Molybdenum (Mo) is widely used as a micronutrient fertilizer to improve plant growth and soil quality. However, the interactions between cell wall biosynthesis and molybdenum have not been explored sufficiently. This study thoroughly investigated the regulatory effects of different concentrations of Mo on tobacco cell wall biosynthesis from physiological and metabolomic aspects. The results indicate that Mo treatment increased the Mo content of tobacco variety K326. Moreover, it significantly up-regulated the gene expression levels of molybdases (NR, AO, SO, XDH) and molybdate transporters in tobacco, whereby the gene expression levels of NR were upregulated by 28.48%, 52.51%, 173.05%, and 246.21%, respectively; and MOT1 and MOT2 were upregulated by 21.49/8.67%, 66.05/30.44%, 93.05/93.26%, and 166.11/114.29%, respectively. Additionally, Mo treatment regulated the synthesis of related enzymes, effectively promoted plant growth, and significantly increased biomass and dry matter accumulation, with the biomass in the leaves increasing significantly by 30.73%, 40.72%, 46.34%, and 12.88%, respectively. The FT-NIR spectroscopy results indicate that after Mo was applied to the soil, the quantity of C-O-C, -COOH, C-H, and N-H functional groups increased. Concurrently, the contents of cellulose, hemicellulose, lignin, protopectin, and soluble pectin in the leaves significantly increased, wherein the content of soluble pectin and hemicellulose increased significantly by 31.01/288.82%, 40.69/343.43%, 69.93/241.73%, and 196.88/223.26%, respectively. Furthermore, the cell walls thickened, increasing the ability of the plant to withstand disturbances. The metabolic network diagrams indicate that Mo regulated galactose metabolism, and arginine and proline acid biosynthesis. The contents of carbohydrates, spermidine, proline, quinic acid, IAA, flavonoids, and other substances were increased, increasing the levels of polysaccharides and pectin within the cell wall, controlling lignin production, and successfully enhancing resistance to abiotic stress. These results offer important perspectives for further investigations into the role of trace elements.

Heavy metal-associated isoprenylated plant proteins (HIPPs) are a class of metallochaperone proteins crucial for plant growth, development, and stress responses. However, a systematic investigation of the tobacco HIPP family has not been conducted. In this study, we identified 68 NtHIPP genes, all of which had conserved heavy metal-associated structural domains and the C-terminal isoprenoid motif CaaX. Phylogenetic analyses categorized NtHIPP genes into five clades, with 37 NtHIPP genes unevenly distributed across 18 tobacco chromosomes. Collinearity relationship analysis revealed that segmental duplication was a major factor in the evolution of these genes, and they exhibit a closer evolutionary relationship to dicots (Arabidopsis thaliana and Solanum lycopersicum) compared to monocots (Triticum aestivum and Oryza sativa). RNA-seq data indicated NtHIPP genes had different expression profiles in the shoot apex, shoot, and root, and some genes showing differential expression in response to drought, cold, cadmium stress, and Ralstonia solanacearum infection. Eight NtHIPP genes, as identified via reverse transcription-quantitative Polymerase Chain Reaction (RT-qPCR), exhibited positive or negative responses to various environmental stimuli under abiotic stress and hormone treatments. This study elucidates the relationship between NtHIPP genes and plant stress responses, providing new candidates for exploring stress-resistant germplasm resources in tobacco.

Salinity stress severely inhibits crop growth and reduces yield. Exogenous selenium (Se) enhances plant abiotic stress tolerance, but how different selenium forms exert their impacts and pathways in mitigating salinity remains ambiguous. Under salt stress, this work compared two Se forms, selenate [Se(VI)] and selenite [Se(IV)], regarding their impacts on development, photosynthetic performance, antioxidative system, osmotic regulators, Se buildup, and stress-related gene expression in Nicotiana tabacum L. Both Se species significantly promoted tobacco growth. (1) Under 150 mmol/L NaCl stress, biomass, net photosynthetic rate and antioxidant enzyme activities decreased significantly, while soluble sugar, free proline, Na⁺/K⁺, Na⁺/Ca²⁺, H₂O₂, MDA contents and NtROS2a, NtLEA5 expression increased significantly. (2) Exogenous Se increased biomass, photosynthetic parameters; antioxidant enzyme activities and NtNAC2, NtCDPK12, NtROS2a expression; elevated Se deposition in roots and leaves; and reduced oxidative damage, ion imbalance and NtLEA5 expression in salt-stressed tobacco, suggesting that Se may improve salt tolerance by regulating these physiological processes and stress-related gene expression. (3) Compared with Se(IV), Se(VI) significantly increased root length, chlorophyll content, stomatal conductance, K⁺ content, SOD/CAT activities, leaf and root Se accumulation as well as and NtNAC2, NtCDPK12 expression, while Se(IV) resulted in higher root diameter, free proline content, Na⁺/K⁺ ratio and NtROS2a expression. In conclusion, both sodium selenate and sodium selenite effectively enhanced tobacco salt tolerance. The salt stress alleviation effect of Se(VI) may be associated with upregulating NtNAC2 and NtCDPK12 to improve antioxidant capacity and photosynthesis, thereby potentially maintaining cell membrane integrity and ion balance, while Se(IV) may exert its effect through upregulating NtROS2a to promote root thickening, reactive oxygen species scavenging and osmotic adjustment. At the tested concentrations, selenate was more effective.