Antifungal Activity of Crude Extracts of <i>Tectona grandis</i> L.f. against Wood Decay Fungi

Wood is mainly made up of cellulose, hemicelluloses, lignin polymers and other organic and inorganic substances, making it susceptible to deteriorate by various biological agents. Tectona grandis L.f. (Teak) is a timber species with high resistance to biological deterioration, valued for its durability, beauty, and mechanical resistance. The purpose of this work was to evaluate the antifungal activity of crude extracts from teak on various fungi that cause wood deterioration. For this, Teak heartwood was obtained, then fragmented and pulverized until obtaining a flour which was used for compounds extraction using the Soxhlet technique coupled to a rotary evaporator through solvents of increasing polarity (hexane, dichloromethane, tetrahydrofuran, and acetone). The extracts obtained were tested against fungal organisms collected in the field, and the LC₅₀ was determined using teak crude extracts on Artemia salina as a biological model. The results obtained showed that a high flour yield was obtained with hexane (0.951 g), followed by tetrahydrofuran (0.446 g), dichloromethane (0.348 g), and acetone (0.152 g). By using nine fungal organisms that predominantly correspond to the genus Aspergillus, the extractable compounds were tested, inhibiting 25% of mycelial growth with tetrahydrofuran (T. versicolor), and 40.9% with dichloromethane (G. trabeum). Likewise, the biological model of A. salina showed an LC₅₀ of 84.9 μg/mL with hexane, 43.3 μg/mL with dichloromethane, 59.6 μg/mL with tetrahydrofuran, and 54.7 μg/mL with acetone. For this reason, it is concluded that Teak wood contains many extractable compounds in relation to its weight, besides having antimicrobial activity when extracted through polar compounds such as dichloromethane and tetrahydrofuran.     

Cloning and Characterization of <i>EuGID1</i> in <i>Eucommia ulmoides</i> Oliver

Gibberellic acid controlled the key developmental processes of the life cycle of landing plants, and regulated the growth and development of plants. In this study, a novel gibberellin receptor gene EuGID1 was obtained from Eucommia ulmoides Oliver. The cDNA of EuGID1 was 1556 bp, and the open reading frame was 1029 bp, which encoded 343 amino acids. EuGID1 had the homology sequence with the hormone-sensitive lipase family. Amino acid sequence alignment confirmed EuGID1 protein had the highest homology with the GID1 protein of Manihot esculenta. EuGID1 was located in the nucleus and cell membrane and had expression in four plant organs. Overexpression of EuGID1 in transgenic Arabidopsis plants promoted plant elongation and increased siliques yield.     

Inhibitory Effect of <i>N</i>, <i>N</i>-Dimethylhexadecylamine on the Growth of White-Rot Fungus <i>Trametes versicolor</i> (L.) in Wood

Wood is an organic material that is a source of carbon of organisms called Wood-decay fungi, and to preserve the wood, various toxic compounds to man and the environment have been used. To analyze the effect of N,N-Dimethylhexadecylamine (DMHDA) on wood attacked by the rotting fungus Trametes versicolor L. We used an in vitro system to expose the fungus T. versicolor to different concentrations of the DMHDA (50, 150 and 450 μM). We quantified the diameter of mycelial growth and laccase activity, also, under these experimental conditions we studied morphological details of the organisms using different scanning equipment including scanning electron microscopy. The growth of T. versicolor exposed to DMHDA for 60 days, showed a concentration-dependent dose behavior, also, the electron microscopy analysis revealed that the morphology and mycelial density was affected by the DMHDA, showing a formation of atypical morphological and thickener folds. Finally, the pieces of wood treated with DMHDA and exposed to the fungus had a lower mass loss, after a period of 60 days of exposure, the values obtained were 0.7, 1.0 and 0.5 g of mass lost for the control, LoC and LoDMHDA treatments respectively. Wood-rot fungi have represented economic losses worldwide, the strategies used have been supported by toxic compounds for the environment. The DMHDA both in the Petri dish system and as a wood preservative was shown to significantly inhibit the growth of T. versicolor.     

