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Sugarcane ( Saccharum ssp., Poaceae) provides enormous metabolites such as sugars, lipid, and other dietary metabolites to humans. Among them, lipids are important metabolites that perform various functions and have promising pharmacological value. However, in sugarcane, few studies are focusing on lipidomics and few lipid compounds were reported, and their pharmacological values are not explored yet. The transcriptomic and widely targeted lipidomics approach quantified 134 lipid compounds from the rind of six sugarcane genotypes. These lipid compounds include 57 fatty acids, 30 lysophosphatidylcholines, 23 glycerol esters, 21 lysophosphatidylethanolamines, 2 phosphatidylcholines, and 1 sphingolipid. Among them, 119 compounds were first time reported in sugarcane rind. Seventeen lipids compounds including 12 fatty acids, 2 glycerol lipids, LysoPC 16:0, LysoPE 16:0, and choline alfoscerate were abundantly found in the rind of sugarcane genotypes. From metabolic and transcriptomic results, we have developed a comprehensive lipid metabolic pathway and highlighted key genes that are differentially expressed in sugarcane. Several genes associated with α-linolenic acid and linoleic acid biosynthesis pathways were highly expressed in the rind of the ROC22 genotype. ROC22 has a high level of α-linolenic acid (an essential fatty acid) followed by ROC16. Moreover, we have explored pharmacological values of lipid compounds and found that the 2-linoleoylglycerol and gingerglycolipid C have strong binding interactions with 3CL pro of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) and these compounds can be utilized against SARS-CoV-2 as therapeutic agents. The transcriptome, metabolome, and bioinformatics analysis suggests that the sugarcane cultivars have a diversity of lipid compounds having promising therapeutic potential, and exploring the lipid metabolism will help to know more compounds that have promising cosmetic and pharmacological value., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rao, Tahir ul Qamar, Wang, Ali, Ma, Han, Duan, Hu and Wang.)

Soil salinity is a major environmental stress that has been negatively affecting the growth and productivity of rice. However, various salt-resistant plant growth-promoting rhizobacteria (PGPR) have been known to promote plant growth and alleviate the damaging effects of salt stress via mitigating physio-biochemical and molecular characteristics. This study was conducted to examine the salt stress potential of Bacillus strains identified from harsh environments of the Qinghai-Tibetan plateau region of China. The Bacillus strains NMTD17, GBSW22, and FZB42 were screened for their response under different salt stress conditions (1, 4, 7, 9, 11, 13, and 16%). The screening analysis revealed strains NMTD17, GBSW22, and FZB42 to be high-salt tolerant, moderate-salt tolerant, and salt-sensitive, respectively. The NMTD17 strain produced a strong biofilm, followed by GBSW22 and FZB42. The expression of salt stress-related genes in selected strains was also analyzed through qPCR in various salt concentrations. Further, the Bacillus strains were used in pot experiments to study their growth-promoting ability and antioxidant activities at various concentrations (0, 100, 150, and 200 mmol). The analysis of growth-promoting traits in rice exhibited that NMTD17 had a highly significant effect and GSBW22 had a moderately significant effect in comparison with FZB42. The highly resistant strain NMTD17 that stably promoted rice plant growth was further examined for its function in the composition of rhizobacterial communities. The inoculation of NMTD17 increased the relative abundance and richness of rhizobacterial species. These outcomes propose that NMTD17 possesses the potential of PGPR traits, antioxidants enzyme activities, and reshaping the rhizobacterial community that together mitigate the harmful effects of salinity in rice plants., Competing Interests: GM was employed by Shenzhen Batian Ecotypic Engineering Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ali, Ayaz, Mu, Hussain, Yuanyuan, Yu, Xu, Manghwar, Gu, Wu and Gao.)

Heavy metal remediation, such as cadmium (Cd 2+ ) by microbial strains is efficient and environment-friendly. In this current study, we exploited the potential of Bacillus strains (Cd 2+ -tolerant; NMTD17, GBSW22, and LLTC96) to regulate Cd 2+ biosorption mechanisms and improve rice seedling growth. The results showed that initial concentration and contact time affected Cd 2+ biosorption, and the kinetic models of pseudo orders were effective in the elaborate biosorption process. Mainly, the bacterial cell wall had the potential for Cd 2+ biosorption, and we found non-significant biosorption alterations among bacterial strains' inner and outer surfaces of cell membranes. Furthermore, the Fourier transform infrared (FTIR) spectroscopy analysis identified the differences in functional groups, such as C-N, PO 2 , -SO 3 , CO, COOH, C-O, C-N, -OH, and -NH that interact in biosorption by Bacillus strains. The scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) examination revealed that the binding of Cd 2+ to microbes was mostly based on ion exchange pathways. Moreover, the Bacillus strains responded to Cd 2+ stress in rice under pot experiment at various concentrations (0, 0.25, and 0.50 mg kg -1 ), and they also influenced the chlorophyll contents and antioxidants activities were studied. The analysis of physio-morphological parameters was observed to be increased, which indicated that all Bacillus strains showed significant effects on rice growth under Cd 2+ stress. These results revealed that the selected strains had the capability for additional use in the development of Cd 2+ bioremediation methods. These strains also provided plant growth-promoting (PGP) traits that can alleviate the harmful effects of Cd 2+ in rice plants., (Copyright © 2022. Published by Elsevier Ltd.)

Salt stress decreases plant growth and is a major threat to crop yields worldwide. The present study aimed to alleviate salt stress in plants by inoculation with halophilic plant growth-promoting rhizobacteria (PGPR) isolated from an extreme environment in the Qinghai-Tibetan Plateau. Wheat plants inoculated with Bacillus halotolerans KKD1 showed increased seedling morphological parameters and physiological indexes. The expression of wheat genes directly involved in plant growth was upregulated in the presence of KKD1, as shown by real-time quantitative PCR (RT-qPCR) analysis. The metabolism of phytohormones, such as 6-benzylaminopurine and gibberellic acid were also enhanced. Mining of the KKD1 genome corroborated its potential plant growth promotion (PGP) and biocontrol properties. Moreover, KKD1 was able to support plant growth under salt stress by inducing a stress response in wheat by modulating phytohormone levels, regulating lipid peroxidation, accumulating betaine, and excluding Na + . In addition, KKD1 positively affected the soil nitrogen content, soil phosphorus content and soil pH. Our findings indicated that KKD1 is a promising candidate for encouraging wheat plant growth under saline conditions., Competing Interests: RB was employed by company Nord Reet UG. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wu, Fan, Wang, Zhao, Ali, Wu, Gu, Borriss, Xie and Gao.)

