Transcriptome analysis of Pennisetum americanum × Pennisetum purpureum and Pennisetum americanum leaves in response to high-phosphorus stress.

Excessive phosphorus (P) levels can disrupt nutrient balance in plants, adversely affecting growth. The molecular responses of Pennisetum species to high phosphorus stress remain poorly understood. This study examined two Pennisetum species, Pennisetum americanum × Pennisetum purpureum and Pennisetum americanum, under varying P concentrations (200, 600 and 1000 µmol·L- 1 KH2PO4) to elucidate transcriptomic alterations under high-P conditions. Our findings revealed that P. americanum exhibited stronger adaption to high-P stress compared to P. americanum× P. purpureum. Both species showed an increase in plant height and leaf P content under elevated P levels, with P. americanum demonstrating greater height and higher P content than P. americanum× P. purpureum. Transcriptomic analysis identified significant up- and down-regulation of key genes (e.g. SAUR, GH3, AHP, PIF4, PYL, GST, GPX, GSR, CAT, SOD1, CHS, ANR, P5CS and PsbO) involved in plant hormone signal transduction, glutathione metabolism, peroxisomes, flavonoid biosynthesis, amino acid biosynthesis and photosynthesis pathways. Compared with P. americanum× P. purpureum, P. americanum has more key genes in the KEGG pathway, and some genes have higher expression levels. These results contribute valuable insights into the molecular mechanisms governing high-P stress in Pennisetum species and offer implications for broader plant stress research.

Impact of LAB Fermentation on the Nutrient Content, Amino Acid Profile, and Estimated Glycemic Index of Sorghum, Pearl Millet, and Kodo Millet.

INTRODUCTION: Millets, owing to their rich nutritional and low-to-moderate glycemic index values, are termed superfoods; however, some anti-nutritional factors, such as tannins, limit the absorption of micro and macronutrients. Non-thermal processing technologies, such as fermentation, can improve nutrient content and reduce these anti-nutritional factors. METHODS: The effect of a controlled submerged fermentation of whole grain sorghum, pearl millet, and dehusked Kodo millet using mixed lactic acid bacteria (LAB) culture in tofu whey-based media on the proximate, antioxidant, tannin content, vitamin B, amino acids profile and estimated glycemic index (eGI) of different millets were evaluated. RESULTS: The protein content (2-12.5%), carbohydrate content (2-13.6%), antioxidant activity (3-49%), vitamin B complex, amino acid profile (89-90%), and eGI of whole grain sorghum, pearl millet, and dehusked Kodo millet improved due to LAB-assisted submerged fermentation. In contrast, fat (4-15%), ash (56-67%), crude fiber (5-34%), minerals, tannin and resistant starch content decreased due to LAB fermentation. CONCLUSION: Controlled LAB fermentation can improve the nutritional quality of sorghum and millets while reducing anti-nutritional factors. This non-thermal process can be adopted industrially to produce more palatable and nutritionally superior millet products.

Native desert plants have the potential for phytoremediation of phytotoxic metals in urban cities: implications for cities sustainability in arid environments.

Arid regions can benefit from using native desert plants, which require minimal freshwater and can aid in remediating soil phytotoxic metals (PTMs) from traffic emissions. In this study, we assessed the ability of three native desert plants-Pennisetum divisum, Tetraena qatarensis, and Brassica tournefortii-to accumulate phytotoxic metals (PTMs) in their different plant organs, including leaves, stems, and roots/rhizomes. The PTMs were analyzed in soil and plant samples collected from Dubai, United Arab Emirates (UAE). The results indicated significantly higher levels of PTMs on the soil surface than the subsurface layer. Brassica exhibited the highest concentrations of Fe and Zn, measuring 566.7 and 262.8 mg kg-1, respectively, while Tetraena accumulated the highest concentration of Sr (1676.9 mg kg-1) in their stems. In contrast, Pennisetum recorded the lowest concentration of Sr (21.0 mg kg-1), while Tetraena exhibited the lowest concentrations of Fe and Zn (22.5 and 30.1 mg kg-1) in their leaves. The roots of Pennisetum, Brassica, and Tetraena demonstrated the potential to accumulate Zn from the soil, with concentration factors (CF) of 1.75, 1.09, and 1.09, respectively. Moreover, Brassica exhibited the highest CF for Sr, measuring 2.34. Pennisetum, however, could not translocate PTMs from its rhizomes to other plant organs, as indicated by a translocation factor (TF) of 1. In contrast, Brassica effectively translocated the studied PTMs from its roots to the stem and leaves (except for Sr in the leaves). Furthermore, Pennisetum exclusively absorbed Zn from the soil into its leaves and stems, with an enrichment factor (EF) greater than 1. Brassica showed the ability to uptake the studied PTMs in its stem and leaves (except for Fe), while Tetraena primarily absorbed Sr and Zn into its stems. Based on the CF and TF results, Pennisetum appears to be a suitable species for phytostabilization of both Fe and Zn, while Brassica is well-suited for Sr and Zn polluted soils. Tetraena shows potential for Zn phytoremediation. These findings suggest that these plants are suitable for PTMs phytoextraction. Furthermore, based on the EF results, these plants can efficiently sequester PTMs.

