SlNAC3 suppresses cold tolerance in tomatoes by enhancing ethylene biosynthesis.

Low temperature stress poses a significant challenge to the productivity of horticultural crops. The dynamic expression of cold-responsive genes plays a crucial role in plant cold tolerance. While NAC transcription factors have been extensively studied in plant growth and development, their involvement in regulating plant cold tolerance remains poorly understood. In this study, we focused on the identification and characterisation of SlNAC3 as the most rapid and robust responsive gene in tomato under low temperature conditions. Manipulating SlNAC3 through overexpression or silencing resulted in reduced or enhanced cold tolerance, respectively. Surprisingly, we discovered a negative correlation between the expression of CBF and cold tolerance in the SlNAC3 transgenic lines. These findings suggest that SlNAC3 regulates tomato cold tolerance likely through a CBF-independent pathway. Furthermore, we conducted additional investigations to identify the molecular mechanisms underlying SINAC3-mediated cold tolerance in tomatoes. Our results revealed that SlNAC3 controls the transcription of ethylene biosynthetic genes, thereby bursting ethylene release in response to cold stress. Indeed, the silencing of these genes led to an augmentation in cold tolerance. This discovery provides valuable insights into the regulatory pathways involved in ethylene-mediated cold tolerance in tomatoes, offering potential strategies for developing innovative approaches to enhance cold stress resilience in this economically important crop species.

Exploring evolution characteristics of eco-environment quality in the Yangtze River Basin based on remote sensing ecological index.

As an important ecological-economic development area in China, scientific understanding of the spatial and temporal changes in eco-environment quality (EEQ) and its drivers in the Yangtze River Basin (YRB) is crucial for the effective implementation of ecological protection projects in the YRB. To address the lack of large-scale EEQ assessment in the YRB, this paper uses the Google Earth Engine (GEE) platform and the Remote Sensing Ecological Index (RSEI) to investigate the spatial and temporal characteristics of EEQ in the YRB from 2000 to 2020, and to analyze the impact of various factors on the EEQ of the YRB. This study showed that: (1) The overall EEQ of YRB was at the 'good' grade over the past 20 years, showing an increasing trend, with the value changing from 0.70 to 0.77. (2) The YRB's EEQ has positive spatial aggregation characteristics, with the northern part of the Jialing River basin and the Han River basin exhibiting a high-high aggregation type and the upper reaches exhibiting a low-low aggregation type. (3) In the past 20 years, the human activities had a greater impact on the EEQ of the YRB; moreover, all factors had a greater impact on the EEQ than a single factor. The interaction between the biological abundance index and population density had the most effect, with a q-value of 0.737 in 2020.

Insights into long-term changes of groundwater levels in the typical region of Zhangjiakou City, China.

Little information is available on the long-term changes of groundwater levels and their associated influencing factors. Zhangjiakou City was chosen as a case to reveal the temporal and spatial dynamics of groundwater level and its driving factors in the long term. Herein, the observation data of groundwater level from 56 wells was investigated from 1981 to 2015, including the collected meteorological data, socio-economic data, and groundwater resource exploitation situation. Results showed that the groundwater level in Zhangjiakou City tended to be decreased, and the decrease rate was gradually accelerated. In the past 35 years, the groundwater level of Bashang Plateau has decreased by 3.59 m < 3.6 m in Yuyang Basin < 7.17 m in Zhuohuai Basin < 20.41 m in Chaixuan Basin. The dynamic changes of groundwater level in four geomorphic units in Zhangjiakou City were significant correlation between the total population and other socio-economic factors, including primary industry production value; common cultivated land area; effective irrigation area; total grain yield; total vegetable yield; total production of pork, beef, and mutton; secondary industry production value; tertiary industry production value; and year-end total population. Furthermore, the principal component analysis of groundwater level variation in Zhangjiakou city showed that the variance contribution rates of the first principal component in the characteristic indicators of the Bashang Plateau, Chaixuan Basin, Zhuohuai Basin, and Yuyang Basin were 75.7%, 83.9%, 66.1%, and 77.8%, respectively, which mainly reflect the information of socio-economic factors. This indicated that socio-economic factors were the main driving factor influencing the continuous decline of groundwater levels in Zhangjiakou City. The obtained findings can provide new insights into the sustainable development of social economy and the rational utilization and allocation of regional water resources.

Elevated nano-α-Fe2O3 enhances arsenic metabolism and dissolved organic carbon release of Microcystis aeruginosa under a phytate environment.

