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.

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.

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.

Mycological and Multiple Mycotoxin Surveillance of Sorghum and Pearl Millet Produced by Smallholder Farmers in Namibia.

Mycological (mycotoxigenic Fusarium and aflatoxigenic Aspergillus spp.) and multiple mycotoxins [aflatoxin B1 (AFB1), fumonisin B (FB), deoxynivalenol and zearalenone] surveillance was conducted on raw whole grain sorghum (Sorghum bicolor) and pearl millet (Pennisetum glaucum) produced on smallholder farms, and processed products sold at open markets in northern Namibia. Fungal contamination was determined with morphological methods as well as with quantitative Real-Time PCR (qPCR). The concentrations of multiple mycotoxins in samples were determined with liquid chromatography tandem mass spectrometry. The incidence of mycotoxigenic Fusarium spp., Aspergillus flavus and A. parasiticus, as well as the concentrations of AFB1 and FB were significantly (P < 0.001) higher in the malts as compared to the raw whole grains, with Aspergillus spp. and AFB1 exhibiting the highest contamination (P < 0.001). None of the analysed mycotoxins were detected in the raw whole grains. Aflatoxin B1 above the regulatory maximum level set by the European Commission was detected in sorghum (2 of 10 samples; 20%; 3-11 µg/kg) and pearl millet (6 of 11 samples; 55%; 4-14 µg/kg) malts. Low levels of FB1 (6 of 10 samples; 60%; 15-245 µg/kg) were detected in sorghum malts and no FB was detected in pearl millet malts. Contamination possibly occurred postharvest, during storage, and/or transportation and processing. By critically monitoring the complete production process, the sources of contamination and critical control points could be identified and managed. Mycotoxin awareness and sustainable education will contribute to reducing mycotoxin contamination. This could ultimately contribute to food safety and security in northern Namibia where communities are exposed to carcinogenic mycotoxins in their staple diet.

Study on the effect of additives on microbial diversity, predicted functional profiles, and fermentation quality of Broussonetia papyrifera and Pennisetum sinese mixed ensilage in the karst region.

In this research, we evaluated the effect of exogenous lactic acid bacteria and Amomum villosum essential oil (AVEO) on the chemical composition, microbial community composition, microbial functional diversity, and fermentation quality of Broussonetia papyrifera (BP) and Pennisetum sinese (PS) mixed silages. The BP:PS mixing ratios were 100:0, 70:30, 50:50, 30:70, and 0:100. After 3 and 30 days of ensiling at 22°C-25°C, microbial diversity and function, and fermentation quality, were assessed. Increasing PS content resulted in decreased ammoniacal nitrogen and pH, increased water-soluble carbohydrate content, increased relative abundance of Lactococcus and Acinetobacter, and reduced relative abundance of Caproiciproducens and Pseudomonas. A 50:50 BP:PS ratio effectively improved the fermentation quality compared to anaerobic fermentation with BP or PS alone, while AVEO treatment further improved fermentation quality by increasing Lactococcus relative abundance. Moreover, as fermentation proceeded, ensiling enhanced the 'Human diseases', 'Environmental information processing', and 'Cellular processes' functions at the first level, as well as the 'Two-component system' and 'ABC transporters' functions at the third level. Different additives affected the fermentation of BP and PS mixed silage by regulating microbial community succession and metabolic pathways during ensiling.

Mycotoxin levels and characterization of natural anti-fungal phytochemicals in pearl millet (Pennisetum glaucum) from Nigeria's six agroecological zones.

