Induced genetic diversity through mutagenesis in wheat gene pool and significant use of SCoT markers to underpin key agronomic traits.

BACKGROUND: This research explores the efficacy of mutagenesis, specifically using sodium azide (SA) and hydrazine hydrate (HZ) treatments, to introduce genetic diversity and enhance traits in three wheat (Triticum aestivum L.) genotypes. The experiment entails subjecting the seeds to different doses of SA and HZ and cultivating them in the field for two consecutive generations: M1 (first generation) and M2 (second generation). We then employed selective breeding techniques with Start Codon Targeted (SCoT) markers to select traits within the wheat gene pool. Also, the correlation between SCoT markers and specific agronomic traits provides insights into the genetic mechanisms underlying mutagenesis-induced changes in wheat. RESULTS: In the study, eleven genotypes were derived from parent varieties Sids1, Sids12, and Giza 168, and eight mutant genotypes were selected from the M1 generation and further cultivated to establish the M2 generation. The results revealed that various morphological and agronomical characteristics, such as plant height, spikes per plant, spike length, spikelet per spike, grains per spikelet, and 100-grain weight, showed increases in different genotypes from M1 to M2. SCoT markers were employed to assess genetic diversity among the eleven genotypes. The bioinformatics analysis identified a correlation between SCoT markers and the transcription factors ABSCISIC ACID INSENSITIVE3 (ABI3) and VIVIPAROUS1 (VP1), crucial for plant development, growth, and stress adaptation. A comprehensive examination of genetic distance and the function identification of gene-associated SCoT markers may provide valuable insights into the mechanisms by which SA and HZ act as mutagens, enhancing wheat agronomic qualities. CONCLUSIONS: This study demonstrates the effective use of SA and HZ treatments to induce gene diversity through mutagenesis in the wheat gene pool, resulting in the enhancement of agronomic traits, as revealed by SCoT markers. The significant improvements in morphological and agronomical characteristics highlight the potential of mutagenesis techniques for crop improvement. These findings offer valuable information for breeders to develop effective breeding programs to enhance wheat quality and resilience through increased genetic diversity.

Potentiating Biosynthesis of Alkaloids and Polyphenolic Substances in Catharanthus roseus Plant Using ĸ-Carrageenan.

Catharanthus roseus is a medicinal plant that produces indole alkaloids, which are utilized in anticancer therapy. Vinblastine and vincristine, two commercially important antineoplastic alkaloids, are mostly found in the leaves of Catharanthus roseus. ĸ-carrageenan has been proven as plant growth promoting substance for a number of medicinal and agricultural plants. Considering the importance of ĸ-carrageenan as a promoter of plant growth and phytochemical constituents, especially alkaloids production in Catharanthus roseus, an experiment was carried out to explore the effect of ĸ-carrageenan on the plant growth, phytochemicals content, pigments content, and production of antitumor alkaloids in Catharanthus roseus after planting. Foliar application of ĸ-carrageenan (at 0, 400, 600 and 800 ppm) significantly improved the performance of Catharanthus roseus. Phytochemical analysis involved determining the amount of total phenolics (TP), flavonoids (F), free amino acids (FAA), alkaloids (TAC) and pigments contents by spectrophotometer, minerals by ICP, amino acids, phenolic compounds and alkaloids (Vincamine, Catharanthine, Vincracine (Vincristine), and vinblastine) analysis uses HPLC. The results indicated that all examined ĸ-carrageenan treatments led to a significant (p ≤ 0.05) increase in growth parameters compared to the untreated plants. Phytochemical examination indicates that the spray of ĸ-carrageenan at 800 mg L-1 increased the yield of alkaloids (Vincamine, Catharanthine and Vincracine (Vincristine)) by 41.85 µg/g DW, total phenolic compounds by 3948.6 µg gallic/g FW, the content of flavonoids 951.3 µg quercetin /g FW and carotenoids content 32.97 mg/g FW as compared to the control. An amount of 400 ppm ĸ-carrageenan treatment gave the best contents of FAA, Chl a, Chl b and anthocyanin. The element content of K, Ca, Cu, Zn and Se increased by treatments. Amino acids constituents and phenolics compounds contents were altered by ĸ-carrageenan.

Constitutive overexpression of GsIMaT2 gene from wild soybean enhances rhizobia interaction and increase nodulation in soybean (Glycine max).

