Recent advancements in the role of histone acetylation dynamics to improve stress responses in plants.

In eukaryotes, transcriptional regulation is determined by the DNA sequence and is facilitated through sophisticated and complex chromatin alterations and histone remodelling. Recent research has shown that the histone acetylation dynamic, an intermittent and reversible substitution, constitutes a prerequisite for chromatin modification. These changes in chromatin structure modulate genome-wide and specific changes in response to external and internal cues like cell differentiation, development, growth, light temperature, and biotic stresses. Histone acetylation dynamics also control the cell cycle. HATs and HDACs play a critical role in gene expression modulation during plant growth and response to environmental circumstances. It has been well established that HATs and HDACs interact with various distinct transcription factors and chromatin-remodelling proteins (CRPs) involved in the transcriptional regulation of several developmental processes. This review explores recent research on histone acyltransferases and histone deacetylases, mainly focusing on their involvement in plant biotic stress responses. Moreover, we also emphasized the research gaps that must be filled to fully understand the complete function of histone acetylation dynamics during biotic stress responses in plants. A thorough understanding of histone acetylation will make it possible to enhance tolerance against various kinds of stress and decrease yield losses in many crops.

Molecular aspects of regeneration in insects.

Regeneration is a fascinating phenomenon observed in various organisms across the animal kingdom. Different orders of class Insecta are reported to possess comprehensive regeneration abilities. Several signalling molecules, such as morphogens, growth factors, and others trigger a cascade of events that promote wound healing, blastema formation, growth, and repatterning. Furthermore, epigenetic regulation has emerged as a critical player in regulating the process of regeneration. This report highlights the major breakthrough research on wound healing and tissue regeneration. Exploring and reviewing the molecular basis of regeneration can be helpful in the area of regenerative medicine advancements. The understanding gathered from this framework can potentially contribute to hypothesis designing with implications in the field of synthetic biology and human health.

Major phenolic compounds, antioxidant, antimicrobial, and cytotoxic activities of Selinum carvifolia (L.) collected from different altitudes in India.

Antibiotic resistance poses a serious threat to public health, raising the number of diseases in the community. Recent research has shown that plant-derived phenolic compounds have strong antimicrobial, antifungal, and cytotoxic properties against a variety of microorganisms and work as great antioxidants in such treatments. The goal of the current work is to evaluate the anticancerous, antibacterial, antifungal, antioxidant, and cytotoxicity activities in the extracts of the different plant parts (leaves, stems, and roots) of S. carvifolia (L.) L. This is a medicinally important plant and has been used for different kinds of diseases and ailments such as hysteria and seizures. The phenolic compounds from the different plant parts were analyzed using HPLC and the following were found to be present: chlorogenic acid, gallic acid, rutin, syringic acid, vanillic acid, cinnamic acid, caffeic acid, and protocatechuic acid. Gallic acid was found to have the highest concentration (13.93 mg/g), while chlorogenic acid (0.25 mg/g) had the lowest. The maximum TPC value, which ranged from 33.79 to 57.95 mg GAE/g dry extract weight, was found in the stem. Root extract with 9.4 mg RE/g had the greatest TFC level. In the leaf and stem extracts, the RSC ranged from 0.747 mg/mL to 0.734 mg/1 mL GE/g dry extract weight, respectively. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used to measure in vitro antioxidant activity. In a concentration-dependent way, promising antioxidant activity was reported. Moreover, 3,5-dinitrosalicylic acid (DNSA) and the Folin-Ciocalteu phenol reagent technique were used to determine reducing sugar content and total phenolic content, respectively. Antibacterial activity against eight strains (MIC: 250-1,000 µg/mL) was analyzed, and the stem extract exhibited maximum activity. Antifungal activity was also assessed, and potent activity was reported especially in the extract obtained from the stem. Cytotoxicity was evaluated using an MTT assay in the A549 cell line, where different doses (0.0625, 0.125, 0.25, 0.5, and 1 mg/mL) of leaf, root, and stem extracts were used. Treatment with these extracts reduced the cell viability, indicating that S. carvifolia may possess anticancer potential, which can be of great therapeutic value.