This study reports levels of multiple mycotoxins across Nigeria's six agro-ecological zones and corresponding levels of natural anti-fungal phytochemicals present in pearl millet (PM). 220 representative composite samples of PM were collected for mycotoxin analysis using ultrahigh performance liquid chromatography-mass spectrometry (UHPLC-MS), and 24 were randomly selected for determination of metabolites using gas chromatography-high resolution time of flight-mass spectrometry (GC-HRTOF-MS). In total, 15 mycotoxins were detected, all with levels below the European Union (EU) permissible limits and level of aflatoxins only up to 1.34 µg/kg. This is in sharp contrast to high levels of mycotoxins reported in maize samples from the same agroecological zones. Phytochemical analysis of the same samples identified a total of 88 metabolites, 30 of which are known anti-fungal properties from other previously published studies. The most common of these include methyl ester, bis (2-ethylhexyl) phthalate, and ç-tocopherol. The number of anti-fungal metabolites recovered from each sample ranged from 3 to 17 and varied widely in both number and composition across the agroecological zones. The anti-fungal metabolites may probably make PM less susceptible to fungal proliferation compared to other grains. Hence, it is worth exploring for possible sources of biological control products from PM.

Plant growth-promoting rhizobacteria Shewanella putrefaciens and Cronobacter dublinensis enhance drought tolerance of pearl millet by modulating hormones and stress-responsive genes.

Drought is a major abiotic stress that affects crop productivity. Endophytic bacteria have been found to alleviate the adverse effects of drought on plants. In the present study, we evaluated the effects of two endophytic bacteria Shewanella putrefaciens strain MCL-1 and Cronobacter dublinensis strain MKS-1 on pearl millet (Pennisetum glaucum (L.) R. Br.) under drought stress conditions. Pearl millet plants were grown under three water levels: field capacity (FC), mild drought stress (MD), and severe drought stress (SD). The effects of inoculation on plant growth, physiological attributes, phytohormone content, and drought stress-responsive genes were assessed. The inoculation of pearl millet seeds with endophytes significantly improved shoot and root dry weight and root architecture of plants grown under FC and drought stress conditions. There was a significant increase in relative water content and proline accumulation in the inoculated plants. Among the phytohormones analyzed, the content of ABA and IAA was significantly higher in endophyte-treated plants under all moisture regimes than in uninoculated plants. C. dublinensis-inoculated plants had higher GA content than uninoculated plants under all moisture regimes. The expression level of genes involved in phytohormone biosynthesis (SbNCED, SbGA20oX, and SbYUC) and coding drought-responsive transcription factors (SbAP2, SbSNAC1 and PgDREB2A) was significantly higher under SD in endophyte-inoculated plants than in uninoculated plants. Thus, these endophytic bacteria presumably enhanced the tolerance of pearl millet to drought stress by modulating root growth, plant hormones, physiology and the expression of genes involved in drought tolerance.

Cultivated and wild pearl millet display contrasting patterns of abundance and co-occurrence in their root mycobiome.

Fungal communities associated with roots play a key role in nutrient uptake and in mitigating the abiotic and biotic stress of their host. In this study, we characterized the roots mycobiome of wild and cultivated pearl millet [Pennisetum glaucum (L.) R. Br., synonym: Cenchrus americanus (L.) Morrone] in three agro-ecological areas of Senegal following a rainfall gradient. We hypothesized that wild pearl millet could serve as a reservoir of endophytes for cultivated pearl millet. We therefore analyzed the soil factors influencing fungal community structure and whether cultivated and wild millet shared the same fungal communities. The fungal communities associated with pearl millet were significantly structured according to sites and plant type (wild vs cultivated). Besides, soil pH and phosphorus were the main factors influencing the fungal community structure. We observed a higher fungal diversity in cultivated compared to wild pearl millet. Interestingly, we detected higher relative abundance of putative pathotrophs, especially plant pathogen, in cultivated than in wild millet in semi-arid and semi-humid zones, and higher relative abundance of saprotrophs in wild millet in arid and semi-humid zones. A network analysis based on taxa co-occurrence patterns in the core mycobiome revealed that cultivated millet and wild relatives had dissimilar groups of hub taxa. The identification of the core mycobiome and hub taxa of cultivated and wild pearl millet could be an important step in developing microbiome engineering approaches for more sustainable management practices in pearl millet agroecosystems.