BACKGROUND: Since the root nodules formation is regulated by specific and complex interactions of legume and rhizobial genes, there are still too many questions to be answered about the role of the genes involved in the regulation of the nodulation signaling pathway. RESULTS: The genetic and biological roles of the isoflavone-7-O-beta-glucoside 6″-O-malonyltransferase gene GsIMaT2 from wild soybean (Glycine soja) in the regulation of nodule and root growth in soybean (Glycine max) were examined in this work. The effect of overexpressing GsIMaT2 from G. soja on the soybean nodulation signaling system and strigolactone production was investigated. We discovered that the GsIMaT2 increased nodule numbers, fresh nodule weight, root weight, and root length by boosting strigolactone formation. Furthermore, we examined the isoflavone concentration of transgenic G. max hairy roots 10 and 20 days after rhizobial inoculation. Malonyldaidzin, malonylgenistin, daidzein, and glycitein levels were considerably higher in GsMaT2-OE hairy roots after 10- and 20-days of Bradyrhizobium japonicum infection compared to the control. These findings suggest that isoflavones and their biosynthetic genes play unique functions in the nodulation signaling system in G. max. CONCLUSIONS: Finally, our results indicate the potential effects of the GsIMaT2 gene on soybean root growth and nodulation. This study provides novel insights for understanding the epistatic relationship between isoflavones, root development, and nodulation in soybean. HIGHLIGHTS: * Cloning and Characterization of 7-O-beta-glucoside 6″-O-malonyltransferase (GsIMaT2) gene from wild soybean (G. soja). * The role of GsIMaT2 gene in the regulation of root nodule development. *Overexpression of GsMaT2 gene increases the accumulation of isoflavonoid in transgenic soybean hairy roots. * This gene could be used for metabolic engineering of useful isoflavonoid production.

Microbial Degradation, Spectral analysis and Toxicological Assessment of Malachite Green Dye by Streptomyces exfoliatus.

Malachite green (MG) dye is a common environmental pollutant that threatens human health and the integrity of the Earth's ecosystem. The aim of this study was to investigate the potential biodegradation of MG dye by actinomycetes species isolated from planted soil near an industrial water effluent in Cairo, Egypt. The Streptomyces isolate St 45 was selected according to its high efficiency for laccase production. It was identified as S. exfoliatus based on phenotype and 16S rRNA molecular analysis and was deposited in the NCBI GenBank with the gene accession number OL720220. Its growth kinetics were studied during an incubation time of 144 h, during which the growth rate was 0.4232 (µ/h), the duplication time (td) was 1.64 d, and multiplication rate (MR) was 0.61 h, with an MG decolorization value of 96% after 120 h of incubation at 25 °C. Eleven physical and nutritional factors (mannitol, frying oil waste, MgSO4, NH4NO3, NH4Cl, dye concentration, pH, agitation, temperature, inoculum size, and incubation time) were screened for significance in the biodegradation of MG by S. exfoliatus using PBD. Out of the eleven factors screened in PBD, five (dye concentration, frying oil waste, MgSO4, inoculum size, and pH) were shown to be significant in the decolorization process. Central composite design (CCD) was applied to optimize the biodegradation of MG. Maximum decolorization was attained using the following optimal conditions: food oil waste, 7.5 mL/L; MgSO4, 0.35 g/L; dye concentration, 0.04 g/L; pH, 4.0; and inoculum size, 12.5%. The products from the degradation of MG by S. exfoliatus were characterized using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). The results revealed the presence of several compounds, including leuco-malachite green, di(tert-butyl)(2-phenylethoxy) silane, 1,3-benzenedicarboxylic acid, bis(2-ethylhexyl) ester, 1,4-benzenedicarboxylic acid, bis(2-ethylhexyl) ester, 1,2-benzenedicarboxylic acid, di-n-octyl phthalate, and 1,2-benzenedicarboxylic acid, dioctyl ester. Moreover, the phytotoxicity, microbial toxicity, and cytotoxicity tests confirmed that the byproducts of MG degradation were not toxic to plants, microbes, or human cells. The results of this work implicate S. exfoliatus as a novel strain for MG biodegradation in different environments.

Domiciliation of Trichoderma asperellum Suppresses Globiosporangium ultimum and Promotes Pea Growth, Ultrastructure, and Metabolic Features.

