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.

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.

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.

DNAJ protein gene expansion mechanism in Panicoideae and PgDNAJ functional identification in pearl millet.

KEY MESSAGE: Analyze the evolutionary pattern of DNAJ protein genes in the Panicoideae, including pearl millet, to identify and characterize the biological function of PgDNAJ genes in pearl millet. Global warming has become a major factor threatening food security and human development. It is urgent to analyze the heat-tolerant mechanism of plants and cultivate crops that are adapted to high temperature conditions. The Panicoideae are the second largest subfamily of the Poaceae, widely distributed in warm temperate and tropical regions. Many of these species have been reported to have strong adaptability to high temperature stress, such as pearl millet, foxtail millet and sorghum. The evolutionary differences in DNAJ protein genes among 12 Panicoideae species and 10 other species were identified and analyzed. Among them, 79% of Panicoideae DNAJ protein genes were associated with retrotransposon insertion. Analysis of the DNAJ protein pan-gene family in six pearl millet accessions revealed that the non-core genes contained significantly more TEs than the core genes. By identifying and analyzing the distribution and types of TEs near the DNAJ protein genes, it was found that the insertion of Copia and Gypsy retrotransposons provided the source of expansion for the DNAJ protein genes in the Panicoideae. Based on the analysis of the evolutionary pattern of DNAJ protein genes in Panicoideae, the PgDNAJ was obtained from pearl millet through identification. PgDNAJ reduces the accumulation of reactive oxygen species caused by high temperature by activating ascorbate peroxidase (APX), thereby improving the heat resistance of plants. In summary, these data provide new ideas for mining potential heat-tolerant genes in Panicoideae, and help to improve the heat tolerance of other crops.

Transcriptomic and physiological analysis provide new insight into seed shattering mechanism in Pennisetum alopecuroides 'Liqiu'.

KEY MESSAGE: Through the histological, physiological, and transcriptome-level identification of the abscission zone of Pennisetum alopecuroides 'Liqiu', we explored the structure and the genes related to seed shattering, ultimately revealing the regulatory network of seed shattering in P. alopecuroides. Pennisetum alopecuroides is one of the most representative ornamental grass species of Pennisetum genus. It has unique inflorescence, elegant appearance, and strong stress tolerance. However, the shattering of seeds not only reduces the ornamental effect, but also hinders the seed production. In order to understand the potential mechanisms of seed shattering in P. alopecuroides, we conducted morphological, histological, physiological, and transcriptomic analyses on P. alopecuroides cv. 'Liqiu'. According to histological findings, the seed shattering of 'Liqiu' was determined by the abscission zone at the base of the pedicel. Correlation analysis showed that seed shattering was significantly correlated with cellulase, lignin, auxin, gibberellin, cytokinin and jasmonic acid. Through a combination of histological and physiological analyses, we observed the accumulation of cellulase and lignin during 'Liqiu' seed abscission. We used PacBio full-length transcriptome sequencing (SMRT) combined with next-generation sequencing (NGS) transcriptome technology to improve the transcriptome data of 'Liqiu'. Transcriptomics further identified many differential genes involved in cellulase, lignin and plant hormone-related pathways. This study will provide new insights into the research on the shattering mechanism of P. alopecuroides.

Pearl millet a promising fodder crop for changing climate: a review.

The agricultural sector faces colossal challenges amid environmental changes and a burgeoning human population. In this context, crops must adapt to evolving climatic conditions while meeting increasing production demands. The dairy industry is anticipated to hold the highest value in the agriculture sector in future. The rise in the livestock population is expected to result in an increased demand for fodder feed. Consequently, it is crucial to seek alternative options, as crops demand fewer resources and are resilient to climate change. Pearl millet offers an apposite key to these bottlenecks, as it is a promising climate resilience crop with significantly low energy, water and carbon footprints compared to other crops. Numerous studies have explored its potential as a fodder crop, revealing promising performance. Despite its capabilities, pearl millet has often been overlooked. To date, few efforts have been made to document molecular aspects of fodder-related traits. However, several QTLs and candidate genes related to forage quality have been identified in other fodder crops, which can be harnessed to enhance the forage quality of pearl millet. Lately, excellent genomic resources have been developed in pearl millet allowing deployment of cutting-edge genomics-assisted breeding for achieving a higher rate of genetic gains. This review would facilitate a deeper understanding of various aspects of fodder pearl millet in retrospect along with the future challenges and their solution. This knowledge may pave the way for designing efficient breeding strategies in pearl millet thereby supporting sustainable agriculture and livestock production in a changing world.

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.

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.

Evaluating Plant Growth-Promoting Rhizobacteria to Improve the Productivity of Forage Pearl Millet.

