Tobacco Cultivars Research Articles

Construction and evaluation of a model for efficient identification of photothermal sensitivity of tobacco cultivars based on agronomic traits.

The photothermal sensitivity of tobacco refers to the degree to which tobacco responds to changes in light and temperature conditions in its growth environment, which is crucial for determining the planting area of cultivars and improving tobacco yield and quality. In order to accurately and effectively evaluate the photothermal sensitivity of tobacco cultivars, this study selected five cultivars and their hybrid combinations with significant differences planted under different ecological conditions from 2021 to 2022 as materials. The experiment was conducted in two locations with significant differences in temperature and light. We measured the agronomic traits and biomass of the experimental materials, and constructed an effective tobacco photothermal sensitivity evaluation model using principal component analysis, membership function, and regression analysis. The reliability of the model was evaluated by utilizing the photosynthetic characteristics, chlorophyll content, and antioxidant enzyme system activity of the experimental materials. The results showed that tobacco biomass is the most important principal component in agricultural traits, and the comprehensive evaluation model for tobacco photothermal sensitivity is: y = 0.4571y1 + 0.2406y2 + 0.1725y3, where the fitting coefficients R2 of y1, y2, and y3 are 0.945, 0.851, and 0.977, respectively; The photothermal sensitivity of the experimental materials was calculated using this model, and the comprehensive ranking of the 11 experimental materials is: G3 < G5 < G10 < G9 < G11 < G6 < G7 < G2 < G4 < G8 < G1. Conventional identification methods have found that G2, G4, G6, G7, G8, and G11 are sensitive materials, G3, G5, and G10 are insensitive materials, and G1 and G9 are intermediate materials. The consistency rate of the evaluation results of the two methods reached 90.91%. And there is a significant correlation between the agronomic traits selected in the model and the physiological indicators selected by conventional evaluation methods, providing a scientific basis for evaluating the light temperature sensitivity of tobacco cultivars using agronomic traits in this study. The results indicate that the photothermal sensitivity evaluation model established in this study provides an efficient, convenient, and reliable method for evaluating the photothermal sensitivity of tobacco.

Characterization of Two Lycopene Beta-cyclases Reveal Their Essential Roles in Photoprotection in Nicotiana Tabacum

Carotenoids are essential isoprenoids with vital roles in photoprotection and antioxidative processes in photosynthetic organisms. Lycopene Beta-cyclase (β-LCY) is a key enzyme in carotenoid biosynthesis; however, the role of β-LCY in photoprotection is still unclear. In this study, we characterized two β-LCY genes in an allotetraploid tobacco cultivar K326, that are primarily expressed in leaves. Reduction of β-LCY expression by RNA interfercnce (RNAi) resulted in decreased carotenoid content, leaf variegation, impaired photosynthetic efficiency, and abnormal chloroplast morphology. β-LCY RNAi plants exhibited hypersensitivity to high light and lower non-photochemical quenching (NPQ) values under both normal and high light conditions. The stability of D1, a core subunit of photosystem II reaction center, was dramatically impaired β-LCY RNAi plants upon exposure to high light, suggesting severe photodamage of photosystem. Furthermore, reactive oxygen species (ROS) content was increased in β-LCY RNAi plants compared to WT plants under both normal and high light conditions. These results indicate that β-LCY is crucial for NPQ, D1 stability and ROS homeostasis, thus protecting photosystem from photooxidative damage. These findings contribute to our understanding on the function of plant β-LCY and its potential implications for plant growth, development, and photoprotection.

Identification, charectrization and genetic transformation of lignin and pectin polysaccharides through CRISPR/Cas9 in Nicotiana tobacum.

