Antifungal potential and defense gene induction in maize against Rhizoctonia root rot by seed extract of Ammi visnaga (L.) Lam.

Methanol extracts from five medicinal plants (Eucalyptus tereticornis Sm., Ammi visnaga (L.) Lam., Azadirachta indica A. Juss., Rheum Palmatum L., and Adansonia digitata L.) were assessed in vitro for antifungal activity against Rhizoctonia solani Kühn, the causal agent of Rhizoctonia root rot of maize. All tested extracts showed antifungal activity with varied extents. Ammi visnaga (khella) extract showed the greatest activity compared with the untreated experimental controls. Observations using transmission electron microscopy showed ultrastructural changes in hyphal cells as a response to exposure to khella extract. Gas chromatography-mass spectrometric analysis of khella extract showed the presence of 69 compounds. The antifungal properties of the extracts were mainly attributed to their content of coumarins and fatty acids. In the greenhouse experiment, treatment of maize plants with khella extract at 15% gave the least incidence of Rhizoctonia root rot. Results of DD-PCR showed up- and down- regulations of some genes in maize as response to the treatment with khela extract. Identification of the randomly selected genes from DD-PCR revealed that they were defense-related, as S-domain class receptor-like kinase 3 and glutathione-S-transferase1. Real-time PCR showed induction in the gene expression of the pathogenesis-related protein chitinases (2.36 fold) and thaumatin-like proteins (8.99 fold) by treatment with khella extract at 15%, which was greater expression than detected at 10 and 20%. This indicates triggering effects from the extract on the maize immune system against the R. solani infection in a concentration-dependent manner. The efficient, low-cost and eco-friendly characteristics of khella extract indicate that it could be used for the control of Rhizoctonia root rot of maize

Biodiversity of Pathogenic and Toxigenic Seed-Borne Mycoflora of Wheat in Egypt and Their Correlations with Weather Variables

Funding Information: Funding: This work was funded by the Egyptian Science, Technology, and Innovation Funding Authority (STIFA) through project No. 30691 (Egypt–UK Grants), the UK Department for Business, Energy, and Industrial Strategy through British Council Newton-Mosharafa (Project No. 332392589) and the UK Biotechnology and Biological Sciences Research Council, Grant/Award Number: BB-SRC/BB/P00489X/1; Innovate UK, Grant/Award Number: 102641. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Simple Summary: Pathogenic fungi cause yield and quality losses and threaten food security. In this study, 198 samples of wheat grains, representing 20 Egyptian wheat cultivars, were collected from 25 wheat-growing governorates across Egypt, and screened for their seed-borne fungi. Twenty genera and 44 species of seed-borne fungi were identified, and their biodiversity indicators and evolutionary relationships were studied based upon similarities in their genetic characteristics. The most frequent fungi were Alternaria alternata and Cladosporium spp., while Tilletia tritici and Ustilago tritici were the most common smut fungi. The highest fungal diversity was recorded for Sinai governorate, while the greatest species richness was recorded in Qena and Sohag governorates. Correlations of the detected fungi with weather variables (temperature, relative humidity, precipitation, wind speed, or solar radiation) were investigated. Our results indicated that the relative humidity was the most influential weather variable, followed by temperature, solar radiation, wind speed, and precipitation, respectively. Despite this study being conducted on the wheat-growing areas in Egypt, our findings are useful for other wheat-growing countries that share the same climatic conditions. The correlation between a given fungus and the climatic variables can be useful in other ecosystems. Abstract: Surveillance investigations for pathogenic and toxigenic fungi are important to refine our understanding of their epidemiology and help in predicting their outbreaks. During 2019, 198 samples of wheat grains were collected from 25 wheat-growing governorates in Egypt to detect and identify seed-borne mycoflora in vitro. Forty-four fungal species belonging to 20 genera were identified. Molecular data for these fungi were analyzed to construct a phylogenetic tree. Occurrence and biodiversity indicators were calculated. Two prevalent pathogens (average incidence > 40%) were Alternaria alternata and Cladosporium spp. Ustilago tritici was present in only seven of the 25 governorates, and less abundant than Tilletia tritici, the causal agent of stinking smut. Sinai governorate recorded the greatest species diversity, while the greatest species richness was in Qena and Sohag governorates. Canonical correspondence analysis of data for 20 fungal genera with temperature, relative humidity, precipitation, wind speed or solar radiation revealed that relative humidity was the most influential weather variable. It showed that occurrence and distribution of the 20 genera corresponded well with three out of four Egyptian climatic regions: Mediterranean, semi-arid, and arid. Knowing pathogen occurrence and distribution in Egypt is the first step to developing future disease management strategies to limit yield losses and improve food security. Despite this study being conducted on the wheat-growing areas in Egypt, our findings are useful for other wheat-growing countries that share the same climatic conditions. The correlation between a given fungus and the climatic variables can be useful in other ecosystems.Peer reviewedFinal Published versio

