Evidence of polyamines mediated 2-acetyl-1-pyrroline biosynthesis in aromatic rice rhizospheric fungal species Aspergillus niger.

Rhizosphere soil of aromatic rice inhabits different fungal species that produce many bioactive metabolites including 2-acetyl-1-pyrroline (2AP). The mechanism for the biosynthesis of 2AP in the fungal system is still elusive. Hence, the present study investigates the role of possible nitrogen (N) precursors such as some amino acids and polyamines as well as the enzymes involved in 2AP synthesis in the fungal species isolated from the rhizosphere of aromatic rice varieties. Three fungal isolates were found to synthesize 2AP (0.32-1.07 ppm) and maximum 2AP was synthesized by Aspergillus niger (1.07 ppm) isolated from rhizosphere of Dehradun Basmati (DB). To determine the N source for 2AP synthesis, various N sources such as proline, glutamate, ornithine putrescine, spermine, and spermidine were used in place of putrescine in the synthetic medium (Syn18). The results showed that maximum 2AP synthesis was found with putrescine (1.07 ppm) followed by spermidine (0.89 ppm) and spermine (0.84 ppm). Further, LC-QTOF-MS analysis revealed the mobilization of spermine and spermidine into the putrescine, indicating that putrescine is the key N source for 2AP synthesis. Moreover, higher enzyme activity of DAO, PAO, and ODC as well as higher content of methylglyoxal metabolite in the A. niger NFCCI 5060 as compared to A. niger NFCCI 4064 (control) suggests the prominent role of these enzymes in the synthesis of 2AP. In conclusion, this study showed evidence of the polyamines mediated 2AP biosynthesis in A. niger NFCCI 5060.

Involvement of abscisic acid in silicon-mediated enhancement of copper stress tolerance in Artemisia annua.

Heavy metal (HM) toxicity is a well-known hazard which causes deleterious impact on the growth and development of plants. The impact of abscisic acid (ABA) in presence of silicon (Si) on plant development and quality traits has largely gone unexplored. The effects of ABA and Si on the growth, yield, and quality characteristics of Artemisia annua L. plants growing under copper (Cu) stress (20 and 40 mg kg-1) were investigated in a pot experiment. During this investigation, Cu stress caused severe damage to the plants but exogenous administration of Si and ABA ameliorated the harmful effects of Cu toxicity, and the plants displayed higher biomass and improved physio-biochemical attributes. Copper accumulated in the roots and shoots and its toxicity caused oxidative stress as demonstrated by the increased 2-thiobarbituric acid reactive substance (TBARS) content. It also resulted in the increased activity of antioxidant enzymes, however, the exogenous Si and ABA supplementation decreased the buildup of reactive oxygen species (ROS) and lipid peroxidation, alleviating the oxidative damage produced by HM stress. Copper toxicity had a considerable negative impact on glandular trichome density, ultrastructure as well as artemisinin production. However, combined Si and ABA enhanced the size and density of glandular trichomes, resulting in higher artemisinin production. Taken together, our results demonstrated that exogenous ABA and Si supplementation protect A. annua plants against Cu toxicity by improving photosynthetic characteristics, enhancing antioxidant enzyme activity, protecting leaf structure and integrity, avoiding excess Cu deposition in shoot and root tissues, and helping in enhanced artemisinin biosynthesis. Our results indicate that the combined application of Si and ABA improved the overall growth of plants and may thus be used as an effective approach for the improvement of growth and yield of A. annua in Cu-contaminated soils.

Suffer or Survive: Decoding Salt-Sensitivity of Lemongrass and Its Implication on Essential Oil Productivity.

