Transcriptome is a collection of mRNA transcripts in a specific physiological state which has become one of the hotspots in biological research to evaluate the genes and networks in different kinds of plants. Transcriptome sequencing technology dates back to 1964 and has nearly 60 years of history. At present, the research mainly focuses on cultivating, breeding, molecular markers, and gene mining. With the increase in oil price, woody oil production can alleviate oil demand, but there are few review articles and molecular biology studies on the transcriptome of woody oil plants. In the past few decades, woody oil plants have made great progress in the development of transcriptome sequencing and bioinformatics. In this review, we reviewed the development history of sequencing technology and the research on the transcriptome of woody oil plants, mainly introducing the germplasm resources, molecular markers, and the application of important functional genes in woody oil plants. This paper not only provides ideas for mining functional genes of oil plants but also provides a reference for molecular breeding research of woody oil plants in the future.

Glandular trichomes (GTs) are one of the epidermal tissues of medicinal plants which function in the synthesis, storage, and secretion of secondary metabolites. The active ingredients of Chinese medicinal materials are mostly secondary metabolites of plants. Accordingly, it is of great research value to explore the quality of medicinal materials using the GTs of medicinal plants as the starting point. However, most of the current studies on GTs of medicinal plants are still at the simple morphological identification stage, and there are few studies on the compounds secreted by GTs and secondary metabolic processes. Here, we reviewed the literature, summarized the morphological types of medicinal plant GTs, separation and purification technology, analysis technology, and biological activities of secondary metabolites, and established a research approach to medicinal plant GTs. We hope to provide a reference for future research on GT inclusions and secondary metabolism.

Transgenic Arabidopsis plants expressing a potato D200 gene encoding a hypothetical protein were subjected to salinity stress and assessed for their tolerance. The D200 Arabidopsis lines exhibited increased chlorophyll content, improved stomatal conductance, less electrolyte leakage, lower accumulation of malondialdehyde (MDA), and a higher amount of proline compared to the wild type (WT) plants under salinity stress. The gene expression analysis revealed that D200 plants accumulated a significantly higher amount of mRNA transcripts of genes encoding three major antioxidant enzymes ascorbate peroxidase (APX), catalase (CAT), and superoxide dismutase (SOD). Chlorophyll a fluorescence kinetics analyses showed the D200 plants were more efficient in terms of primary photochemistry of photosystem II and performance indices. Furthermore, the quantum yields and efficiencies that represent the critical steps of photosynthetic light reactions were analyzed and it was found that D200 plants were photosynthetically more active than the WT plants under salt stress conditions. Overall, these findings suggest that the D200 gene is a potential candidate gene for developing stress-resilient crops in future.

Background: The Viola plants have important value for theoretical and applied research. Agrobacterium-mediated transient expression enables rapid target protein production in plants, yet this system remains poorly established in Viola. Aims: To establish an efficient Agrobacterium-mediated transient expression system in Viola. Methods: By employing two types of A. tumefaciens strains (EHA105 and LBA4404) and green fluorescent protein (GFP) as a reporter, we demonstrated the transient expression in the leaves of four Viola species, including Viola philippica, V. prionantha, V. tricolor, and V. dissecta. Results: GFP transient expression peaked at 4 dpi in all four Viola species. Agrobacterium at OD₆₀₀ = 0.3 yielded optimal expression, and strain EHA105 was superior to LBA4404; V. philippica showed the highest GFP accumulation. In the optimal system, exogenous indole-3-acetic acid (IAA) improves GFP transient expression with maximum promotion at 50 ng/mL, while methyl jasmonate (MeJA) has no facilitating effect. This system enables transient expression of FMDV VP1, whose expression is also markedly increased by 50 ng/mL IAA. Conclusions: This study successfully established a transient expression system for Viola and identified factors affecting Agrobacterium-mediated transient expression efficiency in this genus.

The link between the MtTdp1α (tyrosyl-DNA phosphodiesterase) gene, involved in the repair of DNA topoisomerase I mediated DNA damage, and the plant defense response has been investigated in MtTdp1α-depleted Medicago truncatula transgenic lines obtained by intron-spliced hairpin RNA approach, compared to the control line (CTRL, empty vector). Reduction of cuticle permeability highlighted by chlorophyll efflux assays positively correlated with the level of MtTdp1α gene silencing. The increased cuticle thickness was confirmed by transmission electron microscopy, which revealed an apparent expansion of the epicuticular waxes deposited on the outer surface. RNA-Seq analysis, carried out in the MtTdp1α-depleted plants, revealed the different expression of resistance (R) genes, PAMP (pathogen-associated-molecular pattern) triggered immunity (PTI) genes and transcription factors (TFs) involved in the regulation of the plant defense response.