The beneficial microorganisms represent a new and hopeful solution for a sustainable environment and development. In this investigation, Trichoderma asperellum ZNW, isolated from seeds, was domiciliated within the pea plant for improving growth, disease management, and enhancement of productivity. Globisporangium ultimum NZW was isolated from deformed pea seeds, representing the first record of the pathogen caused by pea damping-off. Both fungi were molecularly identified. T. asperellum ZNW produced several lytic enzymes and bioactive metabolites as detected by GC-MC. The SEM illustrated the mycoparasitic behavior of T. asperellum ZNW on G. ultimum NZW mycelia. In the pot experiment, T. asperellum domiciliated the root and grew as an endophytic fungus, leading to root vessel lignification. Under soil infection, T. asperellum reduced damping-off, by enhancing peroxidase, polyphenol, total phenols, and photosynthetic pigments content. The vegetative growth, yield, and soil dehydrogenase activity were improved, with an enhancement in the numerical diversity of the microbial rhizosphere. This work may enable more understanding of the plant-fungal interaction, yet, working on domiciliation is recommended as a new approach to plant protection and growth promotion under various ecological setups.

Changes in Metabolite Profiling and Expression Levels of Key Genes Involved in the Terpenoid Biosynthesis Pathway in Garden Sage (Salvia officinalis) under the Effect of Hydrazine Hydrate.

Mutagenesis is a highly efficient tool for establishing genetic variation and is widely used for genetic enhancement in various plants. The key benefit of mutation breeding is the prospect of enhancing one or several characteristics of a variety without altering the genetic background. In this study, we exposed the seeds of Salvia officinalis to four concentrations of hydrazine hydrate (HZ), i.e., (0%, 0.1%, 0.2%, and 0.3%) for 6 h. The contents of terpenoid compounds in the S. officinalis plantlets driven from the HZ-treated seeds were determined by GC-MS, which resulted in the identification of a total of 340 phytochemical compounds; 163 (87.48%), 145 (84.49%), 65 (97.45%), and 62 (98.32%), from the four concentrations of HZ (0%, 0.1%, 0.2%, and 0.3%), respectively. Furthermore, we used the qRT-PCR system to disclose the "transcriptional control" for twelve TPS genes related to terpenoid and terpene biosynthesis, namely, SoGPS, SoMYRS, SoNEOD, SoCINS, SoSABS, SoLINS, SoFPPS, SoHUMS, SoTPS6, SoSQUS, SoGGPS, and SoGA2. Altogether, results are likely to ensure some positive relationship between the concentrations of the chemical mutagen HZ used for treating the seeds, the type and amount of the produced terpenes, and the expression of their corresponding genes.

Inhibition Mechanism of Methicillin-Resistant Staphylococcus aureus by Zinc Oxide Nanorods via Suppresses Penicillin-Binding Protein 2a.

Methicillin-resistant Staphylococcus aureus (MRSA) causes life-threatening infections. Zinc oxide is well known as an effective antibacterial drug against many bacterial strains. We investigated the performance of zinc oxide nanorods synthesized by Albmiun as a biotemplate as an antibacterial drug in this study; the fabrication of zinc oxide nanorods was synthesized by sol-gel methods. We performed physicochemical characterization of zinc oxide nanorods by physiochemical techniques such as FTIR spectroscopy, X-ray diffraction, and TEM and investigation of their antimicrobial toxicity efficiency by MIC, ATPase activity assay, anti-biofilm activity, and kill time assays, as well as the mecA, mecR1, blaR1, blaZ, and biofilm genes (ica A, ica D, and fnb A) by using a quantitative RT-PCR assay and the penicillin-binding protein 2a (PBP2a) level of MRSA by using a Western blot. The data confirmed the fabrication of rod-shaped zinc oxide nanorods with a diameter in the range of 50 nm, which emphasized the formation of zinc oxide nanoparticles with regular shapes. The results show that zinc oxide nanorods inhibited methicillin-resistant S. aureus effectively. The MIC value was 23 µg/mL. The time kill of ZnO-NRs against MRSA was achieved after 2 h of incubation at 4MIC (92 µg/mL) and after 3 h of incubation at 2MIC (46 µg/mL), respectively. The lowest concentration of zinc oxide nanorods with over 75% biofilm killing in all strains tested was 32 µg/mL. Also, we examined the influence of the zinc oxide nanorods on MRSA by analyzing mecA, mecR1, blaR1, and blaZ by using a quantitative RT-PCR assay. The data obtained revealed that the presence of 2× MIC (46 µg/mL) of ZnO-NRs reduced the transcriptional levels of blaZ, blaR1, mecA, and mecR1 by 3.4-fold, 3.6-fold, 4-fold, and 3.8-fold, respectively. Furthermore, the gene expression of biofilm encoding genes (ica A, ica B, ica D, and fnb A) was tested using quantitative real-time reverse transcriptase-polymerase chain reaction (rt-PCR). The results showed that the presence of 2× MIC (46 µg/mL) of ZnO-NRs reduced the transcriptional levels of ica A, ica B, ica D, and fnb A. Also, the PBP2a level was markedly reduced after treatment with ZnO-NRs.