Repellent and Insecticidal Activity of Naphthoquinones from the Heartwood of <i>Tectona grandis</i> on <i>Incisitermes marginipennis</i> (Latreille)

Wood is a lignocellulosic material that, because of its organic nature, is a source of nutrients for bacteria, fungi, and insects. Incisitermes marginipennis (Latreille) is an interesting dry wood-boring termite. Because it is an extremely destructive pest, difficult to control due to its cryptic lifestyle, and for its role in the biogeochemical cycle of carbon and other elements; it is a fundamental organism. The use of less toxic and eco-friendly substances is important for protecting domestic, cultural, and historical heritage made of wood or lignocellulosic materials. The aim of this study was to evaluate the biological activity of various naphthoquinones extracted from the heartwood of T. grandis on I. marginipennis. First, heartwood flour was used for Soxhlet-type extractions to isolate and characterize the heartwood compounds by thin-layer chromatography. To identify the heartwood compounds, nuclear magnetic resonance and gas chromatography coupled to a mass spectrometer were used. The information obtained was supported using the Interactive Spectroscopy software. The identified heartwood compounds were lapachol, tectoquinone, deoxylapachol, dehydro-α-lapachone, tectol and tecomaquinone-I. Using nymphal termites of I. marginipennis as a biological model, the isolated and identified molecules were under three different tests (repellent, insecticide and protective) and their biological activity was individually evaluated. The results showed that the molecules obtained have a repellent-protective effect against I. marginipennis, although not in all cases, these molecules have high percentages of mortality and decreased the weight loss of the treated material exposed to I. marginipennis.     

Effects of Barium Stress in <i>Brassica juncea</i> and <i>Cakile maritima</i>: The Indicator Role of Some Antioxidant Enzymes and Secondary Metabolites

Soil contamination by toxic trace metal elements, like barium (Ba), may stimulate various undesirable changes in the metabolic activity of plants. The plant responses are fast and with, direct or indirect, generation of reactive oxygen species (ROS). To cope with the stress imposed by the ROS production, plants developed a dual cellular system composed of enzymatic and non-enzymatic players that convert ROS, and their by-products, into stable nontoxic molecules. To assess the Ba stress response of two Brassicaceae species (Brassica juncea, a glycophyte, and Cakile maritime, a halophyte), plants were exposure to different Ba concentrations (0, 100, 200, 300 and 500 μM). The plants response was evaluated through their morphology and development, the determination of plant leaves antioxidant enzymatic activities and by the production of plants secondary metabolites. Results indicated that the two Brassicaceae species have the ability to survive in an environment containing Ba (even at 500 μM). The biomass production of C. maritima was slightly affected whereas an increase in biomass B. juncea was noticed. The stress imposed by Ba activated the antioxidant defense system in the two species, noticed by the changes in the leaves activity of catalase (CAT), ascorbate peroxidase (APX) and guaicol peroxidase (GPX), and of the secondary metabolites, through the production of total phenols and flavonoids. The enzymatic response was not similar within the two plant species: CAT and APX seem to have a more important role against the oxidative stress in C. maritima while in B. juncea is GPX. Overall, total phenols and flavonoids production was more significant in the plants aerial part than in the roots, of the both species. Although the two Brassicaceae species response was different, in both plants catalytic and non-catalytic transformation of ROS occurs, and both were able to overcome the Ba toxicity and prevent the cell damage.     

Biochemical and Physiological Responses of <i>Arabidopsis thaliana</i> Leaves to Moderate Mechanical Stimulation