Background: Idesia polycarpa var. vestita Diels is an oil-bearing woody plant of significant economic value. Aims: To accelerate the propagation of its elite germplasm, this study aimed to establish an efficient and stable in vitro rapid propagation system using one-year-old stem segments from elite plants. Methods: Young stem explants were disinfected with 75% ethanol for 40 s, followed by 0.1% HgCl₂ for 6 min. MS or 1/2 MS served as basal media, and various combinations of plant growth regulators were tested for axillary bud induction, proliferation, and rooting. Results: The optimal disinfection protocol yielded a 38.10% survival rate. The best axillary bud induction medium was MS + 1.5 mg/L 6-BA + 0.05 mg/L NAA + 0.08 mg/L TDZ, achieving 100% induction. For proliferation, MS + 1.5 mg/L 6-BA + 0.06 mg/L NAA + 0.01 mg/L TDZ (proliferation coefficient 5.81) was optimal. Rooting was best with 1/2 MS + 0.3 mg/L IAA + 0.1 mg/L NAA (100% rooting, 14 roots/plant). Conclusions: This study established a reliable micropropagation system for Idesia polycarpa var. vestita, providing technical support for the efficient production of high-quality seedlings.

NAC (NAM, ATAF1,2, and CUC2) transcription factors play an important role in the responses of plants to various environmental stresses. To investigate the function of SlNAC1, which was found to be a member of the ATAF subfamily in tomato (Solanum lycopersicum L.) plants under heat stress conditions, transgenic tomato plants were generated using an antisense technology. After a treatment at 40 °C for 48 h, in comparison with wild-type (WT) plants, the transgenic plants were severely wilted and exhibited a lower net photosynthetic rate and a maximal photochemical efficiency of photosystem II. Moreover, the transgenic plants displayed a higher ion leakage and malondialdehyde content and a lower proline content. The content of reactive oxygen species (superoxide anion radicals and hydrogen peroxide) were higher, and activities of ascorbate peroxidase and superoxide dismutase lower in the transgenic plants than in the WT plants. The transgenic plants also exhibited a lower accumulation of the transcripts of some heat shock protein genes (Hsp70, Hsp90, sHsp17.4, and sHsp17.6). All of these results suggest that the suppression of SlNAC1 could obviously reduce heat resistance in the tomato plants, and this indicates that SlNAC1 played an important role in the thermal tolerance of the tomato plants.

Proline rich proteins (PRPs), earlier famous as animal salivary proteins, have now been proven as indispensable plant proteins. They are highly rich in proline amino acid residues at the N-terminus whereas a characteristic eight cysteine motif is located at the C-terminus. The PRPs support a number of developmental processes from germination to plant death. Under normal environmental conditions, PRP genes express customarily in different plant parts depending on the specific function to be carried out. During abiotic stresses, PRP genes exhibit an uneven pattern of transcriptional regulation depending on the time and intensity of stress. Transgenic plants overexpressing PRP genes show an enhanced tolerance to abiotic stresses. This review focuses on contemporary functions of PRPs during stresses and proposes that PRPs are involved in the regulation of free cellular proline content during stress in a well synchronized manner.

Bacterial diseases of vegetable crops cause significant losses of yield and substantially decrease food quality. For sustainable development of agriculture, it is highly important to use the most effective strategies for the protection of vegetable crops from bacterial diseases which allows the creation of resistant cultivars and their introduction in regions with an increased risk of damage by phytopathogenic bacteria. This paper reviews the most widespread bacterial diseases of tomatoes, the mechanisms of interaction of plants with phytopathogenic bacteria, and the advantages of the biotechnological strategies over traditional and marker-associated breeding for creation of the resistant tomato cultivars. The current research progress on the use of biotechnological approaches such as cell selection, genetic engineering, genome editing, and gene silencing is summarized, with a special emphasis on the advantages and limitations of these methods.