Induction of Systemic Resistance to Tobacco mosaic virus in Tomato Through Foliar Application of Bacillus amyloliquefaciens Strain TBorg1 Culture Filtrate

The application of microbe-derived products as natural biocontrol agents to boost systemic disease resistance to virus infections in plants is a prospective strategy to make agriculture more sustainable and environmentally friendly. In the current study, the rhizobacterium Bacillus amyloliquefaciens strain TBorg1 was identified based on 16S rRNA, rpoB, and gyrA gene sequences, and evaluated for its efficiency in conferring protection of tomato from infection by Tobacco mosaic virus (TMV). Under greenhouse circumstances, foliar sprays of TBorg1 culture filtrate (TBorg1-CF) promoted tomato growth, lowered disease severity, and significantly decreased TMV accumulation in systemically infected leaves of treated plants relative to untreated controls. TMV accumulation was reduced by 90% following the dual treatment, applied 24 h before and after TMV infection. Significant increases in levels of total soluble carbohydrates, proteins, and ascorbic acid were also found. In addition, a significant rise in activities of enzymes capable of scavenging reactive oxygen species (PPO and POX), as well as decreased levels of non-enzymatic oxidative stress markers (H(2)O(2) and MDA) were observed, compared to untreated plants. Enhanced systemic resistance to TMV was indicated by significantly increased transcript accumulation of polyphenolic pathway (C4H, HCT, and CHI) and pathogenesis-related (PR-1 and PR-5) genes. Out of the 15 compounds identified in the GC-MS analysis, 1,2-benzenedicarboxylic acid mono(2-ethylhexyl) ester and phenol, 2,4-bis(1,1-dimethylethyl), as well as L-proline, N-valeryl-, and heptadecyl ester were present in the highest concentrations in the ethyl acetate extract of TBorg1-CF. In addition, significant amounts of n-hexadecanoic acid, pyrrolo [1,2-a] pyrazine-1,4-dione hexahydro-3-(2-methylpropyl)-, nonane, 5-butyl-, and eicosane were also detected. These compounds may act as inducers of systemic resistance to viral infection. Our findings indicate that the newly isolated B. amyloliquefaciens strain TBorg1 could be a potentially useful rhizobacterium for promoting plant growth and a possible source of biocontrol agents for combating plant virus infections

Comparative Analysis of the Expression Profiles of Pathogenesis-Related Genes in Tomato Systemically Infected with Tobacco Mosaic and Cucumber Mosaic Viruses

In this study, we used RT-qPCR to examine how PR genes were expressed in model tomato (Solanum lycopersicum L.) plants that had been infected with TMV or CMV. Under greenhouse conditions, the indirect ELISA data showed that both viruses were detected for the first time at 6 dpi. Then, the levels of accumulation increased very quickly, reaching a peak of 15 dpi. During the course of the study (1–15 dpi), the Delta CT, NormFinder, BestKeeper, and GeNorm software tools revealed that the β-actin gene was the most informative reference gene in the virally infected tomato tissues. For both the TMV- and CMV-infected tomato plants, the transcriptional expression levels of most tested genes changed between activation and repression, especially in the first 12 dpi. Compared to mock-inoculated plants, the expression levels of PR-1 were induced at all time intervals except at 8 dpi for CMV and at 6, 7, and 8 dpi for TMV infection. Conversely, the greater activation and accumulation of both viruses were associated with the greater up-regulation of PR-2 at 8 dpi, with relative expression levels of 7.28- and 5.84-fold for TMV and CMV, respectively. The up-regulated expression of PR-3, PR-4, and PR-7 was shown at 4 dpi. In contrast, the PR-5 gene was inhibited in TMV at 1 dpi until 9 dpi, and the induction of this gene at 10 dpi increased by 1.72-fold, but PR-5 was observed to up-regulate the expression of CMV at 1 dpi. This study provides the first valuable information on the comparative transcriptional levels of these tomato genes between TMV and CMV infections