Using γ-ray irradiation and epiphytic microbiota inoculation to separate the effects of chemical and microbial factors on fermentation quality and bacterial community of ensiled Pennisetum giganteum.

AIMS: This study aimed to separate the effects of chemical and microbial factors on the fermentation quality and bacterial community of ensiled Pennisetum giganteum. METHODS AND RESULTS: Fresh P. giganteum of two vegetative stages (stage I, GI ; stage II, GII ) was treated as follows: GI epiphytic microbiota + Î³-ray presterilized GI (MI CI ), GII epiphytic microbiota + Î³-ray presterilized GI (MII CI ), GI epiphytic microbiota + Î³-ray presterilized GII (MI CII ), and GII epiphytic microbiota + Î³-ray presterilized GII (MII CII ). Triplicates per treatment were sampled after 30 days of ensiling for chemical and microbial analyses and high-throughput sequencing. Silages made from CII (MI CII and MII CII ) had higher lactic acid concentration and the ratio of lactic to acetic acid, and lower pH and ammonia nitrogen concentration than silages produced by CI (MI CI and MII CI ). Species differential analyses showed that the changes of chemical composition rather than epiphytic microbiota significantly affected the relative abundance of Lactobacillus, Pediococcus and Pantoea in P. giganteum silages. CONCLUSION: These above results manifested that chemical composition was the main factor influencing the fermentation quality and bacterial community of P. giganteum silage in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained results may, therefore, be the first record to provide an in-depth understanding of the relative contributions of chemical and microbial parameters on fermentation quality and bacterial community, which is of great importance for modulating silage fermentation and improving silage quality.

Induction of drought tolerance in Pennisetum glaucum by ACC deaminase producing PGPR- Bacillus amyloliquefaciens through Antioxidant defense system.

Rhizobacteria from pearl millet were screened to produce 1-aminocyclopropane-1-carboxylate (ACC) deaminase and to evaluate its role in alleviating drought stress. Amongst 96 isolates, 28 were positive for ACC deaminase production, with MMR04 offering maximum activity of 2196.23 nmol of α-ketobutyrate produced mg-1 of protein h-1. The ACC deaminase producing rhizobacteria with multiple beneficial properties along with root colonization and non-pathogenic were selected [Bacillus amyloliquefaciens (MMR04), Bacillus subtilis (MMR18) and Stenotrophomonas maltophilia (MMR36)] to confirm the presence of ACC deaminase gene. A significant enhancement in seed germination (91.75%) and seedling vigor (1213.73) was noted upon seed treatment with MMR04 and hence further evaluated for its ability to induce drought stress. The seed treatment with MMR04 improved plant growth parameters and total chlorophyll and RWC in plants grown under severe drought stress (G5) conditions compared to control plants. In addition, MMR04 seed treatment enhanced proline, APX and SOD activity while decreased the MDA content up to 2.3 fold compared to untreated plants (G5). Gene expression studies revealed a significant decrease of 3.3 and 1.8 fold in the relative expression of drought-responsive (DREB-1E) and ethylene-responsive factor (ERF-1B) marker genes, respectively and an increase of 2.2 and 2.9 fold in the relative expression of APX1 and SOD1, respectively in MMR04 treated plants grown under G5 conditions over control. The results confirmed that ACC deaminase producing B. amyloliquefaciens MMR04 could defend the pearl millet plants against drought stress through an antioxidative system, thereby warranting its application in drought stress management.

Trichovariability in rhizosphere soil samples and their biocontrol potential against downy mildew pathogen in pearl millet.