The cultivation of lemongrass (Cymbopogon flexuosus) crop is dominated by its medicinal, food preservative, and cosmetic demands. The growing economy of the lemongrass market suggests the immense commercial potential of lemongrass and its essential oil. Nevertheless, the continuous increase of the saline regime threatens the growth and productivity of most of the plant life worldwide. In this regard, the present experiment explores the salt sensitiveness of the lemongrass crop against five different levels of salt stress. Metabolomic analyses suggest that lemongrass plants can effectively tolerate a salt concentration of up to 80 mM and retain most of their growth and productivity. However, extreme NaCl concentrations (≥160 mM) inflicted significant (α = 0.05) damage to the plant physiology and exhausted the lemongrass antioxidative defence system. Therefore, the highest NaCl concentration (240 mM) minimised plant height, chlorophyll fluorescence, and essential oil production by up to 50, 27, and 45%. The overall data along with the salt implications on photosynthetic machinery and ROS metabolism suggest that lemongrass can be considered a moderately sensitive crop to salt stress. The study, sensu lato, can be used in reclaiming moderately saline lands with lemongrass cultivation converting such lands from economic liability to economic asset.

Exogenous Strigolactone (GR24) Positively Regulates Growth, Photosynthesis, and Improves Glandular Trichome Attributes for Enhanced Artemisinin Production in Artemisia annua.

Artemisia annua is a medicinal plant particularly known for the production of a sesquiterpene lactone artemisinin; a specialty metabolite known for its efficacy in the treatment of malaria by killing different strains of Plasmodium falciparum due to radicals released upon the cleavage of its endoperoxide motif. Considering these facts and the immense medicinal value of artemisinin, the enhancement of in planta production of artemisinin is highly desirable. As strigolactones are known to regulate various aspects of plant growth and development, the effects of foliar spray of different concentrations of synthetic strigolactone analog GR24 (0, 0.5, 1, 2, 4, and 8 µM) on A. annua were studied. As compared to the control group, the foliar application of GR24 had a positive impact on general growth, photosynthesis, and other physiological indices with 4 µM GR24 showing the best results. The results indicate that GR24 application increased the plant biomass and various attributes related to photosynthesis, like total chlorophyll content, chlorophyll fluorescence, stomatal conductance, internal CO2, and net photosynthetic rate. Moreover, the activity of various enzymes related to photosynthesis like carbonic anhydrase, nitrate reductase, and RuBisCO was escalated. The GR24 also improved certain attributes related to glandular trichomes, with a significant enhancement in content and yield of artemisinin as compared to untreated plants.

Parenting practices, stressors and parental concerns during COVID-19 in Pakistan.

BACKGROUND: COVID-19 is a global crisis that has added fear, uncertainty, and stress to parents. Parents are going through several challenges related to school closure, financial insecurity and working remotely. These stressors are affecting the mental health of parents. OBJECTIVES: This study aimed to observe major stressors along with the impact of COVID-19 on parental concerns and practices during lockdown. PARTICIPANTS: Sample (N = 923) was selected through purposive sampling from parents attending Out Patients Departments of hospitals in three provincial capital cities of Pakistan having a high burden of COVID-19, i.e. Lahore, Karachi and Peshawar. Parents having at least one child younger than 18 years were included in the study. METHODS: A quantitative design was used using a COVID-19 Parenting Response Scale (α = 0.74). It was used as a self-administered tool for parents who knew how to read and write Urdu/English language, however it was conducted as a structured interview for those who could not read/write. Data was analyzed by applying descriptive statistics (frequency, mean, percentage), independent sample t-test and Pearson Product Moment Correlation. RESULTS: Findings of the current study showed several stressful factors for parents during COVID-19 pandemic, mainly financial burden, children's education, uncertainty of the situation, and many others. The study also suggests an association of parental concerns during COVID-19 with parenting practices. CONCLUSION: COVID-19 pandemic presents a global crisis not only of the health of the people but also on family relations and mental well-being. Findings of this research indicate the need for targeted and accessible interventions for mental health of parents especially during these challenging circumstances so that they can cope with the challenges in an effective way and be able to take care of their children better.

Efficiency of microbial bio-agents as elicitors in plant defense mechanism under biotic stress: A review.