Foot and mouth disease (FMD) causes severe economic losses to the livestock industry of endemic countries, including Pakistan. Pakistan is part of the endemic pool 3 for foot and mouth disease viruses (FMDV), characterized by co-circulating O, A, and Asia 1 serotypes, as designated by the world reference laboratory for FMD (WRL-FMD). FMDV serotype A lineage ASIA/Iran-05 is widespread in buffalos and cattle populations and was first reported in Pakistan in 2006. This lineage has a high turnover, with as many as 10 sub-lineages reported from Pakistan over the years. In this study, we reconstructed the evolutionary, demographic, and spatial history of serotype A and one of its sub-lineages, A/ASIA/Iran-05/SIS-13, prevalent in Pakistan. We sequenced nearly complete genomes of three isolates belonging to sub-lineage A/ASIA/Iran-05/SIS-13. We estimated recombination patterns and natural selection acting on the serotype A genomes. Source and transmission routes in Pakistan were inferred, and the clustering pattern of isolates of the SIS-13 sub-lineage were mapped on a tree. We hereby report nearly complete genome sequences of isolates belonging to sub-lineage A/ASIA/Iran-05/SIS-13, along with purported recombinant genomes, and highlight that complete coding sequences can better elucidate the endemic history and evolutionary pressures acting on long-term co-circulating FMDV strains.

The plant parasitic nematode, Aphelenchoides besseyi , is a serious pest causing severe damage to various crop plants and vegetables. The Bacillus thuringiensis (Bt) strains, GBAC46 and NMTD81, and the biological strain, FZB42, showed higher nematicidal activity against A. besseyi , by up to 88.80, 82.65, and 75.87%, respectively, in a 96-well plate experiment. We screened the whole genomes of the selected strains by protein-nucleic acid alignment. It was found that the Bt strain GBAC46 showed three novel crystal proteins, namely, Cry31Aa, Cry73Aa, and Cry40ORF, which likely provide for the safe control of nematodes. The Cry31Aa protein was composed of 802 amino acids with a molecular weight of 90.257 kDa and contained a conserved delta-endotoxin insecticidal domain. The Cry31Aa exhibited significant nematicidal activity against A. besseyi with a lethal concentration (LC 50 ) value of 131.80 μg/mL. Furthermore, the results of in vitro experiments (i.e., rhodamine and propidium iodide (PI) experiments) revealed that the Cry31Aa protein was taken up by A. besseyi , which caused damage to the nematode's intestinal cell membrane, indicating that the Cry31Aa produced a pore-formation toxin. In pot experiments, the selected strains GBAC46, NMTD81, and FZB42 significantly reduced the lesions on leaves by up to 33.56%, 45.66, and 30.34% and also enhanced physiological growth parameters such as root length (65.10, 50.65, and 55.60%), shoot length (68.10, 55.60, and 59.45%), and plant fresh weight (60.71, 56.45, and 55.65%), respectively. The number of nematodes obtained from the plants treated with the selected strains (i.e., GBAC46, NMTD81, and FZB42) and A. besseyi was significantly reduced, with 0.56, 0.83., 1.11, and 5.04 seedling mL -1 nematodes were achieved, respectively. Moreover, the qRT-PCR analysis showed that the defense-related genes were upregulated, and the activity of hydrogen peroxide (H 2 O 2 ) increased while malondialdehyde (MDA) decreased in rice leaves compared to the control. Therefore, it was concluded that the Bt strains GBAC46 and NMTD81 can promote rice growth, induce high expression of rice defense-related genes, and activate systemic resistance in rice. More importantly, the application of the novel Cry31Aa protein has high potential for the efficient and safe prevention and green control of plant parasitic nematodes.

Transgenic technology played a crucial role in developing insect-resistant plants resulting in the reduced application of pesticides. This article reports the expression of two cry proteins (Cry3Bb1 and Cry3) in cotton for enhanced resistance against chewing insect pests. The aforementioned genes were synthetically developed and were cloned under appropriate regulatory sequences followed by transformation into Eagle-2 genotype (Gossypium hirsutum) of cotton through shoot apex-cut Agro-infiltration. The transgene integration was validated by polymerase chain reaction using primers flanking the aforementioned cry genes. Transgene expression was assessed by qRT-PCR using GADPH as a reference gene. The relative fold expression analyses revealed the highest expression of the transgene(s) in M1 plants, which is a 4.5-fold expression (Cry3 + Cry3Bb1) followed by M3 (fold expression, 3.0) (Cry3Bb1) and M2 (fold expression, 2.5) (Cry3) transformants of cotton. The confirmed transgenic plants were exposed to insect pests, pink bollworm (Pectinophora gossypiella), and army bollworm (Helicoverpa armigera). Bioassay results revealed that 60% mortality was observed against pink bollworm, and 75% mortality was observed against army bollworm in transgenic plants containing both Cry3Bb1 and Cry3 genes (M1 transgenic plants). In M2 transgenic plants containing only the Cry3Bb1 gene, the mortality was observed to be 40% in the pink bollworm population, whereas 45% mortality was observed in the army bollworm population. In the case of M3 transgenic plants containing single gene-Cry3, the mortality was 20% in the pink bollworm population, whereas 30% mortality was observed in the army bollworm population. Almost no mortality was observed in non-transgenic Eagle-2 control plants. Hence, the developed cotton transformants have improved resistance against chewing insect pests., (© 2022. The Author(s).)

