IntroductionTobacco (Nicotiana tabacum L.) upper leaves constitute approximately one-third of total leaf production, yet their smaller leaf area, compact structure, and imbalanced chemical composition limit suitability for cigarette manufacturing. This study investigated whether gibberellin (GA) and melatonin (MT) could promote upper leaf growth and balance chemical components.MethodsA two-factor randomized block design experiment was conducted to evaluate GA and MT effects on tobacco upper leaves. Morphological parameters, biomass accumulation, and chemical composition were measured. Structural equation modeling and TOPSIS (Technique for Order Preference by Similarity to Ideal Solution) analysis were employed to determine regulatory pathways and optimal hormone combinations.ResultsGibberellin significantly promoted upper leaf morphological development. The 75 mg/L GA treatment increased leaf width and leaf area by 12.96% and 17.69%, respectively, and significantly enhanced both fresh and dry weight (P<0.01). The GA-MT interaction significantly increased biomass accumulation (P<0.05). Structural equation modeling indicated GA primarily drove dry matter accumulation through direct regulation of leaf area (path coefficient, 0.47) and fresh weight (path coefficient, 0.39). Single hormone treatments improved chemical quality, with Chemical Component Usability Index (CCUI) ameliorated under 80 μmol/L MT and 75 mg/L GA treatments. TOPSIS analysis identified 75 mg/L GA + 80 μmol/L MT as the optimal combination (closeness coefficient, 0.644), effectively balancing biomass increase with optimized chemical coordination.DiscussionThis study confirms gibberellin as the dominant factor regulating upper leaf morphology and biomass, while melatonin modulates chemical quality through interaction, providing a theoretical foundation and technical strategy for enhancing growth and chemical quality in tobacco upper leaves.

Nitrogen is a primary factor affecting the yield and quality of tobacco. Insufficient nitrogen results in poor tobacco output and poor leaf quality; in contrast, excessive nitrate can lead to delayed maturity and poor quality of cured tobacco leaves. To elucidate the molecular mechanism underlying the response of tobacco to low nitrogen, we conducted a multiomics analysis of tobacco plants treated with normal (7.5 mM) or low nitrogen (0.75 mM). Our results revealed that after 15 days of low-nitrate treatment, the plants exhibited stunted growth, yellowed leaves, and significant growth inhibition. The total nitrogen levels, and the activities of glutamine synthetase and nitrate reductase in the leaves also decreased significantly. Transcriptome analysis of the tobacco plants under low-nitrogen stress revealed 896 differentially expressed genes, with 404 genes upregulated and 492 genes downregulated in comparison with those in the control. Among the upregulated genes, the expression of Nitab4.5_0000970g0150, which is involved in galactose metabolism, significantly increased. Subsequent metabolome analysis revealed that 28 metabolites significantly changed under low-nitrogen treatment, including 23 metabolites whose levels increased and 6 whose levels decreased compared with those in the control. The increased metabolites were also enriched in galactose metabolism. Integrated transcriptome and metabolome analyses revealed that three pathways significantly changed—amino acid synthesis and metabolism, galactose metabolism and trehalose-6-phosphate metabolism—among which galactose metabolism is important for tobacco in response to low nitrogen stress. These findings provide a theoretical basis for further research on the function of nitrogen regulatory genes in tobacco and for the breeding of tobacco varieties with high nitrogen use efficiency.

Potassium (K+) is the macronutrient that closely related to the growth and development of tobacco plant. The deprivation of K+ increases ethylene production. 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) is the key enzyme in the process of ethylene biosynthesis. To elucidate the relationship between ACO and low potassium (LK) stress, we overexpressed ACO homolog 1 (ACOH1) in tobacco plants. More results showed that overexpression of NtACOH1 in tobacco exhibited improved LK stress tolerance with increased proline, potassium and ACC content in the transgenic plants compared with wild type (WT). Meanwhile, NtACOH1-overexpression significantly decreased K+ efflux rate in transgenic tobacco roots under LK stress. KEGG analysis of transcriptome showed that the spliceosome, biosynthesis of amino acids and MAPK signaling pathway - plant were the three up-regulated pathways. The result of weighted gene co-expression network analysis exhibited that ethylene-responsive transcription factor RAP2-7, MYB transcription factor (MYBAS1), C2H2-13 protein might regulate the expression of NtACOH1. Further researches suggested NtRAP2-7 activated the expression level of NtACOH1 by yeast one-hybrid experiment and Dual-luciferase assays.Our results provide valuable candidate genes and enrich signal transduction network in tobacco in response to LK stress.

