Tobacco is a significant economic crop cultivated in various regions of China. Arbuscular mycorrhizal fungi (AMF) can establish a symbiotic relationship with tobacco and regulate its growth. However, the influences of indigenous AMF on the growth and development of tobacco and their symbiotic mechanisms remain unclear. In this study, a pot inoculation experiment was conducted, revealing that six inoculants - Acaulospora bireticulata(Ab), Septoglomus viscosum(Sv), Funneliformis mosseae(Fm), Claroideoglomus etunicatum(Ce), Rhizophagus intraradices(Ri), and the mixed inoculant (H) - all formed stable symbiotic relationships with tobacco. These inoculants were found to enhance the activities of SOD, POD, PPO, and PAL in tobacco leaves, increase chlorophyll content, IAA content, CTK content, soluble sugars, and proline levels while reducing malondialdehyde content. Notably, among these inoculants, Fm exhibited significantly higher mycorrhizal infection density, arbuscular abundance, and soil spore density in the root systems of tobacco plants compared to other treatments. Membership function analysis confirmed that Fm had the most pronounced growth-promoting effect on tobacco. The transcriptome analysis results of different treatments of CK and inoculation with Fm revealed that 3,903 genes were upregulated and 4,196 genes were downregulated in the roots and stems of tobacco. Enrichment analysis indicated that the majority of these genes were annotated in related pathways such as biological processes, molecular functions, and metabolism. Furthermore, differentially expressed genes associated with auxin, cytokinin, antioxidant enzymes, and carotenoids were significantly enriched in their respective pathways, potentially indirectly influencing the regulation of tobacco plant growth. This study provides a theoretical foundation for the development and application of AMF inoculants to enhance tobacco growth.

Tobacco is one of China's key economic crops, known for its wide distribution, high yield, and renewability. Tobacco stalk fibers (TSFs) share a similar chemical composition to wood fibers, making them a potential reinforcement for plant fiber composites. However, the waste tobacco stalk fibers raw material utilization rate is very low, and wasteful phenomenon is very serious. In this study, we prepared biodegradable TSF/PBAT composites using waste tobacco stalk fibers and polybutylene adipate-co-terephthalate (PBAT) through melt blending and injection molding techniques. The effects of different modifiers on the performance of the composites were systematically investigated, with a particular focus on their influence on the degradation behavior. The results showed that the waste tobacco straw fiber can be used as a reinforcing fiber for PBAT. The addition of modifiers significantly improved the mechanical properties of the composites and effectively slowed down the degradation rate in the soil environment. Among the modifiers, the combined use of maleic anhydride (MA) and hydroxylated multi-walled carbon nanotubes (OM) produced the best results, with the tensile strength and flexural strength of the composite reaching 17.3 MPa and 28.0 MPa, respectively-representing increases of 74.7% and 57.3% compared to the untreated composite. After 16 weeks of soil degradation, the mass loss rate of the MA/OM-modified composite decreased from 10.50 to 6.34%. This study provides a comprehensive exploration of the entire lifecycle of TSF-reinforced PBAT composites and offers important theoretical support for the resource utilization and value-added application of waste tobacco stalks in the field of green composite materials.

Drought and soil salinization significantly constrain agricultural productivity, driving the need for molecular breeding strategies to enhance stress resistance. Zinc finger proteins play a critical role in plant response to abiotic stress. In this study, a gene encoding a C2H2-type zinc finger protein (AfZFP5) was cloned from Amorpha fruticosa, a species known for its strong adaptability. qRT-PCR analysis revealed that AfZFP5 expression is regulated by sorbitol, H2O2, NaCl, and NaHCO3. And all four treatments can cause upregulation of AFZFP5 expression in the roots or leaves of Amorpha fruticosa within 48 h. Transgenic tobacco lines overexpressing AfZFP5 demonstrated enhanced tolerance to drought and salt-alkali stress at germination, seedling, and vegetative stages. Compared to wild-type plants, transgenic lines exhibited significantly higher germination rates, root lengths, and fresh weights when treated with sorbitol, NaCl, and NaHCO3. Under natural drought and salt-alkali stress conditions, transgenic plants showed elevated activities of superoxide dismutase (SOD) and peroxidase (POD), and upregulated expression of oxidative stress-related kinase genes (NtSOD, NtPOD) during the vegetative stage. Additionally, transgenic tobacco displayed lower malondialdehyde (MDA) content and reduced staining levels with 3,3 ' diaminobenzidine (DAB) and Nitro blue tetrazolium (NBT), indicating enhanced reactive oxygen species (ROS) scavenging capacity by AfZFP5 upon salt-alkali stress. Under simulated drought with PEG6000 and salt-alkali stress, chlorophyll fluorescence intensity and Fv/Fm values in transgenic tobacco were significantly higher than in wild-type plants during the vegetative stage, suggesting that AfZFP5 mitigates stress-induced damage to the photosynthetic system. This study highlights the role of AfZFP5 in conferring drought and salt-alkali stress tolerance, providing genetic resources and a theoretical foundation for breeding stress-resistance crops.