Deciphering the kinetics and pathway of lindane biodegradation by novel soil ascomycete fungi for its implication in bioremediation.

Lindane, an organochlorine pesticide, negatively affects living beings and the ecosystem. In this study, the potential of 9 Ascomycetes fungi, isolated from an hexachlorocyclohexane dumpsite soil, was tested for biodegradation of lindane. The strain Pleurostoma richardsiae (FN5) showed lindane biodegradation rate constant (K value) of 0.144 d-1 and a half-life of 4.8d. The formation of intermediate metabolites upon lindane degradation including γ-pentachlorocyclohexene, 2,4-dichlorophenol, phenol, benzene, 1,3- cyclohexadiene, and benzoic acid detected by GC-MS and the potential pathway adopted by the novel fungal strain FN5 for lindane biodegradation has been elucidated. The study of gene profiles with reference to linA and linB in strain FN5 confirmed the same protein family with the reported heterologs from other fungal strains in the NCBI database. This study for the first time provides a thorough understanding of lindane biodegradation by a novel soil-borne Ascomycota fungal strain for its possible application in field-scale bioremediation.

Agrobacterium-mediated Genetic Transformation of Cotton and Regeneration via Somatic Embryogenesis.

Cotton is a significant industrial crop, playing an essential role in the global economy that suffers several setbacks due to biotic and abiotic adversities. Despite such problems, biotechnological advances in cotton are limited because of genetic transformation and regeneration limitations. Here, we present a detailed protocol optimized based on previously published papers, along with our modifications. These involve changes in Agrobacterium concentration, co-cultivation time and temperature, hormones used for regeneration, media manipulation for embryogenic callus production, and efficient rescue of deformed embryos. Further, this protocol has been used in genetic studies on biotic and abiotic stress in cotton. This protocol assures a reproducible stable transgenic cotton development procedure via somatic embryogenesis that can be used by researchers worldwide. This protocol was validated in: Nat Biotechnol (2016), DOI: 10.1038/nbt.3665.

The industrially important genus Kaempferia: An ethnopharmacological review.

Kaempferia, a genus of the family Zingiberaceae, is widely distributed with more than 50 species which are mostly found throughout Southeast Asia. These plants have important ethnobotanical significance as many species are used in Ayurvedic and other traditional medicine preparations. This genus has received a lot of scholarly attention recently as a result of the numerous health advantages it possesses. In this review, we have compiled the scientific information regarding the relevance, distribution, industrial applications, phytochemistry, ethnopharmacology, tissue culture and conservation initiative of the Kaempferia genus along with the commercial realities and limitations of the research as well as missing industrial linkages followed by an exploration of some of the likely future promising clinical potential. The current review provides a richer and deeper understanding of Kaempferia, which can be applied in areas like phytopharmacology, molecular research, and industrial biology. The knowledge from this study can be further implemented for the establishment of new conservation strategies.

Comparative transcriptional analysis of metabolic pathways and mechanisms regulating essential oil biosynthesis in four elite Cymbopogon spp.

Cymbopogon is an important aromatic and medicinal grass with several species of ethnopharmaceutical importance. The genus is extremely rich in secondary metabolites, monoterpenes like geraniol and citral being principal constituents, also used as biomarker for classification and identification of Cymbopogon chemotypes. In the light of this, present study involved RNA sequencing and comparison of expression profiles of four contrasting Cymbopogon species namely C. flexuosus var. Chirharit (citral rich and frost resistant), C. martinii var. PRC-1 (geraniol rich), C. pendulus var. Praman (the most stable and citral-rich genotype), and Jamrosa (a hybrid of C. nardus var. confertiflorus × C. jwarancusa (rich in geraniol and geranyl acetate). The transcriptome profiles revealed marked differences in gene expression patterns of 28 differentially expressed genes (DEGs) of terpenoid metabolic pathways between the four Cymbopogon sp. The major DEGs were Carotenoid Cleavage Dioxygenases (CCD), Aspartate aminotransferase (ASP amino), Mevalonate E-4 hydroxy, AKR, GGPS, FDPS, and AAT. In addition, few TFs related to different regulatory pathways were also identified. The gene expression profiles of DEGs were correlated to the EO yield and their monoterpene compositions. Overall, the PRC-1 (C. martinii) shows distinguished gene expression profiles from all other genotypes. Thus, the transcriptome sequence database expanded our understanding of terpenoid metabolism and its molecular regulation in Cymbopogon species. Additionally, this data also serves as an important source of knowledge for enhancing oil yield and quality in Cymbopogon and closely related taxa. KEY MESSAGE: Unfolding the new secretes surrounding EO biosynthesis and regulation in four contrasting Cymbopogon species.

