Tobacco Nicotiana Research Articles

Bio-organic substitution in tobacco (Nicotiana tabacum L) cultivation: Optimum strategy to lower carbon footprint and boost net ecosystem economic benefit

The partial substitution of organic manure for chemical nitrogen fertilizers, known as organic substitution, is widely regarded as a cleaner and more sustainable production strategy. However, few studies have quantified greenhouse gas emissions, product income and net ecosystem economic benefit (NEEB) using a life cycle assessment (LCA) approach, particularly for typical tobacco (Nicotiana tabacum L.) production. Here, we quantified the yield and quality of a typical tobacco production in Qujing, Yunnan, China, through field experiments and calculated its carbon footprint and NEEB using the LCA approach. Four organic substitution strategies were established with equal nitrogen inputs, including synthesized chemical fertilizer (SN), farmyard organic manure (NF), commercial organic manure (NC), and bio-organic (Trichoderma viride Pers.) manure (NT), each substituting 15% of synthesized nitrogen fertilizer. Compared to the SN strategy, the NT strategy significantly increased yield and income by 10.3% and 9.6%, respectively. In contrast, the NF strategy significantly reduced income, while the NC strategy showed no significant difference. Both the NC and NT strategies significantly reduced N2O cumulative emissions (by 15.9% and 8.0%, respectively), increased δSOC (by 38.4% and 15.0%, respectively), and decreased carbon footprint compared to the SN strategy. However, the NF strategy significantly increased the income-scaled carbon footprint, even though it also notably reduced N2O cumulative emissions (by 22.6%) and increased δSOC (by 7.9%). The NT strategy achieved a win-win scenario of low environmental risk and high economic returns of tobacco production with significantly increased NEEB (by 10.6%) compared to the SN strategy (37.60 × 103 CNY yr−1). This suggests that the bio-organic Trichoderma manure substituting 15% synthesized nitrogen fertilizer is the best organic substitution strategy for sustainable tobacco production.

Transcriptional Modulation of Plant Defense Genes by a Bipartite Begomovirus Promotes the Performance of Its Whitefly Vector

The majority of plant viruses rely on insect vectors for inter-plant transmission. Amid virus transmission, vector-borne viruses such as begomoviruses may significantly modulate host plants in various ways and, in turn, plant palatability to insect vectors. While many case studies on monopartite begomoviruses are available, bipartite begomoviruses are understudied. More importantly, detailed elucidation of the molecular mechanisms involved is limited. Here, we report the mechanisms by which an emerging bipartite begomovirus, the Sri Lankan cassava mosaic virus (SLCMV), modulates plant defenses against whitefly. SLCMV infection of tobacco (Nicotiana tabacum) plants significantly downregulated defenses against whitefly, as whitefly survival and fecundity increased significantly on virus-infected plants when compared to the controls. We then profiled SLCMV-induced transcriptomic changes in plants and identified a repertoire of differentially expressed genes (DEGs). GO enrichment analysis of DEGs demonstrated that the term defense response was significantly enriched. Functional analysis of DEGs associated with defense response revealed that four downregulated DEGs, including putative late blight resistance protein homolog R1B-17 (R1B-17), polygalacturonase inhibitor-like (PGI), serine/threonine protein kinase CDL1-like (CDL1), and Systemin B, directly contributed to plant defenses against whitefly. Taken together, our findings elucidate the role of novel plant factors involved in the modulation of plant defenses against whitefly by a bipartite begomovirus and shed new light on insect vector–virus–host plant tripartite interactions.

Genome-wide analysis of tobacco NtTOM1/TOM3 gene family and identification of NtTOM1a/ NtTOM3a response to tobacco mosaic virus

