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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 KH 2 PO 4 ) 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.

Nanotechnology represents an innovative approach to ameliorating abiotic stress in pearl millet ( Pennisetum glaucum L.) with the application of Copper oxide (CuO − NPs) Titanium dioxide (TiO₂−NPs) and Silicon dioxide (SiO₂−NPs) gaining notable popularity recently. Therefore, we have utilized CuO, TiO₂, and SiO₂ nanoparticles at a concentration of 100 mg L⁻¹ as alleviating agents on P. glaucum grown in soil supplemented with varying levels of lead (Pb) at 0, 100, and 200 mg kg⁻¹ (applied as Pb(NO₃)₂). This study investigates various growth − related attributes, the efficiency of the photosynthetic machinery, indicators of oxidative stress, and responses of both enzymatic and non-enzymatic antioxidants, along with their specific gene expression, sugar content, AsA − GSH cycle, cellular fractionation, proline metabolism and Pb accumulation in different parts of the plant. Our results showed that the Pb toxicity in the soil showed a significant declined in the growth, gas exchange attributes, sugars, AsA − GSH cycle, cellular fractionation, proline metabolism in P. glaucum . However, Pb toxicity significantly increased oxidative stress biomarkers, enzymatic and non-enzymatic antioxidants including their gene expression in P. glaucum seedlings. Although, the application of CuO, TiO₂ and SiO₂ NPs showed a significant increase in the plant growth and biomass, gas exchange characteristics, enzymatic and non-enzymatic compounds and their gene expression and also decreased the oxidative stress. 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.

Downy mildew of pearl millet caused by the biotrophic oomycete Sclerospora graminicola is the most devastating disease which impairs pearl millet production causing huge yield and monetary losses. Chitosan nanoparticles (CNP) were synthesized from low molecular weight chitosan having higher degree of acetylation was evaluated for their efficacy against downy mildew disease of pearl millet caused by Sclerospora graminicola . Laboratory studies showed that CNP seed treatment significantly enhanced pearl millet seed germination percentage and seedling vigor compared to the control. Seed treatment with CNP induced systemic and durable resistance and showed significant downy mildew protection under greenhouse conditions in comparison to the untreated control. Seed treatment with CNP showed changes in gene expression profiles wherein expression of genes of phenylalanine ammonia lyase, peroxidase, polyphenoloxidase, catalase and superoxide dismutase were highly upregulated. CNP treatment resulted in earlier and higher expression of the pathogenesis related proteins PR1 and PR5. Downy mildew protective effect offered by CNP was found to be modulated by nitric oxide and treatment with CNP along with NO inhibitors cPTIO completely abolished the gene expression of defense enzymes and PR proteins. Further, comparative analysis of CNP with Chitosan revealed that the very small dosage of CNP performed at par with recommended dose of Chitosan for downy mildew management.

Main conclusionPearl 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.

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.

Dinanath grass ( Pennisetum pedicellatum Trin. ) is an extensively grown forage grass known for its significant drought resilience. In order to comprehensively grasp the adaptive mechanism of Dinanath grass in response to water deficient conditions, transcriptomic and metabolomics were applied in the leaves of Dinanath grass exposed to two distinct drought intensities (48-hour and 96-hour). Transcriptomic analysis of Dinanath grass leaves revealed that a total of 218 and 704 genes were differentially expressed under 48- and 96-hour drought conditions, respectively. The genes that were expressed differently (DEGs) and the metabolites that accumulated in response to 48-hour drought stress mainly showed enrichment in the biosynthesis of secondary metabolites, particularly phenolics and flavonoids. Conversely, under 96-hour drought conditions, the enriched pathways predominantly involved lipid metabolism, specifically sterol lipids. In particular, phenylpropanoid pathway and brassinosteroid signaling played a crucial role in drought response to 48- and 96-hour water deficit conditions, respectively. This variation in drought response indicates that the adaptation mechanism in Dinanath grass is highly dependent on the intensity of drought stress. In addition, different genes associated with phenylpropanoid and fatty acid biosynthesis, as well as signal transduction pathways namely phenylalanine ammonia-lyase, putrescine hydroxycinnamoyl transferase, abscisic acid 8’-hydroxylase 2, syntaxin-61, lipoxygenase 5, calcium-dependent protein kinase and phospholipase D alpha one, positively regulated with drought tolerance. Combined transcriptomic and metabolomic analyses highlights the outstanding involvement of regulatory pathways related to secondary cell wall thickening and lignin biosynthesis in imparting drought tolerance to Dinanath grass leaves. These findings collectively contribute to an enhanced understanding of candidate genes and key metabolites relevant to drought response in Dinanath grass. Furthermore, they establish a groundwork for the creation of a transcriptome database aimed at developing abiotic stress-tolerant grasses and major crop varieties through both transgenic and genome editing approaches.

Endophytic Trichoderma hamatum UoM 13 isolated from pearl millet roots was evaluated for its efficiency to suppress downy mildew disease. Under laboratory conditions, T. hamatum seed treatment significantly enhanced pearl millet seed germination and seedling vigor. T. hamatum seed treatment resulted in systemic and durable immunity against pearl millet downy mildew disease under greenhouse and field conditions. T. hamatum treated seedlings responded to downy mildew infection with high lignification and callose deposition. Analysis of defense enzymes showed that T. hamatum treatment significantly enhanced the activities of glucanase, peroxidase, phenylalanine ammonia-lyase, and polyphenol oxidase in comparison to untreated control. RT-PCR analysis revealed differentially expressed transcripts of the defense enzymes and PR-proteins in treated, untreated, and checks, wherein PR-1, PR-5, and cell wall defense HRGPs were significantly over expressed in treated seedlings as against their lower expression in controls. T. hamatum treatment significantly stimulated endogenous salicylic acid (SA) levels and significantly upregulated important SA biosynthesis gene isochorismate synthase. The results indicated that T. hamatum UoM13 treatment induces resistance corresponding to significant over expression of endogenous SA, important defense enzymes, PR-proteins, and HRGPs, suggesting that SA biosynthetic pathway is involved in pearl millet for mounting systemic immunity against downy mildew pathogen.