Ascorbic acid (AsA) and glutathione (GSH) contribute to defense responses under abiotic stresses. The present study explored the ascorbate-glutathione cycle and ascorbate regeneration under high concentration (30 mM) of cadmium in the tea plant (Camellia sinensis L.). The tea leaves showed speckles and necrosis from the third day of Cd treatment. The content of superoxide anion (O₂^.-) and hydrogen peroxide (H₂O₂) in the leaves were significantly higher until the seventh day after Cd treatment. The content of O₂^.- and H₂O₂ were the highest on the fifth day (212.7 and 153.6 % of the control, respectively). The AsA content increased (86.9 %) on the first day after Cd treatment and decreased significantly in the subsequent days, while GSH showed a reverse trend. The enzymatic activity assays showed that dehydroascorbate reductase (DHAR) and glutathione reductase (GR) involved in AsA regeneration were downregulated considerably after Cd foliar application. In contrast, the activities of ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR) increased on the first day and then declined. Reverse-transcription quantitative PCR showed upregulation of glutathione synthetase (CsGSHS), γ-glutamylcysteine synthetase (Csγ-ECS), and CsMDHAR of the AsA regeneration pathway and downregulation of CsDHAR and CsGR. The expressions of GDP-L-galactose phosphorylase (CsGGP), L-galactose-1-phosphate phosphatase (CsGPP), and L-galactono- 1,4-lactone dehydrogenase (CsGaILDH) of the L-galactose pathway were also downregulated. The results indicated that AsA, which can respond to Cd stress of plants by increasing antioxidant ability, was consumed to scavenge ROS; moreover, Cd stress inhibited AsA synthesis and regeneration, which made that tea plants suffering severe damage.

Two ginseng species Panax vietnamensis and Panax stipuleanatus are precious medicinal plants restricted in several Vietnam provinces. They are very limited and endangered due to degraded habitats and over-harvesting. To preserve these two species, we used eight nuclear microsatellite markers to investigate genetic variability from the nine populations with 246 individuals for these two ginseng species. Our findings showed a moderate genetic heterozygosity in two species, P. vietnamensis (H_E = 0.386) and P. stipuleanatus (H_E = 0.342). Deficiency of heterozygosity was observed in all the studied populations of P. vietnamensis and three populations of P. stipuleanatus. Some populations had high allelic richness for both species. Private alleles were determined in all the studied populations of P. vietnamensis and two P. stipuleanatus populations. Genetic differentiation was low in two ginseng species. However, habitat loss, over-utilization and over-harvesting can be the main causes of reduced genetic heterozygosity. Neighbor-joining tree and discriminant analysis of principal components detected three major genetic groups. Finally, based on our findings, we propose in situ conservation of populations with high expected heterozygosity, allelic richness, and private alleles. Seed collection should be performed for ex-situ conservation as genetic pools in the future.

The effects of copper and zinc salts on transgenic canola plants expressing rice transcription factor (TF) OsMYB4 were investigated. Transgenic plants (TPs), which showed a high OsMyb4 expression in response to either Cu or to Zn excess, were used for the current study. In leaves of TPs, the content of Cu was equal and the content of Zn was significantly higher than in non-transformed plants (NTPs). The TPs grown on an extremely high concentration of heavy metals (HMs; 150 μМ CuSO₄ or 5 000 μМ ZnSO₄) were able to survive for more than 15 d, while NTPs died after 7 - 9 d of incubation. This indicates that expression of OsMyb4 in canola plants improved their HM tolerance. The TPs tolerance to HMs was confirmed by a higher shoot biomass than that in NTPs. Excess of HMs caused oxidative stress (indicated by increase in malondialdehyde content) especially in leaves of NTPs. This data suggests a protective role of the OsMyb4 TF in oxidative stress. The HMs caused a lower decrease in activities of superoxide dismutase and guaiacol peroxidase in TPs than in NTPs. Higher tolerance of TPs to HMs was also suggested by a considerable increase in the content of low-molecular phenolic compounds, including flavonoids and anthocyanins, as well as proline (a potential antioxidant and chaperone). These data suggest that OsMYB4 may play a role as a positive regulator of phenylpropanoid pathway and proline synthesis. The created canola OsMyb4 TPs may be useful for future applications in phytoremediation of HM-polluted soils.