CRISPR/Cas9 system has been successfully implemented in animals and plants is a second-generation genome editing tool. We are able to optimize a Cas9 system to edited Ntab06050 and Ntab0857410 genes in HD and K326 tobacco cultivars respectively. The gene Ntab06050 is related to lignin synthesis while the gene Ntab0857410 belongs to pectin synthesis by utilizing Agrobacterium-mediated leaf disc method. We have constructed total eight different constructs for the lignin related gene family CCoAMT, out of which three constructs have been selected from Ntab0184090, two constructs from Ntab0392460 while one construct from each Ntab0540120, Ntab0857410 and Ntab0135940 gene. To study the Cas9 system in pectin related genes, total five constructs have been utilized under Cas9 system and multiple target sites were selected by identifying PAM sequences. Out of which three constructs were targeted from NtabGAE1and NtabGAE6 homologous while two were targeted from NtabGAUT4 homologous. Where as, UDP-D-glucuronate 4-epimerase gene family is a Golgi localized, might have a role in the interconvertion of UDP-D-GlcA and UDP-D-GalA in pectin synthesis. We have succeeded in the mutation of pectin related NtabGAUT4 and lignin related NtabCCoAMT genes with 6.2% and 9.4% mutation frequency.

The role of the cytokinin biosynthesis pathways in the rate of tobacco leaf senescence

The regulation of leaf senescence depends on endogenous and exogenous factors, among them phytohormones like cytokinins (CKs). CKs are key players regulating the senescing process, as their endogenous concentration is linked to the onset and rate of senescence progression. Thus, this study aimed to identify the relationship between the activity of endogenous CKs biosynthesis pathways - the cytosolic mevalonate (MVA) and the plastid methyl-erythritol phosphate (MEP) and the rate of leaf senescence. To this end, three distinct tobacco (Nicotiana tabacum L.) cultivars – Xanthi, Golden Virginia and Monte Calme Yellow were analysed. The study involved treatment with exogenous CK – benzyladenine – and two different CK synthesis inhibitors: lovastatin and clomazone. The progression of senescence was induced by light deprivation and monitored with chlorophyll level (SPAD), photosynthetic activity (PAM) and changes in the Rubisco protein profile (SDS-PAGE). Analyses showed that the Xanthi cultivar was characterized by delayed onset of senescence and stay-green phenotype, while Golden Virginia, and particularly Monte Calme Yellow showed rapid leaf senescence. The studies provided valuable information regarding the role of MEP and MVA pathway of CK synthesis in the regulation of tobacco leaf senescence.

Growth, yield and quality variability of flue-cured tobacco in response to soil and climatic factors in Northern Vietnam

Flue-cured tobacco is an annual industrial crop cultivated widely in more than 120 countries, with yield and quality varying between cultivars and with ecological factors. This study investigated variability of four flue-cured tobacco cultivars in response to 11 environmental factors, including soil properties and climatic conditions. The cultivars were grown in four locations in the northern upland region of Vietnam over two years. Growth characteristics, yield, leaf quality, and stability statistics were assessed. Variability and significant effects of cultivar, location, year, and their interactions on plant performance were observed. Positive associations between plant growth characteristics and yield were identified. Fresh and dry yields were significantly positively correlated with soil available nitrogen phosphorus, and temperature. All leaf quality characteristics were significantly correlated with soil available nitrogen, phosphorus, and sunshine hours. Canonical correlation analysis showed that yield was strongly influenced by four ecological factors. Leaf nitrogen and chlorine contents were influenced by seven and nine ecological factors, respectively. Low stability statistics, low average sum of ranks, moderate to high yield and leaf quality indicated that D65 was the most stable cultivar across locations and years. GL2 and GL7 were desired for their stability in leaf quality and yield, respectively.

Integrated transcriptomics and metabolomics provides insights into the Nicotiana tabacum response to heat stress.