Interspecific and intraspecific analysis of Selinum spp. collected from Indian Himalayas using DNA barcoding.

BACKGROUND: DNA barcoding is a powerful method for phylogenetic mapping and species identification. However, recent research has come to a consistent conclusion about the universality of DNA barcoding. We used matK and rbcL markers to test the universality of twelve accessions from different locations belonging to two Selinum species, Selinum tenuifolium Wall. C. B. Clarke and Selinum vaginatum C. B. Clarke, keeping in mind their ability to identify species and establish phylogenetic relationships within and between the accessions. RESULTS: The success rates of PCR amplification using matK and rbcL were 75.26% ± 3.65% and 57.24% ± 4.42%, and the rate of DNA sequencing was 63.84% ± 4.32% and 50.82% ± 4.36%, respectively, suggesting that success rates of species identification of the two fragments were higher than 41.00% (matK, 41.50% ± 2.81%; rbcL, 42.88% ± 2.59%), proving that these fragments might be used to identify species. The best evolutionary tree with good supporting values was produced utilizing combinations of matK + rbcL markers when phylogenetic relationships were built with random fragment combinations. The twelve accessions of Selinum collected from different locations and their molecular sequences of matK and rbcL markers were blasted with other genera of Apiaceae family, and it was found that Selinum is most closely related to Angelica species of Apiaceae family. CONCLUSION: The present study has grouped twelve accessions of Selinum species using molecular markers into phylogenies, which is first-of-its-kind report that established interrelationships within different species of Apiaceae with respect to Selinum.

A Review of Chemistry and Pharmacology of Piperidine Alkaloids of Pinus and Related Genera.

BACKGROUND: Pinus belongs to the family Pinaceae, represented by several species across the globe. Various parts of the plant including needles are rich in biologically active compounds, such as thunbergol, 3-carene, cembrene, α-pinene, quercetin, xanthone. Of all the alkaloids, the piperidine group is one of the important component and holds considerable medicinal importance. METHODS: The group of alkaloids was initially identified from the genus Piper through which a large variety of piperidine molecules have been extracted. The planar structure of this heterocyclic nucleus enables acetamide groups to be added at various ring configurations. RESULTS: Piperidines have gained considerable importance. The broad range of its therapeutic application has paved a way for researchers to implant the nucleus from time to time in diversified pharmacophores and establish new profile. DISCUSSION: Biological functions of piperidine metabolites have been mostly examined on a limited scale, and that most of the findings are preliminary. We have tried to present various clinical applications of piperidine alkaloids in this study that researchers have already attempted to demystify with time. CONCLUSION: We have also illustrated different types of piperidine structures and their sources in different members of the family Pinaceae with special emphasis on Pinus. Given the importance of the piperidine nucleus, the study will enable the researchers to produce scaffolds of highest therapeutic efficacy.

Biomarker-based evaluation of cytogenotoxic potential of glyphosate in Vigna mungo (L.) Hepper genotypes.