India's livestock industry is grappling with a shortage of green fodder, necessitating concerted efforts to boost organized production and ensure a sufficient supply of high-quality forages, crucial for formulating nutritionally balanced, cost-effective, and rumen-healthy animal diets. Hence, this study was conducted to assess the plant growth-promoting characteristics of liquid microbial inoculants and their impact on the yield of forage pearl millet. The bacterial cultures utilized included Sphingobacterium sp., Stenotrophomonas maltophilia, and an isolate from vegetable cowpea, subsequently identified as Burkholderia seminalis. These cultures were initially characterized for their plant growth-promoting traits at different temperature and physiological conditions. All the bacterial cultures were found promising for PGPR traits over varied temperature conditions and the optimum activity was recorded at 40 °C, with tolerance to saline and drought stresses as well as wide pH and temperature ranges. A field experiment was conducted during kharif 2020 at Punjab Agricultural University, Ludhiana and Punjab Agricultural University, Regional Research Station, Bathinda, involving combinations of liquid microbial inoculants along with 100% Recommended Dose of Fertilizer (RDF). It was observed that the treatment including B. seminalis + S. maltophilia along with RDF yielded the highest green fodder and dry matter yield, In conclusion, it is evident that the utilization of these liquid microbial inoculants holds significant potential for playing a pivotal role in the integrated nutrient management of forage pearl millet, thereby contributing to heightened productivity and sustained soil health.

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.

MXene/biochar composites for enhanced wastewater reclamation and bioenergy production: A kinetics and thermodynamics study.

The study explores the synthesis and utilization of biochar (BC) and multi-layer MXene to MXene/biochar (MB) composites for wastewater treatment. Simultaneously, it also investigates their energy generation potential through biomass and soil property assessments. The integrated column and batch treatments have shown significant results, elevating total dissolved solids from 63.7 to 125.5 mg L-1 with column treatment, while reducing them to 6.37 % and 1.35 % with BC and MB treatment, respectively. BC with high carbon content, demonstrated increased hydrophobicity, which was amplified by the integration of MXene, thereby enhancing its potential for advanced wastewater treatment. Treated wastewater exhibited elevated nutrient concentrations (Ca, Cu, Fe, K, Na, and NH4+), promoting the growth of Pennisetum purpureum. WW_B shows promising energy potential, with a higher heating value of 25.03 MJ kg-1 and a lower heating value of 23.57 MJ kg-1. They demonstrated high volatile matter exceeding 70.9 wt %, and a fixed carbon from 10.02 to 27.53 wt %, signifying their potential for efficient conversion and bio-oil yield during pyrolysis. The ultimate analysis emphasized significant carbon, with oxygen content ranging from 43.42 to 47.78 wt %., influencing combustion characteristics. MT_B exhibited its suitability for energy production through thermochemical conversion, underlined by its high flammability and low volatile ignition values. In the absence of BC, the Ea ranged from 24.77 to 77.88 kJ mol-1 in wastewater and from 21.67 to 69.6 kJ mol-1 in MB treated wastewater. Meanwhile, when soil contained BC and was irrigated with wastewater, the Ea varied from 24.66 to 80.91 kJ mol-1. In the case of MB treated wastewater, it ranged from 25.01 to 75.79 kJ mol-1. The research thereby affirms the potential of MB composites to advance water and energy sustainability setting us for broader nexus-based applications.

Omics-driven utilization of wild relatives for empowering pre-breeding in pearl millet.

MAIN CONCLUSION: Pearl millet wild relatives harbour novel alleles which could be utilized to broaden genetic base of cultivated species. Genomics-informed pre-breeding is needed to speed up introgression from wild to cultivated gene pool in pearl millet. Rising episodes of intense biotic and abiotic stresses challenge pearl millet production globally. Wild relatives provide a wide spectrum of novel alleles which could address challenges posed by climate change. Pre-breeding holds potential to introgress novel diversity in genetically narrow cultivated Pennisetum glaucum from diverse gene pool. Practical utilization of gene pool diversity remained elusive due to genetic intricacies. Harnessing promising traits from wild pennisetum is limited by lack of information on underlying candidate genes/QTLs. Next-Generation Omics provide vast scope to speed up pre-breeding in pearl millet. Genomic resources generated out of draft genome sequence and improved genome assemblies can be employed to utilize gene bank accessions effectively. The article highlights genetic richness in pearl millet and its utilization with a focus on harnessing next-generation Omics to empower pre-breeding.

Improvements in fermentation and nutritive quality of elephant grass [Cenchrus purpureus (Schumach.) Morrone] silages: a review.