Little information is available on the effects of nano-α-Fe2O3 on arsenic (As) metabolism of algae and potential associated carbon (C) storage in As-contaminated water with dissolved organic phosphorus (DOP) as a phosphorus (P) source. In this study, Microcystis aeruginosa (M. aeruginosa) was used to investigate impacts of nano-α-Fe2O3 on cell growth and As metabolism of algae under a phytate (PA) environment as well as potential associated C storage. Results showed that nano-α-Fe2O3 had a subtle influence on algal cell growth in a PA environment. Herein, algal cell density (OD680) and chlorophyll a (Chla) were inhibited at elevated nano-α-Fe2O3 levels, which simultaneously limited the decrease of Yield. As suggested, the complexation of PA with nano-α-Fe2O3 could alleviate the negative influence on algal cell growth. Furthermore, the elevated nano-α-Fe2O3 increased As methylation in the PA environment due to higher monomethylarsenic (MMA) and dimethylarsenic (DMA) concentrations in the test media. Additionally, microcystins (MCs) in the media changed consistently with UV254, both of which were relatively lower at 10.0 mg·L-1 nano-α-Fe2O3. Enhanced As(V) methylation of algal cells was found to simultaneously reduce the release risk of As(III) and MC while increasing dissolved organic carbon (DOC) content in media, suggesting unfavorable C storage. Three-dimensional fluorescence analysis revealed that the main DOC constituent was the tryptophan-like component in aromatic proteins. Correlation analysis showed that decreases in pH and the zeta potential and an increase in Chla may lead to metabolic As improvements in M. aeruginosa. The obtained findings highlight the need for greater focus on the potential risks of DOP combined with nano-α-Fe2O3 on algal blooms as well as the biogeochemical cycling processes of As and C storage in As-contaminated water with DOP as the P source.

Psychoactive Comfort Products or Snacks: How Chinese Young Adults Perceive the Potentially Addictive Nature of E-Cigarettes.

The potential health value and pitfalls of e-cigarettes are currently under dispute in the scientific community. Exploring young adult e-cigarette users' perceptions would assist in adding a public dimension of understanding to the literature and in scientific public health decision making. Therefore, in this study, we collected and analyzed data from interviews with young adult (n = 14) e-cigarette users and found that many referred to e-cigarettes as "snacks," indicating that they considered that both their frequency of use and addiction were manageable and that they could stop using e-cigarettes at any time. To further understand the behavior of Chinese young adults in relation to their perception of e-cigarettes as a "snack", the study developed a social context framework (crossroads model) and psychological judgment model to explain how youth e-cigarette users' perception of "controlled addiction and ready cessation" arises. These models can be used to assess the effectiveness of e-cigarette policy.

Genotypic and phenotypic characterization of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Guangzhou, China.

BACKGROUND: G6PD deficiency is a common inherited disorder worldwide and has a higher incidence rate in southern China. Many variants of G6PD result from point mutations in the G6PD gene, leading to decreased enzyme activity. This study aimed to analyse the genotypic and phenotypic characteristics of G6PD deficiency in Guangzhou, China. METHODS: In this study, a total of 20,208 unrelated participants were screened from 2020 to 2022. G6PD deficiency was further analysed by quantitative enzymatic assay and G6PD mutation analysis. The unidentified genotype of the participants was further ascertained by direct DNA sequencing. RESULTS: A total of 12 G6PD mutations were identified. Canton (c.1376G>T) and Kaiping (c.1388G>A) were the most common variants, and different mutations led to varying levels of G6PD enzyme activity. Comparing the enzyme activities of the 6 missense mutations between the sexes, we found significant differences (P < 0.05) in the enzyme activities of both male hemizygotes and female heterozygotes. Two previously unreported mutations (c.1438A>T and c.946G>A) were identified. CONCLUSIONS: This study provided detailed genotypes of G6PD deficiency in Guangzhou, which could be valuable for diagnosing and researching G6PD deficiency in this area.

CUCUME: An RNA methylation database integrating systemic mRNAs signals, GWAS and QTL genetic regulation and epigenetics in different tissues of Cucurbitaceae.

