CRISPR/Cas9 edited SlGT30 improved both drought resistance and fruit yield through endoreduplication.

There is often a trade-off effect between different agronomic traits due to gene pleiotropy, leading to a negative correlation between yield and resistance. Consequently, using gene-editing techniques to develop superior traits becomes challenging. Genetic resources that defy this constraint are scarce but hold great potential as targets for improvement through the utilisation of CRISPR. Transcription factors are critical in modulating numerous gene expressions across diverse biological processes. Here, we found that the trihelix transcription factor SlGT30 plays a role in drought resistance and tomato fruit development. We edited the SlGT30 gene with CRISPR/Cas9 technology and found that the knockout lines showed decreased stomata density in the leaves and large fruits. Subsequent examination revealed that cell ploidy was impacted in the leaves and fruits of SlGT30 knockout lines. SlGT30 knockout affected cell size through the endoreduplication pathway, manifested in decreased stomata density and reduced water loss. Consequently, this resulted in an enhancement of drought resistance. For the fruit, both cell size and cell number increased in the fruit pericarp of knockout lines, improving the fruit size and weight accordingly. Therefore, SlGT30 represents a promising candidate gene for gene editing in breeding practice.

CsHSFA1d Promotes Drought Stress Tolerance by Increasing the Content of Raffinose Family Oligosaccharides and Scavenging Accumulated Reactive Oxygen Species in Cucumber.

Drought is the most severe form of stress experienced by plants worldwide. Cucumber is a vegetable crop that requires a large amount of water throughout the growth period. In our previous study, we identified that overexpression of CsHSFA1d could improve cold tolerance and the content of endogenous jasmonic acid in cucumber seedlings. To explore the functional diversities of CsHSFA1d, we treat the transgenic plants under drought conditions. In this study, we found that the heat shock transcription factor HSFA1d (CsHSFA1d) could improve drought stress tolerance in cucumber. CsHSFA1d overexpression increased the expression levels of galactinol synthase (CsGolS3) and raffinose synthase (CsRS) genes, encoding the key enzymes for raffinose family oligosaccharide (RFO) biosynthesis. Furthermore, the lines overexpressing CsHSFA1d showed higher enzymatic activity of GolS and raffinose synthase to increase the content of RFO. Moreover, the CsHSFA1d-overexpression lines showed lower reactive oxygen species (ROS) accumulation and higher ROS-scavenging enzyme activity after drought treatment. The expressions of antioxidant genes CsPOD2, CsAPX1 and CsSOD1 were also upregulated in CsHSFA1d-overexpression lines. The expression levels of stress-responsive genes such as CsRD29A, CsLEA3 and CsP5CS1 were increased in CsHSFA1d-overexpression lines after drought treatment. We conclude that CsHSFA1d directly targets and regulates the expression of CsGolS3 and CsRS to promote the enzymatic activity and accumulation of RFO to increase the tolerance to drought stress. CsHSFA1d also improves ROS-scavenging enzyme activity and gene expression indirectly to reduce drought-induced ROS overaccumulation. This study therefore offers a new gene target to improve drought stress tolerance in cucumber and revealed the underlying mechanism by which CsHSFA1d functions in the drought stress by increasing the content of RFOs and scavenging the excessive accumulation of ROS.

A Mental Health Management and Cognitive Behavior Analysis Model of College Students Using Multi-View Clustering Analysis Algorithm.

In this new era that is full of social changes, ongoing economic transformation, an abundance of information resources, and a fast pace of life, the pressure that people feel to compete with one another is also increasing day by day. Because of the vast differences in people's states of consciousness and worldviews, interpersonal relationships have become increasingly difficult to navigate. Students in higher education institutions will eventually emerge as the dominant demographic in society. Their mental health has a significant bearing on all aspects of life, including learning and future growth. An objective condition that must be met in order to guarantee that the next generation of talent will have a high level of overall quality is the improvement of the mental health of college students (CSMH) in the new era. One component of public health is the emotional well-being of students in higher education. The state of the public's health is consistently ranked among the most urgent problems facing modern society. However, there is not much hope for the Chinese CSMH. In order to effectively manage their mental health, a variety of educational institutions, including colleges and universities, have proposed a large number of management strategies for CSMH. The vast majority of these strategies are not targeted, and they do not offer a variety of management strategies that are based on the many different psychological states. It is necessary to first be able to accurately predict the mental health status of each individual college student in order to achieve the goal of improving the mental health management of students attending colleges and universities. This study proposes using a multi-view K-means algorithm, abbreviated as MvK-means, to analyze the CSMH's data on mental health. This is possible because the data can be obtained from multiple perspectives. This paper presents a multi-view strategy as well as a weight strategy in light of the fact that each point of view contributes in its own unique way. Different weight values should be assigned to each view's data, which will ultimately result in an improved evaluation effect of the model. The findings of the experiments indicate that the model that was proposed has a beneficial impact on the analysis of the data pertaining to the mental health of college students.