RNA interference (RNAi) triggered by exogenous double-stranded RNA (dsRNA) is a powerful tool to knockdown genetic targets crucial for the growth and development of agriculturally important insect pests. Helicoverpa armigera is a pest feeding on more than 30 economically important crops worldwide and a major threat. Resistance to insecticides and Bt toxins has been gradually increasing in the field. RNAi-mediated knockdown of H. armigera genes by producing dsRNAs homologous to genetic targets in bacteria and plants has a high potential for insect management to decrease agricultural loss. The acetylcholinesterase (AChE), ecdysone receptor (EcR) and v-ATPase-A (vAA) genes were selected as genetic targets. Fragments comprising a coding sequence of < 500 bp were cloned into the L4440 vector for dsRNA production in bacteria and in a TRV-VIGS vector in antisense orientation for transient expression of dsRNA in Solanum tuberosum leaves. After ingesting bacterial-expressed dsRNA, the mRNA levels of the target genes were significantly reduced, leading to mortality and abnormal development in larva of H. armigera. Furthermore, the S. tuberosum plants transformed with TRV-VIGS expressing AChE exhibited higher mortality > 68% than the control plants 17%, recorded ten days post-feeding and significant resistance in transgenic (transient) plants was observed. Moreover, larval lethality and molting defects were observed in larva fed on potato plants expressing dsRNA specific to EcR. Analysis of transcript levels by quantitative RT-PCR revealed that larval mortality was attributable to the knockdown of genetic targets by RNAi. The results demonstrated that down-regulation of H. armigera genes involved in ATP hydrolysis, transcriptional stimulation of development genes and neural conduction has aptitude as a bioinsecticide to control H. armigera population sizes and therefore decreases crop loss., (© 2022. The Author(s).)

The rhizospheric bacterium Pseudomonas protegens Pf-5 can colonize the seed and root surfaces of plants, and can protect them from pathogen infection. Secondary metabolites, including lipopeptides and polyketides produced by Pf-5, are involved in its biocontrol activity. We isolated a crude extract from Pf-5. It exhibited significant surface activity and strong antibacterial activity against Pantoea ananatis DZ-12, which causes maize brown rot on leaves. HPLC analysis combined with activity tests showed that the polyketide pyoluteorin in the crude extract participated in the suppression of DZ-12 growth, and that the lipopeptide orfamide A was the major biosurfactant in the crude extract. Further studies indicated that the pyoluteorin in the crude extract significantly suppressed the biofilm formation of DZ-12, and it induced the accumulation of reactive oxygen species in DZ-12 cells. Scanning electron microscopy and transmission electron microscopy observation revealed that the crude extract severely damaged the pathogen cells and caused cytoplasmic extravasations and hollowing of the cells. The pathogenicity of DZ-12 on maize leaves was significantly reduced by the crude extract from Pf-5 in a dose-dependent manner. The polyketide pyoluteorin had strong antibacterial activity against DZ-12, and it has the potential for development as an antimicrobial agent.

Crop production worldwide is under pressure from multiple factors, including reductions in available arable land and sources of water, along with the emergence of new pathogens and development of resistance in pre-existing pathogens. In addition, the ever-growing world population has increased the demand for food, which is predicted to increase by more than 100% by 2050. To meet these needs, different techniques have been deployed to produce new cultivars with novel heritable mutations. Although traditional breeding continues to play a vital role in crop improvement, it typically involves long and laborious artificial planting over multiple generations. Recently, the application of innovative genome engineering techniques, particularly CRISPR-Cas9-based systems, has opened up new avenues that offer the prospects of sustainable farming in the modern agricultural industry. In addition, the emergence of novel editing systems has enabled the development of transgene-free non-genetically modified plants, which represent a suitable option for improving desired traits in a range of crop plants. To date, a number of disease-resistant crops have been produced using gene-editing tools, which can make a significant contribution to overcoming disease-related problems. Not only does this directly minimize yield losses but also reduces the reliance on pesticide application, thereby enhancing crop productivity that can meet the globally increasing demand for food. In this review, we describe recent progress in genome engineering techniques, particularly CRISPR-Cas9 systems, in development of disease-resistant crop plants. In addition, we describe the role of CRISPR-Cas9-mediated genome editing in sustainable agriculture., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ali, Yu, Hussain, Ali, Ahmar, Sohail, Riaz, Ashraf, Abdalmegeed, Wang, Imran, Manghwar and Zhou.)

Grapefruit (Citrus paradisi) is a widely grown citrus and its fruit is affected by a variety of biotic and abiotic stress. Keeping in view the hazardous effects of synthetic fungicides, the recent trend is shifting towards safer and eco-friendly control of fruit diseases. The present study was aimed to diagnose the fruit rot disease of grapefruit and its control by using zinc oxide green nanoparticles (ZnO NPs). Fruit rot symptoms were observed in various grapefruit growing sites of Pakistan. Diseased samples were collected, and the disease-causing pathogen was isolated. Following Koch's postulates, the isolated pathogen was identified as Rhizoctonia solani. For eco-friendly control of this disease, ZnO NPs were prepared in the seed extract of Trachyspermum ammi and characterized. Fourier transform infrared spectroscopy (FTIR) of these NPs described the presence of stabilizing and reducing compounds such as phenols, aldehyde and vinyl ether, especially thymol (phenol). X-ray diffraction (XRD) analysis revealed their crystalline nature and size (48.52 nm). Energy dispersive X-ray (EDX) analysis elaborated the presence of major elements in the samples, while scanning electron microscopy (SEM) confirmed the morphology of bio fabricated NPs. ZnO NPs exhibited very good anti-fungal activity and the most significant fungal growth inhibition was observed at 1.0 mg/ml concentration of green NPs, in vitro and in vivo. These findings described that the bioactive constituents of T. ammi seed extract can effectively reduce and stabilize ZnO NPs. It is a cost-effective method to successfully control the fruit rot disease of grapefruit., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Cucumber is an important vegetable but highly sensitive to salt stress. The present study was designed to investigate the comparative performance of cucumber genotypes under salt stress (50 mmol L -1 ) and stress alleviation through an optimized level of triacontanol @ 0.8 mg L -1 . Four cucumber genotypes were subjected to foliar application of triacontanol under stress. Different physiological, biochemical, water relations and ionic traits were observed to determine the role of triacontanol in salt stress alleviation. Triacontanol ameliorated the lethal impact of salt stress in all genotypes, but Green long and Marketmore were more responsive than Summer green and 20252 in almost all the attributes that define the genetic potential of genotypes. Triacontanol performs as a good scavenger of ROS by accelerating the activity of antioxidant enzymes (SOD, POD, CAT) and compatible solutes (proline, glycinebetaine, phenolic contents), which lead to improved gas exchange attributes and water relations and in that way enhance the calcium and potassium contents or decline the sodium and chloride contents in cucumber leaves. Furthermore, triacontanol feeding also shows the answer to yield traits of cucumber. It was concluded from the results that the salinity tolerance efficacy of triacontanol is valid in enhancing the productivity of cucumber plants under salt stress. Triacontanol was more pronounced in green long and marketer green than in summer green and 20252. Hence, the findings of this study pave the way towards the usage of triacontanol @ 0.8 mg L -1 , and green long and marketer genotypes may be recommended for saline soil., (© 2021. The Author(s).)