Bacterial-Mediated Salinity Stress Tolerance in Maize (Zea mays L.): A Fortunate Way toward Sustainable Agriculture.

Sustainable agriculture is threatened by salinity stress because of the low yield quality and low crop production. Rhizobacteria that promote plant growth modify physiological and molecular pathways to support plant development and reduce abiotic stresses. The recent study aimed to assess the tolerance capacity and impacts of Bacillus sp. PM31 on the growth, physiological, and molecular responses of maize to salinity stress. In comparison to uninoculated plants, the inoculation of Bacillus sp. PM31 improved the agro-morphological traits [shoot length (6%), root length (22%), plant height (16%), fresh weight (39%), dry weight (29%), leaf area (11%)], chlorophyll [Chl a (17%), Chl b (37%), total chl (22%)], carotenoids (15%), proteins (40%), sugars (43%), relative water (11%), flavonoids (22%), phenols (23%), radical scavenging capacity (13%), and antioxidants. The Bacillus sp. PM31-inoculated plants showed a reduction in the oxidative stress indicators [electrolyte leakage (12%), H2O2 (9%), and MDA (32%)] as compared to uninoculated plants under salinity and increased the level of osmolytes [free amino acids (36%), glycine betaine (17%), proline (11%)]. The enhancement of plant growth under salinity was further validated by the molecular profiling of Bacillus sp. PM31. Moreover, these physiological and molecular mechanisms were accompanied by the upregulation of stress-related genes (APX and SOD). Our study found that Bacillus sp. PM31 has a crucial and substantial role in reducing salinity stress through physiological and molecular processes, which may be used as an alternative approach to boost crop production and yield.

Evaluation of nanoceria on cadmium uptake in Triticum aestivum (L.) and its implications for dietary health risk.

In recent times, significant attention has been directed toward the synthesis and application of nanoparticles (NPs) in agriculture sector. In current study, nanoceria (CeO2 NPs) synthesized by green method were employed to address cadmium (Cd) accumulation in wheat (Triticum aestivum L.) cultivated in field with excess Cd. The application of CeO2 NPs was carried out through foliar spraying, performed twice during the growth of T. aestivum. Four levels of CeO2 NPs were used: T0, T1, T2, and T3 as 0, 50, 75, and 100 mgL-1, respectively. Results highlighted the positive effects of CeO2 NPs on various growth parameters, including plant height, spike length, photosynthetic related attributes, as well as straw and grain of grains in comparison to T1 (control group). Furthermore, CeO2 NPs led to a reduction in oxidative stress in the leaves and enhanced in enzyme activities in comparison to T1. Notably, Cd concentrations in straw, roots, and grains exhibited a decline following the treatment with CeO2 NPs, in contrast to the control group. In terms of health implications, the calculated health risk index associated with dietary consumption of grains by adults remained below the defined threshold with supply of nanoparticles. Foliar application of CeO2 NPs proved to be an effective approach in reducing cadmium content in wheat grains. This reduction holds significant potential for minimizing the risk of cadmium exposure to human health through the food chain. Employing the green synthesis method amplifies the potential for extensive production and a wide array of environmental applications for CeO2 NPs. This dual capacity makes them proficient in tackling environmental stresses while concurrently mitigating adverse ecological effects.

Morphological, Biochemical, and Molecular Characterization of Exotic Brassica Germplasm.