Medicinal plants are a rich source of secondary metabolites, extensively used in traditional health care systems. High altitude biodiversity encompasses the diversified and valuable medicinal plant species. The extreme environmental conditions of high altitude region viz. fluctuating temperatures, high UV radiation, salinity, low oxygen concentration, and high wind velocity limits the plant growth and distribution. Yet, how medicinal plants respond to these extreme conditions is not sufficiently understood. Therefore, addressing plant acclimation to different stresses presents an opportunity to unravel adaptive mechanism of medicinal plants along altitude gradient. This article reviews the recently published research that highlights the major role of proteins in plant adaptation to extreme environmental conditions. In the last few decades, climate change has made a profound impact on high altitude plants. Stress conditions alter cellular homeostasis of plants. With the advent of proteomics, it has become evident that stresses induce changes in proteome by synthesis/expression of novel stress responsive proteins. These proteins constitute a highly organized, complex network that leads to changes in the molecular, biochemical, physiological, and morphological responses of plants. Herein, we comprehensively discuss the proteomics of medicinal plants and its role in adaptation along altitude gradient. This review aims to provide impetus to current research in medicinal plants ranging from developmental to stress biology and to generate basis for genetic engineers and plant breeders to produce next-generation medicinal plants.

Transcription factors are involved in lipid metabolism, and in present study, the transcription factor WRINKLED 1 (GhWRI1) was cloned from Gossypium hirsutum L. cv. Coker 201 by reverse transcription (RT)-PCR and rapid amplification of cDNA ends. The Pro35S:WRI1 vector was constructed and transformed into upland cotton cv. Sumian 20 using the pollen tube pathway method. After PCR and Southern blot verification of the positive transgenic plants, T₂ transgenic lines derived from T₁ individuals with the insertion gene in a single copy were chosen for further dissection. Semi-quantitative and quantitative RT-PCR analyses indicated that GhWRI1 gene expression increased in transgenic plants compared with that in the wild-type. Seed lipid content increased at most transgenic plants, and at the same time, protein content decreased. Compared to the control, major agronomical traits were not affected by overexpression of GhWRI1 in transgenic plants.

Trehalose, which plays important roles in resistance to abiotic stresses and preservation of biological activity in plants, is synthesized by two key enzymes, trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Therefore, the expressions of the TPS and TPP genes directly affect trehalose synthesis and stress resistance of plants. In this study, CkTPS and CkTPP from Caragana korshinskii were identified, and the role of trehalose synthesis in the adaptation of this desert plant to adverse conditions was investigated. Higher CkTPS and CkTPP expressions were observed in the roots, whereas expressions were much lower in leaves and stems, and their expressions were upregulated under drought stress. Histochemical analyses showed that β-glucuronidase expression driven by the CkTPS and CkTPP promoters was strongly induced by abiotic stresses and phytohormones, such as abscisic acid, gibberellin, methyl jasmonate, and mannitol, which suggests that trehalose synthesis may be regulated by various signaling pathways. To determine the functional mechanism underlying the role of trehalose synthesis in regulating drought response in plants, CkTPS and CkTPP were introduced into Arabidopsis. Compared to wild-type (WT) plants, these transgenic plants showed higher germination rate, survival, less damage, better shoot growth, and longer roots under drought stress. Moreover, transgenic plants had a significantly higher content of proline, chlorophyll, trehalose, and activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), and lower malondialdehyde (MDA) content than WT controls. Double-transgenic plants carrying CkTPS and CkTPP showed better growth and stronger drought tolerance than either single transgenic plant line. These results provide a theoretical and experimental basis for further understanding the function and regulatory mechanism of CkTPS and CkTPP, as well as the possibility of their application for improving drought tolerance in crops through genetic engineering.

Although fullerene (C₆₀) has attracted great interest as a carbon-based nanomaterial with unique properties, today, little is known about the interaction of its water-soluble derivates, including fullerenol with higher plants. Here, we investigated how fullerenol [C₆₀(OH)_22-24] affects Zea mays, as a Strategy II plant, depending on its iron status. Iron deficiency chlorosis is a common nutritional disorder affecting plants. Maize plants were grown hydroponically, either with [+Fe^II (ferrous) or +Fe^III (ferric)] or in Fe-free (-Fe^II and -Fe^III) nutrient solution and with or without a fullerenol supply. Fullerenol affected plants differently depending on their Fe status. The beneficial effects of fullerenol were observed in the Fe^II-deprived plants, including successful suppression of plant Fe-deficiency chlorosis mainly in the younger (basal and middle) region of the leaf blade. This region expressed more severe chlorosis as compared with the older (apical) region of the leaf blade. These changes were accompanied by a significant increase in leaf active Fe and lowering the root apoplastic Fe, suggesting that fullerenol may enhance Fe mobilization in the roots, helping to alleviate Fe deficiency chlorosis. By contrast, there were no observable effects in the Fe^III-deprived plants being significantly lower in the root apoplastic Fe as compared with the Fe^II-deficient plants. Additionally, fullerenol did not affect the Fe-sufficient plants, irrespective of the Fe species (Fe^III-EDTA or Fe^II-EDTA) used as Fe-sources. Our results provide new evidence for the beneficial role of Fe-fullerenol interactions in the enhancement of gramineous plant tolerance to Fe deficiency conditions, which are one of the major limiting factors for crop production all over the world.