Numerous harmful microorganisms and insect pests have the ability to cause plant infections or damage, which is mostly controlled by toxic chemical agents. These chemical compounds and their derivatives exhibit hazardous effects on habitats and human life too. Hence, there's a need to develop novel, more effective and safe bio-control agents. A variety of microbes such as viruses, bacteria, and fungi possess a great potential to fight against phytopathogens and thus can be used as bio-control agents instead of harmful chemical compounds. These naturally occurring microorganisms are applied to the plants in order to control phytopathogens. Moreover, practicing them appropriately for agriculture management can be a way towards a sustainable approach. The MBCAs follow various modes of action and act as elicitors where they induce a signal to activate plant defense mechanisms against a variety of pathogens. MBCAs control phytopathogens and help in disease suppression through the production of enzymes, antimicrobial compounds, antagonist activity involving hyper-parasitism, induced resistance, competitive inhibition, etc. Efficient recognition of pathogens and prompt defensive response are key factors of induced resistance in plants. This resistance phenomenon is pertaining to a complex cascade that involves an increased amount of defensive proteins, salicylic acid (SA), or induction of signaling pathways dependent on plant hormones. Although, there's a dearth of information about the exact mechanism of plant-induced resistance, the studies conducted at the physiological, biochemical and genetic levels. These studies tried to explain a series of plant defensive responses triggered by bio-control agents that may enhance the defensive capacity of plants. Several natural and recombinant microorganisms are commercially available as bio-control agents that mainly include strains of Bacillus, Pseudomonads and Trichoderma. However, the complete understanding of microbial bio-control agents and their interactions at cellular and molecular levels will facilitate the screening of effective and eco-friendly bio-agents, thereby increasing the scope of MBCAs. This article is a comprehensive review that highlights the importance of microbial agents as elicitors in the activation and regulation of plant defense mechanisms in response to a variety of pathogens.

Enhancing artemisinin content in and delivery from Artemisia annua: a review of alternative, classical, and transgenic approaches.

MAIN CONCLUSION: This review analyses the most recent scientific research conducted for the purpose of enhancing artemisinin production. It may help to devise better artemisinin enhancement strategies, so that its production becomes cost effective and becomes available to masses. Malaria is a major threat to world population, particularly in South-East Asia and Africa, due to dearth of effective anti-malarial compounds, emergence of quinine resistant malarial strains, and lack of advanced healthcare facilities. Artemisinin, a sesquiterpene lactone obtained from Artemisia annua L., is the most potent drug against malaria and used in the formulation of artemisinin combination therapies (ACTs). Artemisinin is also effective against various types of cancers, many other microbes including viruses, parasites and bacteria. However, this specialty metabolite and its derivatives generally occur in low amounts in the source plant leading to its production scarcity. Considering the importance of this drug, researchers have been working worldwide to develop novel strategies to augment its production both in vivo and in vitro. Due to complex chemical structure, its chemical synthesis is quite expensive, so researchers need to devise synthetic protocols that are economically viable and also work on increasing the in-planta production of artemisinin by using various strategies like use of phytohormones, stress signals, bioinoculants, breeding and transgenic approaches. The focus of this review is to discuss these artemisinin enhancement strategies, understand mechanisms modulating its biosynthesis, and evaluate if roots play any role in artemisinin production. Furthermore, we also have a critical analysis of various assays used for artemisinin measurement. This may help to develop better artemisinin enhancement strategies which lead to decreased price of ACTs and increased profit to farmers.

Endophytic Nanotechnology: An Approach to Study Scope and Potential Applications.

Nanotechnology has become a very advanced and popular form of technology with huge potentials. Nanotechnology has been very well explored in the fields of electronics, automobiles, construction, medicine, and cosmetics, but the exploration of nanotecnology's use in agriculture is still limited. Due to climate change, each year around 40% of crops face abiotic and biotic stress; with the global demand for food increasing, nanotechnology is seen as the best method to mitigate challenges in disease management in crops by reducing the use of chemical inputs such as herbicides, pesticides, and fungicides. The use of these toxic chemicals is potentially harmful to humans and the environment. Therefore, using NPs as fungicides/ bactericides or as nanofertilizers, due to their small size and high surface area with high reactivity, reduces the problems in plant disease management. There are several methods that have been used to synthesize NPs, such as physical and chemical methods. Specially, we need ecofriendly and nontoxic methods for the synthesis of NPs. Some biological organisms like plants, algae, yeast, bacteria, actinomycetes, and fungi have emerged as superlative candidates for the biological synthesis of NPs (also considered as green synthesis). Among these biological methods, endophytic microorganisms have been widely used to synthesize NPs with low metallic ions, which opens a new possibility on the edge of biological nanotechnology. In this review, we will have discussed the different methods of synthesis of NPs, such as top-down, bottom-up, and green synthesis (specially including endophytic microorganisms) methods, their mechanisms, different forms of NPs, such as magnesium oxide nanoparticles (MgO-NPs), copper nanoparticles (Cu-NPs), chitosan nanoparticles (CS-NPs), ß-d-glucan nanoparticles (GNPs), and engineered nanoparticles (quantum dots, metalloids, nonmetals, carbon nanomaterials, dendrimers, and liposomes), and their molecular approaches in various aspects. At the molecular level, nanoparticles, such as mesoporous silica nanoparticles (MSN) and RNA-interference molecules, can also be used as molecular tools to carry genetic material during genetic engineering of plants. In plant disease management, NPs can be used as biosensors to diagnose the disease.

