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.

Genome-based taxonomy and functional prediction of Sphingomonas fuzhouensis sp. nov. and Massilia phyllosphaerae sp. nov. isolated from Pennisetum sp. with plant growth-promoting potential.

Two facultatively aerobic strains, designated SGZ-02T and SGZ-792T, were isolated from plant Pennisetum sp., exhibiting the highest 16S rRNA gene sequence similarities with the type strains of Sphingomonas zeae LMG 28739T (98.6%) and Massilia forsythiae NBRC 114511T (98.4%), respectively. SGZ-02T grew between 5 and 45 °C, pH 5.0-11.0 and tolerated NaCl concentrations of 0-4% (w/v), whereas SGZ-792T thrived at 5-40 °C, pH 5.0-11.0 and NaCl tolerance to 0-3.5% (w/v). The major quinone of SGZ-02T was ubiquinone-10, with the dominant fatty acids being C16:0 (13.5%), Summed Feature 3 (6.3%), C14:02-OH (5.3%) and Summed Feature 8 (66.3%). SGZ-792T predominantly contained ubiquinone-8, with major fatty acids being C16:0 (20.3%), Summed Feature 3 (5.0%) and Summed Feature 8 (54.7%). Average nucleotide identity and digital DNA-DNA hybridization values between two strains and their closest references strains were below the bacterial species threshold. Based on genotypic and phenotypic characteristics, strains SGZ-02T and SGZ-792T are proposed as novel species within the genera Sphingomonas and Massilia, respectively. The suggested names for the new species are Sphingomonas fuzhouensis sp. nov. (SGZ-02T = GDMCC 1.4033T = JCM 36769T) and Massilia phyllosphaerae sp. nov. (SGZ-792T = GDMCC 1.4211T = JCM 36643T), respectively.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Effect of endophytic Bacillus megaterium colonization on structure strengthening, microbial community, chemical composition and stabilization properties of Hybrid Pennisetum.

BACKGROUND: This study was conducted to analyze the effects of endophytic Bacillus megaterium (BM 18-2) colonization on structure strengthening, microbial community, chemical composition and stabilization properties of Hybrid Pennisetum. RESULTS: The BM 18-2 had successfully colonized in the interior tissues in both leaf and stem of Hybrid Pennisetum. During ensiling, the levels of pH, acetic acid (AA), butyric acid (BA), propionic acid (PA), and the population of yeast and aerobic bacteria were significantly (P > 0.05) lower, while lactic acid bacteria (LAB) and lactic acid (LA) were significantly (P < 0.001) higher with the steps forward of ensiling in with BM 18-2 as compared to without BM 18-2 colonized of Hybrid Pennisetum. During the different ensiling days, at days 3, 6, 15, and 30, the genus Brevundimonas, Klebsiella, Lactococcus, Weissella, Enterobacter, Serratia, etc. population were significantly decreased, while genus Pediococcus acidilactici and Lactobacillus plantarum were significantly influenced in treated groups as compared to control. The genus Lactobacillus and Pediococcus were positively correlated with treatment groups. CONCLUSIONS: It is concluded that the endophytic bacteria strain BM 18-2 significantly promoted growth characteristics and biomass yield before ensiling and after ensiling inoculated with or without Lactobacillus plantarum could improve the distinct changes of the undesirable microbial diversity, chemical composition, and stabilization properties in with BM 18-2 as compared to without BM 18-2 colonized Hybrid Pennisetum. © 2019 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.

Diversity and Tissue Preference of Osmotolerant Bacterial Endophytes Associated with Pearl Millet Genotypes Having Differential Drought Susceptibilities.

Genetic and functional diversity of osmotolerant bacterial endophytes colonizing the root, stem, and leaf tissues of pearl millet genotypes differing in their drought susceptibility was assessed. Two genotypes of pearl millet, viz., the drought tolerant genotype TT-1 and the drought susceptible genotype PPMI-69, were used in the present study. Diazotrophs were found to be the predominant colonizers, followed by the Gram positive bacteria in most of the tissues of both the genotypes. Higher proportion of bacterial endophytes obtained from the drought tolerant genotype was found to be osmotolerant. Results of 16S rRNA gene-ARDRA analysis grouped 50 of the highly osmotolerant isolates into 16 clusters, out of which nine clusters had only one isolate each, indicating their uniqueness. One cluster had 21 isolates and remaining clusters were represented by isolates ranging from two to four. The representative isolates from each cluster were identified, and Bacillus was found to be the most prevalent osmotolerant genera with many different species. Other endophytic bacteria belonged to Pseudomonas sp., Stenotrophomonas sp., and Macrococcus caseolyticus. High phylogenetic diversity was observed in the roots of the drought tolerant genotype while different tissues of the drought susceptible genotype showed less diversity. Isolates of Bacillus axarquiensis were present in all the tissues of both the genotypes of pearl millet. However, most of the other endophytic bacteria showed tissue/genotype specificity. With the exception of B. axarquiensis and B. thuringiensis, rest all the species of Bacillus were found colonizing only the drought-tolerant genotype; while M. caseolyticus colonized all the tissues of only the drought susceptible genotype. There was high incidence of IAA producers and low incidence of ACC deaminase producers among the isolates from the root tissues of the drought-tolerant genotype while reverse was the case for the drought-susceptible genotype. Thus, host played an important role in the selection of endophytes based on both phylogenetic and functional traits.

Current Status of Pearl Millet Downy Mildew Prevalence across Agroecological Zones of Senegal.