In this article, we present the response of Medicago truncatula Gaert. cv. Jemalong plants expressing constitutively the Dsp22 gene from Craterostigma plantagineum to water stress and rehydration. The Dsp22 gene encodes an ELIP-like protein thought to protect the chloroplast against photooxidative damage during the dehydration and rehydration. The Dsp22 transgenic homozygous M. truncatula plants showed higher amount of chlorophyll (Chl), lower Chl a/Chl b ratio and higher actual efficiency of energy conversion in photosystem 2 (Φ_PSII) after rehydration, when compared to the wild type. The combined data from the Chl a fluorescence analysis, pigment quantification and biomass accumulation showed that transgenic M. truncatula plants are able to recover from water deprivation better than wild type plants.

Flavonoids are secondary metabolites widely distributed in plants. They not only confer a wide spectrum of pigmentation to plant flowers but also protect plants from various biotic and abiotic stresses. Simultaneously, these compounds also offer health benefits to humans. Significant efforts have been made to correlate specific flavonoid production with biosynthetic pathway gene expression. Some structure genes and transcription factors that regulate the biosynthetic pathway have been identified. However, the diverse and complex control of flavonoid accumulation is still not well understood. In this mini-review, we summarized the improvement of flavonoids by genetic engineering from the aspects of flower colour, plant resistance, and benefits on the human diet. A perspective on flavonoid research in plants is provided.

Wild rice genotypes are rich in genetic diversity. This has potential to improve agronomic rice by allele mining for superior traits. Late embryogenesis abundant (LEA) proteins are often associated with desiccation tolerance and stress signalling. In the present study, a group 3 LEA gene, Wsi18 from the wild rice Oryza nivara was expressed under its own inducible promoter element in stress susceptible cultivated indica rice (cv. IR20). The resulting transgenic plants cultivated in a greenhouse showed enhanced tolerance to soil water deficit. Transgenic plants had higher grain yield, plant survival rate, and shoot relative water content compared to wild type (WT) IR20. Cell membrane stability index, proline and soluble sugar content were also greater in transgenic than WT plants under water stress. These results demonstrate the potential for improving SWS tolerance in agronomically important rice cultivar by incorporating Wsi18 gene from a wild rice O. nivara.

Background: Protocorm-like bodies (PLBs) represent a major morphogenic pathway in orchids and are characterized by cellular reprogramming, tissue differentiation, and developmental reorganization. Although PLB formation is widely explored for orchid micropropagation, endopolyploidy dynamics during PLB morphogenesis remain poorly understood. Aims: This study aimed to characterize endopolyploidy patterns and evaluate nuclear DNA content maintenance during PLB morphogenesis and plant regeneration in Cattleya tigrina. Methods: Basal leaf explants were cultured under PLB-inducing conditions in vitro. Morphological and histological analyses were performed throughout PLB development, while nuclear DNA content was estimated by flow cytometry in leaf regions, PLBs at successive developmental stages, and regenerated plants. Results: The leaf base was the most responsive region for PLB induction, showing intense cell proliferation and meristematic activity. Flow cytometric profiles consistently revealed 2C, 4C, and 8C nuclear DNA peaks, indicating endopolyploidy in somatic tissues. PLBs exhibited transient shifts in ploidy distribution during active proliferation phases, particularly at intermediate developmental stages, when 4C nuclei became predominant. Regenerated plants maintained nuclear DNA contents comparable to the original explants. Conclusions: Endopolyploidy is an intrinsic and developmentally regulated feature of PLB morphogenesis in Cattleya tigrina. PLB-mediated regeneration maintained overall nuclear DNA content under the evaluated in vitro conditions.