The present work is aimed to examine the genetic variability and the distribution pattern of beneficial Trichoderma spp. isolated from rhizosphere samples and their mode of action in improving the plant health. A total of 131 suspected fungi were isolated from the rhizospheric soil and 91 isolates were confirmed as Trichoderma spp. T. asperellum and T. harzianum were found high in the frequency of occurrence. Genetic diversity analysis using RAPD and ISSR revealed the diverse distribution pattern of Trichoderma spp. indicating their capability to adapt to broad agroclimatic conditions. Analysis of genetic diversity using molecular markers revealed intra-species diversity of isolated Trichoderma spp. The frequency of pearl millet (PM) root colonization by Trichoderma spp. was found to be 100%. However, they showed varied results for indole acetic acid, siderophore, phosphate solubilization, ß-1,3-glucanase, chitinase, cellulase, lipase, and protease activity. Downy mildew disease protection studies revealed a strong involvement of Trichoderma spp. in direct suppression of the pathogen (mean 37.41) in the rhizosphere followed by inducing systemic resistance. Our findings highlights the probable distribution and diversity profile of Trichoderma spp. as well as narrate the possible utilization of Trichoderma spp. as microbial fungicides in PM cultivation across different agroclimatic zones of India.

The Flora Compositions of Nitrogen-Fixing Bacteria and the Differential Expression of nifH Gene in Pennisetum giganteum z.x.lin Roots.

The flora compositions of nitrogen-fixing bacteria in roots of Pennisetum giganteum z.x.lin at different growth stages and the expression and copy number of nitrogen-fixing gene nifH were studied by Illumina Miseq second-generation sequencing technology and qRT-PCR. The results showed that there were more than 40,000~50,000 effective sequences in 5 samples from the roots of P. giganteum, with Proteobacteria and Cyanobacteria as the dominant nitrogen-fixing bacteria based on the OTU species annotations for each sample and Bradyrhizobium as the core bacterial genera. The relative expression and quantitative change of nifH gene in roots of P. giganteum at different growth stages were consistent with the changes in the flora compositions of nitrogen-fixing microbia. Both revealed a changing trend with an initial increase and a sequential decrease, as well as changing order as jointing stage>maturation stage>tillering stage>seedling stage>dying stage. The relative expression and copy number of nifH gene were different in different growth stages, and the difference among groups basically reached a significant level (p < 0.05). The relative expression and copy number of nifH gene at the jointing stage were the highest, and the 2-△△CT value was 4.43 folds higher than that at the seedling stage, with a copy number of 1.32 × 107/g. While at the dying stage, it was the lowest, and the 2-△△CT value was 0.67 folds, with a copy number of 0.31 × 107/g.

Evaluation of biological and chemical additives on microbial community, fermentation characteristics, aerobic stability, and in vitro gas production of SuMu No. 2 elephant grass.

BACKGROUND: The study was conducted to evaluate the effects of biological and chemical additives on microbial community, fermentation characteristics, aerobic stability, and in vitro gas production of SuMu No. 2 elephant grass. RESULTS: Aerobic bacteria and yeast were not affected on days 5 and 7 but were significantly (P < 0.224) reduced on days 14, 30, and 60, whereas lactic acid and lactic acid bacteria were significantly (P > 0.001) higher in all ensiling days within all treatment groups. During the ensiling days, the pH, acetic acid, butyric acid, and yeast were decreased in all treatment groups, whereas the Lactobacillus plantarum group and L. plantarum + natamycin group were highly significantly (P > 0.001) decreased. During air exposure, the water-soluble carbohydrates, ammonia nitrogen, lactic acid, and acetic acid were not affected on days 1-4, whereas pH and aerobic bacteria (were significantly (P < 0.05) increased on days 2-4. The addition of Lactobacillus plantarum and natamycin increased the gas production, in vitro dry matter digestibility, and in vitro neutral detergent fiber of SuMu No. 2 elephant grass silages. CONCLUSIONS: The addition of biological and chemical additives, such as L. plantrum alone and the combination with natamycin, affected the undesirable microbial community, fermentation characteristics, aerobic stability, and in vitro gas of SuMu No. 2 elephant grass. © 2021 Society of Chemical Industry.

Effects of re-ensiling on the fermentation quality and microbial community of napier grass (Pennisetum purpureum) silage.