Cadmium (Cd) is a major soil pollutant that threatens plant growth and human health. The plant ATPase associated with various cellular activities (AAA) SKD1 utilizes ATP hydrolysis energy to mediate cellular responses to environmental stress. However, the role and regulatory mechanisms of SKD1 in plant responses to Cd stress are not well understood. This study has demonstrated that the maize SKD1 gene (ZmSKD1) enhanced tobacco's tolerance to Cd stress. Overexpression of ZmSKD1 in tobacco reduced Cd accumulation and improved Cd tolerance. Moreover, ZmSKD1 overexpression enhanced the antioxidant capacity of tobacco, maintaining reactive oxygen species homeostasis and mitigating oxidative damage under Cd stress. The transcription factor AGL8 directly activated ZmSKD1 transcription, which in turn boosted ATPase activity in tobacco. This activation enhanced vesicle trafficking in root cells and accelerated Cd excretion in transgenic tobacco plants. Concurrently, the AGL8-ZmSKD1 module inhibited the expression of several Cd transport-related genes, thereby reducing Cd uptake by tobacco roots. These findings identified the AGL8-ZmSKD1 module as a crucial player in managing Cd stress through the vesicle trafficking pathway, offering valuable insights into strategies for developing crops with reduced Cd accumulation to ensure global food security and human health.

INTRODUCTION Tobacco farming plays a crucial role in the livelihoods of many rural communities in Pakistan, particularly in Khyber Pakhtunkhwa (KPK). However, this agricultural practice is associated with severe environmental degradation and significant health risks to workers during cropping. METHODS This study evaluates the ecological and health impacts of tobacco farming in Pakistan, employing both quantitative (surveys) including 200 respondents (farmers and field workers/laborers) and qualitative methods (in-depth interviews) involving 10 respondents (farmers, policy experts, agriculturist and environmental specialists). The research focuses on Swabi, a key tobacco-growing region, and highlights the negative effects of excessive pesticide use, fertilizer application, and deforestation, which contribute to soil erosion, water pollution, and biodiversity loss RESULTS Regression analysis shows that pesticide use ((3=0.65, p<0.001) and deforestation ((3=0.82, p<0.001) are the leading contributors to ecological degradation. The relationship between tobacco yield and environmental degradation, although showing a trend (p=0.062), is statistically negligible and unlikely to have practical significance ((3=-0.15). Health risks are equally concerning, with farmworkers (labor hired for farming, farmers, landlords) exposed to harmful agrochemicals and nicotine absorption leading to respiratory diseases, skin conditions, and green tobacco sickness (GTS). Pesticide exposure ((3=0.71, p<0.001) and contact with tobacco leaves ((3=0.53, p<0.001) significantly impact workers'health, while using personal protective equipment (PPE) helps mitigate these risks ((3=-0.43, p=0.001). The study also reveals that many farmers are interested in transitioning to alternative crops like maize or cotton, but they face financial and informational barriers. CONCLUSIONS The growing of tobacco in Pakistan entails significant ecological and health dangers, emphasizing the immediate need for the implementation of sustainable farming strategies to mitigate environmental harm and enhance the socio-economic conditions of farmers. Government support through financial incentives, educational programs, and sustainable farming techniques is essential to reduce the environmental damage and improve public health.