Heat stress is a prevalent factor that significantly damages crops, especially with the ongoing global warming and increasing frequency of extreme weather events. Tobacco is particularly sensitive to temperature fluctuations, experiencing reduced yield and quality under high temperatures. However, the underlying molecular mechanisms of heat resistance in tobacco remain poorly understood. This study comprehensively analyzed biochemical, transcriptomic, and metabolomic responses to heat stress on the root and shoot of the tobacco cultivar K326 compared to control conditions. Heat stress significantly increased the activities of antioxidant enzymes (CAT, POD, and SOD) and levels of osmotic mediators (soluble sugars, sucrose, and proline) in the shoot. Furthermore, transcriptome analysis identified 13,176 differentially expressed genes (DEGs) in the root (6,129 up-regulated and 7,047 down-regulated) and 12,283 DEGs (6,621 up-regulated and 5,662 down-regulated) in the shoot. The root had 24 enriched KEGG pathways, including phenylpropanoid metabolism, while the shoot had 32 significant pathways, such as galactose metabolism and MAPK signaling. The metabolomic data identified 647 metabolites in the root and 932 in the shoot, with carbohydrates and amino acids being the main categories. The root had 116 differentially abundant metabolites (DAMs) (107 up-regulated and 9 down-regulated), and the shoot contained 256 DAMs (251 up-regulated and 5 down-regulated). Joint transcriptome and metabolome analysis showed that galactose metabolism and starch and sucrose metabolism were co-enriched in both tissues. In contrast, amino sugar and nucleotide sugar metabolism was enriched in the root, and purine metabolism in the shoot. The purine metabolic pathway in the shoot can modulate the expression of MYB transcription factors by influencing ABA synthesis and signaling, thereby controlling the accumulation of HSPs, raffinose, sucrose, and trehalose to enhance heat tolerance. Furthermore, NtMYB78, an MYB transcription factor, enhances tolerance for heat stress in tobacco. This research offers a foundational framework for investigating and implementing heat-resistant genes and metabolic pathways in the root and shoot of tobacco seedlings.

Overexpression of RpKTI2 from Robinia pseudoacacia Affects the Photosynthetic Physiology and Endogenous Hormones of Tobacco.

Kunitz trypsin inhibitor genes play important roles in stress resistance. In this study, we investigated RpKTI2 cloned from Robinia pseudoacacia and its effect on tobacco. RpKTI2 was introduced into the tobacco cultivar NC89 using Agrobacterium-mediated transformation. Six RpKTI2-overexpressing lines were obtained. Transgenic and wild-type tobacco plants were then compared for photosynthetic characteristics and endogenous hormone levels. Transgenic tobacco showed minor changes in chlorophyll content, fluorescence, and photosynthetic functions. However, the maximum photochemical efficiency (Fv/Fm) increased significantly while intercellular CO2 concentration (Ci) decreased significantly. Stomatal size and hormone content (indole-3-acetic acid, zeatin riboside, gibberellin, and indole-3-propionic acid) were reduced, while brassinosteroid content increased. Random forest regression revealed that RpKTI2 overexpression had the biggest impact on carotenoid content, initial fluorescence, Ci, stomatal area, and indole-3-acetic acid. Overall, RpKTI2 overexpression minimally affected chlorophyll synthesis and photosynthetic system characteristics but influenced stomatal development and likely enhanced the antioxidant capacity of tobacco. These findings provide a basis for future in-depth research on RpKTI2.

Deciphering the impact of nitrogen morphologies distribution on nitrogen and biomass accumulation in tobacco plants.

Nitrogen (N) distribution in plants is intricately linked to key physiological functions, including respiration, photosynthesis, structural development, and nitrogen storage. However, the specific effects of different N morphologies on N accumulation and plant growth are poorly understood. Our research specifically focused on determining how different N morphologies affect N absorption and biomass accumulation. This study elucidated the impact of different application rates (CK: 0 g N/plant; T1: 4 g N/plant; T2: 8 g N/plant) of N fertilizer on N and biomass accumulation in tobacco cultivars Hongda and K326 at different growth stages. Our findings emphasize the critical role of N distribution in various plant parts, including leaves, stems, and roots, in determining the complex mechanisms of N and biomass accumulation in tobacco. We found that in relation to total N, a greater ratio of water-soluble N (N w) in leaves facilitated N accumulation in leaves. In contrast, an increased ratio of SDS (detergent)-insoluble N (N in-SDS) in leaves and non-protein N (N np) in roots hindered this increase. Additionally, our results indicate that a greater proportion of N np in leaves has a negative impact on biomass accumulation in leaves. Furthermore, elevated levels of N in-SDS, N w, and N np in roots, and N np in leaves adversely affected biomass accumulation in tobacco leaves. The Hongda cultivar exhibited greater biomass and N accumulation abilities as compared to K326. Our findings highlight the significant role of distribution of N morphologies on plant growth, as well as N and biomass accumulation in tobacco plants. Understanding N distribution allows farmers to optimize N application, minimizing environmental losses and maximizing yield for specific cultivars. These insights advance sustainable agriculture by promoting efficient resource use and reducing environmental impact.