BACKGROUND: With the rapid development of animal husbandry, the silage trade has increased in frequency. The re-ensiling of materials is often required before or after trading, resulting in the exposure of the silage to air before re-sealing. To develop a re-ensiling technique for silage, different silage exposure periods were simulated to check the possible effects on the fermentation quality and microbial community of silage. RESULTS: Fresh and wilted napier grass (Pennisetum purpureum) were ensiled for 90 days, then exposed to air for 0, 6, 12, 24, 36 or 48 h, before being re-ensiled. As a control, grass was directly ensiled for 180 days. Wilting increased the relative abundance of Klebsiella. The relative abundance of Paenibacillus in the unwilted silage was much higher than that in the wilted silage. Re-ensiling increased the relative abundance of Lactobacillus, but decreased the relative abundances of Klebsiella, Bacillus, and Paenibacillus. In addition, Lactobacillus became the dominant bacteria in the re-ensiled fresh and wilted silages. Re-ensiling within 48 h of exposure did not affect the fermentation quality of the wilted silage, whereas that of the unwilted silage declined when exposed to air for over 24 h. CONCLUSION: Re-ensiling wilted napier grass silage within 48 h of aerobic exposure did not cause the fermentation quality to decline. The unwilted napier grass silage contained a higher relative abundance of Paenibacillus and significantly deteriorated when re-ensiled after over 24 h of aerobic exposure. © 2021 Society of Chemical Industry.

Temporal assessment of root and shoot colonization of elephant grass (Pennisetum purpureum Schum.) host seedlings by Gluconacetobacter diazotrophicus strain LP343.

Gluconacetobacter diazotrophicus is a species of great agronomic potential due to its growth-promotion traits. Its colonization process in different plants has been reported. However, there have been no studies regarding its structural colonization in elephant grass. This is a fast-growing C4-Poaceae plant, and its application in Brazil is mainly aimed at feeding dairy cattle, due to its high nutritional value. Also, in the last decade, this grass has been applied in the production of biofuels. The present study aimed to monitor the colonization process of strain LP343 of G. diazotrophicus inoculated in elephant grass seedlings of PCEA genotype, by using a mCherry-tagged bacterium. Samples of roots and shoots collected at different periods were visualized by confocal laser-scanning microscopy. The colony-counting assay was used to compare the number of cells recovered in different niches and a qPCR was performed for the quantification of endophytic cells in root and shoot tissues. Results suggested that the strain LP343 quickly recognized the PCEA roots as host, attached to the elephant grass roots at 6 h, and 7 days after inoculation were able to colonize the xylem vessels of roots and shoots of elephant grass. This study advances our knowledge about the colonization process of G. diazotrophicus species in elephant grass, contributing to future studies involving the plant-bacteria interaction cultivated under gnotobiotic conditions.

Effects of various epiphytic microbiota inoculation on the fermentation quality and microbial community dynamics during the ensiling of sterile Napier grass.

AIM: To investigate epiphytic microbiota transformation of forages, their adaptation and contributions to fermentation quality of silage. METHODS AND RESULTS: Gamma-irradiated chopped Napier grass were ensiled with distilled water (STR), extracted epiphytic microbiota of Napier grass (NAP), sudan grass (SUD), whole crop corn (WCC) and forage sorghum (FS). Inoculating Napier grass with WCC significantly increased lactic acid (LA) concentrations during the initial ensiling period followed by a decline after 30 days. Relative to other silages (except STR) inoculation with NAP resulted in lower LA and greater pH, ammonia-N and dry matter (DM) losses. Silage inoculated with FS and SUD maintained lower pH as well as higher (P < 0·05) LA concentrations after 60 days of storage. During day 3 of ensilage, WCC, NAP and SUD inoculated silage were dominated by bacterial genera of Lactobacillus, while Lactococcus dominated the FS silage. Final silages were dominated by Lactobacillus in all treatment silages, however Enterobacter (16·3%) in NAP and Acetobacter (25·7%) in WCC silage were also prominent during the final ensiling. CONCLUSION: The inoculation of epiphytic microbiota of forage sorghum and sudan grass positively influenced the microbial community and fermentability of sterile Napier grass silage. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time to investigate the effects of various epiphytic microbiota as silage inoculants which can be used as alternative source of environmental friendly and economically feasible silage additives.

