript levels under spaceflight conditions in comparison with their controls on the ground. The GO terms were enriched in DEGs with up-regulation of the response to temperature, wounding, and protein stabilization and down-regulation of the function in circadian rhythm, gibberellins, and mRNA processes. FT and SOC1 could act as hubs to integrate spaceflight stress signals into the photoperiodic flowering pathway in Arabidopsis in space.

Circumnutation and Growth of Inflorescence Stems of Arabidopsis thaliana in Response to Microgravity under Different Photoperiod Conditions.

Circumnutation is a periodic growth movement, which is an important physiological mechanism of plants to adapt to their growth environments. Gravity and photoperiod are two key environmental factors in regulating the circumnutation of plants, but the coordination mechanism between them is still unknown. In this study, the circumnutation of Arabidopsis thaliana inflorescence stems was investigated on board the Chinese recoverable satellite SJ-10 and the Chinese spacelab TG-2. Plants were cultivated in a special plant culture chamber under two photoperiod conditions [a long-day (LD) light: dark cycle of 16:8 h, and a short-day (SD) light: dark cycle of 8:16 h]. The plant growth and movements were followed by two charge-coupled device (CCD) cameras. The parameter revealed a daily (24 h) modulation on both TG-2 and SJ-10, under both the LD and the SD conditions. The inhibition of circumnutation was more apparent by microgravity under the SD in comparison with that under the LD condition, suggesting the synergistic effects of the combined microgravity and photoperiod on the circumnutation in space. In addition, an infradian rhythm (ca. 21 days long) on the TG-2 was also observed.

Differential protein expression profiling of Arabidopsis thaliana callus under microgravity on board the Chinese SZ-8 spacecraft.

MAIN CONCLUSION: Exposure of Arabidopsis callus to microgravity has a significant impact on the expression of proteins involved in stress responses, carbohydrate metabolism, protein synthesis, intracellular trafficking, signaling, and cell wall biosynthesis. Microgravity is among the main environmental stress factors that affect plant growth and development in space. Understanding how plants acclimate to space microgravity is important to develop bioregenerative life-support systems for long-term space missions. To evaluate the spaceflight-associated stress and identify molecular events important for acquired microgravity tolerance, we compared proteomic profiles of Arabidopsis thaliana callus grown under microgravity on board the Chinese spacecraft SZ-8 with callus grown under 1g centrifugation (1g control) in space. Alterations in the proteome induced by microgravity were analyzed by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry with isobaric tags for relative and absolute quantitation labeling. Forty-five proteins showed significant (p < 0.05) and reproducible quantitative differences in expression between the microgravity and 1g control conditions. Of these proteins, the expression level of 24 proteins was significantly up-regulated and that of 21 proteins was significantly down-regulated. The functions of these proteins were involved in a wide range of cellular processes, including general stress responses, carbohydrate metabolism, protein synthesis/degradation, intracellular trafficking/transportation, signaling, and cell wall biosynthesis. Several proteins not previously known to be involved in the response to microgravity or gravitational stimuli, such as pathogenesis-related thaumatin-like protein, leucine-rich repeat extension-like protein, and temperature-induce lipocalin, were significantly up- or down-regulated by microgravity. The results imply that either the normal gravity-response signaling is affected by microgravity exposure or that microgravity might inappropriately induce altered responses to other environmental stresses.

Modulation of root-skewing responses by KNAT1 in Arabidopsis thaliana.

The KNOTTED1 homeobox (KNOX) family transcription factors are essential for stem cell establishment and maintenance and regulate various aspects of development in all green plants. Expression patterns of the KNOX genes in the roots of plants have been reported, but their role in development remains unclear. Here we show how the KNAT1 gene is specifically involved in root skewing in Arabidopsis. The roots of two mutant alleles of KNAT1 (bp-1 and bp-5) exhibited exaggerated skewing to the right of gravity when grown on both vertical and tilted agar medium surfaces. This skewing phenotype was enhanced by treatments with low concentrations of propyzamide, and required auxin transport. The KNAT1 mutation substantially decreased basipetal auxin transport and increased auxin accumulation in the roots. Using a PIN2-GFP reporter and western blot analysis, we found that this alteration in auxin transport was accompanied by a decrease in PIN2 levels in the root tip. Decreased PIN2 expression in the mutant roots was not accompanied by reduced mRNA levels, suggesting that the KNAT1 mutations affected PIN2 expression at the posttranscriptional level. In vivo visualization of intracellular vacuolar targeting indicated that vacuolar degradation of PIN2-GFP was significantly promoted in the root tips of the bp allelic mutants. Together, these results demonstrate that KNAT1 negatively modulates root skewing, possibly by regulating auxin transport.