Mechanical stimulation of plants can be caused by various abiotic and biotic environmental factors. Apart from the negative consequences, it can also cause positive changes, such as acclimatization of plants to stress conditions. Therefore, it is necessary to study the physiological and biochemical mechanisms underlying the response of plants to mechanical stimulation. Our aim was to evaluate the response of model plant Arabidopsis thaliana to a moderate force of 5 N (newton) for 20 s, which could be compared with the pressure caused by animal movement and weather conditions such as heavy rain. Mechanically stimulated leaves were sampled 1 h after exposure and after a recovery period of 20 h. To study a possible systemic response, unstimulated leaves of treated plants were collected 20 h after exposure alongside the stimulated leaves from the same plants. The effect of stimulation was assessed by measuring oxidative stress parameters, antioxidant enzymes activity, total phenolics, and photosynthetic performance. Stimulated leaves showed increased lipid peroxidation 1 h after treatment and increased superoxide dismutase activity and phenolic oxidation rate after a 20-h recovery period. Considering photosynthetic performance after the 20-h recovery period, the effective quantum yield of the photosystem II was lower in the stimulated leaves, whereas photochemical quenching was lower in the unstimulated leaves of the treated plants. Nonphotochemical quenching was lower in the stimulated leaves 1 h after treatment. Our study suggested that plants sensed moderate force, but it did not induce pronounced change in metabolism or photosynthetic performance. Principal component analysis distinguished three groups–leaves of untreated plants, leaves analysed 1 h after stimulation, while stimulated and unstimulated leaves of treated plants analysed 20 h after treatment formed together the third group. Observed grouping of stimulated and unstimulated leaves of treated plants could indicate signal transduction from the stimulated to distant leaves, that is, a systemic response to a local application of mechanical stimuli.     

<i>DWARF</i> and <i>SMALL SEED1</i>, a Novel Allele of <i>OsDWARF</i>, Controls Rice Plant Architecture,Seed Size,and Chlorophyll Biosynthesis

Plant architecture is a vital agronomic trait to control yield in rice (Oryza sativa L.). A dwarf and small seed 1 (dss1) mutant were obtained from the ethyl methanesulfonate (EMS) mutagenized progeny of a Guizhou glutinous landrace cultivar, Lipingzabianhe. The dss1 mutant displayed phenotypes similar to those of brassinosteroid (BR) deficient mutants, such as dwarfing, dark green and rugose erect leaves, small seeds, and loner neck internode panicles with primary branching. In our previous study, the underlying DSS1 gene was isolated, a novel allele of OsDWARF (OsBR6ox) that encodes a cytochrome P450 protein involved in the BR biosynthetic pathway by MutMap technology. In this work, we confirmed that a Thr335Ile amino acid substitution residing in DSS1/OsDWARF was responsible for the dwarf, panicle architecture, and small seed phenotypes in the dss1 mutants by genetic transformation experiments. The overexpression of OsDWARF in the dss1 mutant background could not only recover dss1 to the normal plant height and panicle architecture but also rescued normal leaf angles, seed size, and leaf color. Thus, the specific mutation in DSS1/OsDWARF influenced plant architecture, seed size, and chlorophyll biosynthesis.     

Phytosulfokine-α Promotes Root Growth by Repressing Expression of <i>Pectin Methylesterase Inhibitor</i> (<i>PMEI</i>) Genes in <i>Medicago truncatula</i>

Phytosulfokine-α (PSK-α), a sulfated pentapeptide with the sequence YIYTQ, is encoded by a small precursor gene family in Arabidopsis. PSK-α regulates multiple growth and developmental processes as a novel peptide hormone. Despite its importance, functions of PSK-α in M. truncatula growth remains unknown. In this study, we identified five genes to encode PSK-α precursors in M. truncatula. All of these precursors possess conserved PSK-α signature motif. Expression pattern analysis of these MtPSK genes revealed that each gene was expressed in a tissue-specific or ubiquitous pattern and three of them were remarkably expressed in root. Treatment of M. truncatula seedlings with synthetic PSK- α peptide significantly promoted root elongation. In addition, expression analysis of downstream genes by RNA-seq and qRT-PCR assays suggested that PSK-α signaling might regulate cell wall structure via PMEI-PME module to promote root cell growth. Taken together, our results shed light on the mechanism by which PSK-α promotes root growth in M. truncatula, providing a new resource for improvement of root growth in agriculture.     