Pearl millet populations characterized by Fusarium prevalence, morphological traits, phenolic content, and antioxidant potential.

BACKGROUND: Pearl millet (Pennisetum glaucum L.) has become increasingly attractive due to its health benefits. It is grown as food for human consumption and fodder for livestock in Africa and Asia. This study focused on five pearl millet populations from different agro-ecological zones from Tunisia, and on characterization by morphological traits, total phenolic and flavonoid content, antioxidant activity, and occurrence of Fusarium. RESULTS: Analysis of variance revealed highly significant differences between populations for the quantitative traits. The highest grain weights occurred in the pearl millet cultivated in Zaafrana and Gergis of Tunisia. Early flowering and early maturing populations cultivated in the center (Zaafrana, Rejiche) and south (Gergis) of Tunisia tended to have a higher grain yield. The Zaafrana population showed the highest value of green fodder potentiel (number and weight of leaves/cultivar and the weight of tillers and total plant/cultivar) followed by Gergis and Rejiche. The Kelibia population showed the highest total phenolic and flavonoid content. Rejiche exhibited the greatest antioxidant activity. Trans-cinnamic, protocatechuic, and hydroxybenzoic acids were the major phenolic compounds in all the extracts. Three Fusarium species were identified in Tunisian pearl millet populations based on morphologic and molecular characterization. Fusarium graminearum and Fusarium culmorum occurred most frequently. The average incidence of the three Fusarium species was relatively low (<5%) in all populations. The lowest infection rate (0.1%) was recorded in the samples from Zaafrana. CONCLUSION: Chemometric analysis confirmed the usefulness of the above traits for discrimination of pearl millet populations, where a considerable variation according to geographical origin and bioclimatic conditions was observed. © 2020 Society of Chemical Industry.

Ensiling of whole-plant hybrid pennisetum with natamycin and Lactobacillus plantarum impacts on fermentation characteristics and meta-genomic microbial community at low temperature.

BACKGROUND: The aim of the current research was to clarify the impacts of the ensiling of whole-plant hybrid pennisetum with natamycin and Lactobacillus plantarum on fermentation characteristics and the meta-genomic microbial community at low temperatures. RESULTS: During the ensiling process, lactic acid (LA) and lactic acid bacteria (LAB) significantly (P < 0.05) increased and acetic acid (AA), water-soluble carbohydrate (WSC), ammonia total nitrogen (NH3-N), and yeast significantly (P < 0.05) reduced in treatments as compared to controls. Different treatments and different ensiling days led to variations in the bacterial community at family and genus levels. The family Lactobacillaceae and genera Lactobacillus and Pediococcus are dominant communities in treatment silage. The family and genus levels bacterial ecology and fermentation quality were analyzed by principal component analysis (PCA). The PCO1, and PCO2 can be explained by 10.81% and 72.14% of the whole variance regularly, similarly in PCO1 and PCO2 can be explained 24.23% and 52.06% regularly. The core bacterial micro-biome operational taxonomic unit (OTU) numbers increased in treatments, as compared to controls, on different hybrid pennisetum ensiling days. CONCLUSIONS: The inoculation of L. plantarum alone and combined with natamycin influenced the fermentation quality and reduced undesirable microorganisms during the fermentation of hybrid pennisetum silage. Natamycin alone did not significantly enhance the concentration of organic acid but numerically enhanced in treatments group as compared to control. © 2020 Society of Chemical Industry.

Effect of alfalfa microbiota on fermentation quality and bacterial community succession in fresh or sterile Napier grass silages.