Tobacco (Nicotiana tabacum L.) shows promise for remediating Cd-contaminated soil due to its significant Cd accumulation capabilities. Although various tobacco varieties exhibit distinct Cd bioaccumulation capacities, a comprehensive understanding of the underlying mechanisms is lacking. This study, conducted using hydroponics, explores differences in Cd accumulation and tolerance mechanisms between two tobacco varieties, Basma and Yunyan 87. The results showed that Cd stress reduced the dry weight, tolerance index, and root morphology for both varieties. Basma exhibited a relatively smaller decline in these indices compared to Yunyan 87. Moreover, Basma demonstrated a higher Cd bioconcentration factor (BCF), concentration, and accumulated content, signifying its superior tolerance and bioaccumulation capacity to Cd compared to Yunyan 87. The Carbonyl Cyanide3-ChloroPhenylhydrazone (CCCP) addition resulted in reduced Cd accumulation and BCFs in both tobacco species. This effect was more pronounced in Basma, suggesting that Basma relies more on an active transport process than Yunyan 87. This could potentially explain its enhanced bioaccumulation ability. Subcellular Cd distribution analysis revealed Basma's preference for distributing Cd in soluble fractions, while Yunyan 87 favoured the cell wall fractions. Transmission electron microscope showed that Basma's organelles were less damaged than Yunyan 87's under Cd stress, possibly contributing to the superior tolerance of Basma. Therefore, these results provided a theoretical foundation for development of Cd-contaminated soil tobacco remediation technology. [GRAPHICS] .

MYB transcription factors exert crucial functions in enhancing plant stress tolerance, which is impacted by soil drought and salinity. In our study, the R2R3-type MYB transcription factor gene LcMYB5 from blue honeysuckle ( Lonicera caerulea L.) was successfully cloned and identified, and confirmed its nuclear localization. LcMYB5 overexpression was vastly enhanced drought and salt tolerance in both blue honeysuckle and tobacco seedlings. After drought stress, transgenic tobacco exhibited an average survival rate of 70.30%, while most wild-type (WT) plants perished, resulting in a survival rate of only 15.33%. Following salt stress, the average survival rate for transgenic tobacco reached 77.24%, compared to just 22.47% for WT plants. Measurements indicated, that transgenic tobacco had higher proline content than WT, as well as higher superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activity. Transgenic tobacco decreased chlorophyll content less dramatically than WT tobacco, despite both tobaccos having decreased chlorophyll content. Furthermore, the level of malondialdehyde (MDA) and relative conductivity were lower in transgenic tobacco compared to WT. Furthermore, LcMYB5 overexpression significantly increased the expression levels of key genes related to drought stress ( NCED1 , NCED2, PYL4, PYL8, and CBL1) and salt stress ( NHX1 , SOD, CAT1, SOS1, and HSP17.8), thus improving transgenic tobacco's stress tolerance. Compared to WT blue honeysuckle, transiently transformed LcMYB5-expressing blue honeysuckle exhibited milder damage under stress conditions, a significant increase in chlorophyll and proline content was observed, the activities of SOD, POD and CAT were also significantly increased, the increase in MDA content and relative conductivity is relatively small. Additionally, In addition, transient expression of LcMYB5 can also positively regulate the expression of these five key genes of drought stress and five key genes of salt stress, so as to improve the resistance of transgenic blue honeysuckle to drought and salt stress. In summary, our study reveals the important regulatory role of LcMYB5 in plant resistance to drought and salt stress, providing theoretical support and potential application value for further improving crop stress resistance.