Molecular Interaction Study to Explore the <i>Nigella sativa</i> Bioactive Components as an Inhibitor of Peptide Deformylase to Inhibit the <i>Xanthomonas oryzae</i> pv. oryzae via Applying Computational Approach

Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most damaging diseases to rice across the world. Various chemicals have been employed so far for the management of bacterial leaf blight. On the other hand, these compounds are damaging to the ecosystem and have an impact on non-target species such as humans and animals. As a result, there is a need to create a new natural inhibitor for BLB management. Deformylase (PDF) enzyme is present in all eubacteria and its necessity in bacterial protein synthesis reveals it as an attractive target for drug development. In this study, the active components of Nigella sativa have been selected based on their previously reported antimicrobial activity and screened on the active site of bacterial PDF by the in silico art of techniques. Among these compounds, dithymoquinone and p-cymene strongly bind with the PDF enzyme with binding energy values of 7.77 kcal/mol and 7.26 kcal/mol, respectively, which is comparatively higher than the control compound (−6.73 kcal/mol). Hence, the “dithymoquinone-PDF” and “p-cymene-PDF” complexes were selected for further study, and their stability was assessed by molecular dynamic (MD) simulation. In MD simulation, both selected compounds exhibited steady-state interaction with PDF for 20 ns. It has been hypothesized that p-cymene and dithymoquinone inhibit peptide deformylase and could be used as antibacterials or pesticides against Xoo against the BLB disease.     

Differential Responses of <i>NHX1</i> and <i>SOS1</i> Gene Expressions to Salinity in two <i>Miscanthus sinensis</i> Anderss. Accessions with Different Salt Tolerance

The lignocellulosic crop Miscanthus spp. has been identified as a good candidate for biomass production. The responses of Miscanthus sinensis Anderss. to salinity were studied to satisfy the needs for high yields in marginal areas and to avoid competition with food production. The results indicated that the relative advantages of the tolerant accession over the sensitive one under saline conditions were associated with restricted Na⁺ accumulation in shoots. Seedlings of two accessions (salt-tolerant ‘JM0119’ and salt-sensitive ‘JM0099’) were subjected to 0 (control), 100, 200, and 300 mM NaCl stress to better understand the salt-induced biochemical responses of genes involved in Na⁺ accumulation in M. sinensis. The adaptation responses of genes encoding for Na⁺ /H⁺ antiporters, NHX1 and SOS1 to NaCl stress were examined in JM0119 and JM0099.The cDNA sequences of genes examined were highly conserved among the relatives of M. sinensis based on the sequencing on approximate 600 bp-long cDNA fragments obtained from degenerate PCR. These salt-induced variations of gene expression investigated by quantitative real-time PCR provided evidences for insights of the molecular mechanisms of salt tolerance in M. sinensis. The expression of NHX1 was up-regulated by salt stress in JM0119 shoot and root tissues. However, it was hardly affected in JM0099 shoot tissue except for a significant increase at the 100 mM salt treatment, and it was salt-suppressed in the JM0099 root tissue. In the root tissue, the expression of SOS1 was induced by the high salt treatment in JM0119 but repressed by all salt treatments in JM0099. Thus, the remarkably higher expression of NHX1 and SOS1 were associated with the resistance to Na⁺ toxicity by regulation of the Na⁺ influx, efflux, and sequestration under different salt conditions.     

Anatomical and Molecular Identification of Ornamental Plant <i>Ficus</i> L. Species

This present study includes twelve species that represent the Ficus genus, namely; aspera, carica, tinctoria subsp. gibbosa, hirta, hispida, neriifolia, palmata, pumila, racemosa, septica, sur, and sycomorus, belonging to the Moraceae family. The species samples were collected from various locations in Egypt. The study focused on the anatomical and molecular characteristics of mature foliage leaves. Since the identification and classification of taxa are highly dependent on the anatomical features of leaves, the anatomical characteristics were recorded in the form of a comparison between the examined plants in the data matrix. This study aims to contribute to the identification of the studied species based on the anatomical details of the matured leaves. Anatomical characterization includes the variations in upper and lower epidermal layers that are covered by a thin or thick cuticle; the number of palisade and spongy layers; crystals; secretory elements; lithocysts; the midrib zone has parenchyma associated with mechanical tissue, vascular system, and investigation of trichomes; on the other hand, in the current study, the phylogenetic analysis was conducted by using the ITS and 5.8 S sequences. From the analysis of all the available data, it could be stated that there is an overall agreement with the anatomical character dendrogram.     