Whether or not hydrogen gas (H 2 ) can reduce cadmium (Cd) toxicity in Ganoderma lucidum has remained largely unknown. Here, we report that Cd-induced growth inhibition in G. lucidum was significantly alleviated by H 2 fumigation or hydrogen-rich water (HRW), evaluated by lower oxidative damage and Cd accumulation. Moreover, the amelioration effects of H 2 fumigation were better than of HRW in an optimum concentration of H 2 under our experimental conditions. Further results showed that H 2 -alleviated growth inhibition in G. lucidum was accompanied by increased nitric oxide (NO) level and nitrate reductase (NR) activity under Cd stress. On the other hand, the mitigation effects were reversed after removing endogenous NO with its scavenger cPTIO or inhibiting H 2 -induced NR activity with sodium tungstate. The role of NO in H 2 -alleviated growth inhibition under Cd stress was proved to be achieved through a restoration of redox balance, an increase in cysteine and proline contents, and a reduction in Cd accumulation. In summary, these results clearly revealed that NR-dependent NO might be involved in the H 2 -alleviated Cd toxicity in G. lucidum through rebuilding redox homeostasis, increasing cysteine and proline levels, and reducing Cd accumulation. These findings may open a new window for H 2 application in Cd-stressed economically important fungi.

Endophytic bacterial communities are beneficial communities for host plants that exist inside the surfaces of plant tissues, and their application improves plant growth. They benefit directly from the host plant by enhancing the nutrient amount of the plant's intake and influencing the phytohormones, which are responsible for growth promotion and stress. Endophytic bacteria play an important role in plant-growth promotion (PGP) by regulating the indirect mechanism targeting pest and pathogens through hydrolytic enzymes, antibiotics, biocontrol potential, and nutrient restriction for pathogens. To attain these benefits, firstly bacterial communities must be colonized by plant tissues. The nature of colonization can be achieved by using a set of traits, including attachment behavior and motility speed, degradation of plant polymers, and plant defense evasion. The diversity of bacterial endophytes colonization depends on various factors, such as plants' relationship with environmental factors. Generally, each endophytic bacteria has a wide host range, and they are used as bio-inoculants in the form of synthetic applications for sustainable agriculture systems and to protect the environment from chemical hazards. This review discusses and explores the taxonomic distribution of endophytic bacteria associated with different genotypes of rice plants and their origin, movement, and mechanism of PGP. In addition, this review accentuates compressive meta data of endophytic bacteria communities associated with different genotypes of rice plants, retrieves their plant-growth-promoting properties and their antagonism against plant pathogens, and discusses the indication of endophytic bacterial flora in rice plant tissues using various methods. The future direction deepens the study of novel endophytic bacterial communities and their identification from rice plants through innovative techniques and their application for sustainable agriculture systems., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Barley ( Hordeum vulgare L.) is a major cereal grain and is known as a halophyte (a halophyte is a salt-tolerant plant that grows in soil or waters of high salinity). We therefore conducted a pot experiment to explore plant growth and biomass, photosynthetic pigments, gas exchange attributes, stomatal properties, oxidative stress and antioxidant response and their associated gene expression and absorption of ions in H. Vulgare . The soil used for this analysis was artificially spiked at different salinity concentrations (0, 50, 100 and 150 mM) and different levels of ascorbic acid (AsA) were supplied to plants (0, 30 and 60 mM) shortly after germination of the seed. The results of the present study showed that plant growth and biomass, photosynthetic pigments, gas exchange parameters, stomatal properties and ion uptake were significantly ( p  < 0.05) reduced by salinity stress, whereas oxidative stress was induced in plants by generating the concentration of reactive oxygen species (ROS) in plant cells/tissues compared to plants grown in the control treatment. Initially, the activity of antioxidant enzymes and relative gene expression increased to a saline level of 100 mM, and then decreased significantly ( P  < 0.05) by increasing the saline level (150 mM) in the soil compared to plants grown at 0 mM of salinity. We also elucidated that negative impact of salt stress in H. vulgare plants can overcome by the exogenous application of AsA, which not only increased morpho-physiological traits but decreased oxidative stress in the plants by increasing activities of enzymatic antioxidants. We have also explained the negative effect of salt stress on H. vulgare can decrease by exogenous application of AsA, which not only improved morpho-physiological characteristics, ions accumulation in the roots and shoots of the plants, but decreased oxidative stress in plants by increasing antioxidant compounds (enzymatic and non-enzymatic). Taken together, recognizing AsA's role in nutrient uptake introduces new possibilities for agricultural use of this compound and provides a valuable basis for improving plant tolerance and adaptability to potential salinity stress adjustment., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