Tobacco (Nicotiana tabacum L.) bacterial wilt, caused by Ralstonia solanacearum, is indeed a highly destructive plant disease, leading to substantial damage in tobacco production. While biological control is considered an effective measure for managing bacterial wilt, related research in this area has been relatively limited compared to other control methods. In order to discover new potential antagonistic bacteria with high biocontrol efficacy against tobacco bacterial wilt, we conducted an analysis of the microbial composition differences between disease-suppressive and disease-conducive soils using Illumina sequencing. As a result, we successfully isolated six strains from the disease-suppressive soil that exhibited antibacterial activity against Ralstonia solanacearum. Among these strains, B4-7 showed the strongest antibacterial activity, even at acidic conditions with a pH of 4.0. Based on genome analysis using Average Nucleotide Identity (ANI), B4-7 was identified as Bacillus velezensis. In greenhouse and field trials, strain B4-7 significantly reduced the disease index of tobacco bacterial wilt, with control efficiencies reaching 74.03% and 46.88% respectively. Additionally, B4-7 exhibited plant-promoting abilities that led to a 35.27% increase in tobacco production in field conditions. Quantitative real-time (qPCR) analysis demonstrated that strain B4-7 effectively reduced the abundance of R. solanacearum in the rhizosphere. Genome sequencing and Liquid Chromatography-Mass Spectrometry (LC-MS) analysis revealed that strain B4-7 potentially produces various lipopeptide metabolites, such as microlactin, bacillaene, difficidin, bacilysin, and surfactin. Furthermore, B4-7 influenced the structure of the rhizosphere soil microbial community, increasing bacterial abundance and fungal diversity, while also promoting the growth of different beneficial microorganisms. In addition, B4-7 enhanced tobacco's resistance to R. solanacearum by increasing the activities of defense enzymes, including superoxide dismutase (SOD), phenylalanine ammonia-lyase (PAL), peroxidase (POD), catalase (CAT), and polyphenol oxidase (PPO). Collectively, these findings suggest that B. velezensis B4-7 holds significant biocontrol potential and can be considered a promising candidate strain for eco-friendly management of tobacco bacterial wilt.

Tobacco black shank (TBS) is a soil-borne fungal disease caused by Phytophthora nicotiana (P. nicotianae), significantly impeding the production of high-quality tobacco. Molybdenum (Mo), a crucial trace element for both plants and animals, plays a vital role in promoting plant growth, enhancing photosynthesis, bolstering antioxidant capacity, and maintaining ultrastructural integrity. However, the positive effect of Mo on plant biotic stress is little understood. This study delves into the inhibitory effects of Mo on P. nicotianae and seeks to unravel the underlying mechanisms. The results showed that 16.32 mg/L of Mo significantly inhibited mycelial growth, altered mycelial morphological structure, damaged mycelial cell membrane, and ultimately led to the leakage of cell inclusions. In addition, 0.6 mg/kg Mo applied in soil significantly reduced the severity of TBS. Mo increased photosynthetic parameters and photosynthetic pigment contents of tobacco leaves, upregulated expression of NtPAL and NtPPO resistance genes, as well as improved activities of SOD, POD, CAT, PPO, and PAL in tobacco plants. Furthermore, Mo could regulate nitrogen metabolism and amino acids metabolism to protect tobacco plants against P. nicotianae infection. These findings not only present an ecologically sound approach to control TBS but also contribute valuable insights to the broader exploration of the role of microelements in plant disease management.

PurposeThe health of rhizosphere soil microorganisms is an important indicator to evaluate soil quality. Therefore, understanding the response of rhizosphere soil microorganisms to tobacco crop succession is crucial for promoting the sustainable development of agriculture.MethodsThe microbial diversity and community structure of rhizosphere soil in continuous cropping and non-cropped tobacco for 7 years were analyzed by the Illumina platform.Result(1) Continuous cropping tobacco cause rhizosphere soil acidification and reduction in alkaline nitrogen (AN) and soil organic matter (SOM). (2) Continuous cropping tobacco reduces the diversity of rhizosphere soil microbial communities, increasing harmful functional microorganisms and declining beneficial ones. (3) The abundance of bacteria that perform nitrification and saprophytic fungi in the rhizosphere soil of continuous cropping areas decreases, inhibiting carbon and nitrogen cycling processes. (4) The composition and diversity of the soil rhizosphere microbial community are affected by the imbalance in the physicochemical property of the rhizosphere.ConclusionContinuous cropping tobacco cause rhizosphere soil acidification and nutrient imbalance, and the carbon and nitrogen cycles involved in microorganisms were damaged. Furthermore, the decreased diversity of rhizosphere soil microorganisms and the increased abundance of pathogenic fungi contribute to the continuous cropping obstacles of tobacco.