Exploration of the pearl millet phospholipase gene family to identify potential candidates for grain quality traits.

BACKGROUND: Phospholipases constitute a diverse category of enzymes responsible for the breakdown of phospholipids. Their involvement in signal transduction with a pivotal role in plant development and stress responses is well documented. RESULTS: In the present investigation, a thorough genome-wide analysis revealed that the pearl millet genome contains at least 44 phospholipase genes distributed across its 7 chromosomes, with chromosome one harbouring the highest number of these genes. The synteny analysis suggested a close genetic relationship of pearl millet phospholipases with that of foxtail millet and sorghum. All identified genes were examined to unravel their gene structures, protein attributes, cis-regulatory elements, and expression patterns in two pearl millet genotypes contrasting for rancidity. All the phospholipases have a high alpha-helix content and distorted regions within the predicted secondary structures. Moreover, many of these enzymes possess binding sites for both metal and non-metal ligands. Additionally, the putative promoter regions associated with these genes exhibit multiple copies of cis-elements specifically responsive to biotic and abiotic stress factors and signaling molecules. The transcriptional profiling of 44 phospholipase genes in two genotypes contrasting for rancidity across six key tissues during pearl millet growth revealed a predominant expression in grains, followed by seed coat and endosperm. Specifically, the genes PgPLD-alpha1-1, PgPLD-alpha1-5, PgPLD-delta1-7a, PgPLA1-II-1a, and PgPLD-delta1-2a exhibited notable expression in grains of both the genotypes while showing negligible expression in the other five tissues. The sequence alignment of putative promoters revealed several variations including SNPs and InDels. These variations resulted in modifications to the corresponding cis-acting elements, forming distinct transcription factor binding sites suggesting the transcriptional-level regulation for these five genes in pearl millet. CONCLUSIONS: The current study utilized a genome-wide computational analysis to characterize the phospholipase gene family in pearl millet. A comprehensive expression profile of 44 phospholipases led to the identification of five grain-specific candidates. This underscores a potential role for at least these five genes in grain quality traits including the regulation of rancidity in pearl millet. Therefore, this study marks the first exploration highlighting the possible impact of phospholipases towards enhancing agronomic traits in pearl millet.

Investigating the role of poly-γ-glutamic acid in Pennisetum giganteum phytoextraction of mercury-contaminated soil.

Farmland mercury (Hg) pollution poses a significant threat to human health, but there is a lack of highly efficient phytoextraction for its remediation at present. This study investigates the impact of poly-γ-glutamic acid (γ-PGA) on the phytoextraction capabilities of Pennisetum giganteum (P. giganteum) in Hg-contaminated soil. Our research indicates that amending γ-PGA to soil markedly enhances the assimilation of soil Hg by P. giganteum and transformation of Hg within itself, with observed increases in Hg concentrations in roots, stems, and leaves by 1.1, 4.3, and 18.9 times, respectively, compared to the control. This enhancement is attributed to that γ-PGA can facilitate the hydrophilic and bioavailable of soil Hg. Besides, γ-PGA can stimulate the abundance of Hg-resistance bacteria Proteobacteria in the rhizosphere of P. giganteum, thus increasing the mobility and uptake of soil Hg by P. giganteum roots. Moreover, the hydrophilic nature of Hg-γ-PGA complexes supports their transport via the apoplastic pathway, across the epidermis, and through the Casparian strip, eventually leading to immobilization in the mesophyll tissues. This study provides novel insights into the mechanisms of Hg phytoextraction, demonstrating that γ-PGA significantly enhances the effectiveness of P. giganteum in Hg uptake and translocation. The findings suggest a promising approach for the remediation of Hg-contaminated soil, offering a sustainable and efficient strategy for environmental management and health risk mitigation.

Impacts of soil type and drought stress on growth and cesium accumulation in Napier grass.