BackgroundTOBAMOVIRUS MULTIPLICATION 1 (TOM1) and its homolog TOBAMOVIRUS MULTIPLICATION 3 (TOM3) play a prominent role in the multiplication of tobacco mosaic virus (TMV) in higher plants. Although homologs of NtTOM1/TOM3 genes have been identified in several plant species, little is known about the characteristics and functions of NtTOM1/TOM3 at the genome-wide level in tobacco (Nicotiana tabacum L.).ResultsIn this study, we performed genome-wide identification and expression pattern analysis of the tobacco NtTOM1/TOM3 gene family. Twelve NtTOM1/TOM3 genes were identified and classified into four groups based on phylogenetic analysis. Sequence and conserved domain analyses showed that all these genes contained a specific DUF1084 domain. Expression pattern analysis showed that NtTOM1a, NtTOM1b, NtTOM1d, NtTOM3a, NtTOM3b, and NtTOM3d were induced by TMV at 1-, 3-, and 9 dpi, whereas the expression of other genes was not responsive to TMV at the early infection stage. TMV virion accumulation showed no obvious difference in either nttom1a or nttom3a mutants compared with the wild type. However, the virus propagation was significantly, but not completely, inhibited in the nttom1atom3a double mutant, indicating that other gene family members may function redundantly, such as NtTOM1b and NtTOM1d. In addition, overexpression of NtTOM1a or NtTOM3a also inhibited the TMV replication to some extent.ConclusionsThe present study performed genome-wide analysis of the NtTOM1/TOM3 gene family in tobacco, and identified NtTOM1a and NtTOM3a as important genes involved in TMV multiplication based on functional analysis. These results provide a theoretical basis for further improving TMV resistance in tobacco.

Identification and characterisation of 'No apical meristem; Arabidopsis transcription activation factor; Cup-shape cotyledon' (NAC) family transcription factors involved in sugar accumulation and abscisic acid signalling in grape (Vitis vinifera).

The 'No apical meristem; Arabidopsis transcription activation factor; Cup-shape cotyledon' (NAC) transcription factors are pivotal in plant development and stress response. Sucrose-non-fermenting-related protein kinase 1.2 (SnRK1) is a key enzyme in glucose metabolism and ABA signalling. In this study, we used grape (Vitis vinifera ) calli to explore NAC's roles in sugar and ABA pathways and its relationship with VvSnRK1.2 . We identified 19 VvNACs highly expressed at 90days after blooming, coinciding with grape maturity and high sugar accumulation, and 11 VvNACs randomly selected from 19 were demonstrated in response to sugar and ABA treatments. VvNAC26 showed significant response to sugar and ABA treatments, and its protein, as a nucleus protein, had transcriptional activation in yeast. We obtained the overexpression (OE-VvNAC26 ) and RNA-inhibition (RNAi-VvNAC26 ) of VvNAC26 in transgenic calli by Agrobacterium tumefaciens -mediated transformation. We found that VvNAC26 negatively influenced fructose content. Under sugar and ABA treatments, VvNAC26 negatively influenced the expression of most sugar-related genes, while positively influencing the expression of most ABA pathway-related genes. Dual-luciferase reporter experiments demonstrated that VvNAC26 significantly upregulates VvSnRK1.2 promoter expression in tobacco (Nicotiana benthamiana ) leaves, although this process in grape calli requires ABA. The levels of sugar content, sugar-related genes, and ABA-related genes fluctuated significantly in OE-VvNAC26 +RNAi-VvSnRK1.2 and OE-VvSnRK1.2 +RNAi-VvNAC26 transgenic calli. These findings indicated that VvNAC26 regulates sugar metabolism and ABA pathway, displaying synergistic interactions with VvSnRK1.2 .

Root endodermal suberization induced by nitrate stress regulate apoplastic pathway rather than nitrate uptake in tobacco (Nicotiana tabacum L.)

Nitrogen levels and distribution in the rhizosphere strongly regulate the root architecture. Nitrate is an essential nutrient and an important signaling molecule for plant growth and development. Hydroponic experiments were conducted to investigate the differences in endodermal suberization in tobacco (Nicotiana tabacum L.) roots at three nitrate levels. Nitrogen accumulation was detected in the roots, shoots, and xylem sap. Nitrate influx on the root surface was also measured using the non-invasive self-referencing microsensor technique (SRMT). RNA-Seq analysis was performed to identify the genes related to endodermal suberization, nitrate transport, and endogenous abscisic acid (ABA) biosynthesis. The results showed that root length, root-shoot ratio, nitrate influx on the root surface, and NiA and NRT2.4 genes were regulated to maintain the nitrogen nutrient supply in tobacco under low nitrate conditions. Low nitrate levels enhanced root endodermal suberization and hence reduced the apoplastic transport pathway, and genes from the KCS, FAR, PAS2, and CYP86 families were upregulated. The results of exogenous fluridone, an ABA biosynthesis inhibitor, indicated that suberization of the tobacco root endodermis had no relevance to radial nitrate transport and accumulation. However, ABA enhances suberization, relating to ABA biosynthesis genes in the CCD family and degradation gene ABA8ox1.