Apospory in Pennisetum squamulatum Fresen is inherited as a single-locus dominant trait. Molecular analysis has identified the apospory-specific genomic region (ASGR), a large segment of one chromosome that is necessary and sufficient to confer apomixis. To transfer apospory from P. squamulatum to pearl millet [P. glaucum (L.) R. Br.], advanced backcrosses were generated. Here we report the characterization of apomictic BC sub(7) and BC sub(8) generations. A BC sub(7)-derived genotype has two P. squamulatum chromosomes, while BC sub(8)-derived apomictic lines inherited only the ASGR chromosome that confers apospory to pearl millet. Morphologically, no significant differences were observed between backcross generations for plant height, leaf length, leaf width, and first internode and inflorescence lengths. However, the BC sub(7) and BC sub(8) lines differed significantly with regard to pollen viability, flowering time, and seed set. Pollen viability increased to 81% in the BC sub(8) lines compared with 37% in BC sub(7). The frequency of aposporous embryo sacs was lower in BC sub(8) (79%) lines compared to BC sub(7) (93%). Meiotic behavior of chromosomes during metaphase I and anaphase I showed that BC sub(8) typically had a single univalent, identified as the ASGR chromosome by FISH (fluorescence in situ hybridization) and GISH (genomic in situ hybridization). Utilizing its unique behavior during meiosis the ASGR chromosome was microdissected and a chromosome-specific library was constructed and sequenced. Analysis of sequences identified both novel and previously known ASGR-linked sequences.

Pearl millet hybrids biofortified with iron (Fe) and zinc (Zn) promise to be part of a long-term strategy to combat micronutrient malnutrition in the arid and semi-arid tropical (SAT) regions of the world. Biofortification through molecular breeding is the way forward to achieving a rapid trait-based breeding strategy. This genome-wide association study (GWAS) was conducted to identify significant marker-trait associations (MTAs) for Fe, Zn, and protein content (PC) for enhanced biofortification breeding. A diverse panel of 281 advanced inbred lines was evaluated for Fe, Zn, and PC over two seasons. Phenotypic evaluation revealed high variability (Fe: 32–120 mg kg −1 , Zn: 19–87 mg kg −1 , PC: 8–16%), heritability (h bs 2  ≥ 90%) and significantly positive correlation among Fe, Zn and PC ( P  = 0.01), implying concurrent improvement. Based on the Diversity Arrays Technology (DArT) seq assay, 58,719 highly informative SNPs were filtered for association mapping. Population structure analysis showed six major genetic groups (K = 6). A total of 78 MTAs were identified, of which 18 were associated with Fe, 43 with Zn, and 17 with PC. Four SNPs viz., Pgl04_64673688, Pgl05_135500493, Pgl05_144482656, and Pgl07_101483782 located on chromosomes Pgl04 (1), Pgl05 (2) and Pgl07 (1), respectively were co-segregated for Fe and Zn. Promising genes, ‘Late embryogenesis abundant protein’, ‘Myb domain’, ‘pentatricopeptide repeat’, and ‘iron ion binding’ coded by 8 SNPs were identified. The SNPs/genes identified in the present study presents prospects for genomics assisted biofortification breeding in pearl millet.

Biofertilizer plays a significant role in crop cultivation that had reduced its inorganic fertilizer use. The effects of inorganic fertilizer reduction combined with Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer on the growth, quality, soil nutrients and diversity of the soil bacterial community in the rhizosphere soil of pakchoi were studied. The experiment composed of 6 treatments, including CK (no fertilization), DL (10% inorganic fertilizer reduction combined with Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer), ZL (25% inorganic fertilizer reduction combined with Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer), SL (50% inorganic fertilizer reduction combined with Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer), FHF (100% inorganic fertilizer) and JZ (100% inorganic fertilizer combined with sterilized Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer). Compared with conventional fertilization, the 25% reduction in chemical fertilizer applied with the Pennisetum giganteum mixed nitrogen-fixing biofertilizer resulted in higher plant height, plant weight, chlorophyll content, soluble protein content, soluble sugar content, vitamin C content, alkali hydrolyzed nitrogen content, available phosphorus content, available potassium content and organic matter content in pakchoi, and these variables increased by 11.81%, 8.54%, 7.37%, 16.88%, 17.05%, 23.70%, 24.24%, 36.56%, 21.09% and 19.72%, respectively. In addition, the 25% reduction in chemical fertilizer applied with the Pennisetum giganteum mixed nitrogen-fixing biofertilizer also had the lowest nitrate content, which was 53.86% lower than that with conventional fertilization. Different fertilizer treatments had a significant effect on the soil bacterial community structure. Compared with conventional fertilization, the coapplication of Pennisetum giganteum z.x.lin mixed nitrogen-fixing biofertilizer and inorganic fertilizer significantly increased the relative abundance of Proteobacteria and Actinobacteria in the soil. The results of the redundancy analysis (RDA) showed that soil organic matter, alkali-hydrolyzed nitrogen, available phosphorus, available potassium, pH and water content had a specific impact on the soil bacterial community. Among the factors, soil water content was the main factor affecting the soil bacterial community, followed by soil organic matter, soil pH, available potassium, soil available phosphorus and soil alkali-hydrolyzed nitrogen.