As an internal modification of transcripts, RNA methylation determines RNA fate by changing RNA-protein binding affinity. In plants, RNA methylation is ubiquitous and is involved in all aspects of RNA post-transcriptional regulation. For instance, long-distance mobile RNAs, strongly influenced by their methylation status, play important roles in plant growth, development and environmental adaptation. Cucumber/pumpkin heterografts are widely used to improve stress tolerance of cucumber and to study mobile RNA signals due to their strong developed vasculature system. Here, we developed the Cucume (Cucurbit RNA methylation, http://cucume.cn/) database for these two important vegetables, cucumber (Cucumis sativus L.) and pumpkin (Cucurbita moschata) with high productivity worldwide. We identified mRNAs harboring 5-methylcytosine (m5C) and N6-methyladenosine (m6A) sites in pumpkin and cucumber at the whole genome level via Methylated RNA Immunoprecipitation sequencing (MeRIP-seq) of different tissues and the vascular exudates. In addition to RNA methylation sites, the Cucume database includes graft-transmissible systemic mRNAs identified in previous studies using cucumber/pumpkin heterografts. The further integration of cucumber genome-wide association analysis (GWAS) and quantitative trait loci (QTL) allows the study of RNA methylation-related genetic and epigenetic regulation in cucurbits. Therefore, the here developed Cucume database will promote understanding the role of cucurbit RNA methylation in RNA mobility and QTL, ultimately benefitting future breeding of agronomic crop germplasms.

The role of shoot-derived RNAs transported to plant root in response to abiotic stresses.

A large number of RNA molecules are transported over long-distance between shoots and roots via phloem in higher plants. Mobile RNA signals are important for plants to tackle abiotic stresses. Shoot-derived mobile RNAs can be involved in the response to different developmental or environmental signals in the root. Some environmental conditions such as climate change, water deficit, nutrient deficiency challenge modern agriculture with more expeditious abiotic stress conditions. Root architecture determines the ability of water and nutrient uptake and further abiotic stress tolerance, and shoot tissue also determines the balance between shoot-root relationship in plant growth and adaptations. Thus, it is necessary to understand the roles of shoot-derived RNA signals and their potential function in roots upon abiotic stresses in the model plants (Arabidopsis thaliana and Nicotiana benthamiana) and agricultural crops. In this review, we summarize the so-far discovered shoot-derived mobile RNA transportation to the root under abiotic stress conditions, e.g. drought, cold stress and nutrient deficiencies. Furthermore, we will focus on the biological relevance and the potential roles of these RNAs in root development and stress responses which will be an asset for the future breeding strategies.

The E3 ubiquitin ligase CHIP protects against sepsis-induced myocardial dysfunction by inhibiting NF-κB-mediated inflammation via promoting ubiquitination and degradation of karyopherin-α 2.

Cardiac dysfunction has been recognized as a major contributor to mortality in sepsis, which is closely associated with inflammatory reactions. The carboxy terminus of Hsc70-interacting protein (CHIP), a U-box E3 ubiquitin ligase, defends against cardiac injury caused by other factors, but its role in sepsis-induced cardiac dysfunction has yet to be determined. The present study was designed to investigate the effects of CHIP on cardiac dysfunction caused by sepsis and the molecular mechanisms underlying these processes. We discovered that the CHIP level decreased gradually in the heart at different time points after septic model construction. The decline in CHIP expression of lipopolysaccharide (LPS)-stimulated cardiomyocytes was related to c-Jun activation that inhibited the transcription of CHIP. Functional biology experiments indicated that CHIP bound directly to karyopherin-α 2 (KPNA2) and promoted its degradation through polyubiquitination in cardiomyocytes. CHIP overexpression in cardiomyocytes obviously inhibited LPS-initiated release of TNF-α and IL-6 by promoting KPNA2 degradation, reducing NF-κB translocation into the nucleus. Consistent with the in vitro results, data obtained from animal experiments indicated that septic transgenic mice with heart-specific CHIP overexpression showed a weaker proinflammatory response and reduced cardiac dysfunction than septic control mice. Furthermore, we found that the therapeutic effect of compound YL-109 on cardiac dysfunction in septic mice was due to the upregulation of myocardial CHIP expression. These findings demonstrated that sepsis-initiated the activation of c-Jun suppressed CHIP transcription. CHIP directly promoted ubiquitin-mediated degradation of KPNA2, which reduced the production of proinflammatory cytokines by inhibiting the translocation of NF-κB from the cytoplasm into the nucleus in myocardium, thereby attenuating sepsis-induced cardiac dysfunction.

Pursuing individualism without feminism: Leisure life and gender politics of young female bar-goers in urban China.