Membrane protein provision controls prothylakoid biogenesis in tobacco etioplasts.

The cytochrome b559 heterodimer is a conserved component of photosystem II whose physiological role in photosynthetic electron transfer is enigmatic. A particularly puzzling aspect of cytochrome b559 has been its presence in etiolated seedlings, where photosystem II is absent. Whether or not the cytochrome has a specific function in etioplasts is unknown. Here, we have attempted to address the function of cytochrome b559 by generating transplastomic tobacco (Nicotiana tabacum) plants that overexpress psbE and psbF, the plastid genes encoding the two cytochrome b559 apoproteins. We show that strong overaccumulation of the PsbE apoprotein can be achieved in etioplasts by suitable manipulations of the promoter and the translation signals, while the cytochrome b559 level is only moderately elevated. The surplus PsbE protein causes striking ultrastructural alterations in etioplasts; most notably, it causes a condensed prolamellar body and a massive proliferation of prothylakoids, with multiple membrane layers coiled into spiral-like structures. Analysis of plastid lipids revealed that increased PsbE biosynthesis strongly stimulated plastid lipid biosynthesis, suggesting that membrane protein abundance controls prothylakoid membrane biogenesis. Our data provide evidence for a structural role of PsbE in prolamellar body formation and prothylakoid biogenesis, and indicate that thylakoid membrane protein abundance regulates lipid biosynthesis in etioplasts.

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.

Attraction of Nicotiana benthamiana to Bemisia tabaci is related to a chemical signal in plant volatile, undecane.

The whitefly Bemisia tabaci is one of the world's foremost agricultural pests. Recently, we found that a wild relative of tobacco (Nicotiana benthamiana) demonstrates remarkable attractiveness and nearly 100% lethality towards whiteflies. Therefore, it can act as a dead-end trap crop for whitefly control in the field. However, the underlying mechanism of the significant attractiveness of N. benthamiana towards whiteflies is unclear. Binary-choice assays and olfactory experiments showed that compared to common tobacco (N. tabacum), the volatile of N. benthamiana has a greater attraction to whiteflies. Then we analyzed and compared volatiles from these two Nicotiana species by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). We identified 16 chemical compounds that are more abundant in N. benthamiana than in N. tabacum. Seven compounds were further tested with olfactometer assays and we found that, among them, undecane strongly attracted whiteflies. Further experiments revealed that even 0.005 μg mL-1 undecane is attractive to whiteflies. We also silenced the genes that may influence the biosynthesis of undecane and found the production of undecane decreased after silencing NbCER3, and that N. benthamiana plants with less undecane lost their attraction to whiteflies. In addition, we found that applying 0.005 μg mL-1 undecane on yellow sticky traps can increase the number of stuck insects on the traps by ≈40%. Undecane from the volatile of N. benthamiana is a critical chemical signal that attracts whiteflies and NbCER3 involved in the biosynthesis of undecane. Undecane may be used to improve the efficiency of yellow sticky traps for whitefly control. © 2024 Society of Chemical Industry.

Metabolic and Molecular Insights Into Nicotiana benthamiana Trichome Exudates: An Ammunition Depot for Plant Resistance Against Insect Pests.