The aim of this study was to reveal the reconstitution dynamics of alfalfa microbiota and their contribution to the fermentation quality of Napier grass silages. Napier grass was harvested at approximately 20% dry matter content, chopped to a theoretical length of cut of 2 to 3 cm, and ensiled in laboratory polyethylene plastic bags (400 × 250 mm) for 30 d. The Napier grass was treated with the following: natural fermentation and distilled water (NG), inoculum of alfalfa microbiota (AM), gamma-ray irradiation and distilled water (IR), and gamma-ray radiation and alfalfa microbiota (IR+AM). Three milliliters of inoculum (containing 8.93 log cfu/mL lactic acid bacteria, 9.76 log cfu/mL Enterobacteriaceae, 5.94 log cfu/mL yeast, and 6.53 log cfu/mL mold) eluted from equivalent fresh alfalfa (450 g) was added to each silo of AM and IR+AM treatments, and 3 mL of distilled water was added to the silo of the NG and IR treatments. Three triplicate silos per treatment were opened on d 1, 3, 5, 7, 14, and 30 for sampling and analysis of fermentation quality and bacterial community. Relative to the NG silages, IR+AM silages exhibited a higher lactic acid concentration. The higher acetic acid concentration in NG than in IR+AM silages after 7 d of ensiling was attributed to the dominant genus of Leuconostoc (64.29-49.04%). Adding alfalfa microbiota to sterile Napier grass could increase ammonia-N concentration compared with NG and IR silages after 3 d of ensiling. Leuconostoc was the most predominant genus in NG silages, followed by Lactobacillus. Pediococcus had a greater relative abundance than the indigenous microorganisms and was exclusively found in AM and IR+AM silages, whereas Lactobacillus exhibited a slight increase after 30 d of ensiling (relative abundance in each silage: 6.29 vs. 3.82%, respectively). Lactobacillus was the predominant genus in IR silages since the onset of the ensiling. These results suggest that alfalfa microbiota affected bacterial community succession in Napier grass silages, which in turn affected the fermentation products. Adding alfalfa microbiota to sterile Napier grass could decrease acetic acid concentration compared with NG silages; however, it increased ammonia-N concentration compared with IR silages after 3 d of ensiling.

Screening of natural lactic acid bacteria with potential effect on silage fermentation, aerobic stability and aflatoxin B1 in hot and humid area.

AIM: To effectively make high-quality silage in hot and humid area. METHODS AND RESULTS: The natural lactic acid bacteria (LAB) strains CZ149, XH358, XH753 and XH761 isolated from corn and Napier grass were screened by the potential of low pH growth and high lactic acid production, and their effect on silage quality, aerobic stability and aflatoxin B1 production of whole-crop corn was also studied in Sichuan, China. Four selected strains were Gram-positive and catalase-negative, rod-shaped strains that are able to grow at pH 3·5 and at 45°C. Strains CZ149, XH358, XH753 and XH761 were identified as Lactobacillus plantarum, L. salivarius, L. rhamnosus and L. paracasei, respectively. After 60 days of fermentation, all LAB strains showed no significant relationship with the quality of corn silage, whereas the lowest aflatoxin B1 and lactic-to-acetic ratio were detected in strain XH753-treated silage. Strain CZ149-treated silage showed worse aerobic stability and higher aflatoxin B1 concentration than control, whereas strain XH753-treated silage had better aerobic stability and lowest aflatoxin B1 concentration after aerobic exposure in hot and humid condition for 5 days. CONCLUSIONS: The three L. plantarum strains used in this study are not suitable as inoculants for local whole-crop corn silage, whereas L. rhamnosus 753 could prolong the aerobic stability and inhibit the accumulation of aflatoxin B1 at hot and humid condition. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new information of LAB inoculants for corn silage in hot and humid areas. Especially, a screened strain of Lactobacillus rhamnosus 753 can be used as a candidate strains to make high-quality silage in tropical and subtropical areas.