Oilseed rape (Brassica napus L.) is an important oilseed crop. We examined the diversity of germplasm expressed at three distinct levels (i.e., morphological, biochemical, and DNA levels). In this study, 150 B. napus L. accessions with three check varieties were provided by Bioresources Conservation Institute. The germplasm was grown in field conditions for data collection of 15 quantitative and nine qualitative agro-morphological traits. The result indicated that for 15 quantitative agro-morphological traits, the highest coefficient of variation was recorded for plant height and days to flowering initiation. For nine qualitative traits, most of the accessions have a spatulate leaf, brown color seeds, yellow flowers, and erect silique attitude. The best adoptable genetically diverse exotic Brassica germplasms were selected, i.e., accessions 24178, 24881, 24199, 24214, 24242, and 24192. Based on biochemical analysis for high oil content and high oleic acid content, chakwal sarsoon and accession 24192 were selected. For high oleic and linoleic acids, accession 24181 performed best, for low erucic acid accessions 24177 and 24195. Based on molecular (SSR) markers, the top 50 selected genotypes were evaluated with 30 SSR markers. The 47 genotypes with three check varieties were clustered in six major groups; the coefficient of similarity ranged between 0.18 and 1.00. Based on SSR data, the germplasms accession 24178 and Abasin were the most diverse genotypes. These genotypes have the capacity and could be used in future breeding programs. High genetic variations were investigated through the SSR among the studied genotypes of Brassica napus L. The present study also concluded that SSR is a better technique for intraspecific genetic diversity. Other modern techniques should be applied such as SNIP for the investigation of a high level of genetic diversity among crop plants in the future.

Correction: Helal et al. Improving Yield Components and Desirable Eating Quality of Two Wheat Genotypes Using Si and NanoSi Particles under Heat Stress. Plants 2022, 11, 1819.

In the original publication [...].

Evaluating the genotoxicity of salinity stress and secondary products gene manipulation in lime, Citrus aurantifolia, plants.

Salinity is a significant abiotic stress that has a profound effect on growth, the content of secondary products, and the genotoxicity of cells. Lime, Citrus aurantifolia, is a popular plant belonging to the family Rutaceae. The interest in cultivating this plant is due to the importance of its volatile oil, which is included in many pharmaceutical industries, but C. aurantifolia plants are affected by the NaCl salinity levels. In the present study, a comet assay test has been applied to evaluate the genotoxic impact of salinity at 0, 50, 100, and 200 mM of NaCl on C. aurantifolia tissue-cultured plants. Furthermore, terpene gene expression was investigated using a semi-quantitative real-time polymerase chain reaction. Results from the two analyses revealed that 200 mM of NaCl stress resulted in high levels of severe damage to the C. aurantifolia plants' DNA tail 21.8%, tail length 6.56 µm, and tail moment 3.19 Unit. The relative highest expression of RtHK and TAT genes was 2.08, and 1.693, respectively, when plants were exposed to 200 mM of NaCl, whereas pv4CL2RT expressed 1.50 in plants subjected to 100 mM of NaCl. The accumulation of transcripts for the RTMYB was 0.951 when plants were treated with NaCl at 50 mM, and RtGPPS gene was significantly decreased to 0.446 during saline exposure at 100 mM. We conclude that the comet assay test offers an appropriate tool to detect DNA damage as well as RtHK, TAT, and pv4CL2RT genes having post-transcriptional regulation in C. aurantifolia plant cells under salinity stress. Future studies are needed to assess the application of gene expression and comet assay technologies using another set of genes that show vulnerability to different stresses on lime and other plants.

Cloning and Characterization of 1,8-Cineole Synthase (SgCINS) Gene From the Leaves of Salvia guaranitica Plant.

Monoterpenes are one of the most common groups belonging to the terpenoid family, with a C10 structure comprising of two isoprene units. Most of monoterpenes are volatile plant compounds, and they act as signaling molecules between plants and the environment, particularly as defensive compounds against herbivores and pathogens. In this study, 1,8-cineole synthase (SgCINS) gene was identified and cloned from the leaves of Salvia guaranitica plant. To examine the role of SgCINS in insect resistance, we transformed and expressed this gene into tobacco leaves. The metabolic analysis revealed that the production of various types and amount of terpenoid was increased and decreased in SgCINS overexpression and control lines, respectively, suggesting that overexpressing SgCINS in transgenic tobacco plants lead to an increase in the production of various types of terpenoids and other phytochemical compounds. These results indicated why transgenic tobacco was highly resistant against cotton worm than the highly susceptible control plants. Our results demonstrate that the SgCINS gene can play an important role in plants against cotton worm insect attack, and pave the way for using terpenoids genes for improving resistance to insect attack in higher plants.