Aphelenchoides besseyi Parasitizing Tobacco Cultivars in Brazil.

Foliar nematodes (Aphelenchoides spp.) are known to be parasites of tobacco in restricted areas, but symptoms caused by A. besseyi in tobacco are not well characterized, despite the great importance of this nematode worldwide. This study aimed to evaluate the host reaction of four Nicotiana tabacum cultivars (Comum, Xanthi, Samsun, and TNN) and N. benthamiana cultivar Comum to A. besseyi and to characterize the symptoms and the parasitism of this nematode. Two experiments were conducted under greenhouse conditions with controlled humidity and temperature, in which the plants were inoculated with 600 A. besseyi. At 30 days after inoculation (DAI), nematodes present in the soil, roots, and shoot parts were extracted, and roots and shoot tissues were stained with acid fuchsin. A high number of A. besseyi was obtained per gram of shoot tissues (125 to 2,169 nematodes), and severe symptoms were observed in leaves and inflorescences of all cultivars. The symptoms included foliar distortion and deformation, necrotic spots delimited by the veins, flower abortion, and poor development of plants. In addition, A. besseyi was observed to penetrate tobacco roots at 30 DAI, and nematodes were also observed in the foliar mesophyll, inflorescences, and stems, a parasitism that has not been previously reported in tobacco plants. The disease caused by A. besseyi in tobacco could be a concern for growers in southern and northeastern Brazil because this nematode can cause severe damage to the marketable leaves of tobacco, reducing its commercial value.

Overexpression of SQUAMOSA promoter binding protein-like 4a (NtSPL4a) alleviates Cd toxicity in Nicotiana tabacum

Squamosa Promoter Binding Protein-Like (SPL) plays a crucial role in regulating plant development and combating stress, yet its mechanism in regulating resistance to Cd toxicity remains unclear. In this study, we cloned a nuclear-localized transcription factor, NtSPL4a, from the tobacco cultivar TN90. Transient co-expression results showed that miR156 significantly reduced the expression of NtSPL4a by binding to the 3′-UTR of its transcript. We obtained transgenic tobacco overexpressing NtSPL4a (including the 3′-UTR) and NtSPL4aΔ (lacking the 3′-UTR) through Agrobacterium-mediated genetic transformation. Compared to the wild type (WT), overexpression of NtSPL4a/NtSPL4aΔ shortened the flowering time and exhibited a more developed root system. The transgenic tobacco showed significantly reduced Cd content, being 85.1% (OE-NtSPL4a) and 46.7% (OE-NtSPL4aΔ) of WT, respectively. Moreover, the upregulation of NtSPL4a affected the mineral nutrient homeostasis in transgenic tobacco. Additionally, overexpression of NtSPL4a/NtSPL4aΔ effectively alleviated leaf chlorosis and oxidative stress induced by Cd toxicity. One possible reason is that the overexpression of NtSPL4a/NtSPL4aΔ can effectively promote the accumulation of non-enzymatic antioxidants. A comparative transcriptomic analysis was performed between transgenic tobacco and WT to further unravel the global impacts brought by NtSPL4a. The tobacco overexpressing NtSPL4a had 183 differentially expressed genes (77 upregulated, 106 downregulated), while the tobacco overexpressing NtSPL4aΔ had 594 differentially expressed genes (244 upregulated, 350 downregulated) compared to WT. These differentially expressed genes mainly included transcription factors, metal transport proteins, flavonoid biosynthesis pathway genes, and plant stress-related genes. Our study provides new insights into the role of the transcript factor SPL in regulating Cd tolerance.