Drawing upon fieldwork data collected among young female bar-goers in China, we examine the gender politics manifested by their leisure activities and argue that these young women are pursuing individualism without feminism. By analyzing the act of bar hopping, we reveal the internal tension of individuation, which urges women to engage in gender labor in order to obtain preferential treatment regarding consumption and a competitive advantage in esthetic femininity. This indicates that the central axis of Chinese women's individualization is a hybrid of neoliberal individuals and essentialist gender roles. It encourages women to see themselves voluntarily as subordinate subjects in market transactions and feminized objects under male scrutiny. The pursuit of individualism without feminism prevents women from overcoming the highly gendered settings of bar scenarios, or imagining the female as a whole group with communal solidarity. The Chinese female individual manifests herself as a hostage of the collusion between the commodification of social life and a conservative gender regime. The case of China brings the individualization thesis out of its original compressed-modernity framework and indicates that East Asia's quest for modernity must address the compressed conditions of individualism and feminism simultaneously.

Retinal Vessel Segmentation Based on B-COSFIRE Filters in Fundus Images.

Retinal vessel extraction plays an important role in the diagnosis of several medical pathologies, such as diabetic retinopathy and glaucoma. In this article, we propose an efficient method based on a B-COSFIRE filter to tackle two challenging problems in fundus vessel segmentation: (i) difficulties in improving segmentation performance and time efficiency together and (ii) difficulties in distinguishing the thin vessel from the vessel-like noise. In the proposed method, first, we used contrast limited adaptive histogram equalization (CLAHE) for contrast enhancement, then excerpted region of interest (ROI) by thresholding the luminosity plane of the CIELab version of the original RGB image. We employed a set of B-COSFIRE filters to detect vessels and morphological filters to remove noise. Binary thresholding was used for vessel segmentation. Finally, a post-processing method based on connected domains was used to eliminate unconnected non-vessel pixels and to obtain the final vessel image. Based on the binary vessel map obtained, we attempt to evaluate the performance of the proposed algorithm on three publicly available databases (DRIVE, STARE, and CHASEDB1) of manually labeled images. The proposed method requires little processing time (around 12 s for each image) and results in the average accuracy, sensitivity, and specificity of 0.9604, 0.7339, and 0.9847 for the DRIVE database, and 0.9558, 0.8003, and 0.9705 for the STARE database, respectively. The results demonstrate that the proposed method has potential for use in computer-aided diagnosis.

"Returning to Ordinary Citizenship": A Qualitative Study of Chinese PWUD's Self-Management Strategies and Disengagement Model of Identity.

How PWUD (people who use drugs) live under drug governance is an important research question. This study adopts a qualitative research method to explore how PWUD in China self-manage after perceiving the dilemma of incomplete citizenship and the social pressure brought by drug control arrangements. Through analysis of 130 PWUD's files and in-depth interviews with 10 interviewees (from the 24 preliminary interviews), this study found that PWUD developed action strategies of hidden mobility (spatial isolation), disconnection of past experiences (time isolation), instrumental actions, as well as narrative strategies of reframing themselves as ordinary citizens with attempts of reversing identity disadvantages. Further, PWUD's self-management strategies manifest as a disengagement model in which the actors (PWUD, not rehabilitation agencies) do not intend to develop integrative positive identities through dispersed, practiced behavioral strategies, but attempt to return to pre-addiction, non-socially exclusionary citizenship experiences. The disengagement model and its negative effect on PWUD's social integration help us reflect on the current implementation of rehabilitation projects and institutional settings of drug governance.

Rootstock-scion exchanging mRNAs participate in the pathways of amino acids and fatty acid metabolism in cucumber under early chilling stress.

Cucumber (Cucumis sativus L.) often experiences chilling stress that limits their growth and productivity. Grafting is widely used to improve abiotic stress resistance by alternating a vigorous root system, suggesting there exists systemic signals communication between distant organs. mRNAs are reported to be evolving in fortification strategies by long-distance signaling when plants suffering from chilling stress. However, the potential function of mobile mRNAs alleviating chilling stress in grafted cucumber is still unknown. Here, the physiological changes, mobile mRNAs profiling, transcriptomic and metabolomic changes in above- and underground tissues of all graft combinations of cucumber and pumpkin responding to chilling stress were established and analyzed comprehensively. The co-relationship between the cluster of chilling-induced pumpkin mobile mRNAs with Differentially Expressed Genes (DEGs) and Differentially Intensive Metabolites (DIMs) revealed that four key chilling-induced pumpkin mobile mRNAs were highly related to glycine, serine and threonine synthesis and fatty acid ß-oxidative degradation metabolism in cucumber tissues of heterografts. The verification of mobile mRNAs, potential transport of metabolites and exogenous application of key metabolites of glycerophospholipid metabolism pathway in cucumber seedlings confirmed that the role of mobile mRNAs in regulating chilling responses in grafted cucumber. Our results build a link between the long-distance mRNAs of chilling-tolerant pumpkin and the fatty acid ß-oxidative degradation metabolism of chilling-sensitive cucumber. It helps to uncover the mechanism of signaling interaction between scion and stock responding to chilling tolerant in grafted cucumber.