The potato (Solanum tuberosum L.) plays an important role in ensuring global food security. Potato tubers store abundant nutrients and are also reproductive organs. The adjustment of tuber sprouting plays a vital role in timely sowing and improving tuber product quality. CBL-interacting protein kinases (CIPKs) exert an important function in the entire life cycle of plants and in coping with stress. In our present study, we found that the StCIPK23 expression level increased during storage and that overexpression of StCIPK23 can accelerate tuber sprouting. Physiological assays indicated that overexpressing StCIPK23 altered carbohydrate metabolism and antioxidant-related enzyme activities during storage. Starch branching enzyme (SBEI) gene expression was upregulated, while sucrose synthase (SS), 3-phosphoglyceric phosphokinase (PGK), and glyceraldehyde-3-phosphate dehydrogenase 1 (GAPC1) gene expression were downregulated in StCIPK23-overexpressing potato. High gibberellin (GA) content and low abscisic acid (ABA) content were also detected in transgenic tubers. We conducted metabolomics analysis on bud eyes, and the results showed a total of 94 differential metabolites were found. Among them, 61 metabolites were increased, the top three metabolites were coumaryl alcohol, glutathione and quercetin–glucoside–glucoside–rhamnoside. Our results suggest that StCIPK23 is a positive regulator of potato tuber sprouting.

Cigar leaf is a special type of tobacco plant, which is the raw material for producing high-quality cigars. The content and proportion of nicotine and other composite substances of cigar leaves have a crucial impact on their quality and vary greatly with the time of harvest. Hyperspectral remote sensing technology has been widely used in the field of crop monitoring because of its advantages of large area coverage, fast information acquisition, short cycle turnover, strong real-time performance and high efficiency. Therefore, it is important to accurately monitor nicotine content of field crops in a timely manner in the production of high-quality cigar leaf. To this end, this study set out to measure crop reflectance spectra acquired by UAV drones from tobacco field crops by hyperspectral image acquisition. MSC, SG, and SNV were combined and applied to the raw data. The output of these operations was then further processed by CARS, SPA, and UVE algorithms to determine the nicotine sensitive bands. Three machine learning algorithms were then used to analyze the data: PLS, BP, RF, and the SVM. An inversion model of the content of nicotine was established, and the model was evaluated for accuracy. The main research conclusions are as follows: (1) With the increase in the rate of application of nitrogen fertilizer, the nicotine content of cigar leaves increased; (2) Processing data by the CARS, SPA, and UVE methods reduces the degree of data redundancy and information co-linearity in the screening of the content of nicotine sensitive bands; (3) The MSC-SNV-SG-CARS-BP model has the best predictive accuracy on the nicotine content. The prediction accuracy of the testing set was R2 = 0.797, RMSE = 0.078,RPD = 2.182.

Nitrogen is an important nutrient required for plant growth and development, but most of the time plants face nitrogen deficiency, all it is important to study the mechanism of low nitrogen tolerance in plants. This study addresses this gap by investigating the role of the StDWF1 gene through the generation and analysis of transgenic potato lines overexpressing StDWF1 (OE1, OE2, OE3). Exogenous BL treatment showed that the StDWF1 gene responded to oleuropein lactone. Phenotypic assessments under normal nitrogen (NN) and low nitrogen (LN) conditions demonstrated that OE2 consistently outperformed WT, showing a 43% increase in root vitality and a 23% retention of chlorophyll under LN. Additionally, OE2 transgenics accumulated significantly higher levels of nitrate nitrogen (64.1% increase) and ammonium nitrogen (53% increase) compared to WT. Enzymatic assays further confirmed elevated activities of glutamine synthetase and nitrate reductase in both OE1 and OE2 lines. Hormone analyses showed that BL content of StDWF1 overexpression lines was significantly increased under LN conditions, higher Oleandrin lactone (BL) content of OE2 improved plant stress tolerance, and WT was more affected by low nitrogen stress than OE2, resulting in higher levels of stress hormones than OE2. Temporal gene expression analysis showed significant upregulation of key nitrogen metabolism-related genes (NR, NiR, AT, NRT2.1) in OE2, with StDWF1 expression reaching 79% higher than WT at 3 h. Protein–protein interaction assays, including yeast two-hybrid and BiLC assays, verified the interaction between StDWF1 and StGRP1, suggesting the existence of a functional network to enhance low-nitrogen tolerance in potato plants. In conclusion, these findings suggest that overexpression of StDWF1 significantly enhances low-nitrogen tolerance in transgenic potato lines, providing a promising strategy for improving crop performance under nitrogen-limited conditions.