An Interspecies Recombinant Sida Yellow Mosaic China Virus Isolate and Betasatellite Cause a Leaf Curl Disease in Tobacco in Hainan, China.

Tobacco (Nicotiana tabacum) is an herbaceous crop. Cigar tobacco, a group of tobacco cultivars, has recently been planted in a few provinces in China. Since its introduction, symptoms such as leaf curling and vein thickening have appeared. Here we report a begomovirus, Sida yellow mosaic China virus-Hainan isolate (designated SiYMCNV-HN), associated with the betasatellite (designated SiYMCNB-HN) as the causal agent of a leaf curl disease in cigar tobacco (N. tabacum cv. Haiyan101) in Hainan Province, China. Phylogenetic and recombination analyses indicate that SiYMCNV-HN is an interspecies recombinant with a SiYMCNV isolate as the major parent and a Sida yellow vein Vietnam virus isolate as the minor parent. Full-length infectious clones of SiYMCNV-HN and SiYMCNB-HN were generated, which were highly infectious and induced high pathogenicity through agroinfiltration in Nicotiana benthamiana and N. tabacum. This newly reported recombinant begomovirus poses potential threats to tobacco plantations in the region.

Quasi-targeted metabolomics revealed isoliquiritigenin and lauric acid associated with resistance to tobacco black shank

ABSTRACT Tobacco black shank (TBS), caused by Phytophthora nicotianae, is a severe disease. Plant root exudates play a crucial role in mediating plant-pathogen interactions in the rhizosphere. However, the specific interaction between key secondary metabolites present in root exudates and the mechanisms of disease resistance remains poorly understood. This study conducted a comprehensive comparison via quasi-targeted metabolomic analysis on the root exudate metabolites from the tobacco cultivar Yunyan87 and K326, both before and after inoculation with P. nicotianae. The results showed that the root exudate metabolites changed after P. nicotianae inoculation, and the root exudate metabolites of different tobacco cultivar was significantly different. Furthermore, homovanillic acid, lauric acid, and isoliquiritigenin were identified as potential key compounds for TBS resistance based on their impact on the mycelium growth of the pathogens. The pot experiment showed that isoliquiritigenin reduced the incidence by 55.2%, while lauric acid reduced it by 45.8%. This suggests that isoliquiritigenin and lauric acid have potential applications in the management of TBS. In summary, this study revealed the possible resistance mechanisms of differential metabolites in resistance of commercial tobacco cultivar, and for the first time discovered the inhibitory effects of isoliquiritigenin and homovanillic acid on P. nictianae, and attempt to use plants secondary metabolites of for plant protection.

Identification and Biological Characterization of a New Cyst Nematode, Heterodera glycines sbsp.n. tabacum, Parasitizing Tobacco in China.

In an investigation of diseases from plant-parasitizing nematodes in Henan Province, a cyst nematode was found on tobacco roots and in rhizosphere soil. We identified this strain as a new cyst nematode subspecies, Heterodera glycines sbsp.n. tabacum. The cysts and second-stage juveniles (J2s) parasitizing Henan tobacco were larger than those of H. glycines. A single 345-bp fragment was amplified from H. glycines sbsp.n. tabacum, whereas the 345- and 181-bp fragments were amplified from the soybean cyst nematode. Thus, H. glycines sbsp.n. tabacum was distinct from H. glycines. There were base transversions at 504 sites and base transitions at 560, 858, 920, and 921 sites in the rDNA-ITS sequences of H. glycines sbsp.n. tabacum compared with H. glycines, and there were base transitions at 41, 275, 278, and 380 sites in the mtDNA-COI sequences. In the phylogenetic tree based on the rDNA-ITS and mtDNA-COI regions, H. glycines sbsp.n. tabacum was clustered on a single branch. Based on the randomly amplified polymorphic DNA (RAPD) technique, sequence characterized amplified region (SCAR)-PCR primers were designed. A single 1,113-bp fragment was amplified by specific primers (HtF1/HtR1) from H. glycines sbsp.n. tabacum, while no fragments were obtained from H. glycines. The H. glycines sbsp.n. tabacum can infect soybean plants but cannot complete its life cycle on soybean. Eleven tested tobacco cultivars were infected, with an average reproduction factor (Rf) of 9.74 and a maximum of 64.2 in 'K326'. The cumulative egg hatching rate of H. glycines sbsp.n. tabacum in the presence of tobacco root exudates was 42.6% at 32 days posthatching, which was significantly greater than that in the presence of soybean root exudates (30.3%) or sterile water (33.1%). In summary, the cyst nematode population parasitizing Henan tobacco was identified as a new subspecies, H. glycines sbsp.n. tabacum.