Nicotiana benthamiana, a widely acknowledged laboratory model plant for molecular studies, exhibits lethality to certain insect pests and can serve as a dead-end trap plant for pest control in the field. However, the underlying mechanism of N. benthamiana'sresistance against insects remains unknown. Here, we elucidate that the lethal effect of N. benthamiana on the whitefly Bemisia tabaci arises from the toxic glandular trichome exudates. By comparing the metabolite profiles of trichome exudates, we found that 51 metabolites, including five O-acyl sugars (O-AS) with medium-chain acyl moieties, were highly accumulated in N. benthamiana. Silencing of two O-AS biosynthesis genes, branched-chain keto acid dehydrogenase (BCKD) and Isopropyl malate synthase-C (IPMS-C), significantly reduced the O-AS levels in N. benthamiana and its resistance against whiteflies. Additionally, we demonstrated that the higher expression levels of BCKD and IPMS-C in the trichomes of N. benthamiana contribute to O-AS synthesis and consequently enhance whitefly resistance. Furthermore, overexpression of NbBCKD and NbIPMS-C genes in the cultivated tobacco Nicotiana tabacum enhanced its resistance to whiteflies. Our study revealed the metabolic and molecular mechanisms underlying the lethal effect of N. benthamiana on whiteflies and presents a promising avenue for improving whitefly resistance.

A conserved protein family in mirid bug Riptortus pedestris plays dual roles in regulating plant immunity.

The mirid bug (Riptortus pedestris), a major soybean pest, migrates into soybean fields during the pod filling stage and causes staygreen syndrome, which leads to substantial yield losses. The mechanism by which R. pedestris elicits soybean (Glycine max) defenses and counter-defenses remains largely unexplored. In this study, we characterized a protein family from R. pedestris, designated Riptortus pedestris HAMP 1 (RPH1) and its putative paralogs (RPH1L1, 2, 3, 4, and 5), whose members exhibit dual roles in triggering and inhibiting plant immunity. RPH1 and RPH1L1 function as herbivore-associated molecular patterns (HAMPs), activating pattern-triggered immunity (PTI) in tobacco (Nicotiana benthamiana) and G. max. Furthermore, RPH1 stimulates jasmonic acid and ethylene biosynthesis in G. max, thereby enhancing its resistance to R. pedestris feeding. Additionally, RPH1 homologs are universally conserved across various herbivorous species, with many homologs also acting as HAMPs that trigger plant immunity. Interestingly, the remaining RPH1 putative paralogs (RPH1L2-5) serve as effectors that counteract RPH1-induced PTI, likely by disrupting the extracellular perception of RPH1. This research uncovers a HAMP whose homologs are conserved in both chewing and piercing-sucking insects. Moreover, it unveils an extracellular evasion mechanism utilized by herbivores to circumvent plant immunity using functionally differentiated paralogs.

Cloning and Functional Study of AmGDSL1 in Agropyron mongolicum.

Agropyron mongolicum Keng is a diploid perennial grass of triticeae in gramineae. It has strong drought resistance and developed roots that can effectively fix the soil and prevent soil erosion. GDSL lipase or esterases/lipase has a variety of functions, mainly focusing on plant abiotic stress response. In this study, a GDSL gene from A. mongolicum, designated as AmGDSL1, was successfully cloned and isolated. The subcellular localization of the AmGDSL1 gene (pCAMBIA1302-AmGDSL1-EGFP) results showed that the AmGDSL1 protein of A. mongolicum was only localized in the cytoplasm. When transferred into tobacco (Nicotiana benthamiana), the heterologous expression of AmGDSL1 led to enhanced drought tolerance. Under drought stress, AmGDSL1 overexpressing plants showed fewer wilting leaves, longer roots, and larger root surface area. These overexpression lines possessed higher superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and proline (PRO) activities. At the same time, the malondialdehyde (MDA) content was lower than that in wild-type (WT) tobacco. These findings shed light on the molecular mechanisms involved in the GDSL gene's role in drought resistance, contributing to the discovery and utilization of drought-resistant genes in A. mongolicum for enhancing crop drought resistance.