Fructans play vital roles in enhancing plant abiotic stress tolerance by reducing oxidative damage, stabilizing cell membranes, improving the osmotic adjustment capacity, and lowering the freezing point. In this study, a sucrose: fructan-6-fructosyltransferase (6-SFT) gene involved in the synthesis of fructans was isolated from Dasypyrum villosum, Dv-6-SFT, using genomic walking and reverse transcription (RT)-PCR. Alignment of the cDNA sequence with its genomic counterpart showed that no introns were present in the Dv-6-SFT gene, and thus it differs from all other plant 6-SFTs that have been cloned previously. Sequence analysis showed that the cDNA of the Dv-6-SFT sequence comprised 2 175 bp with a 1 863 bp open reading frame, and its deduced protein comprised 620 amino acids with a predicted molecular mass of 68.47 kDa. The Dv-6-SFT gene was transferred into tobacco (Nicotiana tabacum L.) cv. W38 via Agrobacterium-mediated transformation. The screened plants were tested by PCR and semi-quantitative RT-PCR, and the transgenic plants were evaluated under drought, cold, and salt stresses. The Dv-6-SFT transgenic tobacco plants had higher resistance to drought, cold, and salt stress than the non-transgenic plants. Further analysis showed that the transgenic plant expressing Dv-6-SFT had increased content of saccharides and proline, but reduced content of malondialdehyde in leaves. The results of this study demonstrate that the Dv-6-SFT gene is a potential candidate for conferring abiotic stress tolerance in plants and it could be used in crop improvement breeding programs.

Heat shock and almost all types of stresses associated with oxidative stress are accompanied by heat shock protein (HSP) expression. HSPs are involved in refolding denatured proteins and directing unrepairable proteins for degradation. Thus, under stress conditions, HSPs help to restore cellular balance. However, in virus-infected plants, HSP70 can have both positive and negative effects because viruses usually recruit HSP70. HSP70 can promote the replication and translation of the viral genome, the formation of viral replication complexes, and the propagation of viral particles from cell to cell and throughout the plant. HSP gene silencing in various virus-host plants systems and the comparison of susceptible and resistant species have shown that HSPs70 accelerate the development of infection. Conversely, during the process known as thermotherapy, the temperature increase inhibits viral replication in some host and virus systems. The success of thermotherapy depends not only on the temperature and treatment period or duration but also on the plant species and viral strain. In this review, we discuss the ambiguous role that HSPs70 play during viral infections in plants towards weighing the balance between their positive and negative functions.

Background: The tomato brown rugose fruit virus (Tobamovirus fructirugosum, ToBRFV) is an emerging tobamovirus that has quickly become a significant obstacle to the production of tomatoes and peppers worldwide. It is now classified as a regulated quarantine pathogen. Effective containment requires rapid, reliable, inexpensive, and safe diagnostic protocols for routine screening in laboratories and production systems.
Aims: We aimed to thoroughly evaluate integrated molecular and serological diagnostic methods for ToBRFV and develop biosafe positive controls suitable for high-throughput and decentralized applications.
Methods: We evaluated the following methods: conventional RT-PCR, one-enzyme RTX-PCR, immunocapture RT-PCR, recombinase polymerase amplification, loop-mediated isothermal amplification with colorimetric detection, Western blotting, dot blot, and tissue blot immunoassay. The non-infectious positive control was prepared using the GoldenBraid 3.0 cloning system. We developed a single-seed assay that enables direct testing of tomato seed stocks.
Results: Among the evaluated molecular methods, RTX-PCR was particularly advantageous due to its minimal sample handling, reduced cost, and ability to bypass RNA extraction. The tissue blot immunoassay enabled high-throughput, low-cost screening of hundreds of samples per day using only basic equipment. Although ToBRFV was frequently detected in seeds harvested from infected plants, no systemic infection was observed in progeny seedlings, confirming the low rate of true vertical transmission. A non-infectious positive control was prepared and successfully employed in molecular methods.
Conclusions: Our findings provide an integrated diagnostic framework combining molecular, serological, and biosafety tools to effectively monitor and contain ToBRFV in commercial production and phytosanitary settings.