Novel Mutations in Putative Nicotinic Acid Phosphoribosyltransferases of Mycobacterium tuberculosis and Their Effect on Protein Thermodynamic Properties.

pncB1 and pncB2 are two putative nicotinic acid phosphoribosyltransferases, playing a role in cofactor salvage and drug resistance in Mycobacterium tuberculosis. Mutations have been reported in first- and second-line drug targets, causing resistance. However, pncB1 and pncB2 mutational data are not available, and neither of their mutation effects have been investigated in protein structures. The current study has been designed to investigate mutations and also their effects on pncB1 and pncB2 structures. A total of 287 whole-genome sequenced data of drug-resistant Mycobacterium tuberculosis isolates from Khyber Pakhtunkhwa of Pakistan were retrieved (BioSample PRJEB32684, ERR2510337-ERR2510445, ERR2510546-ERR2510645) from NCBI. The genomic data were analyzed for pncB1 and pncB2 mutations using PhyResSE. All the samples harbored numerous synonymous and non-synonymous mutations in pncB1 and pncB2 except one. Mutations Pro447Ser, Arg286Arg, Gly127Ser, and delTCAGGCCG1499213>1499220 in pncB1 are novel and have not been reported in literature and TB databases. The most common non-synonymous mutations exhibited stabilizing effects on the pncB1 structure. Moreover, 36 out of 287 samples harbored two non-synonymous and 34 synonymous mutations in pncB2 among which the most common was Phe204Phe (TTT/TTC), present in 8 samples, which may have an important effect on the usage of specific codons that may increase the gene expression level or protein folding effect. Mutations Ser120Leu and Pro447Ser, which are present in the loop region, exhibited a gain in flexibility in the surrounding residues while Gly429Ala and Gly127Ser also demonstrated stabilizing effects on the protein structure. Inhibitors designed based on the most common pncB1 and pncB2 mutants may be a more useful strategy in high-burden countries. More studies are needed to elucidate the effect of synonymous mutations on organism phenotype.

Combined effect of Zinc lysine and biochar on growth and physiology of wheat (Triticum aestivum L.) to alleviate salinity stress.

Globally, Food security main threaten by abiotic stress like salinity and levels amongst the majority serious environmental stressors which reduce crop yield mass production. Biochar application has received much attention in agricultural practices as it enhances crop quality and production. The present study was carried out to analyze the role of lysine zinc and biochar on growth enhancement of wheat (Triticum aestivum L. cv. PU-2011) under saline stress (EC 7.17 dSm-1). Seeds were sown in pots containing saline soil with and without 2% biochar, and foliar application of Zn-lysine (0, 1.0, and 2.0 mM) was made at different time intervals during plant growth. A combined application of biochar and Zn-lysine 2.0 mM highly improved the physiological attributes such as chlorophyll a (37%), chlorophyll b (60%), total chlorophyll (37%), carotenoids (16%), photosynthesis rate (Pn) 45%, stomatal conductance (gs) 53%, transpiration rate (Tr) 56%, and water use efficiency (WUE) 55%. The levels of malondialdehyde (MDA) 38%, hydrogen peroxide (H2O2) 62%, and electrolyte leakage (EL) 48% were decreased with the combined application of biochar and Zn-lysine 2.0 mM as compared with other treatments. The activities of catalase (CAT) 67%, superoxide dismutase (SOD) 70%, and ascorbate peroxidase (APX) 61% as well as catalase (CAT) 67% were regulated with the combined biochar and Zn-lysine 2.0 mM treatment. Similarly, the combined application of biochar and zinc-lysine (2.0 mM) enhanced the growth and yield attributes such as shoot length (79%), root fresh weight (62%), shoot fresh weight (36%), root dry weight (86%), shoot dry weight (39%), grain weight (57%), and spike length (43%) as compared with untreated control. The concentrations of sodium (Na) decreased whereas potassium (K), iron (Fe), and zinc (Zn) concentrations were enhanced in plants with the combined application of Zn-lysine and biochar. Overall, results showed that the combined application of Zn-lysine (2.0 mM) and biochar significantly inhibited the negative effect of salinity and improved the growth and physiological performance of wheat plants. The combined use of Zn-lysine and biochar might be a practical solution to tackle salt stress in plants, but field studies by growing various crops under varied environmental conditions are needed before any recommendation to farmers.