Development of a stable attenuated double-mutant of tobacco mosaic virus for cross-protection

Tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum) are two major economic crops in China. Tobacco mosaic virus (TMV; genus Tobamovirus) is the most prevalent virus infecting both crops. Currently, some widely cultivated tobacco and tomato cultivars are susceptible to TMV and there is no effective strategy to control this virus. Cross-protection can be a safe and environmentally friendly strategy to prevent viral diseases. However, stable attenuated TMV mutants are scarce. In this study, we found that the substitutions in the replicase p126, arginine at position 196 (R196) with aspartic acid (D), glutamic acid at position 614 (E614) with glycine (G), serine at position 643 (S643) with phenylalanine (F), or D at position 730 (D730) with S, significantly reduced the virulence and replication of TMV. However, only the mutation of S643 to F reduced the RNA silencing suppression activity of TMV p126. A double-mutant TMV-E614G-S643F induced no visible symptom and was genetically stable through six successive passages in tobacco plants. Furthermore, our results showed that TMV-E614G-S643F double-mutant could provide effective protection against the wild-type TMV infection in tobacco and tomato plants. This study reports a promising mild mutant for cross-protection to control TMV in tobacco and tomato plants.

Comparative field adaptability of some potential cultivars of cigar tobacco in Hanzhong, Shaanxi province: growth and gas exchange attributes

Comparative field adaptability of some potential cultivars of cigar tobacco in Hanzhong, Shaanxi province: growth and gas exchange attributes

Ralstonia solanacearum differentially modulates soil physicochemical properties and rhizospheric bacteriome of resistant and susceptible tobacco cultivars

Ralstonia solanacearum is a devastating soilborne pathogen which poses significant yield and economic losses to tobacco production globally. The impact of R. solanacearum on rhizosphere bacteriome and soil physicochemical characteristics in resistant and susceptible tobacco cultivars is poorly understood. This study aims to determine the effect of R. solanacearum on soil physicochemical parameters and rhizosphere bacteriome of resistant (K326) and susceptible (Hongda) tobacco cultivars at various growth stages. Results demonstrated that the contents of available potassium and phosphorus, as well as soil pH were significantly increased in K326 soils (CK and T2) compared with Hongda (T1) after 21, 42, and 63 days post-inoculation (dpi) of R. solanacearum except for available nitrogen which showed an opposite trend. The qPCR results showed a significant decrease in R. solanacearum population in rhizosphere of K326 (T2) compared to the Hongda (T1) at 21 and 63 dpi than that after 42 dpi. The rhizosphere bacteriome analysis through 16S rRNA amplicon sequencing revealed that rhizosphere bacterial community composition was significantly different between two tobacco cultivars (Hongda and K326) and this effect was more prominent after 63 dpi (93 days after post-transplantation), suggesting that each cultivar recruits a unique set of bacterial communities. There was no obvious difference observed in the rhizosphere bacteriome of CK (K326) and T2 (K326), which might be attributed to the same genetic makeup and inherent resistance of K326 to bacterial wilt infection. Analysis of co-occurrence networks revealed that the microbial network in T1 (Hongda) was more complex than those in T2 (K326) and CK (K326), while the networks in CK and T2 were almost identical. The present research highlights the time-course relationship between environmental factors and rhizosphere bacteriome of tobacco cultivars showing different levels of resistance against R. solanacearum. Conclusively, studying the plant-soil-microbe interaction system in susceptible and resistant tobacco cultivars may enable us to develop effective integrated disease control plans for the healthy production of tobacco crops.