Providing sufficient food to burgeoning population from the steadily shrinking arable land seems to be very difficult in near future and is one of the foremost challenges for plant scientists. In addition, there are several biotic and abiotic stresses which frequently encounter crop plants during various stages of life cycle, resulting in considerable yield losses. Environmental stresses, including drought, flooding, salinity, temperature (both low and high), high radiation, and xenobiotics induce toxicity, membrane damage, excessive reactive oxygen species (ROS) production, reduced photosynthesis, and altered nutrient acquisition. Several indigenous defence mechanisms (physiological and molecular) are triggered in plants on exposure to environmental cues. Enhancement of resistance of crop plants to environmental stresses has been the topic of prime interest for agriculturalists and plant scientists since long. Development of water and salinity stress-tolerant crops through genetic engineering provides an avenue towards the reclamation of farmlands that have been lost due to salinity and lack of irrigation water/rainfall. Understanding the complexity of stress tolerance mechanisms in orthodox or model plants at the genetic and molecular levels improves feasibility of enhancing tolerance of sensitive crop plants.

Background: In the last decades, vacuum infiltration had been applied for the Agrobacterium-mediated transient expression of foreign gene in plants. However, the relevant influencing factors have not been fully studied yet. Aims: This study aimed to evaluate the effects of vacuum infiltration pressure, time of vacuum infiltration, concentration of Agrobacterium tumefaciens (A. tumefaciens), and incubation time post infiltration on the transient expression of foreign gene in leaves of Nicotiana benthamiana (N. benthamiana). Methods: Two A. tumefaciens strains, LBA4404 and EHA105, carrying a reporter gene of green fluorescent protein (GFP), were used to infiltrate leaves of N. benthamiana via vacuum infiltration. The changes of GFP expression with changes of vacuum infiltration pressure, time of vacuum infiltration, concentration of A. tumefaciens, and incubation time post infiltration were measured. Results: Increase of vacuum pressure from 0.03 to 0.07 MPa increased the GFP expression in the leaves infiltrated with either LBA4404 or EHA105, and the vacuum pressure at 0.07 MPa made almost all of area of the infiltrated leaf express GFP. The increase of the time of vacuum infiltration from 10 to 20 min significantly enhanced the GFP expression in either the LBA4404 or EHA105-infected leaves. Among the different concentrations of Agrobacterium (from OD600 0.1 to 0.9), the concentrations of LBA4404 strain suspension at OD600 0.5 and EHA105 strain suspension at OD600 0.3, respectively, were most effective concentrations for enhancing the GFP expression in the infected leaves. The increase of incubation time post infiltration from 2 to 4 days largely enhanced the intensity of GFP expression in the either LBA4404 or EHA105-infected leaves. However, further increase of incubation time post infiltration from 4 to 6 days decreased the GFP expression in the infected leaves. Conclusions: Vacuum infiltration pressure/time, concentration of A. tumefaciens, and the incubation time post infiltration were important factors affecting the level of transient expression by vacuum infiltration. Optimizing these factors is essential for improving the level of transient expression of foreign gene by vacuum infiltration.

Cold stress is one of the major limitations to crop productivity worldwide. We investigated the effects of multiple gene expression from cold tolerant Capsella bursa-pastoris in transgenic tobacco (Nicotiana tabaccum) plants. We combined CblCE53 and CbCBF into a reconstruct vector by isocaudomers. Plant overexpression of CbICE53 under the stress inducible CbCOR15b promoter and CbCBF under a constitutive promoter showed increased tolerance to both chilling and freezing temperatures in comparison to wild-type plants, according to the electrolyte leakage and relative water content. The expressions of endogenous cold-responsive genes in transgenic tobacco (NtDREB1, NtDREB3, NtERD10a and NtERD10b) were obviously upregulated under normal and low temperature conditions. These results suggest that the CbICE53 + CbCBF transgenic plants showed a much greater cold tolerance as well as no dwarfism and delayed flowering. Thus they can be considered as a potential candidate for transgenic engineering for cold tolerant tobacco.

Background: Many concerns have been raised about the extensive use of agrochemicals and their impacts on environment and soil. Some serious issues include increased soil salinization, toxicity and decrease in soil microbial biodiversity. Biostimulants have emerged as a promising ecofriendly alternative to agrochemicals. Humic substances (HSs) are biostimulants with many beneficial effects in plants physiology and productivity. Aims: This review aims to synthesize current evidence on the role of HSs in supporting sustainable agriculture that improves crop productivity while maintaining good soil health in changing environmental conditions. Methods: Relevant studies related to HSs and their effects on soil fertility, plant growth, stress tolerance and hormonal regulation were collected from major scientific databases including Scopus, Web of Science, and Google Scholar. The main outcomes of these studies were extracted and analyzed to provide an overview of the available evidence. Results: The literature shows that the integration of HSs into agricultural practices improves soil structure, increases water retention, maintains healthy microbial diversity and enhances plant productivity. Conclusions: This review highlights the importance of using HSs in enhancing soil health and plant physiology. However, further studies are required to integrating new technologies and diversify HSs sources, plants species, and different environments.