With the promotion of sustainable agricultural practices, reducing fertilizer application has become a necessary trend. Potassium is an essential mineral element for potato growth and development, and GABA, as a plant growth regulator, is considered to be able to alleviate abiotic stress. In this study, the effect of this method on potato growth was explored by simulating the environment of potassium fertilizer application reduction and combining GABA treatment. To reduce potassium fertilizer application, an appropriate concentration of GABA was sprayed on the leaves of potted potato plants, which significantly alleviated potassium deficiency stress. In the potted potato experiment, three potassium fertilizer rates were applied: A1 (225 kg/ha), A2 (112.5 kg/ha, 1/2K), and A3 (56.25 kg/ha, 1/4K), along with two foliar treatments: B1 (water) and B2 (5 mM GABA). Under different potassium conditions(A1, A2, A3), after foliar application of 5 mM GABA: the tuber starch content increased by 5.00

CBL-interacting protein kinases (CIPKs) play important regulatory roles in plant growth development and abiotic stress tolerance. However, the biological roles of these genes in response to low-nitrate (LN) stress in potato plants have not been determined. Here, we reported that StCIPK23 was expressed mainly in roots and leaves. StCIPK23 was located mainly in the cell membrane, nucleus, and cytoplasm. Further research suggested that, compared with wild-type (WT) plants, StCIPK23-overexpressing plants were taller and had significantly greater nitrate and ammonium nitrogen contents under LN stress. StCIPK23 overexpression can increase StAT, StNRT2.1, StNR, StGS1-3, and StGOGAT expression levels in StCIPK23 transgenic seedlings compared to those in WT plants under LN stress. The results of yeast two-hybrid and luciferase complementation imaging experiments suggested that StCIPK23 could interact with StCBL3. Real-time reverse transcription–PCR revealed the StCIPK23 expression level peaked at 6 h and the StCBL3 expression level peaked at 9 h in the roots under LN stress. In conclusion, we found that StCIPK23 and StCBL3 form a complex to regulate the expression of key genes in the nitrogen metabolism pathway to improve LN tolerance in potato plants.

Unwinding enzymes are a group of highly conserved enzymes that play important roles in plant growth and development as well as in adverse and stress conditions. The DEAD-box family represents the largest subfamily of deconjugating enzymes. However, little research has been conducted on the DEAD-box RNA gene family (StRH) in potato. Identifying and screening members of the DEAD-box gene family in potato could help us to better understand the response of potatoes to adverse and stress conditions and the related tolerance mechanisms. We identified a total of 73 DEAD-box gene family members from the potato genome-wide database. A comprehensive bioinformatics study of the potato DEAD-box family was carried out by a range of analytical methods. The evolutionary tree demonstrated that the potato DEAD-box family was divided into five distinct taxa. Three pairs of segmental repeats were identified on the chromosomes. In addition, we selected nine DEAD-box genes for qPCR(Quantitative Real-time PCR), and the results indicated that all of these genes were involved in both the ABA hormone stress and drought stress. These findings may help to increase our understanding of the functional diversity of the potato DEAD-box gene family under different environmental stresses.

Potatoes are widely planted worldwide and are the third most important food crop. Mutation breeding involves the artificial use of various physical and chemical factors to induce plants to produce new genotypes. In this study, we established a potato radiation mutation system and analysed the features of a radiation mutant. The LD50 of the potato callus was 24.8 Gy after linear regression analysis. The radiation mutant 1 (RM1) showed significant dwarfism and strong growth; RM1 plants decreased in height by 31%. The root and leaf fresh weights of the RM1 increased 1.8-fold compared to the wild-type (WT, Chuanyu 10) cultivar. Leaf microstructure results showed that the thicknesses of the upper epidermis and lower epidermis of RM1 plants were greater than those of the WT cultivar. Transcriptome analysis of seeding revealed 1179 upregulated and 1641 downregulated differentially expressed genes (DEGs) in RM1 plants. Further analysis showed that the expression levels of WRKY and MYB transcription factors, CIPK and MAPK protein kinases, ABC transporters, hormones, and ROS pathway genes were altered. These results provide theoretical support for potato breeding research and enrich the functional network of vital breeding-related genes.

Potassium (K+) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of Nt GF14e, a 14-3-3 gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of Nt GF14e transgenic tobacco lines compared with those in the wild type plants. The net K+ efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between Nt GF14e overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (CIPK2), Nt CIPK23, Nt CIPK25, H+-ATPase isoform 2 a (AHA2a), Nt AHA4a, Stelar K+ outward rectifier 1(SKOR1), and high affinity K+ transporter 5 (HAK5). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.