Effects of biochar treatments on the elemental composition of tobacco (Nicotiana tabacum L.) leaves based on the priming period

This study determined the effects of different doses of biochars (B) on Virginia tobacco (Nicotiana tabacum L.) cultivar, on first and second harvest dependent change in plant nutrients (N, P, K, Ca, Mg, Cl, Zn, Fe, Mn, Cu, and B), leaf color parameters (L*, a*, and b*), chlorophyll value (SPAD), electrolyte leakage (EL), crude ash, number of leaves, and plant height. Pot experiments were conducted with biochar treatments of 10 tons ha−1 (B1), 20 tons ha−1 (B2), 40 tons ha−1 (B3), and 80 tons ha−1 (B4). Tobacco leaf macroelement (N, P, K, Ca, and Mg) levels increased with increasing biochar doses. The highest values were obtained for B4 treatments (80 tons ha−1) and the lowest for control (B0) treatments. Microelements (Fe, Zn, Mn, and B) exhibited a non linear change, while Cl and Cu exhibited a linear change. Color parameters (L*, a*, and b*) for the first and second priming showed the highest L* and b* values for B2 and B3 treatments, respectively, and the highest a* values for the B2 treatment. Leaf SPAD values increased with increasing biochar doses; further, the obtained SPAD values were ordered as B4 > B3 > B2 > B1 > B0. Leaf electrolyte leakage values were 25.90 %–37.25 % in the first priming and 26.90 %–40.59 % in the second priming. For both the primings, the highest crude ash values (21.94 % and 19.05 %) were observed for the B4 treatments, whereas the lowest values (17.89 % and 17.01 %) were observed for the B0 treatments. the tallest plant height (121.9 cm) and the highest number of leaves (45.3) were determined in B4 applications. Overall, considering the nutrition and quality of tobacco, B2 application is recommended.

Effects of Cadmium Toxicity on Chemical Contents and Leaf Yield of Three Sun-Cured Tobacco Varieties

ABSTRACT Tobacco plants grown in cadmium (Cd)-contaminated soils accumulate high concentration of Cd in leaves and the Cd in leaves can readily be transferred to the human body through smoking of cigarettes. This study was conducted to investigate the effects of four soil Cd concentrations (0, 5, 10, and 20 mg kg−1) on leaf Cd, zinc (Zn), and nitrogen (N) concentrations and nicotine, reducing sugar and phenolic compounds of sun-cured Canik 190/5, Xanthi 81, and Birlik 124 varieties grown in a semi-arid condition of Turkey. Increasing doses of Cd did not significantly decrease leaf yield of tobacco cultivars, while significant (p < .05) increase and decrease were recorded in leaf Cd and Zn concentrations, respectively. The highest leaf Cd concentration was accumulated in Birlik 124 (60.5 mg kg−1), and the lowest in Canik 190/5 (55.35 mg kg−1) variety. Nicotine concentration of Xanthi 81 and Birlik 124 cultivars in the highest Cd dose (Cd20) increased compared to the nicotine in control application, while nicotine concentration of Canik 190/5 cultivars decreased in Cd20 dose. The results revealed that all three tobacco varieties were able to accumulate Cd in their leaves under high soil Cd levels. The effects of Cd on N uptake and nicotine, reducing sugar and phenolic compounds were different among the tobacco varieties. The result may be associated with differences in responses of tobacco cultivars to soil Cd concentration.

First report of Tobacco Fusarium Root Rot Caused by Fusarium meridionale in China.