Allantoin Alleviates Seed Germination Thermoinhibition in <i>Arabidopsis</i>

Allantoin as the metabolite of purine catabolism can store and remobilize nitrogen for plant growth and development. However, emerging evidence suggests it also contributes to plant tolerance to stress response through altering abscisic acid (ABA) and reducing reactive oxygen species (ROS) level. 1-CYS PEROXIREDOXIN (PER1) is a seed-specific antioxidant that enhances seed longevity through scavenging ROS over-accumulation. High temperature (HT) suppresses seed germination and induces seed secondary dormancy, called as seed germination thermoinhibition. However, the mechanism that allantoin and PER1 regulate seed germination thermoinhibition remains unknown. In this study, we reported that allantoin treatment enhances seed germination under HT stress. Consistently, the aln mutants displayed higher seed germination, as well as more accumulation of endogenous allantoin, than that of wild-type control. Further biochemical and genetic analyses showed that allantoin reduces ABA content under HT, and allantoin targets PER1 to efficiently scavenge HT-induced ROS accumulation, meanwhile, the function of allantoin requires PER1 during seed gemination thermotolerance. Collectively, our finding proposes a novel function of allantoin in enhancing seed germination tolerance to HT, and uncovers the underlying mechanism by which allantoin regulates seed germination through altering ABA metabolism and PER1-mediated ROS level under HT stress.     

Techniques and applications of three‐dimensional electron microscopy in entomological research

Structural studies using two‐dimensional (2D) images show limitations in understanding the structure and functions of cellular organelle and protein. To overcome the difficulty, over the last few years 3D reconstruction techniques using electron microscopy have been developed at extremely high speed. In this paper, currently available 3D reconstruction techniques of electron microscopy (such as electron tomography, serial section analysis and single particle analysis) are introduced using our data as examples of the application. The 3D structure of mitochondria with the defect of mitochondrial protein in round worm, Caenorhabditis elegans, through electron tomography, the cell–cell interaction in lamina of Drosophila melanogaster by serial‐section using ultramicrotome and high‐voltage electron microscopy and a thin filament related to muscle contraction in Drosophila melanogaster were used for examples of the application. These results through 3D reconstruction reveal the structural changes in a cellular organelle and protein that had not been shown by 2D structure.     

Genome-Wide Identification,Evolution and Expression Analyses of <i>GA2ox</i> Gene Family in <i>Brassica napus</i> L.

Gibberellin 2-oxidases (GA2ox) are important enzymes that maintain the balance of bioactive GAs in plants. GA2ox genes have been identified and characterized in many plants, but these genes were not investigated in Brassica napus. Here, we identified 31 GA2ox genes in B. napus and 15 of these BnaGA2ox genes were distributed in the A and C subgenomes. Subcellular localization predictions suggested that all BnaGA2ox proteins were localized in the cytoplasm, and gene structure analysis showed that the BnaGA2ox genes contained 2–4 exons. Phylogenetic analysis indicated that BnGA2ox family proteins in monocotyledons and dicotyledons can be divided into four groups, including two C₁₉-GA2ox and two C₂₀-GA2ox clades. Group 4 is a C₂₀-GA2ox Class discovered recently. Most BnaGA2ox genes had a syntenic relationship with AtGA2ox genes. BnaGA2ox genes in the C subgenome had experienced stronger selection pressure than genes in the A subgenome. BnaGA2ox genes were highly expressed in specific tissues such as those involved in growth and development, and most of them were mainly involved in abiotic responses, regulation of phytohormones and growth and development. Our study provided a valuable evolutionary analysis of GA2ox genes in monocotyledons and dicotyledons, as well as an insight into the biological functions of GA2ox family genes in B. napus.     