Influence of Weather Conditions and the Aphid Population on the Potato Virus Y Infection of Tobacco in the Field

Potato virus Y (PVY) is a major tobacco (Nicotiana tabacum L.) pathogen that causes severe crop losses. We studied the influence of meteorological factors and a population of twelve aphid species on the development of PVY in field-grown tobacco from 1996 to 2010 in Poland. Three PVY-susceptible tobacco varieties were used in the study. The mean virus incidence ranged from 18% in 2010 to almost 99% in 1996, 2004, and 2009. For determining the relationship between tobacco plant infection and meteorological conditions and aphid populations, logistic regression analysis was used. It was found that the probability of PVY infection is significantly dependent on the average air temperature, relative humidity, number of days with an average temperature of at least 25 °C, and the abundance of Aphis fabae and Brachycaudus helichrysi. The probability of infection of tobacco plants with potato virus Y decreased with increasing air temperature and relative humidity. In addition, with each subsequent day with a temperature of at least 25 °C, the risk of infection decreased by 24%. Furthermore, it was often observed that high populations of Aphis fabae and Brachycaudus helichrysi were associated with a high incidence of virus infection in tobacco plants.

Identification and Functional Studies on the Role of PlSPL14 in Herbaceous Peony Stem Development.

Stem strength plays a crucial role in the growth and development of plants, as well as in their flowering and fruiting. It not only impacts the lodging resistance of crops, but also influences the ornamental value of ornamental plants. Stem development is closely linked to stem strength; however, the roles of the SPL transcription factors in the stem development of herbaceous peony (Paeonia lactiflora Pall.) are not yet fully elucidated. In this study, we obtained and cloned the full-length sequence of PlSPL14, encoding 1085 amino acids. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression level of PlSPL14 gradually increased with the stem development of P. lactiflora and was significantly expressed in vascular bundles. Subsequently, utilizing the techniques of virus-induced gene silencing (VIGS) and heterologous overexpression in tobacco (Nicotiana tabacum L.), it was determined that PlSPL14-silenced P. lactiflora had a thinner xylem thickness, a decreased stem diameter, and weakened stem strength, while PlSPL14-overexpressing tobacco resulted in a thicker xylem thickness, an increased stem diameter, and enhanced stem strength. Further screening of the interacting proteins of PlSPL14 using a yeast two-hybrid (Y2H) assay revealed an interactive relationship between PlSPL14 and PlSLR1 protein, which acts as a negative regulator of gibberellin (GA). Additionally, the expression level of PlSLR1 gradually decreased during the stem development of P. lactiflora. The above results suggest that PlSPL14 may play a positive regulatory role in stem development and act in the xylem, making it a potential candidate gene for enhancing stem straightness in plants.

Leaf grey spot caused by Arcopilus aureus on tobacco in China.

Tobacco (Nicotiana tabacum L.) is one of the most widely cultivated industrial crops worldwide. From April to July 2023, about 40% of tobacco seedlings in the greenhouse exhibited irregular taupe lesions in Zhengzhou, Henan Province, China. At an early stage of the lesion development, light grey spots with the diameter of 1-2 mm were observed, these spots gradually expanded and connected into large irregular lesions causing leaf wrinkling or withered. A total of 12 infected leaf tissues were sterilized with 75% ethanol for 45 s, rinsed three times in sterilized water and then plated on potato dextrose agar (PDA) medium for 10 days at 28°C in darkness. Seven fungal colonies that show the similar appearance were isolated and three of them (MB-1, MB-2 and MB-3) were used for subsequent identification. Colonies of these strains on PDA with loose mycelium and orange-red pigment on the underside, white aerial in the center and light yellow hyphae near the periphery, formed in the shape of a concentric ring pattern. Ascomata appeared from the 14th day, were black, spherical or ellipsoid with walls of textura angularis, and size was 53.8-101.1 μm × 50.3-104.3 μm (n=30). Terminal hairs were brown and straight, gradually tapering toward the tips. Asci clavate or fusiform, spore bearing part 16.2-29.2 × 7.3-11.4 μm (n=21), with 8 irregularly arranged ascospores, evanescent. Ascospores are brown at maturity, biapiculate, navicular or fusiform shapes with size of 8.7-12.8 μm × 4.8-6.9 μm (n=100), and more or less inaequilateral. Single spore strains derived from these strains exhibited the morphological features consistent with the original strains. The morphological characteristics of the fungus were consistent with the description of Arcopilus aureus (Chivers) X.W. Wang & Samson (= Chaetomium aureum Chivers) (Lee et al. 2019). Furthermore, the sequences of RPB2 region were amplified from these strains and the result sequences (GenBank accession no. OR513105-OR513108) all showed a 100.00% identity with A. aureus strain CBS 538.73 (GenBank accession no. KX976807.1). It was reported that the RPB2 gene was efficient in discriminating Arcopilus species (Tavares et al. 2022), thus a maximum likelihood (ML) phylogenetic tree based on the RPB2 gene sequences were constructed using MEGA 7.0 with 1000 replications of bootstrapping (Kumar et al. 2016), which revealed that these strains formed a well-supported clade with A. aureus strains of (CBS 153.52 and CBS538.73) (Wang et al. 2022). Pathogenicity analysis were performed on healthy flue-cured tobacco seedlings leaves (cv Y85) by using mycelial agar plugs (5 mm in diameter) and spore suspension (1×106 spores/mL), and the PDA plugs and sterile water were used for control group, respectively. Tobacco seedlings were incubated in a 25°C and 70% RH growth chamber. After seven days, the leaves showed obvious symptoms, with taupe lesions and yellow halos on the periphery, whereas no symptoms were found on the control leaves. The A. aureu was then reisolated from inoculated diseased leaves. Previously, A. aureus has been only reported to cause leaf black disease on Pseudostellaria heterophylla in China (Yuan et al. 2021). To our knowledge, this is the first reported of A. aureus causing tobacco leaf grey spot worldwide. Arcopilus aureus has been reported as a plant biocontrol fungus (Wang et al. 2013). However, due to the potential serious damage in tobacco seedlings caused by this fungus, the use of A. aureus as a plant biocontrol agent needs to be given more attention, and disease control measures of this pathogen should be developed.