In our previous study, the decontamination efficiency of cesium-137 (137Cs) by Napier grass (Pennisetum purpureum Schum.) in the field was shown to be variable and often influenced by natural environmental factors. To elucidate the factors influencing this variable 137Cs-decontamination efficiency, we investigated the influences of soil type and drought stress on Cs accumulation using cesium-133 (133Cs) in Napier grass grown in plastic containers. The experiment was performed using two soil types (Soil A and B) and three different soil moisture conditions: well-watered control (CL), slight drought stress (SD), and moderate drought stress (MD). Overall, our results indicate that soil type and drought have a significant impact on plant growth and 133Cs accumulation in Napier grass. Plant height (PH), tiller number (TN), leaf width (Wleaf), and dry matter weight of aboveground parts (DWabove) and root parts (DWroot) in Soil B were greater than those in Soil A. Drought stress negatively affected chlorophyll fluorescence parameters (maximal quantum efficiency of photosystem (PS) II photochemistry and potential activity of PS II), PH, TN, Wleaf, DWabove, DWroot, and total 133Cs content (TCs), but it had a positive effect on 133Cs concentration. The 133Cs concentration in the aboveground parts (Csabove) was increased by MD approximately 1.62-fold in Soil A and 1.11-fold in Soil B compared to each CL counterpart. The TCs in the aboveground parts (TCsabove) decreased due to drought by approximately 19.9%-39.0% in Soil A and 49.9%-62.7% in Soil B; however, there was no significant effect on TCsabove due to soil type. The results of this study indicate that soil moisture is a key factor in maintaining Napier grass 137Cs-decontamination efficiency.

Introgression of Δ1-pyrroline-5-carboxylate synthetase (PgP5CS) confers enhanced resistance to abiotic stresses in transgenic tobacco.

Δ1-pyrroline-5-carboxylate synthetase (P5CS) is one of the key regulatory enzymes involved in the proline biosynthetic pathway. Proline acts as an osmoprotectant, molecular chaperone, antioxidant, and regulator of redox homeostasis. The accumulation of proline during stress is believed to confer tolerance in plants. In this study, we cloned the complete CDS of the P5CS from pearl millet (Pennisetum glaucum (L.) R.Br. and transformed into tobacco. Three transgenic tobacco plants with single-copy insertion were analyzed for drought and heat stress tolerance. No difference was observed between transgenic and wild-type (WT) plants when both were grown in normal conditions. However, under heat and drought, transgenic plants have been found to have higher chlorophyll, relative water, and proline content, and lower malondialdehyde (MDA) levels than WT plants. The photosynthetic parameters (stomatal conductance, intracellular CO2 concentration, and transpiration rate) were also observed to be high in transgenic plants under abiotic stress conditions. qRT-PCR analysis revealed that the expression of the transgene in drought and heat conditions was 2-10 and 2-7.5 fold higher than in normal conditions, respectively. Surprisingly, only P5CS was increased under heat stress conditions, indicating the possibility of feedback inhibition. Our results demonstrate the positive role of PgP5CS in enhancing abiotic stress tolerance in tobacco, suggesting its possible use to increase abiotic stress-tolerance in crops for sustained yield under adverse climatic conditions.

Comprehensive genomic analysis of Bacillus subtilis and Bacillus paralicheniformis associated with the pearl millet panicle reveals their antimicrobial potential against important plant pathogens.

BACKGROUND: Plant microbiome confers versatile functional roles to enhance survival fitness as well as productivity. In the present study two pearl millet panicle microbiome member species Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36 found to have beneficial traits including plant growth promotion and broad-spectrum antifungal activity towards taxonomically diverse plant pathogens. Understanding the genomes will assist in devising a bioformulation for crop protection while exploiting their beneficial functional roles. RESULTS: Two potential firmicute species were isolated from pearl millet panicles. Morphological, biochemical, and molecular characterization revealed their identities as Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36. The seed priming assays revealed the ability of both species to enhance plant growth promotion and seedling vigour index. Invitro assays with PBs 12 and PBl 36 showed the antibiosis effect against taxonomically diverse plant pathogens (Magnaporthe grisea; Sclerotium rolfsii; Fusarium solani; Alternaria alternata; Ganoderma sp.) of crops and multipurpose tree species. The whole genome sequence analysis was performed to unveil the genetic potential of these bacteria for plant protection. The complete genomes of PBs 12 and PBl 36 consist of a single circular chromosome with a size of 4.02 and 4.33 Mb and 4,171 and 4,606 genes, with a G + C content of 43.68 and 45.83%, respectively. Comparative Average Nucleotide Identity (ANI) analysis revealed a close similarity of PBs 12 and PBl 36 with other beneficial strains of B. subtilis and B. paralicheniformis and found distant from B. altitudinis, B. amyloliquefaciens, and B. thuringiensis. Functional annotation revealed a majority of pathway classes of PBs 12 (30) and PBl 36 (29) involved in the biosynthesis of secondary metabolites, polyketides, and non-ribosomal peptides, followed by xenobiotic biodegradation and metabolism (21). Furthermore, 14 genomic regions of PBs 12 and 15 of PBl 36 associated with the synthesis of RiPP (Ribosomally synthesized and post-translationally modified peptides), terpenes, cyclic dipeptides (CDPs), type III polyketide synthases (T3PKSs), sactipeptides, lanthipeptides, siderophores, NRPS (Non-Ribosomal Peptide Synthetase), NRP-metallophone, etc. It was discovered that these areas contain between 25,458 and 33,000 secondary metabolite-coding MiBiG clusters which code for a wide range of products, such as antibiotics. The PCR-based screening for the presence of antimicrobial peptide (cyclic lipopeptide) genes in PBs 12 and 36 confirmed their broad-spectrum antifungal potential with the presence of spoVG, bacA, and srfAA AMP genes, which encode antimicrobial compounds such as subtilin, bacylisin, and surfactin. CONCLUSION: The combined in vitro studies and genome analysis highlighted the antifungal potential of pearl millet panicle-associated Bacillus subtilis PBs12 and Bacillus paralicheniformis PBl36. The genetic ability to synthesize several antimicrobial compounds indicated the industrial value of PBs 12 and PBl 36, which shed light on further studies to establish their action as a biostimulant for crop protection.