[The role of SnRK2 in the response to stress, the growth and development of plants].

Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) is a specific Ser/Thr protein kinase in plants. SnRK2 can regulate the expression of downstream genes or transcription factors through phosphorylation of substrates to achieve stress resistance regulation in different tissue parts, and make plants adapt to adverse environment. SnRK2 has a small number of members and a molecular weight of about 40 kDa, and contains a conserved N-terminal kinase domain and a divergent C-terminal regulatory domain, which plays an important role in the expression of enzyme. This review summarized the recent research progresses on the discovery, structure, and classification of SnRK2, and its function in response to various stresses and in regulating growth and development, followed by prospecting the future research direction of SnRK2. This review may provide a reference for genetic improvement of crop stress resistance.

Research on a Rice Counting Algorithm Based on an Improved MCNN and a Density Map.

The thousand grain weight is an index of size, fullness and quality in crop seed detection and is an important basis for field yield prediction. To detect the thousand grain weight of rice requires the accurate counting of rice. We collected a total of 5670 images of three different types of rice seeds with different qualities to construct a model. Considering the different shapes of different types of rice, this study used an adaptive Gaussian kernel to convolve with the rice coordinate function to obtain a more accurate density map, which was used as an important basis for determining the results of subsequent experiments. A Multi-Column Convolutional Neural Network was used to extract the features of different sizes of rice, and the features were fused by the fusion network to learn the mapping relationship from the original map features to the density map features. An advanced prior step was added to the original algorithm to estimate the density level of the image, which weakened the effect of the rice adhesion condition on the counting results. Extensive comparison experiments show that the proposed method is more accurate than the original MCNN algorithm.

A novel method for credit scoring based on feature transformation and ensemble model.

Credit scoring is a very critical task for banks and other financial institutions, and it has become an important evaluation metric to distinguish potential defaulting users. In this paper, we propose a credit score prediction method based on feature transformation and ensemble model, which is essentially a cascade approach. The feature transformation process consisting of boosting trees (BT) and auto-encoders (AE) is employed to replace manual feature engineering and to solve the data imbalance problem. For the classification process, this paper designs a heterogeneous ensemble model by weighting the factorization machine (FM) and deep neural networks (DNN), which can efficiently extract low-order intersections and high-order intersections. Comprehensive experiments were conducted on two standard datasets and the results demonstrate that the proposed approach outperforms existing credit scoring models in accuracy.

Transcriptomic and Physiological Analysis Reveal That α-Linolenic Acid Biosynthesis Responds to Early Chilling Tolerance in Pumpkin Rootstock Varieties.

Climate changes especially chilling stress affects cucurbit crops during winter seasonal production. Grafting to pumpkin rootstocks is widely used to improve the vigor of cucurbits, especially cucumber (Cucumis sativus L.) plants, in the face of chilling stress. In our study, multi-disciplinary aspect approaches were used to investigate growth changes of pumpkin under chilling stress. Firstly, the morphological and physiological characteristics of 14 pumpkin (Cucurbita moschata) varieties following different periods of chilling stress was analyzed by using physiological means. Mathematical results of principal component analysis (PCA) with chlorophyll-a, chlorophyll-b, carotenoid contents, chilling injury index and relative electrolyte permeability indicated that relative electrolyte permeability as the primary judgment index was best associated with the comparison of chilling tolerance in pumpkin rootstock varieties. Then, transcriptomic and DCMU (Diuron) application and chlorophyll fluorescence examination analysis of pumpkin leaves revealed that 390 Cucurbita moschata differentially expressed genes (CmoDEGs) that affect photosynthesis were upregulated in leaves. 127 CmoDEGs both in leaves and roots were enriched for genes involved in unsaturated fatty acid metabolism, suggesting that plasma membrane lipids are involved in chilling perception. The results of increased composition of unsaturated fatty acid in leaves and qRT-PCR analysis of relative mRNA abundance confirmed that α-linolenic acid biosynthesis was responding to pumpkin chilling tolerance. The integration of physiological, mathematical bioinformatical and biological analysis results contributes to our understanding of the molecular mechanisms underlying chilling tolerance and its improvement in cucumber grafted on pumpkin rootstocks. It provided an important theoretical basis and reference for further understanding on the impact of climate change on plant physiological changes.