Elephant grass [Pennisetum purpureum Schumach. syn. Cenchrus purpureus (Schumach.) Morrone], also known as Napier grass and King grass, includes varieties Taiwán, Gigante, Merkerón, Maralfalfa, and others. The grass achieves high biomass production in tropical-subtropical, temperate, and arid areas. The high-water concentration of elephant grass suggests that ensiling could offer an alternative way to preserve the nutritional quality of the grass during storage, however, some considerations should be addressed because of the particularities of the grass. Ensiling elephant grass may produce adequate fermentation but could suffer effluent losses and subsequent losses of nutrients due to leaching. To improve fermentation and nutrient characteristics of elephant grass silages, several studies were conducted with the inclusion of additives. Lactic acid bacteria inocula have reduced pH and increased crude protein content of elephant grass silage, but aerobic stability of silages could be affected by the bacterial inoculation. There is limited information, however, on the potential of different silage inoculants to reduce growth of spoilage microorganisms during the aerobic phase of silage prepared with elephant grass. Exogenous fibrolytic enzymes also may improve elephant grass silage quality by enhancing microbial fiber-degradation with subsequent increase in lactic acid and its associated pH reduction. Another study approach to improve fermentation and nutritional quality of elephant grass silages involved the addition of different feeds at ensiling, including conventional feeds such corn, wheat, rice bran, and molasses or alternative feeds such as different dehydrated by-products obtained from the food industries of juice and jelly. In the manuscript, the presented scientific information shows the great potential of the different manipulations to improve the quality of elephant grass silages and with possible enhance of the economic profit and sustainability of livestock farming in the tropical areas.

The impact of addition of pearl millet starch-germ complex in white bread on nutritional, textural, structural, and glycaemic response: Single blinded randomized controlled trial in healthy and pre-diabetic participants.

The rise of pre-diabetes at the global level has created a significant interest in developing low glycaemic index food products. The pearl millet is a cheaper source of starch and its germ contains significant amount of protein and fat. The complexing of pearl millet starch and germ by dry heat treatment (PMSGH) resulted an increase in the resistant starch content upto 45.09 % due to formation of amylose-glutelin-linoleic acid complex. The resulting pearl millet starch germ complex was incorporated into wheat bread at 20, 25, and 30 %. The PMSGH incorporated into bread at 30 % reduced the glycaemic index to 52.31. The PMSGH incorporated bread had significantly (p < 0.05)increased in the hardness with a reduction in springiness and cohesiveness. The structural attributes of the 30 % PMSGH incorporated bread revealed a significant (p < 0.05)increase in 1040/1020 cm-1 ratio and relative crystallinity. The consumption of functional bread incorporated with pearl millet starch germ complex reduced blood glucose levels and in vivo glycaemic index in healthy and pre-diabetic participants when compared to white bread. Hence, the study showed that the incorporation of pearl millet starch-germ complex into food products could be a potential new and healthier approach for improving dietary options in pre-diabetes care.

Genome-wide characterization of the NBLRR gene family provides evolutionary and functional insights into blast resistance in pearl millet (Cenchrus americanus (L.) Morrone).

MAIN CONCLUSION: The investigation is the first report on genome-wide identification and characterization of NBLRR genes in pearl millet. We have shown the role of gene loss and purifying selection in the divergence of NBLRRs in Poaceae lineage and candidate CaNBLRR genes for resistance to Magnaporthe grisea infection. Plants have evolved multiple integral mechanisms to counteract the pathogens' infection, among which plant immunity through NBLRR (nucleotide-binding site, leucine-rich repeat) genes is at the forefront. The genome-wide mining in pearl millet (Cenchrus americanus (L.) Morrone) revealed 146 CaNBLRRs. The variation in the branch length of NBLRRs showed the dynamic nature of NBLRRs in response to evolving pathogen races. The orthology of NBLRRs showed a predominance of many-to-one orthologs, indicating the divergence of NBLRRs in the pearl millet lineage mainly through gene loss events followed by gene gain through single-copy duplications. Further, the purifying selection (Ka/Ks < 1) shaped the expansion of NBLRRs within the lineage of pear millet and other members of Poaceae. Presence of cis-acting elements, viz. TCA element, G-box, MYB, SARE, ABRE and conserved motifs annotated with P-loop, kinase 2, RNBS-A, RNBS-D, GLPL, MHD, Rx-CC and LRR suggests their putative role in disease resistance and stress regulation. The qRT-PCR analysis in pearl millet lines showing contrasting responses to Magnaporthe grisea infection identified CaNBLRR20, CaNBLRR33, CaNBLRR46 CaNBLRR51, CaNBLRR78 and CaNBLRR146 as putative candidates. Molecular docking showed the involvement of three and two amino acid residues of LRR domains forming hydrogen bonds (histidine, arginine and threonine) and salt bridges (arginine and lysine) with effectors. Whereas 14 and 20 amino acid residues of CaNBLRR78 and CaNBLRR20 showed hydrophobic interactions with 11 and 9 amino acid residues of effectors, Mg.00g064570.m01 and Mg.00g006570.m01, respectively. The present investigation gives a comprehensive overview of CaNBLRRs and paves the foundation for their utility in pearl millet resistance breeding through understanding of host-pathogen interactions.