The Endosperm-Specific Expression of <i>YUCCA</i> Genes Enhances Rice Grain Filling

Grain filling is a crucial process that affects yield in rice (Oryza sativa L.). Auxin biosynthesis and signaling are closely related to rice yield; therefore, it is important to understand the effects of auxin biosynthesis on rice grain filling to improve crop yield. In this study, we used physiological and molecular strategies to identify the roles of auxin in rice grain filling. Exogenous application of auxin (IAA) or auxin analogues (2, 4-D) to young spikelets and flag leaves improved the seed-setting rate and yield per spike. Furthermore, real-time quantitative PCR assays confirmed that nine members of the OsYUCCA family of auxin biosynthetic genes were upregulated during grain filling, implication that auxin biosynthesis plays a major role in grain development. The specific expression of either Arabidopsis AtYUCCA1 or OsYUCCA2 in the endosperm or leaves resulted in increased expression of OsIAA genes and auxin content of seeds, as well as increased grain filling and seed-setting rate. This result establishes that the auxin content in grains and leaves is important for grain development. Our findings further highlight the potential applications for improving rice yield by elevating targeted gene expression in specific tissues.     

First Report of <i>Fusarium striatum</i> Causing Root Rot Disease of <i>Panax notoginseng</i> in Yunnan,China

Panax notoginseng is a traditional Chinese medicinal plant. Root rot of P. notoginseng is one of the most serious diseases affecting P. notoginseng growth and causes wilted leaves, fewer lateral roots and rotten roots. Root rot is a soil-borne disease, and mainly occurs from June to August in Yunnan Province when the temperatures are high and the air is humid. In this study, the endophytic fungal genus Fusarium isolate E-2018.1.22-#3.2 was obtained from a P. notoginseng embryo. Fusarium isolate E-2018.1.22-#3.2 was identified as Fusarium striatum based on morphological characteristics and molecular analysis. The fungus was found to have conidiophores and macroconidia, and its ITS, LSU and TEF-1α genes shared 100%, 99.2% and 99% identities with the homologous genes of Fusarium striatum, respectively. Isolate F. striatum E-2018.1.22-#3.2 can cause root rot symptoms, including black, soft roots, fewer lateral roots and leaf wilt, in 93% of the experimental P. notoginseng plants, and could be re-isolated, fulfilling Koch’s postulates. When the P. notoginseng plants were treated with the fungicide pyraclostrobin, isolate F. striatum E-2018.1.22-#3.2 was unable to cause root rot. We have therefore demonstrated that F. striatum E-2018.1.22-#3.2 is able to cause root rot disease in P. notoginseng. This is the first report of root rot disease caused by F. striatum on P. notoginseng in China.     

Efficient Evergreen Plant Regeneration of <i>Cinnamomum japonicum</i> Sieb. through <i>in vitro</i> Organogenesis

Cinnamomum japonicum Sieb. is an excellent roadside tree and medicinal tree species with considerable ornamental and economic value. In this study, we successfully developed a large-scale micropropagation protocol for C. japonicum for the first time. Sterilized shoots were excised and used as explants for shoot induction on several basal media, supplemented with different concentrations of plant growth regulators (PGRs), such as Thidiazuron (TDZ), N⁶ -Benzyladenine (6-benzylaminopurine) (BA), α-naphthaleneacetic acid (NAA) and Gibberellic acid (GA₃). After comparison, the most efficient medium for shoot regeneration was 1/2 Murashige and Skoog (MS) medium containing 0.5 mg L^–1 BA, 0.05 mg L^–1 NAA and 0.2 mg L^–1 GA₃, which resulted in an average number of induced shoots per explant and shoot length of 5.2 and 1.62 cm at 28 d, respectively. Then, elongated adventitious shoots were transferred to induce roots. 86.7% of shoots was able to root on 1/2 MS medium supplemented with 0.5 mg L^–1 NAA and 0.1 mg L^–1 BA. The earliest rooting time observed was after 21 d and the average root length was up to 3.3 cm after 28 d. Our study shows that C. japonicum can be successfully regenerated through de novo organogenesis, which lays a foundation for future transformation research on this tree.