Abiotic stress tolerance in pearl millet: Unraveling molecular mechanisms via transcriptomics.

Pearl millet (Pennisetum glaucum (L.)) is a vital cereal crop renowned for its ability to thrive in challenging environmental conditions; however, the molecular mechanisms governing its salt stress tolerance remain poorly understood. To address this gap, next-generation RNA sequencing was conducted to compare gene expression patterns in pearl millet seedlings exposed to salt stress with those grown under normal conditions. Our RNA sequencing analysis focused on shoots from 13-day-old pearl millet plants subjected to either salinity stress (150 mmol of NaCl for 3 days) or thermal stress (50°C for 60 s). Of 36,041 genes examined, 17,271 genes with fold changes ranging from 2.2 to 19.6 were successfully identified. Specifically, 2388 genes were differentially upregulated in response to heat stress, whereas 4327 genes were downregulated. Under salt stress conditions, 2013 genes were upregulated and 4221 genes were downregulated. Transcriptomic analysis revealed four common abiotic KEGG pathways that play crucial roles in the response of pearl millet to salt and heat stress: phenylpropanoid biosynthesis, photosynthesis-antenna proteins, photosynthesis, and plant hormone signal transduction. These metabolic pathways are necessary for pearl millet to withstand and adapt to abiotic stresses caused by salt and heat. Moreover, the pearl millet shoot heat stress group showed specific transcriptomics related to KEEG metabolic pathways such as cytochrome P450, cutin, suberine, and wax biosynthesis, zeatin biosynthesis, crocin biosynthesis, ginsenoside biosynthesis, saponin biosynthesis, and biosynthesis of various plant secondary metabolites. In contrast, pearl millet shoots exposed to salinity stress exhibited transcriptomic changes associated with KEEG metabolic pathways related to carbon fixation in photosynthetic organisms, mismatch repair, and nitrogen metabolism. Our findings underscore the remarkable cross-tolerance of pearl millet to simultaneous salt and heat stress, elucidated through the activation of shared abiotic KEGG pathways. This study emphasizes the pivotal role of transcriptomics analysis in unraveling the molecular responses of pearl millet under abiotic stress conditions.

Phytostabilization of fly ash from a coalmine in Botswana and biovalorisation of the recovered Napier grass (Pennisetum purpureum Schumach.).

The disposal of fly ash (FA) from coal power plants polluting the air, soil, and groundwater is a major environmental concern. Phytoremediation to rehabilitate fly ash dumpsites is a promising alternative but has practical concerns about the disposal of harvested biomass. This study investigated the effect of supplementing fly ash with fresh sewage sludge (FSS), aged sewage sludge, food waste, and compost (COM) to enhance the phytoremediation potential of Napier grass and its subsequent utilization for ethanol production. The highest removal of Mn (1196.12 g ha-1) and Ni (128.06 g ha-1) from FA could be obtained when Napier is grown in the presence of FSS and inorganic fertilizer (NPK). In addition, the highest bioethanol yield (19.31 g L-1) was obtained from Napier grown in fly ash with COM + NPK, thus providing additional economic benefits aside from the remediation process. Given the significant levels of heavy metals present in the pulp and bio-slurry after ethanol production, further research is required in this area to determine the best ways to utilize this waste such as converting it into biochar.