Salicylic acid-mediated alleviation of soil boron toxicity in Mentha arvensis and Cymbopogon flexuosus: Growth, antioxidant responses, essential oil contents and components.

Boron (B) toxicity is a notable abiotic hindrance that restricts crop productivity by disturbing several physiological and biochemical processes in plants. This study was aimed to elucidate the role of salicylic acid (SA) in conferring tolerance to B stress in Mentha arvensis and Cymbopogon flexuosus. Boron toxicity led to a considerable decrease in shoot height and root length, fresh and dry mass of shoot and root, and physiological and biochemical parameters. However, exogenously applied SA relieved the adverse effects caused by B toxicity and led to an increase in growth parameters under B stress and non-stress conditions. The treatment of B resulted in its increased accumulation in roots and shoots of both the plants which, in turn, caused oxidative damage as evident by increased content of malondialdehyde and catalase, peroxidase, superoxide dismutase and glutathione reductase enzyme activities. However, exogenous SA supply significantly affected antioxidant enzyme activities and protected the plants from excess B. Moreover, the essential oil content of two selected plants declined under B toxicity and significantly enhanced in SA-treated stressed plants. The contents of menthol and menthyl acetate in M. arvensis were lowered in B stressed plants which significantly improved in SA treated B-stressed and in their respective SA alone treatment. Similarly, citral-A and citral-B content of C. flexuosus declined under B toxicity, however, SA reversed the negative effects of B toxicity on essential oil components. This assessment stipulated the promising role of exogenously applied SA in alleviating B toxicity in M. arvensis and C. flexuosus by improving antioxidant machinery and limiting B uptake which protects the structural integrity of leaves and also helps in increasing essential oil content.

Professional team response to violence against children: From experts to teamwork.

BACKGROUND: Prevention and management of violence against children requires a multidisciplinary approach. Countries and regions tackle this problem according to their understanding and capacity, and therefore these teams work differently in different countries and regions. OBJECTIVES: Current study is a bird's eye view looking at the response to violence against children through exploring multidisciplinary approaches and challenges in child protection in high, upper-middle, and lower-middle-income countries. PARTICIPANTS: Participants for this study were selected through purposive sampling from eleven countries, one key informant from each country (N = 11). METHODS: Current study is qualitative in nature and phenomenological design was used. Key informants were approached through email, and semi-structured online interviews were conducted. Thematic analysis was used to analyze the data and several themes emerged. RESULTS: Capacity building systems are established in upper-middle and high-income countries, whereas lower-income countries are struggling with these systems, the Philippines having a better structured system. The process of child protection services is mostly similar except the availability of multidisciplinary teams, which lacks in lower-middle and even in upper-middle-income countries. These teams function relatively well in high-income countries although the lead role in these teams differs from country to country. There are many challenges in managing these teams in low-middle income countries. Child protection systems are also reasonably well established in upper and upper-middle-income countries whereas gaps are visible between policies and implementation in lower-middle-income countries. CONCLUSION: It is the responsibility of each country to protect children as per their commitment to SDGs. Although the functioning of multidisciplinary teams seems better in high and upper-middle countries, the low-middle income countries are trying to combat violence against children according to their priorities, some are doing better than others.

Understanding youth participation to volunteer for child rights.