Overexpression of Terpenoid Biosynthesis Genes Modifies Root Growth and Nodulation in Soybean (Glycine max).

Root nodule formation in many leguminous plants is known to be affected by endogen ous and exogenous factors that affect formation, development, and longevity of nodules in roots. Therefore, it is important to understand the role of the genes which are involved in the regulation of the nodulation signaling pathway. This study aimed to investigate the effect of terpenoids and terpene biosynthesis genes on root nodule formation in Glycine max. The study aimed to clarify not only the impact of over-expressing five terpene synthesis genes isolated from G. max and Salvia guaranitica on soybean nodulation signaling pathway, but also on the strigolactones pathway. The obtained results revealed that the over expression of GmFDPS, GmGGPPS, SgGPS, SgFPPS, and SgLINS genes enhanced the root nodule numbers, fresh weight of nodules, root, and root length. Moreover, the terpene content in the transgenic G. max hairy roots was estimated. The results explored that the monoterpenes, sesquiterpenes and diterpenes were significantly increased in transgenic soybean hairy roots in comparison with the control. Our results indicate the potential effects of terpenoids and terpene synthesis genes on soybean root growth and nodulation. The study provides novel insights for understanding the epistatic relationship between terpenoids, root development, and nodulation in soybean.

Improving Yield Components and Desirable Eating Quality of Two Wheat Genotypes Using Si and NanoSi Particles under Heat Stress.

Global climate change is a significant challenge that will significantly lower crop yield and staple grain quality. The present investigation was conducted to assess the effects of the foliar application of either Si (1.5 mM) or Si nanoparticles (1.66 mM) on the yield and grain quality attributes of two wheat genotypes (Triticum aestivum L.), cv. Shandweel 1 and cv. Gemmeiza 9, planted at normal sowing date and late sowing date (heat stress). Si and Si nanoparticles markedly mitigated the observed decline in yield and reduced the heat stress intensity index value at late sowing dates, and improved yield quality via the decreased level of protein, particularly glutenin, as well as the lowered activity of α-amylase in wheat grains, which is considered a step in improving grain quality. Moreover, Si and nanoSi significantly increased the oil absorption capacity (OAC) of the flour of stressed wheat grains. In addition, both silicon and nanosilicon provoked an increase in cellulose, pectin, total phenols, flavonoid, oxalic acid, total antioxidant power, starch and soluble protein contents, as well as Ca and K levels, in heat-stressed wheat straw, concomitant with a decrease in lignin and phytic acid contents. In conclusion, the pronounced positive effects associated with improving yield quantity and quality were observed in stressed Si-treated wheat compared with Si nanoparticle-treated ones, particularly in cv. Gemmeiza 9.

Molecular Identification of Zantedeschia Culture with Determination of Its Morphometric and Metabolic Activities for Mediterranean Acclimatization.

Calla lily (Zantedeschia albomaculata (Hook.) Baill.) is an herbaceous or semi-evergreen perennial grown from rhizomes. It is commonly named "Spotted Arum". Ribosomal RNAs (rRNAs) are found in all known organisms and are known for being functionally equivalent in all of them. A completely new in vitro culture protocol was applied to Z. albomaculata with two hormones, 6-Benzylaminopurine (BAP) and kinetin, to obtain full growth and multiplication. Due to their highly conserved sequences, the analysis of small-subunit rRNAs (16S-18S rRNAs) can provide precise statistical evaluation of a wide variety of phylogenetic connections. As a result, the plant's 18S rRNA gene allowed for identification and partial sequencing. Also, the traditional floral method and the novel application technique for identification were applied to Z. albomaculata. In this paper we systemically describe the structural strategies of the plant's adaptation to the surroundings at the morphological, physiological, and anatomical scale. Most the essential oils and fatty acids found in Z. albomaculata are omega fatty acids, octadecenoic acid, linoleic acid, and palmitic acid. All these fatty acids have industrial, medicinal, and pharmaceutical applications. The significant findings are the spadix sheathing leaves, and the precipitation of raphides calcium oxalate. The mitotic index showing the division activity was recorded, and it was 17.4%. The antimicrobial activity of Z. albomaculata ethanol extract was performed via the well diffusion method. This extract has shown high activity against Escherichia coli and Pseudomonas aeruginosa, compared to its lower activity against Bacillus cereus. By defining these characteristics and in vitro culture conditions, we will be able to acclimatize the plant in greenhouses, and then transfer it to the open field. The findings of this work identified the general characteristics of Zantedeschia albomaculata as an ornamental and medicinal plant in order to acclimatize this plant for cultivation in the Mediterranean climate.