<i>Trichoderma viride</i> Isolate Tvd44 Enhances Potato Growth and Stimulates the Defense System against Potato Virus Y

Biological treatments may be employed to combat viral plant infections. In this study, Trichoderma viride was applied as a biocontrol agent to enhance the systemic resistance of potato plants against potato virus Y (PVY). T. viride isolate Tvd44 (OQ991378) was isolated and molecularly characterized before being used as an agent against PVY. The foliar application of Tvd44 on PVY-inoculated potatoes significantly promoted plant growth, height, roots, and number of leaves. Results also showed that the levels of peroxidase (POX), polyphenol oxidase (PPO), total proteins, and chlorophyll increased in potato leaves 21 days post-inoculation compared to untreated plants. Results of qPCR assays conducted on Tvd44-treated plants exhibited a reduction in PVY-CP accumulation levels up to 18.76-fold compared to untreated plants (101.82-fold). qPCR results also showed that defense-related genes (PR-1, POD, PAL, CHS, and HQT) were highly expressed in all Tvd44-treated plants. Three compounds: thiocarbamic acid, N,N-dimethyl, S-1,3-diphenyl-2-butenyl ester; 1,1-dicyano-2-methyl-4-(p-cyanophenyl) propane; and trans-[(2,3-diphenylcyclopropyl)methyl] phenyl sulfide were the most abundant compounds detected in the ethyl acetate extract of Tvd44-culture filtrate using GC–MS analysis. Our finding supports the efficacy of T. viride isolate Tvd44 as a potential agent that can successfully control PVY infections in potatoes and increase the productivity of the crop

Molecular, serological and biological characterizations of Potato Leaf Roll Virus in infected potato plants in Egypt, and its effects on plant cell organelles

Potato Leaf Roll Virus (PLRV) is one of the most serious viruses infecting potato plant (Solanum tuberosum L.) in Egypt. Indirect Enzyme Linked Immunosorbent Assay (Indirect- ELISA) results revealed that 70 % of the collected samples were infected with PLRV. A multiplex Polymerase Chain Reaction (PCR) was carried out using three different sets of primers, specific for both PLRV and Potato virus Y (PVY) isolates. For confirmation; the movement coat protein (MP) gene was isolated from the infected plant tissues, and a band with molecular size 336 bp was obtained using Reverse transcription-Polymerase chain reaction (RT-PCR). The DNA sequence of the Egyptian PLRV-Banha -MP gene was deposited in GenBank under an accession number of KR002119. Moreover, sequence analysis revealed that the Egyptian PLRV isolate was closely related to a New Zealand isolate of PLRV (GU002341), with identity of 100%. Transmission electron microscope (TEM) examination of PLRV showed isometric particles, with approximate size of 24-30 nm. The cytopathological examination of the potato plant infected with PLRV revealed many cellular effects such as; partially degraded and deformed chloroplast, starch with an increased size and color change, in addition to nucleus and cytoplasmic bridge. It could be concluded that PLRV is present in Egypt, infecting most of the potato cultivars. Moreover, four different strains of PLRV were detected based on the Single strand conformation polymorphism (SSCP) of the MP gene. The aims of the current study were to identify the PLRV infection of potato plant in Egypt using; molecular, serological and biological methods, in addition to studying the effect of this virus on potato cell organelles. This is the first record of the presence of four different strains of PLRV infecting potato in Egypt, using SSCP assay

Foliar Application of Nanoclay Promotes Potato (Solanum tuberosum L.) Growth and Induces Systemic Resistance against Potato Virus Y