BACKGROUND: Today, Pakistan has the highest percentage of young people in its history, who can become champions of child rights through channelizing their energy. Voices for Children's Aspirations and Needs (VCAN) student society at the University of Lahore is one such student body, working to promote child rights and eliminate violence against children. OBJECTIVES: The study identifies factors contributing to young people's motivation to join VCAN student society working for children's rights and influence their participation in activities, along with attitudinal shift. The current study also attempts to find out effective mediums to engage youth in volunteering activities for the prevention of violence against children. PARTICIPANTS: Sample (N = 40) was selected through purposive sampling from the VCAN Student Society at the University of Lahore. METHOD: The data were collected through a questionnaire developed by the researchers (α = 0.86). Collected data were analyzed by finding descriptive statistics (mean, percentage and frequency) and paired samplet-test using statistics software, SPSS, version 21.0. RESULTS: Motivation for joining student society included wanting to work for the betterment of children (57.5 %) and to volunteer for the prevention of violence against children (47.5 %). There was a statistically significant difference (t=-4.16, p < .001) in the perception of participants about child rights before and after joining the society. Factors such as lack of required knowledge, unclear roles, and contrast between expected role and skillset (45.0 %, 42.5 %, and 40.0 % respectively) discouraged youth participation. 37.5 % of the participants considered WhatsApp as an extremely effective way to engage youth whereas 32.5 % reported Facebook as an extremely effective medium for the same purpose. CONCLUSION: Student societies provide a great platform for students to be active for a social cause. Educational institutions should encourage and facilitate youth participation for such causes.

Exogenous abscisic acid mediates ROS homeostasis and maintains glandular trichome to enhance artemisinin biosynthesis in Artemisia annua under copper toxicity.

One of the major abiotic stresses that cause environmental pollution is heavy metal stress. In the present investigation, copper (Cu) toxicity caused morphological and cellular damages to the Artemisia annua L. plants but supplementation of abscisic acid (ABA) ameliorated the damaging effect of Cu. Copper toxicity significantly reduced the shoot and root lengths; fresh and dry weights of shoot. However, exogenous application of ABA to Cu-treated plants significantly attenuated the damaging effects on plants caused by Cu toxicity. Copper stress also reduced the physiological and biochemical parameters, but ABA application ameliorated the negative effects of Cu in the affected plant. Accumulation of Cu in plant tissues significantly increased the membrane damage and oxidative enzyme activities such as catalase (CAT), peroxidase (POX) and superoxide dismutase (SOD). Further, the impact of high concentration of Cu on density, area and ultrastructure of glandular trichomes and artemisinin content was studied. Moreover, the foliar application of ABA improved the area, density of glandular trichomes and secured the plant cells from Cu toxicity. Therefore, this investigation indicated that the exogenous application of ABA protects A. annua plant by increasing antioxidant enzymes activity, which helps in maintaining cell integrity of leaves and results in increased artemisinin production.

PGPR-mediated induction of systemic resistance and physiochemical alterations in plants against the pathogens: Current perspectives.

Plant growth-promoting rhizobacteria (PGPR) are diverse groups of plant-associated microorganisms, which can reduce the severity or incidence of disease during antagonism among bacteria and soil-borne pathogens, as well as by influencing a systemic resistance to elicit defense response in host plants. An amalgamation of various strains of PGPR has improved the efficacy by enhancing the systemic resistance opposed to various pathogens affecting the crop. Many PGPR used with seed treatment causes structural improvement of the cell wall and physiological/biochemical changes leading to the synthesis of proteins, peptides, and chemicals occupied in plant defense mechanisms. The major determinants of PGPR-mediated induced systemic resistance (ISR) are lipopolysaccharides, lipopeptides, siderophores, pyocyanin, antibiotics 2,4-diacetylphoroglucinol, the volatile 2,3-butanediol, N-alkylated benzylamine, and iron-regulated compounds. Many PGPR inoculants have been commercialized and these inoculants consequently aid in the improvement of crop growth yield and provide effective reinforcement to the crop from disease, whereas other inoculants are used as biofertilizers for native as well as crops growing at diverse extreme habitat and exhibit multifunctional plant growth-promoting attributes. A number of applications of PGPR formulation are needed to maintain the resistance levels in crop plants. Several microarray-based studies have been done to identify the genes, which are associated with PGPR-induced systemic resistance. Identification of these genes associated with ISR-mediating disease suppression and biochemical changes in the crop plant is one of the essential steps in understanding the disease resistance mechanisms in crops. Therefore, in this review, we discuss the PGPR-mediated innovative methods, focusing on the mode of action of compounds authorized that may be significant in the development contributing to enhance plant growth, disease resistance, and serve as an efficient bioinoculants for sustainable agriculture. The review also highlights current research progress in this field with a special emphasis on challenges, limitations, and their environmental and economic advantages.