Ureide permeases (UPSs) mediate the transport of ureides, including allantoin and allantoate, which act as nitrogen-transporting compounds in plants and have recently been found to play a role in cellular signaling. To date, UPSs have not been reported in potato, and their identification is important for further function studies and for understanding molecular mechanisms of plant adverse responses. Based on potato genomic data, we identified 10 StUPS genes in potato (Solanum tuberosum L.). Then, we conducted a comprehensive study of the identified StUPS genes using bioinformatics methods. Genome phylogenetic and genomic localization analyses revealed that StUPSs can be classified into four categories, are highly homologous to Arabidopsis thaliana UPS members, and are distributed on three chromosomes. The six StUPS genes were investigated by RT–qPCR, and the findings indicated that all of these genes are involved in the response to several stresses, including low nitrogen, cold, ABA, salt, H2O2, and drought. This study establishes a strong theoretical framework for investigating the function of potato UPS genes, as well as the molecular mechanisms underlying the responses of these genes to various environmental stresses.

The BTB/POZ (broad-complex, tramtrack, and bric-a-brac) family of proteins is widespread in plants and animals and plays important roles in growth, development, metabolism, and environmental responses. There are few reports on BTB family genes in potato. In this study, 34 sequences containing conserved BTB domains were obtained from the potato gene database, and the phylogenetic, physical, and chemical properties, gene structure, conserved motif, domain, and chromosomal localization of the potato BTB protein family were analyzed via bioinformatics methods. In addition, we used qRT-PCR to detect 12 selected StBTB genes. The results confirmed that these genes are involved in cold, ABA, salt, hydrogen peroxide (H2O2), drought, and low-nitrogen stress, which is highly important for elucidating BTB family members and studying stress response and tolerance mechanisms. This study provides a theoretical basis for the study of the function and expression of potato BTB and lays a solid foundation for further understanding the molecular mechanism of the potato BTB gene under various environmental stresses.

Potato tubers are rich in starch, vitamins, protein, minerals, and other nutrients. However, tuber sprouting produces solanine and reduces the commodity value of potatoes during storage. At present, it is known that some plant essential oils can inhibit tuber sprouting. It has been reported that grape seed oil (GSO) has antioxidant, anti-inflammatory, and anticancer characteristics, reducing blood lipids and delaying aging. In this study, we found for the first time that GSO delayed tuber sprouting, and the soluble sugar content and peroxidase activity changed after 60 days of GSO treatment. Furthermore, a comparative proteomic analysis of tuber bud eyes showed that after 30 days of GSO treatment, there were 206 and 129 differentially abundant proteins (DAPs) with increased and decreased abundance levels, respectively. After analysis, we found that 15 ROS-related proteins and 14 proteins involved in energy metabolism were DAPs. Among them, gamma aminobutyrate transaminase 1 had decreased abundance after GSO treatment. Meanwhile, the transcription level of genes related to GABA synthesis increased significantly according to qRT-PCR analysis. Our results provide new approaches to the proteomic mechanism of potato sprouting after GSO treatment and provide a theoretical basis for the application of GSO in inhibiting potato seed sprouts.

雪茄烟叶的标准化生产,对于提高烟叶质量、提升生产规模、增加烟农收入具有十分重要的意义.为了提升德阳雪茄烟叶生产的标准化水平,进行了德阳雪茄烟叶生产综合标准体系的建设.所建立的标准体系,由基础标准、试验与分析标准、环境条件与生产资料标准、生产技术标准、产品标准与质量监控标准、管理与服务标准等六大板块构成,包含各类标准共计63项,涵盖了雪茄烟叶生产的育苗、整地、机耕、覆膜、移栽、田间管理、采收以及调制等主要环节.开展了德阳雪茄烟叶生产综合标准体系在雪茄烟叶生产中的贯彻与应用,取得了良好的成效.