Potato virus Y (PVY) is one of the most harmful phytopathogens. It causes big problems for potatoes and other important crops around the world. Nanoclays have been extensively studied for various biomedical applications. However, reports on their interactions with phytopathogens, particularly viral infections, are still limited. In this study, the protective activity of Egyptian nanoclay (CE) and standard nanoclay (CS) against PVY was evaluated on potato (Solanum tuberosum L.) plants. Their physicochemical and morphological properties were examined with scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and energy dispersive spectrometer (EDS). SEM and TEM analyses revealed that CE has a spherical and hexagonal structure ranging from 20 to 80 nm in size, while CS has boulder-like and tubular structures of about 320 nm in size. FTIR and EDS showed that both nanoclay types have different functional groups and contain many vital plant nutrients that are necessary for every stage and process of the plant, including development, productivity, and metabolism. Under greenhouse conditions, a 1% nanoclay foliar application enhanced potato growth, reduced disease symptoms, and reduced PVY accumulation levels compared with non-treated plants. Significant increases in levels of antioxidant enzymes (PPO and POX) and considerable decreases in oxidative stress markers (MDA and H2O2) were also reported. Moreover, a significant increase in the transcriptional levels of defense-related genes (PAL-1, PR-5, and CHI-2) was observed. All experiment and analysis results indicate that the CE type is more effective than the CS type against PVY infection. Based on these results, the foliar applications of nanoclay could be used to manage plant viral infections in a way that is both effective and environmentally friendly. To our knowledge, this is the first report of the antiviral activity of the foliar application of nanoclay against PVY infection

Mapping archaeal diversity in soda lakes by coupling 16S rRNA PCR-DGGE analysis with remote sensing and GIS technology

Abstract: The haloarchaeal diversity of four hypersaline alkaline lakes from the Wadi El-Natrun depression (Northern Egypt) was investigated using culture-independent polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rRNA gene phylotypes, which was combined with remote sensing and geographic information system (GIS) data to highlight the distribution pattern of the microbial diversity in water and sediment samples. The majority of archaeal sequences identified in all four lakes belonged to the phyla Euryarchaeota and Crenarchaeota. Sediment samples from Beida Lake and water samples from El-Hamra Lake showed the highest levels of archaeal diversity. Sequence similarities ≥ 95% were found between six of the acquired clones and uncultured Halorhabdus, Euryarchaeota, and archaeon clones. In addition, two clones shared a high level of sequence similarity (97%) with unclassified archaea, while other nine clones exhibited 96% to 99% sequence similarity with uncultured archaeon clones, and only one clone showed 97% identity with an uncultured Crenarchaeota. Likewise, 7 DGGE bands presented a sequence similarity of 90 to 98% to Halogranum sp., Halalkalicoccus tibetensis, Halalkalicoccus jeotgali, uncultured Halorubrum, Halobacteriaceae sp., or uncultured haloarchaeon. In conclusion, while the variety of alkaliphilic haloarchaea in the examined soda lakes was restricted, the possibility of uncovering novel species for biotechnological applications from these extreme habitats remains promising

Global economic burden of unmet surgical need for appendicitis

Background There is a substantial gap in provision of adequate surgical care in many low- and middle-income countries. This study aimed to identify the economic burden of unmet surgical need for the common condition of appendicitis. Methods Data on the incidence of appendicitis from 170 countries and two different approaches were used to estimate numbers of patients who do not receive surgery: as a fixed proportion of the total unmet surgical need per country (approach 1); and based on country income status (approach 2). Indirect costs with current levels of access and local quality, and those if quality were at the standards of high-income countries, were estimated. A human capital approach was applied, focusing on the economic burden resulting from premature death and absenteeism. Results Excess mortality was 4185 per 100 000 cases of appendicitis using approach 1 and 3448 per 100 000 using approach 2. The economic burden of continuing current levels of access and local quality was US $92 492 million using approach 1 and$73 141 million using approach 2. The economic burden of not providing surgical care to the standards of high-income countries was $95 004 million using approach 1 and$75 666 million using approach 2. The largest share of these costs resulted from premature death (97.7 per cent) and lack of access (97.0 per cent) in contrast to lack of quality. Conclusion For a comparatively non-complex emergency condition such as appendicitis, increasing access to care should be prioritized. Although improving quality of care should not be neglected, increasing provision of care at current standards could reduce societal costs substantially