Melatonin (N-acetyl-5-methoxytryptamine), a naturally occurring small molecule, can protect plants against abiotic stress after exogenous treatmenting with it. It is not known if melatonin homologs, such as 5-methoxytryptamine and 5-methoxyindole, that are easy and more cost-effective to synthesize can stimulate the plant immune system in the same manner as melatonin. In the present study, we assessed the biological activity of the melatonin homologs, 5-methoxytryptamin and 5-methoxyindole. The results showed that melatonin and its homologs all induced disease resistance against Phytophthora nicotianae in Nicotiana benthamiana plants. The application of all three compounds also induced stomatal closure and the production of reactive oxygen species. Gene expression analysis indicated that the expression of genes involved in hydrogen peroxide (H 2 O 2 ), nitric oxide (NO) production, and salicylic acid (SA) biosynthesis was significantly upregulated by all three compounds. Four homologs of the melatonin receptors were identified by blasting search with the phytomelatonin receptor in Arabidopsis . Molecular docking studies were also used to identify four putative melatonin receptors in N. benthamiana . Further experimentation revealed that silencing of the melatonin receptors trP47363 and trP13076 in N. benthamiana compromised the induction of stomatal closure, PR-1a gene expression and SA accumulation by all three compounds. Collectively, our data indicate that the induction of defense responses in N. benthamiana by melatonin, 5-methoxytryptamine, and 5-methoxyindole involves the melatonin receptors trP47363 and trP13076., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kong, Sheng, Liang, Ali, Gu, Wu, Chen, Liu and Gao.)

Bacillus volatiles to control plant nematodes is a topic of great interest among researchers due to its safe and environmentally friendly nature. Bacillus strain GBSC56 isolated from the Tibet region of China showed high nematicidal activity against M. incognita , with 90% mortality as compared with control in a partition plate experiment. Pure volatiles produced by GBSC56 were identified through gas chromatography and mass spectrometry (GC-MS). Among 10 volatile organic compounds (VOCs), 3 volatiles, i.e., dimethyl disulfide (DMDS), methyl isovalerate (MIV), and 2-undecanone (2-UD) showed strong nematicidal activity with a mortality rate of 87%, 83%, and 80%, respectively, against M. incognita . The VOCs induced severe oxidative stress in nematodes, which caused rapid death. Moreover, in the presence of volatiles, the activity of antioxidant enzymes, i.e., SOD, CAT, POD, and APX, was observed to be enhanced in M. incognita -infested roots, which might reduce the adverse effect of oxidative stress-induced after infection. Moreover, genes responsible for plant growth promotion SlCKX1 , SlIAA1 , and Exp18 showed an upsurge in expression, while AC01 was downregulated in infested plants. Furthermore, the defense-related genes ( PR1 , PR5 , and SlLOX1 ) in infested tomato plants were upregulated after treatment with MIV and 2-UD. These findings suggest that GBSC56 possesses excellent biocontrol potential against M. incognita . Furthermore, the study provides new insight into the mechanism by which GBSC56 nematicidal volatiles regulate antioxidant enzymes, the key genes involved in plant growth promotion, and the defense mechanism M. incognita -infested tomato plants use to efficiently manage root-knot disease.

Plant diseases significantly impact the global economy, and plant pathogenic microorganisms such as nematodes, viruses, bacteria, fungi, and viroids may be the etiology for most infectious diseases. In agriculture, the development of disease-free plants is an important strategy for the determination of the survival and productivity of plants in the field. This article reviews biosensor methods of disease detection that have been used effectively in other fields, and these methods could possibly transform the production methods of the agricultural industry. The precise identification of plant pathogens assists in the assessment of effective management steps for minimization of production loss. The new plant pathogen detection methods include evaluation of signs of disease, detection of cultured organisms, or direct examination of contaminated tissues through molecular and serological techniques. Laboratory-based approaches are costly and time-consuming and require specialized skills. The conclusions of this review also indicate that there is an urgent need for the establishment of a reliable, fast, accurate, responsive, and cost-effective testing method for the detection of field plants at early stages of growth. We also summarized new emerging biosensor technologies, including isothermal amplification, detection of nanomaterials, paper-based techniques, robotics, and lab-on-a-chip analytical devices. However, these constitute novelty in the research and development of approaches for the early diagnosis of pathogens in sustainable agriculture.

Copper (Cu) is an abundant essential micronutrient element in various rocks and minerals and is required for a variety of metabolic processes in both prokaryotes and eukaryotes. However, excess Cu can disturb normal development by adversely affecting biochemical reactions and physiological processes in plants. The present study was conducted to explore the potential of gibberellic acid (GA 3 ) on fibrous jute (Corchorus capsularis L.) seedlings grown on Cu mining soil obtained from Hubei Province China. Exogenous application of GA 3 (10, 50, and 100 mg/L) on 60-day-old seedlings of C. capsularis which was able to grow in highly Cu-contaminated soil (2221 mg/kg) to study different morphological, physiological, and Cu uptake and accumulation in different parts of C. capsularis seedlings. According to the results, increasing concentration of GA 3 (more likely 100 mg/L) alleviates Cu toxicity in C. capsularis seedlings by increasing plant growth, biomass, photosynthetic pigments, and gaseous exchange attributes. The results also showed that exogenous application of GA 3 reduced oxidative stress in C. capsularis seedlings by the generation of extra reactive oxygen species (ROS). The reduction in oxidative stress in C. capsularis seedlings is because that plant has strong enzymatic antioxidants [superoxidase dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT)], which ultimately increased their activities to overcome oxidative damage in the cells/tissues. In addition to the plant growth, biomass, and photosynthesis, foliar application of GA 3 also helps to increase metal (Cu) concentration in different parts of the plants when compared to 0 mg/L of application of GA 3 . From these findings, we can conclude that foliar application of GA 3 plays a promising role in reducing ROS generation in the plant cells/tissues and increased phytoextraction of Cu in different plant parts. However, more investigation is needed on field experiments to find a combination of GA 3 with a very higher concentration of Cu using fibrous C. capsularis.