Folic Acid Reinforces Maize Tolerance to Sodic-Alkaline Stress through Modulation of Growth, Biochemical and Molecular Mechanisms.

The mechanism by which folic acid (FA) or its derivatives (folates) mediates plant tolerance to sodic-alkaline stress has not been clarified in previous literature. To apply sodic-alkaline stress, maize seedlings were irrigated with 50 mM of a combined solution (1:1) of sodic-alkaline salts (NaHCO3 and Na2CO3; pH 9.7). Maize seedlings under stressed and non-stressed conditions were sprayed with folic acid (FA) at 0 (distilled water as control), 0.05, 0.1, and 0.2 mM. Under sodic-alkaline stress, FA applied at 0.2 mM significantly improved shoot fresh weight (95%), chlorophyll (Chl a (41%), Chl b (57%), and total Chl (42%)), and carotenoids (27%) compared to the untreated plants, while root fresh weight was not affected compared to the untreated plants. This improvement was associated with a significant enhancement in the cell-membrane stability index (CMSI), relative water content (RWC), free amino acids (FAA), proline, soluble sugars, K, and Ca. In contrast, Na, Na/K ratio, H2O2, malondialdehyde (MDA), and methylglycoxal (MG) were significantly decreased. Moreover, seedlings treated with FA demonstrated significantly higher activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), and ascorbate peroxidase (APX) compared to the untreated plants. The molecular studies using RT-qPCR demonstrated that FA treatments, specifically at 0.2 mM, enhanced the K+/Na+ selectivity and the performance of photosynthesis under alkaline-stress conditions. These responses were observed through up-regulation of the expression of the high-affinity potassium-transporter protein (ZmHKT1), the major core protein of photosystem II (D2-Protein), and the activity of the first enzyme of carbon fixation cycle in C4 plants (PEP-case) by 74, 248, and 225% over the untreated plants, respectively. Conversely, there was a significant down-regulation in the expression ZmSOS1 and ZmNHX1 by 48.2 and 27.8%, respectively, compared to the untreated plants.

Alpha Lipoic Acid as a Protective Mediator for Regulating the Defensive Responses of Wheat Plants against Sodic Alkaline Stress: Physiological, Biochemical and Molecular Aspects.

Recently, exogenous α-Lipoic acid (ALA) has been suggested to improve the tolerance of plants to a wide array of abiotic stresses. However, there is currently no definitive data on the role of ALA in wheat plants exposed to sodic alkaline stress. Therefore, this study was designed to evaluate the effects of foliar application by ALA at 0 (distilled water as control) and 20 µM on wheat seedlings grown under sodic alkaline stress (50 mM 1:1 NaHCO3 & Na2CO3; pH 9.7. Under sodic alkaline stress, exogenous ALA significantly (p ≤ 0.05) improved growth (shoot fresh and dry weight), chlorophyll (Chl) a, b and Chl a + b, while Chl a/b ratio was not affected. Moreover, leaf relative water content (RWC), total soluble sugars, carotenoids, total soluble phenols, ascorbic acid, K and Ca were significantly increased in the ALA-treated plants compared to the ALA-untreated plants. This improvement was concomitant with reducing the rate of lipid peroxidation (malondialdehyde, MDA) and H2O2. Superoxide dismutase (SOD) and ascorbate peroxidase (APX) demonstrated greater activity in the ALA-treated plants compared to the non-treated ones. Conversely, proline, catalase (CAT), guaiacol peroxidase (G-POX), Na and Na/K ratio were significantly decreased in the ALA-treated plants. Under sodic alkaline stress, the relative expression of photosystem II (D2 protein; PsbD) was significantly up-regulated in the ALA treatment (67% increase over the ALA-untreated plants); while Δ pyrroline-5-carboxylate synthase (P5CS), plasma membrane Na+/H+ antiporter protein of salt overly sensitive gene (SOS1) and tonoplast-localized Na+/H+ antiporter protein (NHX1) were down-regulated by 21, 37 and 53%, respectively, lower than the ALA-untreated plants. These results reveal that ALA may be involved in several possible mechanisms of alkalinity tolerance in wheat plants.