NtZIP5A/B is involved in the regulation of Zn/Cu/Fe/Mn/Cd homeostasis in tobacco.

Plants grow in soils with varying concentrations of microelements, often in the presence of toxic metals e.g. Cd. To cope, they developed molecular mechanisms to regulate metal cross-homeostasis. Understanding underlying complex relationships is key to improving crop productivity. Recent research suggests that the Zn and Cd uptake protein NtZIP5A/B [Zinc-regulated, Iron-regulated transporter-like Proteins (ZIPs)] from tobacco (Nicotiana tabacum L. v. Xanthi) is involved in the regulation of a cross-talk between the two metals. Here, we support this conclusion by showing that RNAi-mediated silencing of NtZIP5A/B resulted in a reduction of Zn accumulation and that this effect was significantly enhanced by the presence of Cd. Our data also point to involvement of NtZIP5B in regulating a cross-talk between Cu, Fe, and Mn. Using yeast growth assays, Cu (but not Fe or Mn) was identified as a substrate for NtZIP5B. Furthermore, GUS-based analysis showed that the tissue-specific activity of the NtZIP5B promoter was different in each of the Zn-/Cu-/Fe-/Mn deficiencies applied with/without Cd. The results indicate that NtZIP5B is involved in maintaining multi-metal homeostasis under conditions of Zn, Cu, Fe, and Mn deficiency, and also in the presence of Cd. It was concluded that the protein regulates the delivery of Zn and Cu specifically to targeted different root cells depending on the Zn/Cu/Fe/Mn status. Importantly, in the presence of Cd, the activity of the NtZIP5B promoter is lost in meristematic cells and increased in mature root cortex cells, which can be considered a manifestation of a defense mechanism against its toxic effects.

Genome-wide identification and expression analysis of the U-box E3 ubiquitin ligase gene family related to bacterial wilt resistance in tobacco (Nicotiana tabacum L.) and eggplant (Solanum melongena L.).