Contamination of soil and water with metals and metalloids is one of the most serious problems worldwide due to a lack of a healthy diet and food scarcity. Moreover, the cultivation of oilseed crops such as rapeseed ( Brassica napus L.) with tannery wastewater could contain a large amount of toxic heavy metals [e.g., chromium (Cr)], which ultimately reduce its yield and directly influence oilseed quality. To overcome Cr toxicity in B. napus , a pot experiment was conducted to enhance plant growth and biomass by using newly introduced role of micronutrient-amino chelates [Zinc-lysine (Zn-lys)], which was irrigated with different levels [0% (control), 33%, 66%, and 100%] of tannery wastewater. According to the results of present findings, very high content of Cr in the wastewater directly affected plant growth and composition as well as gas exchange parameters, while boosting up the production of reactive oxygen species (ROS) and induced oxidative damage in the roots and leaves of B. napus . However, activities of antioxidants initially increased (33% of wastewater), but further addition of tannery wastewater in the soil caused a decrease in antioxidant enzymes, which also manifested by Zn content, while the conscious addition of wastewater significantly increased Cr content in the roots and shoots of B. napus . To reduce Cr toxicity in B. napus plants, exogenous supplementation of Zn-lys (10 mg/L) plays an effective role in increasing morpho-physiological attributes of B. napus and also reduces the oxidative stress in the roots and leaves of the oilseed crop ( B. napus ). Enhancement in different growth attributes was directly linked with increased in antioxidative enzymes while decreased uptake and accumulation of Cr content in B. napus when cultivated in wastewater with the application of Zn-lys. Zn-lys, therefore, plays a protective role in reducing the Cr toxicity of B. napus through an increase in plant growth and lowering of Cr uptake in various plant organs. However, further studies at field levels are required to explore the mechanisms of Zn-lys mediated reduction of Cr and possibly other heavy metal toxicity in plants.

The use of chemicals around the globe in different industries has increased tremendously, affecting the health of people. The modern world intends to replace these noxious chemicals with environmental friendly products for the betterment of life on the planet. Establishing enzymatic processes in spite of chemical processes has been a prime objective of scientists. Various enzymes, specifically microbial proteases, are the most essentially used in different corporate sectors, such as textile, detergent, leather, feed, waste, and others. Proteases with respect to physiological and commercial roles hold a pivotal position. As they are performing synthetic and degradative functions, proteases are found ubiquitously, such as in plants, animals, and microbes. Among different producers of proteases, Bacillus sp. are mostly commercially exploited microbes for proteases. Proteases are successfully considered as an alternative to chemicals and an eco-friendly indicator for nature or the surroundings. The evolutionary relationship among acidic, neutral, and alkaline proteases has been analyzed based on their protein sequences, but there remains a lack of information that regulates the diversity in their specificity. Researchers are looking for microbial proteases as they can tolerate harsh conditions, ways to prevent autoproteolytic activity, stability in optimum pH, and substrate specificity. The current review focuses on the comparison among different proteases and the current problems faced during production and application at the industrial level. Deciphering these issues would enable us to promote microbial proteases economically and commercially around the world.

Glycosylation, a posttranslational modification, has a major role in recombinant anticancer therapeutic proteins, as most of the approved recombinant therapeutics are glycoproteins. The constant amino acid sequence of therapeutics determines the enzymatic activity, while the presence of glycans influences their pharmacokinetics, solubility, distribution, serum half-life, effector function, and binding to receptors. Glycoproteins expressed in different expression systems acquire their own oligosaccharides, which increases the protein diversity. The heterogeneity of glycans creates hurdles in downstream processing, ultimately leading to variable anticancer therapeutic efficacy. Therefore, glycoproteins require an appropriate expression system to obtain structurally and functionally identical glycans, as in humans. In many expression systems, the N -glycosylation pathway remains conserved in the endoplasmic reticulum, but divergence is observed when the protein enters the Golgi complex. Hence, in recent decades, numerous approaches have been adopted to engineer the Golgi's N -glycosylation pathway to attain human-like glycans. Several researchers have tried to engineer the N -glycosylation pathway of expression systems. In this review, we examine the glycosylation pattern in various expression systems, along with emerging technologies for glycosylation engineering of anticancer therapeutic drugs. Cancer Res; 78(11); 2787-98. ©2018 AACR ., (©2018 American Association for Cancer Research.)

Background: Cancer is a genetic disease and mainly arises due to a number of reasons include activation of onco-genes, malfunction of tumor suppressor genes or mutagenesis due to external factors., Methods: This article was written from the data collected from PubMed, Nature, Science Direct, Springer and Elsevier groups of journals., Results: Oncogenes are deregulated form of normal proto-oncogenes required for cell division, differentiation and regulation. The conversion of proto-oncogene to oncogene is caused due to translocation, rearrangement of chromosomes or mutation in gene due to addition, deletion, duplication or viral infection. These oncogenes are targeted by drugs or RNAi system to prevent proliferation of cancerous cells. There have been developed different techniques of molecular biology used to diagnose and treat cancer, including retroviral therapy, silencing of oncogenes and mutations in tumor suppressor genes., Conclusion: Among all the techniques used, RNAi, zinc finger nucleases and CRISPR hold a brighter future towards creating a Cancer Free World., Competing Interests: Conflict of interests The authors declare that there is no conflict of interest.

As long as the genetically modified crops are gaining attention globally, their proper approval and commercialization need accurate and reliable diagnostic methods for the transgenic content. These diagnostic techniques are mainly divided into two major groups, i.e., identification of transgenic (1) DNA and (2) proteins from GMOs and their products. Conventional methods such as PCR (polymerase chain reaction) and enzyme-linked immunosorbent assay (ELISA) were routinely employed for DNA and protein based quantification respectively. Although, these Techniques (PCR and ELISA) are considered as significantly convenient and productive, but there is need for more advance technologies that allow for high throughput detection and the quantification of GM event as the production of more complex GMO is increasing day by day. Therefore, recent approaches like microarray, capillary gel electrophoresis, digital PCR and next generation sequencing are more promising due to their accuracy and precise detection of transgenic contents. The present article is a brief comparative study of all such detection techniques on the basis of their advent, feasibility, accuracy, and cost effectiveness. However, these emerging technologies have a lot to do with detection of a specific event, contamination of different events and determination of fusion as well as stacked gene protein are the critical issues to be addressed in future.