Tobacco (Nicotiana tabacum L.) was an important economic crop in China. A survey in Yunnan Province in the last several years showed that the incidence of tobacco root rot was 3 to 30%. In July 2021, root rot symptoms were observed with an average incidence of 5% on tobacco (cultivar Yunyan 87) in Dali (25.61° N, 100.27° E). Typical disease symptoms included plants stunted at early stages, brown-colored withering lower leaves and roots that became brown. Under high humidity conditions, symptoms of rot expanded in the roots, also the whole plant became wilted and stunted, and some plants ultimately died. Infected pieces of stem tissues and root were dissected and then sterilized with 2% NaOCl for 30 s, rinsed three times with sterile distilled water, and dried with sterilized filter paper. Three pieces were plated onto potato dextrose agar (PDA) for 3 days at 25°C with a 12-h light period. Colonies on PDA were characterized by white to pale yellow flocculent aerial mycelium, and a pink to red pigment in the agar. To induce sporulation, mycelium on PDA was transferred to carnation leaf agar (CLA) medium. After incubation for 7 days, a single spore was isolated from representative isolate 21DL16 for morphological and molecular analyses. Macroconidia observed on CLA were falcate, slightly curved, three to five septate, measured 33.1 to 53.7 × 3.2 to 4.6 μm (n=50), with a typical foot shaped basal cell. Morphological characteristics of the fungus were in agreement with the description of Fusarium graminearum (Leslie and Summerell 2006). For further identification, the internal transcribed spacer (ITS) region rDNA, translation elongation factor 1ɑ (EF-1α) and RNA polymerase II second largest subunit (RPB2) gene were amplified and sequenced using primers ITS1/ITS4 (White et al. 1990), EF1/EF2 (O'Donnell et al. 2015) and RPB2-5F/RPB2-7cR (Reeb et al. 2004), respectively. Although the ITS sequence (GenBank accession no. OM392025) cannot distinguish F. meridionale from F. graminearum, combined phylogenetic analysis of the sequence of TEF1 (ON062055) and RPB2 (ON211932) clearly showed that the pathogen is F. meridionale that the sequences were 100% similarity, 0.0e-value and 100% query coverage to F. meridionale. Pathogenicity studies were conducted on six-leaf-stage tobacco seedlings cultivar Yunyan 87. A conidial suspension (1×105 spores/mL) was poured over the roots of tobacco seedlings. Three seedlings were treated with sterile water that served as controls. All 10 seedlings were maintained at 25°C at 70% relative humidity. After 5 days, the lower leaves showed symptoms of wilting and the roots of all inoculated seedlings become discolored, that were similar with the original symptoms, whereas the control seedlings did not develop symptoms. The fungus reisolated from the inoculated seedlings was identical to F. meridionale using the EF-1α gene sequence. To date, Fusarium root rot on tobacco in China was caused by F. oxysporium (Chen 2013). However, to the best of our knowledge, this is the first report of F. meridionale causing root rot on tobacco in China. Identification of F. meridionale as a root rot agent might provide important insight for disease management practices on tobacco caused by Fusarium species.

Developing an efficient and visible prime editing system to restore tobacco 8-hydroxy-copalyl diphosphate gene for labdane diterpene Z-abienol biosynthesis.

Prime editing (PE) is a versatile CRISPR-Cas based precise genome-editing platform widely used to introduce a range of possible base conversions in various organisms. However, no PE systems have been shown to induce heritable mutations in tobacco, nor in any other dicot. In this study, we generated an efficient PE system in tobacco that not only introduced heritable mutations, but also enabled anthocyanin-based reporter selection of transgene-free T1 plants. This system was used to confer Z-abienol biosynthesis in the allotetraploid tobacco cultivar HHDJY by restoring a G>T conversion in the NtCPS2 gene. High levels of Z-abienol were detected in the leaves of homozygous T1 plants at two weeks after topping. This study describes an advance in PE systems and expands genome-editing toolbox in tobacco, even in dicots, for use in basic research and molecular breeding. And restoring biosynthesis of Z-abienol in tobacco might provide an efficient way to obtain Z-abienol in plants.