A zinc finger protein SlSZP1 protects SlSTOP1 from SlRAE1-mediated degradation to modulate aluminum resistance.

In acidic soils, aluminum (Al) toxicity is the main factor inhibiting plant root development and reducing crops yield. STOP1 (SENSITIVE TO PROTON RHIZOTOXICITY 1) was a critical factor in detoxifying Al stress. Under Al stress, STOP1 expression was not induced, although STOP1 protein accumulated, even in the presence of RAE1 (STOP1 DEGRADATION E3-LIGASE). How the Al stress triggers and stabilises the accumulation of STOP1 is still unknown. Here, we characterised SlSTOP1-interacting zinc finger protein (SlSZP1) using a yeast-two-hybrid screening, and generated slstop1, slszp1 and slstop1/slszp1 knockout mutants using clustered regularly interspaced short palindromic repeats (CRISPR) in tomato. SlSZP1 is induced by Al stress but it is not regulated by SlSTOP1. The slstop1, slszp1 and slstop1/slszp1 knockout mutants exhibited hypersensitivity to Al stress. The expression of SlSTOP1-targeted genes, such as SlRAE1 and SlASR2 (ALUMINUM SENSITIVE), was inhibited in both slstop1 and slszp1 mutants, but not directly regulated by SlSZP1. Furthermore, the degradation of SlSTOP1 by SlRAE1 was prevented by SlSZP1. Al stress increased the accumulation of SlSTOP1 in wild-type (WT) but not in slszp1 mutants. The overexpression of either SlSTOP1 or SlSZP1 did not enhance plant Al resistance. Altogether, our results show that SlSZP1 is an important factor for protecting SlSTOP1 from SlRAE1-mediated degradation.

Heat shock-induced cold acclimation in cucumber through CsHSFA1d-activated JA biosynthesis and signaling.

Cucumber (Cucumis sativus) originated in tropical areas and is very sensitive to low temperatures. Cold acclimation is a universal strategy that improves plant resistance to cold stress. In this study, we report that heat shock induces cold acclimation in cucumber seedlings, via a process involving the heat-shock transcription factor HSFA1d. CsHSFA1d expression was improved by both heat shock and cold treatment. Moreover, CsHSFA1d transcripts accumulated more under cold treatment after a heat-shock pre-treatment than with either heat shock or cold treatment alone. After exposure to cold, cucumber lines overexpressing CsHSFA1d displayed stronger tolerance for cold stress than the wild type, whereas CsHSFA1d knockdown lines obtained by RNA interference were more sensitive to cold stress. Furthermore, both the overexpression of CsHSFA1d and heat-shock pre-treatment increased the endogenous jasmonic acid (JA) content in cucumber seedlings after cold treatment. Exogenous application of JA rescued the cold-sensitive phenotype of CsHSFA1d knockdown lines, underscoring that JA biosynthesis is key for CsHSFA1d-mediated cold tolerance. Higher JA content is likely to lead to the degradation of CsJAZ5, a repressor protein of the JA pathway. We also established that CsJAZ5 interacts with CsICE1. JA-induced degradation of CsJAZ5 would be expected to release CsICE1, which would then activate the ICE-CBF-COR pathway. After cold treatment, the relative expression levels of ICE-CBF-COR signaling pathway genes, such as CsICE1, CsCBF1, CsCBF2 and CsCOR1, in CsHSFA1d overexpression lines were significantly higher than in the wild type and knockdown lines. Taken together, our results help to reveal the mechanism underlying heat shock-induced cold acclimation in cucumber.

Research Advances in Heterotrimeric G-Protein α Subunits and Uncanonical G-Protein Coupled Receptors in Plants.

As crucial signal transducers, G-proteins and G-protein-coupled receptors (GPCRs) have attracted increasing attention in the field of signal transduction. Research on G-proteins and GPCRs has mainly focused on animals, while research on plants is relatively rare. The mode of action of G-proteins is quite different from that in animals. The G-protein α (Gα) subunit is the most essential member of the G-protein signal cycle in animals and plants. The G-protein is activated when Gα releases GDP and binds to GTP, and the relationships with the GPCR and the downstream signal are also achieved by Gα coupling. It is important to study the role of Gα in the signaling pathway to explore the regulatory mechanism of G-proteins. The existence of a self-activated Gα in plants makes it unnecessary for the canonical GPCR to activate the G-protein by exchanging GDP with GTP. However, putative GPCRs have been found and proven to play important roles in G-protein signal transduction. The unique mode of action of G-proteins and the function of putative GPCRs in plants suggest that the same definition used in animal research cannot be used to study uncanonical GPCRs in plants. This review focuses on the different functions of the Gα and the mode of action between plants and animals as well as the functions of the uncanonical GPCR. This review employs a new perspective to define uncanonical GPCRs in plants and emphasizes the role of uncanonical GPCRs and Gα subunits in plant stress resistance and agricultural production.