A proteomic approach to analyzing responses of Arabidopsis thaliana root cells to different gravitational conditions using an agravitropic mutant, pin2 and its wild type.

BACKGROUND: Root gravitropsim has been proposed to require the coordinated, redistribution of the plant signaling molecule auxin within the root meristem, but the underlying molecular mechanisms are still unknown. PIN proteins are membrane transporters that mediate the efflux of auxin from cells. The PIN2 is important for the basipetal transport of auxin in roots and plays a critical role in the transmission of gravity signals perceived in the root cap to the root elongation zone. The loss of function pin2 mutant exhibits a gravity-insensitive root growth phenotype. By comparing the proteomes of wild type and the pin2 mutant root tips under different gravitational conditions, we hope to identify proteins involved in the gravity-related signal transduction. RESULTS: To identify novel proteins involved in the gravity signal transduction pathway we have carried out a comparative proteomic analysis of Arabidopsis pin2 mutant and wild type (WT) roots subjected to different gravitational conditions. These conditions included horizontal (H) and vertical (V) clinorotation, hypergravity (G) and the stationary control (S). Analysis of silver-stained two-dimensional SDS-PAGE gels revealed 28 protein spots that showed significant expression changes in altered gravity (H or G) compared to control roots (V and S). Whereas the majority of these proteins exhibited similar expression patterns in WT and pin2 roots, a significant number displayed different patterns of response between WT and pin2 roots. The latter group included 11 protein spots in the H samples and two protein spots in the G samples that exhibited an altered expression exclusively in WT but not in pin2 roots. One of these proteins was identified as annexin2, which was induced in the root cap columella cells under altered gravitational conditions. CONCLUSIONS: The most interesting observation in this study is that distinctly different patterns of protein expression were found in WT and pin2 mutant roots subjected to altered gravity conditions. The data also demonstrate that PIN2 mutation not only affects the basipetal transport of auxin to the elongation zone, but also results in an altered expression of proteins in the root columella.

Protein storage vacuoles are transformed into lytic vacuoles in root meristematic cells of germinating seedlings by multiple, cell type-specific mechanisms.

We have investigated the structural events associated with vacuole biogenesis in root tip cells of tobacco (Nicotiana tabacum) seedlings preserved by high-pressure freezing and freeze-substitution techniques. Our micrographs demonstrate that the lytic vacuoles (LVs) of root tip cells are derived from protein storage vacuoles (PSVs) by cell type-specific sets of transformation events. Analysis of the vacuole transformation pathways has been aided by the phytin-dependent black osmium staining of PSV luminal contents. In epidermal and outer cortex cells, the central LVs are formed by a process involving PSV fusion, storage protein degradation, and the gradual replacement of the PSV marker protein α-tonoplast intrinsic protein (TIP) with the LV marker protein γ-TIP. In contrast, in the inner cortex and vascular cylinder cells, the transformation events are more complex. During mobilization of the stored molecules, the PSV membranes collapse osmotically upon themselves, thereby squeezing the vacuolar contents into the remaining bulging vacuolar regions. The collapsed PSV membranes then differentiate into two domains: (1) vacuole "reinflation" domains that produce pre-LVs, and (2) multilamellar autophagosomal domains that are later engulfed by the pre-LVs. The multilamellar autophagosomal domains appear to originate from concentric sheets of PSV membranes that create compartments within which the cytoplasm begins to break down. Engulfment of the multilamellar autophagic vacuoles by the pre-LVs gives rise to the mature LVs. During pre-LV formation, the PSV marker α-TIP disappears and is replaced by the LV marker γ-TIP. These findings demonstrate that the central LVs of root cells arise from PSVs via cell type-specific transformation pathways.

Changes in gravitational forces induce the modification of Arabidopsis thaliana silique pedicel positioning.

The laterals of both shoots and roots often maintain a particular angle with respect to the gravity vector, and this angle can change during organ development and in response to environmental stimuli. However, the cellular and molecular mechanisms of the lateral organ gravitropic response are still poorly understood. Here it is demonstrated that the young siliques of Arabidopsis thalinana plants subjected to 3-D clinostat rotation exhibited automorphogenesis with increased growth angles between pedicels and the main stem. In addition, the 3-D clinostat rotation treatment significantly influenced the development of vascular bundles in the pedicel and caused an enlargement of gap cells at the branch point site together with a decrease in KNAT1 expression. Comparisons performed between normal and empty siliques revealed that only the pedicels of siliques with normally developing seeds could change their growth angle under the 3-D clinostat rotational condition, while the pedicels of the empty siliques lost the ability to respond to the altered gravity environment. These results indicate that the response of siliques to altered gravity depends on the normal development of seeds, and may be mediated by vascular bundle cells in the pedicel and gap cells at branch point sites.