Pearl millet is a dominant staple cereal crop for smallholder farmers in Senegal. However, the crop is constrained by various nonbiotic and biotic stresses such as downy mildew disease. To assess the prevalence of this disease in Senegal, a field survey was conducted during the rainy season of 2017 across eight main pearl millet production regions following latitudinal gradient with different climatic conditions. Results showed that downy mildew prevalence was higher in Kaolack (incidence = 68.19%), Kaffrine (incidence = 77.19%), Tambacounda (incidence = 97.03%), Sedhiou (incidence = 82.78%), and Kolda (incidence = 98.01%) than Thies (incidence = 28.21%), Diourbel (incidence = 24.46%), and Fatick (incidence = 37.75%) regions. The field survey revealed an incidence as high as 98% and 28% of infected area in surveyed fields. Significant correlations between geographic coordinates, disease incidence, and infected areas were also observed. This study provided information that could help to understand the prevalence of downy mildew in pearl millet in Senegal.

Comparative genomic analysis revealed rapid differentiation in the pathogenicity-related gene repertoires between Pyricularia oryzae and Pyricularia penniseti isolated from a Pennisetum grass.

BACKGROUND: A number of Pyricularia species are known to infect different grass species. In the case of Pyricularia oryzae (syn. Magnaporthe oryzae), distinct populations are known to be adapted to a wide variety of grass hosts, including rice, wheat and many other grasses. The genome sizes of Pyricularia species are typical for filamentous ascomycete fungi [~ 40 Mbp for P. oryzae, and ~ 45 Mbp for P. grisea]. Genome plasticity, mediated in part by deletions promoted by recombination between repetitive elements [Genome Res 26:1091-1100, 2016, Nat Rev Microbiol 10:417-430,2012] and transposable elements [Annu Rev Phytopathol 55:483-503,2017] contributes to host adaptation. Therefore, comparisons of genome structure of individual species will provide insight into the evolution of host specificity. However, except for the P. oryzae subgroup, little is known about the gene content or genome organization of other Pyricularia species, such as those infecting Pennisetum grasses. RESULTS: Here, we report the genome sequence of P. penniseti strain P1609 isolated from a Pennisetum grass (JUJUNCAO) using PacBio SMRT sequencing technology. Phylogenomic analysis of 28 Magnaporthales species and 5 non-Magnaporthales species indicated that P1609 belongs to a Pyricularia subclade, which is genetically distant from P. oryzae. Comparative genomic analysis revealed that the pathogenicity-related gene repertoires had diverged between P1609 and the P. oryzae strain 70-15, including the known avirulence genes, other putative secreted proteins, as well as some other predicted Pathogen-Host Interaction (PHI) genes. Genomic sequence comparison also identified many genomic rearrangements relative to P. oryzae. CONCLUSION: Our results suggested that the genomic sequence of the P. penniseti P1609 could be a useful resource for the genetic study of the Pennisetum-infecting Pyricularia species and provide new insight into evolution of pathogen genomes during host adaptation.

Large elevation and small host plant differences in the arbuscular mycorrhizal communities of montane and alpine grasslands on the Tibetan Plateau.

Understanding the diversity and community structure of arbuscular mycorrhizal fungi (AMF) in extreme conditions is fundamental to predict the occurrence and evolution of either symbiotic partner in alpine ecosystems. We investigated the AMF associations of three plant species at elevations ranging between 3105 and 4556 m a.s.l. on Mount Segrila on the Tibetan Plateau. Three of four locations were studied in two consecutive years. The AMF diversity and community composition in the roots of Carex pseudofoetida, Pennisetum centrasiaticum, and Fragaria moupinensis differed little. However, at high elevations, the abundance of members of Acaulosporaceae increased relative to that of Glomeraceae. Plants at lower elevation sites, where Glomeraceae predominated as root symbionts, had higher leaf nitrogen and phosphorus concentrations than plants at higher elevation sites, where Acaulosporaceae predominated. The overall phylogenetic relatedness of the AMF increased with increasing elevation. This suggests that abiotic filtering may play an important role in the structuring of symbiotic AMF communities along elevational gradients. The functional role of Acaulosporaceae whose relative abundance was found to increase with elevation in alpine environments needs to be clarified in future studies.

Inheritance and Allelic Relationship Among Downy Mildew Resistance Genes in Pearl Millet.

Pearl millet downy mildew (DM), caused by Sclerospora graminicola, is of serious economic concern to pearl millet farmers in the major crop-growing areas of the world. To study the inheritance and allelic relationship among genes governing resistance to this disease, three DM-resistant pearl millet lines (834B, IP 18294-P1, and IP 18298-P1) and one susceptible line (81B) were selected on the basis of disease reaction under greenhouse conditions against two isolates of S. graminicola (Sg 526-1 and Sg 542-1). Three resistant parents were crossed with the susceptible parent to generate F1, F2, and backcross BC1P1 (susceptible parent × F1) and BC1P2 (resistant parent × F1) generations for inheritance study. To carry out a test for allelism, the three resistant parents were crossed with each other to generate F1 and F2 generations. The different generations of these crosses were screened for disease reaction against two isolates (Sg 526-1 and Sg 542-1) by artificial inoculation under greenhouse conditions. The segregation pattern of resistance in the F2 and corresponding backcross generations revealed that resistance to DM is controlled by a single dominant gene in 834B and IP 18294-P1 and by two dominant genes in IP 18298-P1. A test for allelism inferred that a single dominant gene for resistance in 834B is nonallelic to that which governs resistance in IP 18294-1, whereas one of the two dominant genes for DM resistance in IP 18298-P1 against the test isolates is allelic to the gene for DM resistance in 834B and a second gene is allelic to the resistance gene present in IP 18294-P1.