Using energy crops as a phytoremediation agent for fly ash dumpsites has the potential to remediate heavy metal contamination and provide additional economic benefits. Napier grass was able to tolerate high concentrations of heavy metals and yield high biomass in fly ash in the presence of organic amendments. The harvested biomass was successfully converted into substrate for bioethanol production using heavy metal-tolerant yeast. This is the first report on the production of ethanol from the phytoremediation biomass of Napier grass.

Rhizosphere microbiome of plants used in phytoremediation of mine tailing dams.

Rhizospheric microbial communities improve the effectiveness of hyperaccumulators in the phytoremediation of heavy metals. However, limited access to tailing dams and inadequate assessment of plants' phytoremediation potential limit the characterization of native accumulators, hindering the effectiveness of local remediation efforts. This study evaluates the heavy metal sequestration potentials of Pennisetum purpureum, Leucaena leucocephala, and Pteris vittata and their associated rhizospheric microbial communities at the Marlu and Pompora tailing dams in Ghana. The results indicate shoot hyperaccumulation of Cd (334.5 ± 6.3 mg/kg) and Fe (10,647.0 ± 12.6 mg/kg) in P. purpureum and L. leucocephala, respectively. Analysis of rhizospheric bacterial communities revealed the impact of heavy metal contamination on bacterial community composition, associating Fe and Cd hyperaccumulation with Bacillus, Arthrobacter, and Sphingomonas species. This study reports the hyperaccumulation potentials of L. leucocephala and P. purpureum enhanced by associated rhizosphere bacterial communities, suggesting their potential application as an environmentally friendly remediation process of heavy metals contaminated lands.

The novelty of this study is the report of the heavy metal accumulation potentials of the fast-growing herbaceous plants Leucaena leucocephala and Pennisetum purpureum at abandoned gold mine tailing dams in Ghana. Substantial amounts of Cd (334.5 mg/kg) and Fe (10,647.0 mg/kg) were accumulated in P. purpureum and L. leucocephala shoots, indicating their hyperaccumulation potentials. The concentration of heavy metal levels in the rhizosphere influenced the bacterial community structure, with the abundance of Bacillus, Arthrobacter, and Sphingomonas linked to hyperaccumulation. The outcome of this study supports the utilization of L. leucocephala and P. purpureum in the phytoremediation of lands contaminated with heavy metals.

Combined exposure of PVC-microplastic and mercury chloride (HgCl2) in sorghum (Pennisetum glaucum L.) when its seeds are primed titanium dioxide nanoparticles (TiO2-NPs).

The present work studied the impact of different levels of PVC-microplastics (PVC-MPs), namely 0 (no PVC-MPs), 2, and 4 mg L-1, along with mercury (Hg) levels of 0 (no Hg), 10, and 25 mg kg-1 in the soil, while concurrently applying titanium dioxide-nanoparticles (TiO2-NPs) at 0 (no TiO2-NPs), 50, and 100 µg mL-1 to sorghum (Pennisetum glaucum L.) plants. This study aimed to examine plant growth and biomass, photosynthetic pigments and gas exchange characteristics, oxidative stress indicators, and the response of various antioxidants (enzymatic and non-enzymatic) and their specific gene expression, proline metabolism, the AsA-GSH cycle, and cellular fractionation in the plants. The research outcomes indicated that elevated levels of PVC-MPs and Hg stress in the soil notably reduced plant growth and biomass, photosynthetic pigments, and gas exchange attributes. However, PVC-MPs and Hg stress also induced oxidative stress in the roots and shoots of the plants by increasing malondialdehyde (MDA), hydrogen peroxide (H2O2), and electrolyte leakage (EL) which also induced increased compounds of various enzymatic and non-enzymatic antioxidants and also the gene expression and sugar content. Furthermore, a significant increase in proline metabolism, the AsA-GSH cycle, and the pigmentation of cellular components was observed. Although, the application of TiO2-NPs showed a significant increase in plant growth and biomass, gas exchange characteristics, enzymatic and non-enzymatic compounds, and their gene expression and also decreased oxidative stress. In addition, the application of TiO2-NPs enhanced cellular fractionation and decreased the proline metabolism and AsA-GSH cycle in P. glaucum plants. These results open new insights for sustainable agriculture practices and hold immense promise in addressing the pressing challenges of heavy metal contamination in agricultural soils.

Purple Napiergrass (Pennisetum purpureum Schumach) Hot Water Extracts Ameliorate High-Fat Diet-Induced Obesity and Metabolic Disorders in Mice.