Herbicides have proven to be a boon for agricultural fields. Their inherent property to kill weeds and unwanted vegetation makes them an essential biological tool for farmers and agricultural systems. Besides being capable of destroying weeds, they also exhibit certain effects on non-target crop plants. In the present study, a laboratory experiment was performed to assess the effect of glyphosate on Vigna mungo root meristem cells. Seeds of five different genotypes of V. mungo were treated with a series of concentrations of glyphosate ranging from 1 to 10 mM, and their effects on mitotic cell division were studied. Healthy and uniform-sized seeds were selected and were allowed to grow in Petri plates for 3 days, and all the doses were maintained in triplicates. Roots were fixed at day 3 after treatment (DAT) for cytological microscopic slide preparation. The results obtained indicate the dose-dependent reduction in the mitotic index in all the genotypes and an increase in the percentage of chromosomal aberrations (CAs) and relative abnormality rate (RAR). Most commonly observed chromosome aberrations at lower doses (< 6 mM) were fragments, stickiness, and disoriented metaphase, while at higher doses (6 to 10 mM) bridges, laggards, spindle disorientation, and clumping were obvious. The increase in the percentage of CAs and RAR indicates the inhibitory effect of glyphosate on cell cycle progression at various stages in root tip cells. The present study is a fine example of a biomarker-based genotoxic assessment of mitotic damage caused by glyphosate.

RNAi-based gene silencing in Phenacoccus solenopsis and its validation by in planta expression of a double-stranded RNA.

BACKGROUND: Cotton is a cash crop majorly affected by many hemipteran pests, among them the cotton mealybug, Phenacoccus solenopsis. Cotton mealybug attack has a devastating effect on cotton production and causes huge yield losses. RESULTS: In this study, 25 potential RNA interference (RNAi) target genes were selected from the iBeetle database and a transcriptome data set for P. solenopsis. To assess the effectiveness of the selected target genes, three methods were utilized to deliver double-stranded (ds)RNA (ingestion, artificial diet bioassay and transient gene silencing). dsRNA molecules at different concentrations were fed to insects and insect mortality was recorded for each target gene. Based on the mortality data, three genes, Krüppel homologue-1, ADP-ATP/Translocase and IDGF-1, were selected for further gene expression studies using a reduced concentration of dsRNA (5 µg/ml). Of the three genes, Krüppel homologue-1 showed significantly downregulated expression (by 70.81% and 84.33%) at two different time points (8 and 14 days). An RNAi silencing construct was designed for Krüppel homologue-1 under control of the double enhancer CamV35S promoter in the plant binary vector. Significant downregulation of gene expression, by 66.69% and 81.80%, was found for Krüppel homologue-1 using transient gene silencing at the same time intervals. CONCLUSION: This work provides the first evidence for targeting the Krüppel homologue-1 gene in a hemipteran pest, P. solenopsis, using RNAi technology through oral delivery and in planta-based transient gene silencing methods. © 2020 Society of Chemical Industry.

Effect of virus infection on the secondary metabolite production and phytohormone biosynthesis in plants.

Plants have evolved according to their environmental conditions and continuously interact with different biological entities. These interactions induce many positive and negative effects on plant metabolism. Many viruses also associate with various plant species and alter their metabolism. Further, virus-plant interaction also alters the expression of many plant hormones. To overcome the biotic stress imposed by the virus's infestation, plants produce different kinds of secondary metabolites that play a significant role in plant defense against the viral infection. In this review, we briefly highlight the mechanism of virus infection, their influence on the plant secondary metabolites and phytohormone biosynthesis in response to the virus-plant interactions.

Ethnobotany and Pharmacology of Pinus Species Growing Naturally in Indian Himalayas: A Plant Review.

Pinus species are amongst the most commonly used medicinal plants in Indian Northwestern Himalayas. Nearly all parts of the plant especially needles are believed to possess medicinal and clinical values in traditional systems of medicine such as Ayurvedic, Unani or Homeopathic. They are used to heal numerous diseases, including afflictions of the skin, eyes, throats and ears. Various parts of the plant including needles are rich in biologically active compounds, such as thunbergol, 3-carene, cembrene, α-pinene, quercetin, xanthone, etc. The scientific data available for the biological potential of this plant and its constituents have been found to be scanty and also does not satisfy the basis of their age-old folklore and local uses. This review focuses on a detailed analysis of ethnopharmacognosical studies on the needles of different Pinus species growing naturally in northwestern region of Indian Himalayas including investigations and findings that have come up with a concrete view of the abilities of plant components for example phenolics, flavonoids, tannins and other constituents to treat oxidative, inflammatory and microbial responses. Careful evaluation of the data presented may be helpful for the researchers to discover and evaluate the specific chemical entities responsible for the traditional medicinal uses of P. roxburghii, P.wallichiana, P. merkusii, P.kesiya and P.thunbergii.