Effect of dietary inclusion of Pennisetum purpureum (Napier) grass on growth performance, rumen fermentation and meat quality of feedlot sussex red steers.

The aim of this study was to evaluate the growth performance, fermentation indices and meat quality of Sussex steers fed totally mixed rations that composed of graded inclusion levels of Napier grass (NP). Three experimental diets designated as diet 1 (0.0 g kg-1 NP: Control), diet 2 (300 g kg-1 NP grass) and diet 3 (600 g kg-1 NP) were formulated. Twenty-four male steers aged 8 months with an average body weight of 185.0 ± 30 kg were used. In a completely randomized design, the animals were allocated to the diets and fed for 120 days. Dietary NP inclusion reduced (P < 0.05) the animals' average daily gain and increased the feed efficiency. The steers' daily feed intake and final body weight decreased (P < 0.05) with a 600 g kg-1 inclusion level. The fermentation indices were not affected (P > 0.05) by the inclusion. While the inclusion reduced (P < 0.05) warm muscle temperature, it had no effect (P > 0.05) on carcass dressing percentage, warm and cold initial and ultimate pH. However, 600 g kg-1 inclusion level reduced (P > 0.05) warm and cold carcass weights. Meat physical attributes, moisture characteristics and tenderness were not affected (P > 0.05) by dietary treatments, except for the 7-days aged meat thaw loss, which increased at 600 g kg-1 inclusion level. Inclusion of 300 g kg-1 increased meat protein and fat, but dry and organic matter contents decreased with increasing inclusion levels. Dietary inclusion of NP grass up to 300 g kg-1 in steers' diets improved feed intake, carcass traits and yielded meat high in protein and fat.

Pearl millet flour and green gram milk based probiotic beverage.

The probiotic beverage was developed using germinated and ungerminated pearl millet flour and green gram milk. The germinated and ungerminated pearl millet flour was added to green gram milk at different concentrations (0.5-2.5 %) along with sugar and cardamom. The mixtures were then inoculated with probiotic bacteria Lactobacillus acidophilus incubated at 37 °C for 6 h. Characterization of probiotic beverages was carried out during storage at (4 ± 1)°C for 21 days. The germinated flour beverage had high acidity as compared to the ungerminated flour beverage. The probiotic count in germinated and ungerminated flour beverages ranged from 8.19 to 8.77 × 107 and 8.04 to 8.52 × 107 log CFU/mL, respectively. Antioxidant activity, polyphenol content increased with an increase in the concentration of flour in the beverage. The LC-MS analysis found the existence of vitexin and isovitexin as the main polyphenolic compounds in the probiotic beverage. Non-dairy probiotic beverage prepared with 0.5 % germinated millet flour gave the best taste, color, texture, and rheological properties.

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.

Genome-Wide Identification and Transcriptional Analysis of AP2/ERF Gene Family in Pearl Millet (Pennisetum glaucum).

The apetala2/ethylene response factor (AP2/ERF) gene family plays a crucial role in regulating plant growth and development and responding to different abiotic stresses (e.g., drought, heat, cold, and salinity). However, the knowledge of the ERF family in pearl millet remains limited. Here, a total of 167 high-confidence PgERF genes are identified and divided into five subgroups based on gene-conserved structure and phylogenetic analysis. Forty-one pairs of segmental duplication are found using collinear analysis. Nucleotide substitution analysis reveals these duplicated pairs are under positive purification, indicating they are actively responding to natural selection. Comprehensive transcriptomic analysis reveals that PgERF genesare preferentially expressed in the imbibed seeds and stem (tilling stage) and respond to heat, drought, and salt stress. Prediction of the cis-regulatory element by the PlantCARE program indicates that PgERF genes are involved in responses to environmental stimuli. Using reverse transcription quantitative real-time PCR (RT-qPCR), expression profiles of eleven selected PgERF genes are monitored in various tissues and during different abiotic stresses. Transcript levels of each PgERF gene exhibit significant changes during stress treatments. Notably, the PgERF7 gene is the only candidate that can be induced by all adverse conditions. Furthermore, four PgERF genes (i.e., PgERF22, PgERF37, PgERF88, and PgERF155) are shown to be involved in the ABA-dependent signaling pathway. These results provide useful bioinformatic and transcriptional information for understanding the roles of the pearl millet ERF gene family in adaptation to climate change.

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.