Purple Pennisetum (Pennisetum purpureum Schumach), a hybrid between Taihucao No. 2 and the local wild species of purple Pennisetum, has dark red stems and leaves due to its anthocyanin content. This study explores the potential of purple napiergrass extracts (PNE) in alleviating obesity and metabolic disorders induced by a high-fat diet in mice, where 50% of the caloric content is derived from fat. Mice were orally administered low-dose or high-dose PNE alongside a high-fat diet. Experimental findings indicate that PNE attenuated weight gain, reduced liver, and adipose tissue weight, and lowered blood cholesterol, triglyceride, low-density lipoprotein, and blood sugar levels. Stained sections showed that PNE inhibited lipid accumulation and fat hypertrophy in the liver. Immunoblotting analysis suggested that PNE improved the inflammatory response associated with obesity, dyslipidemia, and hyperglycemia induced by a high-fat diet. Furthermore, PNE potentially functions as a PPAR-γ agonist, increasing the adiponectin (ADIPOQ) concentration and suppressing inflammatory factors, while elevating the anti-inflammatory factor interleukin-10 (IL-10) in the liver. PNE-treated mice showed enhanced activation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) and AMP-activated protein kinase (AMPK) pathways and increased fatty acid oxidation and liver lipolysis. In conclusion, this study elucidated the mechanisms underlying the anti-inflammatory, PI3K/Akt, and AMPK pathways in a high-fat diet-induced obesity model. These findings highlight the potential of PNE in reducing weight, inhibiting inflammation, and improving blood sugar and lipid levels, showing the potential for addressing obesity-related metabolic disorders in humans.

[Effects of Combined Pollution of Microplastics and Cadmium on Microbial Community Structure and Function of Pennisetum hydridum Rhizosphere Soil].

The combined pollution of microplastics and heavy metals can potentially interact. This may have an important impact on the growth and development of plants and the rhizosphere microbial community and function. In this study, the effects of heavy metal cadmium combined with different types of microplastics(PE and PS), different particle sizes(13 µm and 550 µm), and different concentrations(0.1% and 1%) on Pennisetum hydridum growth were studied under pot conditions. The results showed that the effects of the combined pollution of MPs and Cd on plant dry weight and Cd accumulation varied with different types, concentrations, and particle sizes of MPs, and the combined pollution stress increased, whereas the Cd content and Cd accumulation decreased. Metagenomic analysis showed that the combined contamination of MPs and Cd could change the composition of the bacterial community and reduce bacterial diversity, among which the ACE index and Chao1 index in the 550 µm 0.1% PE+Cd treatment group were the most significant. Metagenomic analysis of microbial species function showed that the main functional groups were metabolism, amino acid transport and metabolism, energy generation and conversion, and signal transduction mechanisms. Compared with that under single Cd pollution, the addition of MPs could change the gene abundance of functional groups such as metabolism, amino acid transport and metabolism, and energy generation and conversion, and the effects of different MPs types, concentrations, and particle sizes varied. In this study, metagenomics and amplification sequencing were used to analyze the effects of the combined pollution of MPs and Cd on the bacterial community and function in P. hydridum in order to provide basic data and scientific basis for the ecotoxicological effects of the combined heavy metal pollution of MPs and its biological remediation.

Iron and zinc biofortification of rice by synergistic expression of OsNAS2 gene with monocot (Pennisetum glaucum) and dicot (Phaseolus vulgaris) ferritins.

Iron and zinc deficiencies are the most prevalent cause of global hidden hunger. Rice, being one of the most consumed crops worldwide, is suitable to target for Fe and Zn biofortification. In present study, we generated rice transgenic lines to meet the recommended dietary requirement of iron and zinc through endosperm specific expression of dicot (kidney bean) and monocot (pearl millet) Ferritins along with constitutive expression of rice nicotianamine synthase 2 (OsNAS2) gene. Visualization through perls' prussian staining and quantification by ICP-MS showed significant improvement in grain iron content in all the transgenic lines. The transgenic lines expressing any of the three selected gene combinations (PvFerrtin-OsNAS2, feedPgFerrtin-OsNAS2 and foodPgFerritin-OsNAS2), showed the potential to surpass the 30% of the estimated average requirement (13 µg/g Fe and 28 µg/g Zn) proposed for rice in HarvestPlus breeding program. Though the expression of PvFerritin along with OsNAS2 gene in IET10364 (indica) variety showed the best result, providing up to 4.2- and 3.5-fold increase in iron (30.56 µg/g) and zinc (60.1 µg/g) content, respectively; in polished grains compared to non-transgenic control. Thus, the lines developed in our study can be used for further breeding purpose to enhance the iron and zinc content in commercial rice varieties.

Effects of pin-to-plate atmospheric cold plasma for modification of pearl millet (Pennisetum glaucum) starch.