Status of Selinum spp. L. a Himalayan Medicinal Plant in India: A Review of Its Pharmacology, Phytochemistry and Traditional Uses.

There is a growing trend of herbal medicines in India as well as other parts of the world. The use of herbal medicines alone or alongside prescription drugs for disease management is quite common now. We hereby carry out a review of dominant species of Selinum mainly S. vaginatum C.B. Clarke and S. wallichianum (DC.) Raizada & H.O. Saxena which are potent source of herbal medicine and whose potential is still not exploited. They are found in Northern Himalayas in the states of Kashmir, Himachal Pradesh and Uttarakhand in India, and also distributed in adjoining countries like China, Nepal, and West Pakistan. It is a perennial, primitive, high altitude, therapeutic herbal plant belonging to family Apiaceae (Umbelliferae) traditionally used for various diseases like epilepsy, seizures, and hysteria, etc. The aim of this review was to provide a summary on botanical characterization, traditional uses, phytochemistry and pharmacological activity of Selinum spp. The available information will be further tapped for commercial use of this plant without endangering its status. Based on the available evidence on the species pharmacology and chemistry, we highlight in which their therapeutic potential can be properly harnessed for possible integration into the country's healthcare system and be further passed on globally.

Recombinant Plant Engineering for Immunotherapeutic Production.

PURPOSE OF REVIEW: The requirement for large quantities of therapeutic proteins has fueled a great interest in the production of recombinant proteins in plant bioreactors. The vaccines and bio-therapeutic protein production in plants hold the promise of significantly lowering the cost of manufacturing life-saving drugs. This review will reflect the current status and challenges that the molecular farming platform faces becoming a strategic solution for the development of low-cost bio-therapeutics for developing countries. RECENT FINDINGS: Different plant parts have been successfully identified as suitable expression systems for the commercial production of therapeutic proteins for some human and animal diseases ranging from common cold to AIDS. The processed therapeutics from such sources are devoid of any toxic components. The large-scale cultivation of these transgenic plants would be possible anywhere in the world including developing countries, which lack sophisticated drug manufacturing units. A couple of such commercially generated products have already hit the market with success. Newer methods using suitable plant viruses and recombinant gene expression systems have already been devised for producing therapeutic proteins and peptides. SUMMARY: Plants are promising bio-factories for therapeutic protein production because of their several advantages over the other expression systems especially the advanced mechanisms for protein synthesis and post-translational modification which are very much similar to animal cells. Plant biotechnologists are much attracted to the bio-farming because of its flexibility, scalability, low manufacturing cost, as well as the lack of risk of toxic or pathogenic contamination. A number of projects on bio-farming are designed and are at various developmental stages but have not yet become available to the pharmaceutical industry. Therefore, we need further advancement in the optimization of lab protocols for up-scaling the production of such therapeutics at commercial level with a promise to offer their best clinical use.

Arsenic mediated modifications in Bacillus aryabhattai and their biotechnological applications for arsenic bioremediation.

The present study reports the arsenic (As) tolerance mechanism of bacteria Bacillus aryabhattai (NBRI014). The data explores the intracellular accumulation and volatilization of As from the culture medium after 48 h of exposure to 25,000 mg l-1 arsenate As(V). The study also provides the evidence of presence of ars operon in bacteria, which may have played an important role in reducing As toxicity. Additionally, we found 7 differentially expressed proteins to be up-regulated in bacterial cells upon As exposure which may have role in reducing As toxicity inside bacterial cells. Furthermore, Fourier transform infrared (FTIR) spectroscopic techniques were useful to describe the structural and compositional alterations in bacterial cells after As treatment. It showed the changes in peak positions of the spectrum pattern when NBRI014 was grown in medium containing As, indicating that these functional groups viz. (amino, alkyl halides and hydroxyl) present on bacterial surface, which may be involved in As binding. The above results signify that biotechnological application of the isolate NBRI014 could be helpful in removal of As from polluted sites.

Expression of an insecticidal fern protein in cotton protects against whitefly.