Impact of Long-Term Copper Exposure on Growth, Photosynthesis, Antioxidant Defence System and Artemisinin Biosynthesis in Soil-Grown Artemisia annua Genotypes.

The effects of copper (Cu) exposure on growth and physiological characteristics of three genotypes (CN-12, Cim-Sanjeevani and Cim-Arogya) of Artemisia annua L. were elucidated. The plants were grown under naturally illuminated greenhouse conditions and were harvested after physiological maturity (120 days after sowing). Results suggest that 10 mg kg- 1 Cu significantly enhanced the growth and physiological parameters like enzyme activities, photosynthesis. At higher concentrations, Cu inhibited the growth, biomass, photosynthetic parameters; while increased lipid peroxidation in all the genotypes. The activities of antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase were upregulated by the Cu stress. The highest applied concentration of Cu (60 mg kg- 1) proved most toxic for plants. Moreover, artemisinin content was increased upto 10 mg kg- 1 of Cu treatment, compared with control, however, the artemisinin accumulation decreased at higher doses of Cu in all the genotypes. On the basis of studied parameters, Cim-Arogya was found to be most tolerant among all for Cu toxicity.

Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions.

BACKGROUND: In response to various environmental stresses, many plant species synthesize L-proline in the cytosol and accumulates in the chloroplasts. L-Proline accumulation in plants is a well-recognized physiological reaction to osmotic stress prompted by salinity, drought and other abiotic stresses. L-Proline plays several protective functions such as osmoprotectant, stabilizing cellular structures, enzymes, and scavenging reactive oxygen species (ROS), and keeps up redox balance in adverse situations. In addition, ample-studied osmoprotective capacity, L-proline has been also ensnared in the regulation of plant improvement, including flowering, pollen, embryo, and leaf enlargement. SCOPE AND CONCLUSIONS: Albeit, ample is now well-known about L-proline metabolism, but certain characteristics of its biological roles are still indistinct. In the present review, we discuss the L-proline accumulation, metabolism, signaling, transport and regulation in the plants. We also discuss the effects of exogenous L-proline during different environmental conditions. L-Proline biosynthesis and catabolism are controlled by several cellular mechanisms, of which we identify only very fewer mechanisms. So, in the future, there is a requirement to identify such types of cellular mechanisms.

Trichoderma erinaceum Bio-Priming Modulates the WRKYs Defense Programming in Tomato Against the Fusarium oxysporum f. sp. lycopersici (Fol) Challenged Condition.

The beneficial association and interaction of rhizocompetent microorganisms are widely used for plant biofertilization and amelioration of stress-induced damage in plants. To explore the regulatory mechanism involved in plant defense while associating with beneficial microbial species, and their interplay when co-inoculated with pathogens, we evaluated the response of tomato defense-related WRKY gene transcripts. The present study was carried out to examine the qRT-PCR-based relative quantification of differentially expressed defense-related genes in tomato (Solanum lycopersicum L.; variety S-22) primed with Trichoderma erinaceum against the vascular wilt pathogen (Fusarium oxysporum f. sp. lycopersici). The tissue-specific and time-bound expression profile changes under the four different treatments "(unprimed, Fol challenged, T. erinaceum primed and Fol+ T. erinaceum)" revealed that the highest upregulation was observed in the transcript profile of SlWRKY31 (root) and SlWRKY37 (leaf) in T. erinaceum bioprimed treated plants at 24 h with 16.51- and 14.07-fold increase, respectively. In contrast, SlWRKY4 showed downregulation with the highest repression in T. erinaceum bioprimed root (24 h) and leaf (48 h) tissue samples with 0.03 and 0.08 fold decrease, respectively. Qualitative expression of PR proteins (chitinases and glucanases) was found elicited in T. erinaceum primed plants. However, the antioxidative activity of tomato superoxide dismutase and catalase increased with the highest upregulation of SOD and SlGPX1 in Fol + T. erinaceum treatments. We observed that these expression changes were accompanied by 32.06% lesser H2O2 production in T. erinaceum bioprimed samples. The aggravated defense response in all the treated conditions was also reflected by an increased lignified stem tissues. Overall, we conclude that T. erinaceum bio-priming modulated the defense transcriptome of tomato after the Fol challenged conditions, and were accompanied by enhanced accumulation of defense-related WRKY transcripts, increased antioxidative enzyme activities, and the reinforcements through a higher number of lignified cell layers.