Tobacco is a critical cash crop in China, so its growing status has received more and more attention. How to acquire accurate plant area, row spacing, and plant spacing at the same time have been key points for its grow status monitoring and yield prediction. However, accurately detecting small and densely arranged tobacco plants during the rosette stage poses a significant challenge. In Sichuan Province, the contours of scattered tobacco fields with different shapes are not well-extracted. Additionally, there is a lack of simultaneous methods for extracting crucial tobacco planting information, including area, row spacing, and plant spacing. In view of the above scientific problems, we proposed a method to extract the planting information of tobacco at the rosette stage with Unmanned Aerial Vehicle (UAV) remote sensing images. A detection model, YOLOv8s-EFF, was constructed for the small and weak tobacco in the rosette stage. We proposed an extraction algorithm for tobacco field area based on extended contours for different-shaped fields. Meanwhile, a planting distance extraction algorithm based on tobacco coordinates was presented. Further, four experimental areas were selected in Sichuan Province, and image processing and sample label production were carried out. Four isolated tobacco fields with different shapes in four experimental areas were used to preliminarily verify the effectiveness of the model and algorithm proposed. The results show that the precision ranges of tobacco field area, row spacing, and plant spacing were 96.51~99.04%, 90.08~99.74%, and 94.69~99.15%, respectively. And another two experimental areas, Jiange County, Guangyuan, and Dazhai County, Gulin County, and Luzhou, were selected to evaluate the accuracy of the method proposed in the research in practical application. The results indicate that the average accuracy of tobacco field area, row spacing, and plant spacing extracted by this method reached 97.99%, 97.98%, and 98.31%, respectively, which proved the extraction method of plant information is valuable.

Tobacco is one of the most important cash crops in China, and accurate acquisition of its planting information is of great significance for tobacco management and yield prediction. In this paper, a tobacco information extraction method based on UAV high-resolution images is proposed. The method uses the RtinaNet target detection model to achieve the identification and localization of tobacco in UAV high-resolution images, and generates tobacco coordinate information. Based on the tobacco coordinates, we designed two algorithms for extracting plant spacing, row spacing and area of tobacco fields, respectively. The experimental results showed that the accuracy of the method reached 98.49%-99.77%, 97.08%-99.98% and 94.48%-99.63% for area, row spacing and plant spacing extracted from the seven experimental tobacco fields, respectively.

Drought is an important environmental factor affecting plant growth and development. Tobacco is a widely cultivated and economically important crop worldwide. Drought can decrease the yield and quality of tobacco. Peroxidase III can maintain the balance of peroxy free radicals and H2O2 in plants and participate in a variety of physiological processes, including lignin biosynthesis and auxin catabolism. In this study, we first found that a CRISPR/Cas9-mediated peroxidase 63-like (NtPOD63 L) knockout mutant had high drought tolerance, such as reduced stomatal apertures, increased lignin content and interfascicular fiber number, and induced ABA-related gene expression. Then, a quantitative phosphoproteomic analysis of tobacco mutant leaves was conducted after PEG treatment. Phosphoproteomic analysis revealed 609 phosphorylated proteins, among which 266 were upregulated and 343 were downregulated. KEGG pathway analysis indicated that the cluster with "Pyruvate metabolism" contained the most upregulated pathways. The cluster with "RNA degradation", "Cysteine and methionine metabolism," and "Tryptophan metabolism contained the three primary downregulated pathways. Sixty-seven kinases were found to change phosphorylation levels. Our study provides new information for understanding NtPOD63 L function in drought stress and describes the molecular regulation mechanisms of the tobacco response to drought at the posttranslational level.

Adenylate kinase (ADK) is a key enzyme that is widely distributed in animals and plants. It plays an important role in growth and stress response. However, ADK genes in potato (StADK) have been little reported. It is of great significance to identify ADK members and understand the molecular mechanism of stress response and tolerance. Based on the potato genome data, 23 StADK genes were identified at a genome-wide level. We then performed a comprehensive study using a bioinformatics method. The results of the evolutionary tree showed that StADK proteins were divided into four groups, and they were highly homologous to the Arabidopsis thaliana ADK members. Meanwhile, our study found that they existed on eight chromosomes, and we obtained three pairs of fragment duplications. Furthermore, we detected the six selected StADK genes using qRT-PCR, and the results confirmed that the genes are involved in the regulation of cold, ABA, salt, H2O2 and drought stresses. Our study provides a theoretical basis for studying the function of the potato ADK genes and lays a solid foundation for further understanding the molecular mechanism of the potato ADK genes under various environmental stresses.

In order to compare the effects of multiple linear regression and BP neural network in the SPAD inversion modeling of tobacco leaves, the multispectral image of the tobacco canopy was collected and the vegetation index was extracted. Then, using multiple linear regression and BP neural network algorithm, the inverse regression model was constructed respectively. The results showed that among themodels, the R~2 of the model built by BP neural network was 0.85, and the RMSE was 2.21. By contrast, the R~2 of the model constructed by the multiple linear regression method was only 0.51, and the RMSE was 1.52. The regression inversion precision of BP neural network was the better. The results of this study show that the BP neural network can be used to construct the SPAD inversion modeling of tobacco leaves.