Whitefly (Bemisia tabaci) damages field crops by sucking sap and transmitting viral diseases. None of the insecticidal proteins used in genetically modified (GM) crop plants to date are effective against whitefly. We report the identification of a protein (Tma12) from an edible fern, Tectaria macrodonta (Fee) C. Chr., that is insecticidal to whitefly (median lethal concentration = 1.49 µg/ml in in vitro feeding assays) and interferes with its life cycle at sublethal doses. Transgenic cotton lines that express Tma12 at ∼0.01% of total soluble leaf protein were resistant to whitefly infestation in contained field trials, with no detectable yield penalty. The transgenic cotton lines were also protected from whitefly-borne cotton leaf curl viral disease. Rats fed Tma12 showed no detectable histological or biochemical changes, and this, together with the predicted absence of allergenic domains in Tma12, indicates that Tma12 might be well suited for deployment in GM crops to control whitefly and the viruses it carries.

Molecular Characterization of Vitellogenin and Vitellogenin Receptor of Bemisia tabaci.

Vitellogenin (Vg) plays vital role in oocytes and embryo development in insects. Vg is synthesized in the fat body, moves through haemolymph and accumulates in oocytes. Vitellogenin receptors (VgR) present on the surface of oocytes, are responsible for Vg transportation from haemolymph to oocytes. Here, we cloned and characterized these genes from Bemisia tabaci Asia1 (BtA1) species. The cloned BtA1Vg and BtA1VgR genes consisted of 6,330 and 5,430 bp long open reading frames, which encoded 2,109 and 1,809 amino acid (AA) residues long protein. The BtA1Vg protein comprised LPD_N, DUF1943 and VWFD domains, typical R/KXXR/K, DGXR and GL/ICG motifs, and polyserine tracts. BtA1VgR protein contained 12 LDLa, 10 LDLb and 7 EGF domains, and a trans-membrane and cytoplasmic region at C-terminus. Phylogenetic analyses indicated evolutionary association of BtA1Vg and BtA1VgR with the homologous proteins from various insect species. Silencing of BtA1VgR by siRNA did not affect the transcript level of BtA1Vg. However, BtA1Vg protein accumulation in oocytes was directly influenced with the expression level of BtA1VgR. Further, BtA1VgR silencing caused significant mortality and reduced fecundity in adult whiteflies. The results established the role of BtA1VgR in transportation of BtA1Vg in oocytes. Further, these proteins are essential for fecundity, and therefore these can be potential RNAi targets for insect control in crop plants.

Oral Rabies Vaccine Design for Expression in Plants.

Vaccination is the sensitization process of the immune system against any pathogen. Generally, recombinant subunit vaccines are considered safer than attenuated vaccines. As whole pathogenic organisms are used in the immunization process, the attenuated vaccines are considered more risky than subunit vaccines. Rabies is the oldest known zoonosis which spreads through a neurotropic Lyssavirus primarily mediated through infected canine bites. Rabies causes worldwide loss of more than 60,000 human lives every year. Animal vaccination is equally important to check the transmission of rabies into humans. Rabies oral vaccination can be a good alternative where multiple booster and priming regimens are required while the painful vaccination process can continue for long durations. Introduction of oral vaccines was made to ease the discomfort associated with the mode of introduction of conventional vaccines into the body. Although the rabies oral vaccine can substantially reduce the cost of vaccination in the developing countries, mass immunization programs need larger quantities of vaccines which should be delivered at nominal cost. Expression of recombinant antigen proteins in E. coli is often not viable because of lack of post-translational modifications and folding requirements. Though yeast and insect cell line expression systems have post-translational processing and modifications, significantly different immunological response against their post-translational modification pattern limits their deployment as an expression system. As an alternative, plants are emerging as a promising system to express and deliver wide range of functionally active biopharmaceutical product at lower cost for mass immunization programs. As generation of vaccine antigenic proteins in plant systems are cheaper, the strategy will benefit developing countries where this disease causes thousands of deaths every year. In this chapter, we will discuss about our efforts toward development of oral rabies vaccine and the methodological steps involved during this procedure in detail.