Structural and functional dissection of differentially expressed tomato WRKY transcripts in host defense response against the vascular wilt pathogen (Fusarium oxysporum f. sp. lycopersici).

The WRKY transcription factors have indispensable role in plant growth, development and defense responses. The differential expression of WRKY genes following the stress conditions has been well demonstrated. We investigated the temporal and tissue-specific (root and leaf tissues) differential expression of plant defense-related WRKY genes, following the infection of Fusarium oxysporum f. sp. lycopersici (Fol) in tomato. The genome-wide computational analysis revealed that during the Fol infection in tomato, 16 different members of WRKY gene superfamily were found to be involved, of which only three WRKYs (SolyWRKY4, SolyWRKY33, and SolyWRKY37) were shown to have clear-cut differential gene expression. The quantitative real time PCR (qRT-PCR) studies revealed different gene expression profile changes in tomato root and leaf tissues. In root tissues, infected with Fol, an increased expression for SolyWRKY33 (2.76 fold) followed by SolyWRKY37 (1.93 fold) gene was found at 24 hrs which further increased at 48 hrs (5.0 fold). In contrast, the leaf tissues, the expression was more pronounced at an earlier stage of infection (24 hrs). However, in both cases, we found repression of SolyWRKY4 gene, which further decreased at an increased time interval. The biochemical defense programming against Fol pathogenesis was characterized by the highest accumulation of H2O2 (at 48 hrs) and enhanced lignification. The functional diversity across the characterized WRKYs was explored through motif scanning using MEME suite, and the WRKYs specific gene regulation was assessed through the DNA protein docking studies The functional WRKY domain modeled had ß sheets like topology with coil and turns. The DNA-protein interaction results revealed the importance of core residues (Tyr, Arg, and Lys) in a feasible WRKY-W-box DNA interaction. The protein interaction network analysis revealed that the SolyWRKY33 could interact with other proteins, such as mitogen-activated protein kinase 5 (MAPK), sigma factor binding protein1 (SIB1) and with other WRKY members including WRKY70, WRKY1, and WRKY40, to respond various biotic and abiotic stresses. The STRING results were further validated through Predicted Tomato Interactome Resource (PTIR) database. The CELLO2GO web server revealed the functional gene ontology annotation and protein subcellular localization, which predicted that SolyWRKY33 is involved in amelioration of biological stress (39.3%) and other metabolic processes (39.3%). The protein (SolyWRKY33) most probably located inside the nucleus (91.3%) with having transcription factor binding activity. We conclude that the defense response following the Fol challenge was accompanied by differential expression of the SolyWRKY4(↓), SolyWRKY33(↑) and SolyWRKY37(↑) transcripts. The biochemical changes are occupied by elicitation of H2O2 generation and accumulation and enhanced lignified tissues.

Synergistic effects of plant defense elicitors and Trichoderma harzianum on enhanced induction of antioxidant defense system in tomato against Fusarium wilt disease.