The present study was done to analyze the effect of atmospheric pressure non-thermal pin-to-plate plasma at a range of different voltages (170, 200, and 230V) at different time intervals (10, 20, and 30 mins) on under-utilized pearl millet starch. The untreated and treated starches were analyzed for amylose content, pH, carbonyl, and carboxyl group, reducing sugar, turbidity, water, and oil binding property, pasting property, DSC, FTIR, XRD, and molecular weight. As cold plasma contains highly reactive species and free radicals, it is expected to cause noticeable modifications in the attributes of treated starch. There has been a significant reduction (p < 0.05) in turbidity value by 38.97% and pH value of starch from 6.49 to 4.05. Plasma-treated samples produced clearer pastes with higher stability over storage time. Cold plasma treatment also led to an increase in the ζ potential. However, there has been no significant change in the water activity and oil-binding capacity of the starch. Reducing sugar content, average molecular weight, degree of polymerization, pasting property, XRD, and FTIR data confirmed that cross-linking takes place in samples treated at lower voltages and lesser time followed by depolymerization occurring in harshly treated plasma samples. The study thus points out the possible use of cold plasma for starch modification to produce starches with altered properties.

Dynamics of Phyllosphere Microbiota and Chemical Parameters at Various Growth Stages and Their Contribution to Anaerobic Fermentation of Pennisetum giganteum.

This work evaluated the dynamic changes of phyllosphere microbiota and chemical parameters at various growth stages of Pennisetum giganteum and their effects on the bacterial community, cooccurrence networks, and functional properties during anaerobic fermentation. P. giganteum was collected at two growth stages (early vegetative stage [PA] and late vegetative stage [PB]) and was naturally fermented (NPA and NPB) for 1, 3, 7, 15, 30, and 60 days, respectively. At each time point, NPA or NPB was randomly sampled for the analysis of chemical composition, fermentation parameter, and microbial number. In addition, the fresh, 3-day, and 60-day NPA and NPB were subjected to high-throughput sequencing and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional prediction analyses. Growth stage obviously affected the phyllosphere microbiota and chemical parameters of P. giganteum. After 60 days of fermentation, NPB had a higher lactic acid concentration and ratio of lactic acid to acetic acid but a lower pH value and ammonia nitrogen concentration than NPA. Weissella and Enterobacter were dominant in 3-day NPA and Weissella was dominant in 3-day NPB, while Lactobacillus was the most abundant genus in both 60-day NPA and NPB. The complexity of bacterial cooccurrence networks in the phyllosphere decreased with P. giganteum growth. The ensiling process further decreased the complexity of bacterial networks, with the simplest bacterial correlation structures in NPB. There were great differences in the KEGG functional profiles of PA and PB. Ensiling promoted the metabolism of lipid, cofactors, vitamins, energy, and amino acids but suppressed the metabolism of carbohydrates and nucleotides. Storage time had a greater influence than growth stage on bacterial community diversity, cooccurrence networks, and functional profiles of P. giganteum silage. Differences in bacterial diversity and functionality of P. giganteum silage caused by growth stage appear to be offset by long-term storage. IMPORTANCE The phyllosphere microbiota consists of various and complex microbes, including bacteria with crucial relevance to the quality and safety of fermented food and feed. It initially derives from soil and becomes specific to its host after interaction with plants and climate. Bacteria associated with the phyllosphere are highly abundant and diverse, but we know little about their succession. Here, the phyllospheric microbiota structure was analyzed within the growth of P. giganteum. We also evaluated the effects of phyllosphere microbiota and chemical parameter changes on the anaerobic fermentation of P. giganteum. We observed remarkable differences in bacterial diversity, cooccurrence, and functionality of P. giganteum at various growth stages and storage times. The obtained results are important for understanding the fermentation mechanism and may contribute to high-efficient production without additional cost.

Reduction in the cadmium (Cd) accumulation and toxicity in pearl millet (Pennisetum glaucum L.) by regulating physio-biochemical and antioxidant defense system via soil and foliar application of melatonin.