RNA Motifs and Modification Involve in RNA Long-Distance Transport in Plants.

A large number of RNA molecules have been found in the phloem of higher plants, and they can be transported to distant organelles through the phloem. RNA signals are important cues to be evolving in fortification strategies by long-distance transportation when suffering from various physiological challenges. So far, the mechanism of RNA selectively transportation through phloem cells is still in progress. Up to now, evidence have shown that several RNA motifs including Polypyrimidine (poly-CU) sequence, transfer RNA (tRNA)-related sequence, Single Nucleotide Mutation bound with specific RNA binding proteins to form Ribonucleotide protein (RNP) complexes could facilitate RNA mobility in plants. Furthermore, some RNA secondary structure such as tRNA-like structure (TLS), untranslation region (UTR) of mRNA, stem-loop structure of pre-miRNA also contributed to the mobility of RNAs. Latest researchs found that RNA methylation such as methylated 5' cytosine (m5C) played an important role in RNA transport and function. These studies lay a theoretical foundation to uncover the mechanism of RNA transport. We aim to provide ideas and clues to inspire future research on the function of RNA motifs in RNA long-distance transport, furthermore to explore the underlying mechanism of RNA systematic signaling.

Pneumocystis pneumonia in liver transplant recipients.

The clinical course of Pneumocystis pneumonia in liver transplant recipients has not been well investigated. Therefore, we collected and analyzed the clinical, epidemiological, and molecular data from patients with Pneumocystis pneumonia as well as paired controls (Chinese Clinical Trial Registry, ChiCTR2100046028; www.chictr.org.cn). There were a total of ten patients diagnosed with Pneumocystis pneumonia containing prospectively included six patients and retrospectively collected four patients, of which seven were transferred to the surgical intensive care unit and four died. The transmission map revealed that inter-patient transmission of Pneumocystis jirovecii was impossible; P. jirovecii detection was negative in all air samples. It was positive only in one sample from the twelve healthcare workers who had close contact with diseased patients. Five out of 79 liver transplant recipients during the outbreak were colonized with Pneumocystis jirovecii compared to 2 out of 94 after the outbreak upon admission (P>0.05). Liver transplant recipients with Pneumocystis pneumonia had totally different genotypes based on multilocus sequence typing. Additionally, we found an unreported mutation in the cytochrome b gene. The absolute CD19+ B-cell counts (odds ratio: 1.028; 95% confidence interval: 1.000-1.057; P=0.049) were defined to be the only significant independent risk factor. At a cut-off value of 117.16/µL, the sensitivity and specificity were 100% and 70%, respectively. Pneumocystis pneumonia is a severe complication following liver transplantation. The outbreak may not be caused by nosocomial transmission. A decrease in absolute CD19+ B-cell counts may be associated with the development of Pneumocystis pneumonia.

Optimization of RNA In Situ Hybridization for mRNA Localization Detection in Mature Tissue of Cucumber Seedlings.

Cucumber (Cucumis sativus L.) is one of the main vegetable crops in China. The physiological cultivation mechanism and gene function characteristics of cucumber are of great significance to the construction of modern agriculture. Due to the low genetic transformation rate of cucumber, only in situ hybridization, which does not involve the progress of gene modified transformation, is convenient to study mRNA localization, so it is more suitable for determination on mRNA localization in the mature tissue of cucumber. At present, the existing in situ hybridization technology system is more suitable for cucumber meristem than for the mature tissue of cucumber seedlings. Therefore, we optimized the traditional plant in situ hybridization protocol. Taking a known gene CsNPF7.2 (Nitrate Transporter Families protein) as an example, we then optimized the steps of plant tissue culture, gene probe preparation, plant material sampling and fixation, preparation of cross section, hybridization pretreatment, hybridization incubation, chromogenic reaction, microscopy examination, and treatment after reaction termination in order to obtain a new RNA in situ hybridization technique suitable for identification on mRNA localization in mature tissues of cucumber seedlings. This optimized technique will ensure the yield of probes, the integrity of RNA molecules, and the clarity and integrity of plant tissue structure, which is conducive to the study of gene function and screening of key genes in cucumber.