Plant defense against their pathogens can be induced by a complex network of different inducers. The present study investigates the synergistic effect of Trichoderma harzianum, exogenous salicylic acid (SA) and methyl jasmonate (MeJA) over the response and regulation of the antioxidant defense mechanisms and lipid peroxidation in tomato plants against Fusarium wilt disease. In the present work, tomato plants were infected by Fusarium oxysporum f. sp. lycopersici 3 days after inoculated with T. harzianum and/or sprayed daily for 3 days with chemical inducers (SA and MeJA). Plants were analysed at 0, 24, 48, 72 and 96 h after inoculation with Fusarium oxysporum f. sp. lycopersici. Infection of tomato plants by pathogen led to strong reduction in the dry weight of roots and shoots with the enhanced concentration of H2O2 and varying degree of lipid peroxidation. Concurrently, exogenous SA, when applied with pathogen greatly enhanced H2O2 content as well as activities of antioxidant enzymes except catalase (CAT) and ascorbate peroxidase (APx). The pathogen challenged plants pretreated with T. harzianum and MeJA together exhibited less lipid peroxidation and as well as the elevated level of ascorbic acid and enhanced activities of antioxidant enzymes. All applied treatments protected tomato seedlings against Fusarium wilt disease but the percentage of protection was found higher in plants pretreated with the combination of T. harzianum and chemical inducers.

Alternaria Toxins: Potential Virulence Factors and Genes Related to Pathogenesis.

Alternaria is an important fungus to study due to their different life style from saprophytes to endophytes and a very successful fungal pathogen that causes diseases to a number of economically important crops. Alternaria species have been well-characterized for the production of different host-specific toxins (HSTs) and non-host specific toxins (nHSTs) which depend upon their physiological and morphological stages. The pathogenicity of Alternaria species depends on host susceptibility or resistance as well as quantitative production of HSTs and nHSTs. These toxins are chemically low molecular weight secondary metabolites (SMs). The effects of toxins are mainly on different parts of cells like mitochondria, chloroplast, plasma membrane, Golgi complex, nucleus, etc. Alternaria species produce several nHSTs such as brefeldin A, tenuazonic acid, tentoxin, and zinniol. HSTs that act in very low concentrations affect only certain plant varieties or genotype and play a role in determining the host range of specificity of plant pathogens. The commonly known HSTs are AAL-, AK-, AM-, AF-, ACR-, and ACT-toxins which are named by their host specificity and these toxins are classified into different family groups. The HSTs are differentiated on the basis of bio-statistical and other molecular analyses. All these toxins have different mode of action, biochemical reactions and signaling mechanisms to cause diseases. Different species of Alternaria produced toxins which reveal its biochemical and genetic effects on itself as well as on its host cells tissues. The genes responsible for the production of HSTs are found on the conditionally dispensable chromosomes (CDCs) which have been well characterized. Different bio-statistical methods like basic local alignment search tool (BLAST) data analysis used for the annotation of gene prediction, pathogenicity-related genes may provide surprising knowledge in present and future.

Improvement Strategies, Cost Effective Production, and Potential Applications of Fungal Glucose Oxidase (GOD): Current Updates.

Fungal glucose oxidase (GOD) is widely employed in the different sectors of food industries for use in baking products, dry egg powder, beverages, and gluconic acid production. GOD also has several other novel applications in chemical, pharmaceutical, textile, and other biotechnological industries. The electrochemical suitability of GOD catalyzed reactions has enabled its successful use in bioelectronic devices, particularly biofuel cells, and biosensors. Other crucial aspects of GOD such as improved feeding efficiency in response to GOD supplemental diet, roles in antimicrobial activities, and enhancing pathogen defense response, thereby providing induced resistance in plants have also been reported. Moreover, the medical science, another emerging branch where GOD was recently reported to induce several apoptosis characteristics as well as cellular senescence by downregulating Klotho gene expression. These widespread applications of GOD have led to increased demand for more extensive research to improve its production, characterization, and enhanced stability to enable long term usages. Currently, GOD is mainly produced and purified from Aspergillus niger and Penicillium species, but the yield is relatively low and the purification process is troublesome. It is practical to build an excellent GOD-producing strain. Therefore, the present review describes innovative methods of enhancing fungal GOD production by using genetic and non-genetic approaches in-depth along with purification techniques. The review also highlights current research progress in the cost effective production of GOD, including key advances, potential applications and limitations. Therefore, there is an extensive need to commercialize these processes by developing and optimizing novel strategies for cost effective GOD production.