Cadmium (Cd) is among the toxic pollutants that harms the both animals and plants. The natural antioxidant, melatonin can improve Cd-stress tolerance but its potential role in reducing Cd stress and resilience mechanisms in pearl millet (Pennisetum glaucum L.) is remain unclear. The present study suggests that Cd causes severe oxidative damage by decreasing photosynthesis, and increasing reactive oxygen species (ROS), malondialdehyde content (MDA), and Cd content in different parts of pearl millet. However, exogenous melatonin (soil application and foliar treatment) mitigated the Cd toxicity and enhanced the growth, antioxidant defense system, and differentially regulated the expression of antioxidant-responsive genes i. e superoxide dismutase SOD-[Fe] 2, Fe-superoxide dismutase, Peroxiredoxin 2C, and L-ascorbate peroxidase-6. The results showed that foliar melatonin at F-200/50 significantly increased the plant height, chlorophyll a, b, a+b and carotenoids by 128%, 121%, 150%, 122%, and 69% over the Cd treatment, respectively. The soil and foliar melatonin at S-100/50 and F-100/50 reduced the ROS by 36%, and 44%, and MDA by 42% and 51% over the Cd treatment, respectively. Moreover, F200/50 significantly boosted the activities of antioxidant enzymes i. e SOD by 141%, CAT 298%, POD 117%, and APX 155% over the Cd treatment. Similarly, a significant reduction in Cd content in root, stem, and leaf was found on exposure to higher concentrations of exogenous melatonin. These findings suggest that exogenous melatonin may significantly and differentially improve the tolerance to Cd stress in crop plants. However, field applications, type of plant species, concentration of dose, and type of stress may vary with the degree of tolerance in crop plants.

Silicon-mediated regulation of cadmium transport and activation of antioxidant defense system enhances Pennisetum glaucum resistance to cadmium stress.

Pennisetum glaucum is an important forage grass for livestock. However, the large accumulation of cadmium (Cd) in plant tissues increases the risk of heavy metals entering the food chain in Cd-contaminated soils. Silicon (Si) can inhibit cadmium (Cd) uptake and enhance tolerance of plant to Cd toxicity, but whether and how Si alleviates Cd toxicity in grass and the underlying mechanisms are unclear. The present study explored the differential mechanisms of silicon-induced Cd transport in apoplast and symplast, Cd distribution in root tissue and antioxidant defense system in P. glaucum under Cd stress through hydroponic and pot experiments. The present results showed that exogenous Si supply significantly reduced Cd concentrations in apoplast and symplast; Si treatment increased monosilicic acid concentration in apoplast and symplast of the roots and shoots under Cd stress. Elemental analysis of root microdomains showed that Si treatment increased the distribution of Cd and Si in the endodermis by 42.6% and 14.0%, respectively. Si alleviated the adverse influences of Cd on plant growth, which were manifested in root morphological traits and root activity. In addition, Si addition significantly increased the activities of catalase and superoxide dismutase by 37.0% and 72.7%, and improved the efficiency of the ascorbate-glutathione cycle in Cd-stress shoots. Furthermore, Si significantly reduced the contents of hydrogen peroxide and superoxide anion in Cd-stressed shoots by 16.6% and 48.7%, respectively. These findings demonstrate that Si enhances the resistance of P. glaucum to Cd stress through regulating Cd transport pathways and activating antioxidant defense systems.

Physiological and transcriptome analyses demonstrate the silver nanoparticles mediated alleviation of salt stress in pearl millet (Pennisetum glaucum L).

Pearl millet (Pennisetum glaucum L.) is a highly nutritive-value summer-annual forage crop used for hay, silage, grazing, and green chop. However, abiotic stresses including salinity negatively affect its growth and productivity. Furthermore, the nanotechnology is attaining greater consideration to reduce the impact of environmental stresses in plants. In the present study, transcriptome responses of silver nanoparticles (AgNPs) in pearl millet under salinity were investigated. The treatments were given as Control, NaCl (250 mM), AgNPs (20 mg/L), and NaCl + AgNPs to pearl millet seedlings after thirteen days of seed sowing. After 1 h of given treatments, leaf samples were collected and subjected to physio-chemical examination and transcriptome analyses. Salt stress increased the hydrogen peroxide (H2O2), malondialdehyde (MDA) content, and proline as compared to other treatments. In addition, the combined applications of NaCl + AgNPs ameliorated the oxidative damage by increasing antioxidant enzymes activities including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). Furthermore, RNA sequencing data showed 6016 commonly annotated Differentially Expressed Transcripts (DETs) among various treated combinations. Among them, 427 transcripts were upregulated, and 136 transcripts were downregulated at nanoparticles vs control, 1469 upregulated and 1182 downregulated at salt vs control, 494 upregulated and 231 downregulated at salt + nanoparticles vs control, 783 upregulated and 523 downregulated at nanoparticles vs salt. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that Mitogen-activated protein kinase (MAPK) signaling pathway, biosynthesis of secondary metabolites, and plant hormonal signal transduction pathway were the enriched among all identified pathways. In addition, Reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR) showed that salinity up regulated the relative expression of DETs in pearl millet while, AgNPs optimized their expression that are associated with various molecular and metabolic functions. Overall, AgNPs treatments effectively improved the morphology, physiology, biochemistry, and gene expression pattern under salinity which could be attributed to positive impacts of AgNPs on pearl millet.