The E3 enzyme in the UPS pathway is a crucial factor for inhibiting substrate specificity. In Solanaceae, the U-box E3 ubiquitin ligase has a complex relationship with plant growth and development, and plays a pivotal role in responding to various biotic and abiotic stresses. The analysis of the U-box gene family in Solanaceae and its expression profile under different stresses holds significant implications. A total of 116 tobacco NtU-boxs and 56 eggplant SmU-boxs were identified based on their respective genome sequences. Phylogenetic analysis of U-box genes in tobacco, eggplant, tomato, Arabidopsis, pepper, and potato revealed five distinct subgroups (I-V). Gene structure and protein motifs analysis found a high degree of conservation in both exon/intron organization and protein motifs among tobacco and eggplant U-box genes especially the members within the same subfamily. A total of 15 pairs of segmental duplication and 1 gene pair of tandem duplication were identified in tobacco based on the analysis of gene duplication events, while 10 pairs of segmental duplication in eggplant. It is speculated that segmental duplication events are the primary driver for the expansion of the U-box gene family in both tobacco and eggplant. The promoters of NtU-box and SmU-box genes contained cis-regulatory elements associated with cellular development, phytohormones, environment stress, and photoresponsive elements. Transcriptomic data analysis shows that the expression levels of the tobacco and eggplant U-box genes in different tissues and various abiotic stress conditions. Using cultivar Hongda of tobacco and cultivar Yanzhi of eggplant as materials, qRT-PCR analysis has revealed that 15 selected NtU-box genes and 8 SmU-box may play important roles in response to pathogen Ras invasion both in tobacco and eggplant.

Enhancing rooting tobacco (Nicotiana tabacum) plant by loaded indole-3-butyric acid in alginate/chitosan nanocapsule.

Recently, nanocarriers have been utilized for encapsulating and sustained release of agrochemicals specifically auxins. Due to their potential applications such as increased bioavailability and improved crop yield and nutritional quality. Herein, the efficacy of alginate/chitosan nanocapsules as a nanocarrier for the hormone indole-3-butyric acid (IBA) loading and its effect on rooting tobacco plants has been carried out in the present study. The average particle size of IBA-alginate/chitosan nanocapsules was measured by Dynamic light scattering analysis at 321 nm. Scanning electron microscope studies revealed the spherical shape of nanoparticles with an average size of 97 nm. The average particle size of IBA-alginate/chitosan nanocapsules was measured by Dynamic light scattering analysis at 321 nm. The characteristic peaks of IBA on alginate/chitosan nanocapsules were identified by Fourier transform infrared spectroscopic analysis. Also, high efficiency (35%) of IBA hormone loading was observed. The findings indicated that the concentration of 3 mgL-1 of IBA-alginate/chitosan nanocapsules has the highest efficiency in increasing the rooting in tobacco (Nicotiana tabacum) plants compared to other treatments. According to our results, we can introduce alginate/chitosan nanocapsules as an efficient nanocarrier in IBA hormone transfer applications and their use in agriculture.

ELONGATED HYPOCOTYL 5 regulates steroidal glycoalkaloid biosynthesis and fungal tolerance in tomato.

Tomato (Solanum lycopersicum L.) is rich in nutrients and has been an important target for enhancing the accumulation of various metabolites. Tomato also contains cholesterol-derived molecules, steroidal glycoalkaloids (SGAs), which contribute to pathogen defense but are toxic to humans and considered antinutritional compounds. Previous studies suggest the role of various transcription factors in SGA biosynthesis; however, the role of light and associated regulatory factors has not been studied in tomatoes. Here, we demonstrated that SGA biosynthesis is regulated by light through the ELONGATED HYPOCOTYL 5 homolog, SlHY5, by binding to light-responsive G-boxes present in the promoters of structural and regulatory genes. SlHY5 complemented Arabidopsis (Arabidopsis thaliana) and tobacco (Nicotiana tabacum) hy5 mutants at molecular, morphological, and biochemical levels. CRISPR/Cas9-based knockout tomato plants, SlHY5CR, showed downregulation of SGA and phenylpropanoid pathway genes, leading to a significant reduction in SGA (α-tomatine and dehydrotomatine) and flavonol contents, whereas plants overexpressing SlHY5 (SlHY5OX) showed the opposite effect. Enhanced SGA and flavonol levels in SlHY5OX lines provided tolerance against Alternaria solani fungus, while SlHY5CR lines were susceptible to the pathogen. This study advances our understanding of the HY5-dependent light-regulated biosynthesis of SGAs and flavonoids and their role in biotic stress in tomatoes.