Phenotypic stability under stress environment facilitate the fitness of genotype and opens new horizons to explore the cryptic genetic variation. Variation in tolerance to drought stress, a major grain yield constraint to global maize production, was identified, at the phenotypic and genotypic level. Here we found a prominent hybrid H 9 that showed fitness over four growing seasons for grain yield under water stress conditions. Genotypic and phenotypic correlation of yield attributing traits over four seasons demonstrated that cobs per plant, 100 seed weight, number of grains rows per cob, total dry matter, cob diameter had positive association ( r 2 = 0.3-0.9) to grain yield. The perturbation was found for chlorophyll content as it showed moderate to strong association ( P < 0.01) over four seasons, might be due to environment or genotype dependent. Highest heritability (95%) and genetic advance (79%) for grain yield was found in H 9 over four consecutive crop growing seasons. Combined analysis over four seasons showed that studied variables together explained 85% of total variation in dependent structure (grain yield) obtained by Principal component analysis. This significant finding is the best example of phenotypic stability of grain yield in H 9 and made it best fitted for grain yield under drought stress scenario. Detailed genetic analysis of H 9 will help us to identify significant loci and alleles that made H 9 the best fitted and it could serve as a potential source to generate novel transgressive levels of tolerance for drought stress in arid/semiarid regions.

Maize Chlorotic Mottle Virus (MCMV) is a deleterious pathogen which causes Maize Lethal Necrosis Disease (MLND) that results in substantial yield loss of Maize crop worldwide. The positive-sense RNA genome of MCMV (4.4 kb) encodes six proteins: P32 (32 kDa protein), RNA dependent RNA polymerases (P50 and P111), P31 (31 kDa protein), P7 (7 kDa protein), coat protein (25 kDa). P31, P7 and coat protein are encoded from sgRNA1, located at the 3'end of the genome and sgRNA2 is located at the extremity of the 3'genome end. The objective of this study is to locate the possible attachment sites of Zea mays derived miRNAs in the genome of MCMV using four diverse miRNA target prediction algorithms. In total, 321 mature miRNAs were retrieved from miRBase (miRNA database) and were tested for hybridization of MCMV genome. These algorithms considered the parameters of seed pairing, minimum free energy, target site accessibility, multiple target sites, pattern recognition and folding energy for attachment. Out of 321 miRNAs only 10 maize miRNAs are predicted for silencing of MCMV genome. The results of this study can hence act as the first step towards the development of MCMV resistant transgenic Maize plants through expression of the selected miRNAs.

Human immunodeficiency virus (HIV) is the chief contributor to global burden of disease. In 2010, HIV was the fifth leading cause of disability-adjusted life years in people of all ages and leading cause for people aged 30-44 years. It is classified as a member of the family Retroviridae and genus Lentivirus based on the biological, morphological, and genetic properties. It infects different cells of the immune system, such as CD4+ T cells (T-helper cells), dendritic cells, and macrophages. HIV has two subtypes: HIV-1 and HIV-2. Among these strains, HIV-1 is the most virulent and pathogenic. Advanced diagnostic methods are exploring new ways of treatment and contributing in the reduction of HIV cases. The diagnostic techniques like PCR, rapid test, EIA, p24 antigen, and western blot have markedly upgraded the diagnosis of HIV. Antiretroviral therapy and vaccines are promising candidates in providing therapeutic and preventive regimes, respectively. Invention of CRISPR/Cas9 is a breakthrough in the field of HIV disease management.

Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell's serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n = 5), aborted fetuses (n = 13), and vaginal swabs (n = 12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.

The present study describes the frequency of Foot and Mouth Disease (FMD) virus serotypes (O, A and Asia-1) in major regions (all provinces) of Pakistan using Indirect Sandwich ELISA. Also, spatial distribution of various FMD serotypes and their comparison is discussed. A total of 590 samples (Epithelial tissue) have been analyzed during a period of five years (2005-2009). Out of 590 samples, 180 were found positive, giving an overall confirmation of FMDV about 33.2 %. Of the prevalent serotypes, FMDV 'O' serotype caused most outbreaks (20.7 %), followed by serotype A (6.6 %) and serotype Asia-1 (4.6 %) while there was no positive case of type 'C'. The study clearly showed that the disease was more frequent in the agro-climatic zones than in hilly areas. Based on the data of 590 samples (>50 outbreaks), the overall prevalence of FMDV in cattle and buffaloes in Pakistan was 33.2 %, while in cattle alone, it was 37.1 %, higher than in buffalo (28.7 %). There were eight cases of mixed serotypes infection, indicating the presence of endemic state of disease. Another significant feature was the change over time. In phase-I (2005-2007), there was an overall prevalence of 29.4 %, while the occurrence of the serotype O, A and Asia-1 was 20.4 %, 2.9 % and 4.7 %, respectively. During phase-II (2008-2009), the overall prevalence was 59.21 %, while those of serotype O, A and Asia-1 were 22.4 %, 31.6 % and 4.0 %, respectively. This clearly indicated a shift from serotype O to A, which may help to explain the occurrence of more severe outbreaks, despite vaccination.