The gene of Zinnia elegans L. coding for S-like extracellular ribonuclease (ZRNase II) was used to produce transgenic tobacco plants with an increased ribonuclease activity. The protein-coding part of ZRNase II included the signal peptide sequence so the transgenic protein was located extracellularly. The cDNA of ZRNase II was cloned under the control of 2'-promoter of the mannopine synthase (MAS 2') gene from Ti-plasmid of Agrobacterium tumefaciens. It was shown that the resultant transgenic plants had an increased ribonuclease activity of the crude extracts and the induction of MAS 2' promoter by wounding additionally increased the activity. The plants of two transforming lines characterized by different ribonuclease activities were used to analyze the transgene influence on plant resistance to tobacco mosaic virus. The plants demonstrated either absence of disease symptoms or a significant delay in their appearance, depending on the virus content in the inoculum and ribonuclease activity.

Salt stress is one of the abiotic stresses limiting the yield of crops worldwide. However, the molecular mechanisms underlying the regulation of plant response to salt stress are not completely elucidated. Ethylene response factors (ERFs) are a subfamily of the AP2 (APETALA2)/ERF transcription factor family that regulates multiple aspects of plant growth and development, and plant responses to biotic and abiotic stresses. ERF96 is one of the small ERFs that is involved in plant defense response and abscisic acid signaling in Arabidopsis. By using real time quantitative PCR, we found that the expression of ERF96 in the wild type Arabidopsis thaliana (cv. Col-0) seedlings was induced by NaCl treatment. The transgenic plants overexpressing ERF96 were more tolerant to salt stress in terms of NaCl inhibited seed germination, early seedling development, and fresh mass. Consistent with these observations, elevated expressions of some NaCl-responsive genes including responsive drought 29 (RD29A), Δ¹-pyrroline-5-carboxylate synthetase (P5CS), cold responsive 15A (COR15A), and kinase 1 (KIN1) were observed in the transgenic plants in the presence of NaCl. We also found that the Na⁺ and K⁺ content and expressions of genes related to Na⁺/K⁺ homeostasis including stelar K⁺ outward rectifier (SKOR) and potassium transport 2/3 (AKT2/3) were altered in the ERF96 transgenic plants in response to NaCl treatment. Taken together, these results showed that overexpression of ERF96 enhanced plant tolerance to salt stress, indicating that ERF96 is a positive regulator of salt tolerance in Arabidopsis.

Stilbene synthases (STS) are plant enzymes that are responsible for the biosynthesis of stilbenes, which are plant phenolic compounds with valuable biological properties. Stilbenes also play important roles in plant tolerance to biotic and abiotic stresses. Therefore, plants that overexpress STS genes can be more resistant to various stresses. This paper investigated the effects of STS gene overexpression in Arabidopsis thaliana (L.) Heynh. Columbia-0 plants on stilbene content and tolerance to the following abiotic stresses: low and high temperatures, salinity, drought, and ultraviolet irradiation (UV-B and UV-C). We used VaSTS1 and VaSTS7 genes from grapevine (Vitis amurensis Rupr.) expressed under the double cauliflower mosaic virus 35S (CaMV35S) promoter. This study firstly demonstrated that overexpression of the VaSTS1 and VaSTS7 genes in A. thaliana plants considerably increased plant tolerance to UV-B and UV-C, while the tolerance to the low and high temperatures, salinity, and drought was not affected. We showed that the highest trans-piceid and trans-resveratrol total content was in ST1 A. thaliana plants that overexpressed the VaSTS1 gene in the range 8.28 - 22.66 µg g^-1(f.m.). ST7 plants that overexpressed the VaSTS7 gene showed only trans-resveratrol at 0.02 - 0.08 µg g^-1(f.m). Stilbene content and UV tolerance in transgenic A. thaliana plants correlated with STS transgene expression. STS expression, UV tolerance, and stilbene content was higher in VaSTS1 transgenic plants compared with that in VaSTS7 transgenic plants.