In vitro assessment of Ethanolic extracts from specific herbs on controlling aquatic leech (Tyrannobdella rex) and land leech (Philobdella sp.) infestation in Mithun (Bos frontalis)

Leech infestation poses a significant threat to Mithun (Bos frontalis) in the north-eastern region of India, leading to various health issues and potential fatality. To address this pressing concern, we conducted a comprehensive research study aimed at assessing the efficacy of herbal plant extracts against aquatic leeches, particularly Tyrannobdella rex, and land leeches of the Philobdella sp. Our investigation involved the evaluation of six distinct plant extracts, with a focus on their ability to combat leech infestation. The results of our study revealed that among the various plant extracts tested, only the ethanolic extracts of soapnut (Sapindus mukorossi) and tobacco (Nicotiana tabacum) exhibited notable effectiveness in combating aquatic leeches. At a concentration of 5%, these extracts displayed significant lethality, with soapnut extract achieving a remarkable kill time of 6.0 ± 0.40 min, while tobacco extract showed a kill time of 31.5 ± 1.32 min. In the case of land leeches, tobacco extract proved to be highly efficient, with an average kill time of 1.5 ± 0.28 min at a 5% concentration. Soapnut extract also exhibited effectiveness against land leeches, albeit with a slightly longer kill time of 14.25 ± 1.10 min at the same concentration. Additionally, Litsea grass oil (Litsea cubeba) demonstrated promising efficacy against both aquatic and land leeches, suggesting its potential as a versatile leech control agent. These compelling findings have significant implications for the management and control of leech infestation among Mithun populations. By identifying and harnessing the leech-repelling properties of soapnut, tobacco, and Litsea grass oil, this research offers practical and environmentally friendly solutions for mitigating the adverse effects of leech infestation. Furthermore, the insights gained from this study pave the way for the development of innovative strategies to safeguard the health and well-being of Mithun in the future.

Benefits of Canavalia ensiformis, arbuscular mycorrhizal fungi, and mineral fertilizer management in tobacco production

Tobacco (Nicotiana tabacum L.) has long been vital to Cuban agriculture, with its products renowned for their quality. Cuban tobacco is grown in soils with a long history of continuous farming using traditional fertilization methods characterized by recommended doses of mineral fertilizers. This study aims to improve the nutrition resource strategy in tobacco cultivation to ensure high yields of superior-grade tobacco leaves with adequate quality and increase fertilization efficiency. With this goal, a field experiment evaluated the traditional method of fallow with alternatives of nutrient supply systems for the production of black tobacco in Ultic Paleustalf soils. The experiment utilized Canavalia ensiformis (Can) treated with a mycorrhizal inoculum (AMF) based on the Glomus cubense strain (INCAM-4) as a preceding green manure, combined with successive mineral fertilizations for tobacco during four growing seasons in a randomized block design with factorial arrangement. Canavalia presented a positive response to mycorrhizal inoculation, significantly increasing dry biomass production (87.34%, 129.96%), mycorrhizal colonization (26.90%, 103.66%), and spore production (26.79%, 52.52%) for Can and Can+AMF treatments respectively. A biplot analysis established a strong relationship between the biomass and mycorrhizal performance of Canavalia and the growth, yield, and mycorrhizal colonization of tobacco. The results indicate that inoculated Canavalia enhances mycorrhizal performance in successional tobacco, with Can+AMF significantly increasing mycorrhization of tobacco roots by (110.06%). Moreover, the combination of Can inoculate with AMF and 75% of the recommended mineral fertilization dose consistently produced the highest tobacco yields (42.06%), growth, and mycorrhizal activity across the four years while maintaining satisfactory combustibility. In this nutrition supply system, variations of the recommended fertilizer dose significantly decreased the percentage of mycorrhizal colonization. After four growing seasons using Can + AMF and Canavalia without inoculations, soil organic matter, and availability of exchangeable calcium, magnesium, and pH increased slightly without decreasing available phosphorus and potassium contents. Consequently, we conclude that Canavalia ensiformis, with an inoculum based on the Glomus cubense strain and 75% of the recommended dose of mineral fertilizers, provides an enhanced nutrition alternative system for black tobacco production.