An outbreak resulting in mortality in Sindh Ibex (Capra aegagrus blythi) was investigated. There was a history of about 36 deaths (both young and adult) during the period of 1 month. Disease appeared in a generalized form, affecting the respiratory and digestive systems. Major lesions were respiratory distress, pustules on and in the mouth, ocular-nasal discharges, and severe diarrhea. The most significant lesion was the oculonasal discharges and diarrhea. Deaths were mainly due to blindness, anorexia, diarrhea, and respiratory arrest. Both adult (mortality = 21) and young (mortality = 15) animals were affected with the disease. Peste des petits ruminants virus (PPRV) antigen was detected in the spleen, lung, lymph node, and swab samples by immunocapture enzyme-linked immunosorbent assay. Spleen and lung samples were also tested and found positive for the presence of F-gene of PPRV by polymerase chain reaction. Thirteen of 20 serum samples from nearby sheep and goats were found positive for antibodies to PPRV. The disease threatened the huge population of ibex in the wild life park, which was spread over a large area, but vaccination of the domestic population of sheep and goats in the surrounding villages appeared to control the disease.

The present investigation included a detailed description of the factors directly associated with PPRV infection in Pakistan. A total of 1,056 suspected serum samples were analyzed for the presence of antibodies to PPRV with no history of vaccination against PPR. The samples were collected from sixty two (62) suspected outbreaks from twenty five (25) major regions of the country. Samples were collected from the animals suffering from diarrhea and showing severe respiratory signs. Competitive enzyme linked immuno-sorbant assay (cELISA) was performed to detect the presence of antibodies in the serum against PPRV. Findings suggested that the overall PPR antibody sero-prevalence recorded in sheep was 54.09% as compared to 44.15% in goats. Geography, species, sex, age and season are the major factors associated with PPRV infection. Among various age groups, the animals showed the higher prevalence (67.48% and 52.28% in sheep and goats, respectively) at >2 years as compared with the other age groups. The area-wise highest sero-prevalence was in Sindh province which was 55.10%. It was highest in the plains of Punjab and Sindh province and also in hilly areas (AJK, northern areas and northern Punjab). These findings may be correlated with variations in the sheep and goats husbandry practices within different geographic regions and the topography of different areas. The study also indicated the scenario of antibodies circulation in the population and proving that PPR is now becoming endemic and is one of the major emerging TAD in Pakistan.

This project was conducted to investigate the comparative efficiency of competitive ELISA (cELISA), standard Agar Gel Immunodiffusion Test (AGID) and Precipitinogen Inhibition Test (PIT) for the diagnosis of Peste des Petits Ruminants (PPR) in Pakistan. To deal with this, serum samples from 198 sheep and 82 goats were collected from three different government livestock farms and all the samples were run simultaneously with the three serological tests. The samples found positive for PPR antibodies through cELISA, AGID and PIT were 96 (34.2%), 60 (21.4%) and 72 (25.7%), respectively. Kappa statistics were applied to evaluate the concordance between the laboratory-based test (cELISA) and field-based tests (AGID and PIT). Kappa statistics scores for cELISA versus AGID and PIT were 0.6343 (95% Confidence Interval CI 0.5231-0.7456) and 0.7134 (95% Confidence Interval CI 0.5987-0.8281), respectively, which indicate a "substantial" agreement between cELISA and AGID and "significant" agreement between cELISA and PIT. AGID and PIT revealed relative diagnostic sensitivities with cELISA of 59.3% and 69.7% and relative diagnostic specificities of 98.3% and 97.2%, respectively. The data suggested that for mass screening and control of PPR, these serological tests proved practical in the absence of cELISA since they have high relative diagnostic specificities and a satisfactory relative diagnostic sensitivities.

Present study was designed to investigate the prevalence and mortality (%) caused by Peste des Petitis Ruminant (PPR) and its possible association with abortion in goat flocks at different areas of Pakistan. A total of 140 animals were samples in the population of 650 which was having 185 deaths (Mortality rate = 28 %) from three different regions of the country. There were 58 abortions in the 140 pregnant goats of above said population One hundred & ten (110) serum samples from diseased, recovered and apparently healthy animals were tested for the presence of PPR antibodies by competitive ELISA (c ELISA). Eighty-four (84) animals were positive for PPR antibodies whereas in apparently healthy adult goats in the same flock, no PPR antibodies were detected. Twenty-four (24) tissue samples collected from the dead animals and six samples from aborted fetus were tested for the presence of PPR antigen by Immuno-capture ELISA (Ic ELISA). Nineteen (19) out of thirty (30) organ samples mainly from lung, spleen, lymph node were found positive for PPR antigen but negative from lungs of aborted fetus. There was a high rate of abortions (28-45%) in each of the outbreak and it was highest in the outbreak of Golra Sharif, Islamabad (No. = 21 in total population of 100). As the serum samples from the aborted dams were found positive for PPR antibodies so the study provides the possible association of mortality and prevalence of PPR disease with high rate of abortions in goat.

Objective: To determine the prognosis of seizures in epileptic children and identify early predictors of intractable childhood epilepsy., Study Design: Case-control study., Place and Duration of Study: The Epilepsy Centre of the Children's Hospital Lahore, from February 2005 to April 2007., Patients and Methods: All children (aged 1 month to 16 years) with idiopathic or cryptogenic epilepsy who were treated and followed at the centre during the study period were included. The patients who had marked seizures even after two years of adequate treatment were labeled as intractable epileptics (cases). Children who had no seizure for more than one year at last follow-up visit were the controls. Adequate treatment was described as using at least three anti-epileptic agents either alone or in combination with proper compliance and dosage. Records of these patients were reviewed to identify the variables that may be associated with seizure intractability., Results: Of 442 epileptic children, 325 (74%) intractable and 117 (26%) control epileptics were included in the study. Male gender (OR=3.92), seizures onset in infancy (OR=5.27), = 10 seizures before starting treatment (OR=3.76), myoclonic seizures (OR=1.37), neonatal seizures (OR=3.69), abnormal EEG (OR=7.28) and cryptogenic epilepsy (OR=9.69) and head trauma (OR=4.07) were the factors associated with intractable epilepsy. Seizure onset between 5-7 years of age, idiopathic epilepsy, and absence seizures were associated with favourable prognosis in childhood epilepsy., Conclusion: Intractable childhood epilepsy is expected if certain risk factors such as type, age of onset, gender and cause of epilepsy are found. Early referral of such patients to the specialized centres is recommended for prompt and optimal management.