[Analysis of tobacco regeneration plants from the protoplasts produced by electrofusion [corrected] in space].

Vacuolated mesophyll protoplasts of Nicotiana rustica L. were electrically fused with evacuolated protoplasts of the same genus (N. tabacum cv. 'Gexin No.1') during a 7-day space flight in the Chinese spacecraft "SZ-4". The initial cell division leading to micro-callus formation took place after landing (Fig.1). Higher plating efficiencies were observed in the flight samples than the control culture, but the frequency of plantlets regeneration reduced by about 20% of the control (Table 1). The hybrid characters were tested by chromosome counting, isozyme analysis and comparison of morphological characteristics (Figs.2-4). About 32% of the regenerates showed hybrid character. Leaf morphological modifications were found in 3 hybrids, i.e., H23, H25 and H27. After backcrossing with N. rustica, alterations in flower color and leaf shape occurred in the somatic hybrid H23 (Fig.5). These results demonstrate that the hybrids formed under microgravity condition could regenerate fertile plants.

Effects of Lantana camara leaf extract on the activity of superoxide dismutase and accumulation of H2O2 in water hyacinth leaf.

Water hyacinth (Eichhornia crassipes) is one of the most productive plants, but is also a troublesome weed in the world. In order to protect the public water system from chemical herbicides pollution, biological method has been suggested to control the growth and the reproduction of this weed. Lantana (Lantana camara L.) is an important weed of the family Verbenaceae and its leaf extract is highly toxic to water hyacinth. The results of this study showed that the extract of lantana leaves suppressed the emergence of leaf buds of water hyacinth plant, and caused the decay of its leaves by foliar spraying. In addition, the increase of SOD activity in water hyacinth leaves was in accordance with the accumulation of H(2)O(2) and the increase in degree of membrane peroxidation, while the activity of catalase, which might remove the excessive H(2)O(2) in water hyacinth leaves, was inhibited by treatment with lantana extract. At tissue level, high H(2)O(2) histochemical labeling was detected in guard cells after treatment with lantana extract. This overproduction of H(2)O(2) could kill the leaf cells and cause leaf necrosis in the treated plant. Therefore, the high toxicity of lantana leaf extract to water hyacinth might be due to oxidative stress.

The GAOLAOZHUANGREN2 gene is required for normal glucose response and development of Arabidopsis.

Recent studies of glucose (Glc) sensing and signaling have revealed that Glc acts as a critical signaling molecule in higher plants. Several Glc sensing-defective Arabidopsis mutants have been characterized in detail, and the corresponding genes encoding Glc-signaling proteins have been isolated. However, the full complexity of Glc signaling in higher plants is not yet fully understood. Here, we report the identification and characterization of a new Glc-insensitive mutant, gaolaozhuangren2 (glz2), which was isolated from transposon mutagenesis experiments in Arabidopsis. In addition to its insensitivity to Glc, the glz2 plant exhibits several developmental defects such as short stature with reduced apical dominance, short roots, small and dark-green leaves, late flowering and female sterility. Treatment with 4% Glc blocked expression of the OE33 gene in wild-type plants, whereas expression of this gene was unchanged in the glz2 mutant plants. Taken together, our results suggest that the GLZ2 gene is required for normal glucose response and development of Arabidopsis.

The GAOLAOZHUANGREN1 gene encodes a putative glycosyltransferase that is critical for normal development and carbohydrate metabolism.

Glycosyltransferases are enzymes that catalyze the attachment of a sugar molecule to specific acceptor molecules. These enzymes have been shown to play important roles in a number of biological processes. Whereas a large number of putative glycosyltransferase genes have been identified by genomic sequencing, the functions of most of these genes are unknown. Here we report the characterization of an Arabidopsis mutant, designated gaolaozhuangren1 (glz1), which is allelic to parvus characterized recently. The glz1 mutant exhibited a reduced plant stature, reduced size of organs in the shoot and dark-green leaves, indicating an important role of GLZ1 gene in normal development. The earliest GLZ1 expression appears at the shoot apical region of 4-d-old seedlings, which coincides with the onset of the glz1 morphological phenotypes. GLZ1 is expressed in a tissue-specific and developmentally regulated manner, predominantly in the stem and silique, and moderately in the flower. GLZ1 expression is strong in the midrib of rosette and cauline leaves; however, its expression was not detectable in the midrib of the cotyledon. Further analyses revealed that carbohydrate composition and distribution were aberrant in the glz1 mutant. These, together with the GLZ1 expression pattern, suggest a requirement for the GLZ1 function in normal sink-source transition during plant development.