To further explore the effects of heterofermentative lactic acid bacteria (LAB) on silage fermentation and aerobic stability, whole-plant corn at around the 1/2 milk-line stage was freshly chopped and ensiled in laboratory silos with deionized water (control), Lactobacillus buchneri (LB), or L. rhamnosus (LR). Each treatment was prepared in triplicate for 3, 14, and 60 d of fermentation, followed by 3 and 7 days of aerobic exposure. The dynamic changes in microbial community were studied by single molecule real-time (SMRT) sequencing. The results showed that the two LAB inoculants altered the microbial communities in different ways. Succession from L. plantarum to L. buchneri and L. rhamnosus was observed in LB- and LR-treated silage, respectively. Both silages improved aerobic stability (82 and 78 h vs. 44 h) by occupying the microbial niche to produce higher levels of acetic acid at terminal fermentation. Because Acetobacter fabarum dominated in the silages after aerobic exposure, beta diversity dramatically decreased. In this study, a. fabarum was reported for the first time in silage and was related to aerobic spoilage. The two heterofermentative LAB produced acetic acid and improved the aerobic stability of the corn silage by occupying the microbial niche at terminal fermentation. Inoculated L. rhamnosus had a greater pH for a longer period of time after opening and less DM loss at day 7.

Soil health is seriously threatened by the overuse of chemical fertilizers in agricultural management. Biogas slurry is often seen as an organic fertilizer resource that is rich in nutrients, and its use has the goal of lowering the amount of chemical fertilizers used while preserving crop yields and soil health. However, the application of continuous biogas slurry has not yet been studied for its long-term impact on soil nutrients and microbial communities in a rotation system of annual ryegrass-silage maize (Zea mays). This study aimed to investigate the impacts on the chemical properties and microbial community of farmland soils to which chemical fertilizer (NPK) (225 kg ha⁻¹), biogas slurry (150 t ha⁻¹), and a combination (49.5 t ha⁻¹ biogas slurry + 150 kg ha⁻¹ chemical fertilizer) were applied for five years. The results indicated that compared to the control group, the long-term application of biogas slurry significantly increased the SOC, TN, AP, and AK values by 45.93%, 39.52%, 174.73%, and 161.54%, respectively; it neutralized acidic soil and increased the soil pH. TN, SOC, pH, and AP are all important environmental factors that influence the structural composition of the soil’s bacterial and fungal communities. Chemical fertilizer application significantly increased the diversity of the bacterial community. Variation was observed in the composition of soil bacterial and fungal communities among the different treatments. The structure and diversity of soil microbes are affected by different methods of fertilization; the application of biogas slurry not only increases the contents of soil nutrients but also regulates the soil’s bacterial and fungal community structures. Therefore, biogas slurry can serve as a sustainable management measure and offers an alternative to the application of chemical fertilizers for sustainable intensification.

Wilted (around 35% DM) or un-wilted (around 35% DM) Italian ryegrass treated with three additives (formic acid, FA; Lactobacillus plantarum, LP; Lactobacillus buchneri, LB) was utilized to evaluate the effects of the dry matter (DM) contents on the microbial community and fermentation characteristics, which was ensiled for 60 days in a laboratory-scale silo, followed by 3 days of aerobic exposure. Significantly lower pH and higher lactic acid (LA) contents were observed in the LP-treated group ensiled at both DM contents (differences were significant when p < 0.05). The contents of LA, acetic acid (AA), numbers of lactic acid bacteria (LAB) and ammonia nitrogen (NH₃-N) in the FA-treated group were significantly lower than those in other treatments (p < 0.05). L. buchneri was the dominant bacteria after 60 days fermentation, while Enterobacteria became prevalent after 3 days of aerobic exposure. L. buchneri was found in the LB-treated group with higher acetic acid. Although the best fermentation quality was observed in the LP-treated silages, the aerobic stability was lowest compared to other groups (p < 0.05). In conclusion, our findings suggest that the DM content of Italian ryegrass affected its epiphytic microbial community and the effectiveness of the different type of additives. Formic acid was more suitable for un-wilted Italian ryegrass silage, L. plantarum had a better effect in wilted Italian ryegrass silage, and L. buchneri prolonged the aerobic stability of Italian ryegrass. DM content and purpose of ensiling should be the key factors for choosing different types of additives for Italian ryegrass silage.

Miscanthus sinensis is an ornamental grass, non-food bioenergy crop, and forage with high feeding value. It can adapt to many kinds of soil conditions due to its high level of resistance to various abiotic stresses. However, a low level of seed germination restricts the utilization and application of M. sinensis. It is reported that the Homeodomain-leucine zipper (HD-Zip) gene family participates in plant growth and development and ability to cope with outside environment stresses, which may potentially regulate seed germination and stress resistance in M. sinensis. In this study, a complete overview of M. sinensis HD-Zip genes was conducted, including gene structure, conserved motifs, chromosomal distribution, and gene duplication patterns. A total of 169 members were identified, and the HD-Zip proteins were divided into four subgroups. Inter-chromosomal evolutionary analysis revealed that four pairs of tandem duplicate genes and 72 segmental duplications were detected, suggesting the possible role of gene replication events in the amplification of the M. sinensis HD-Zip gene family. There was an uneven distribution of HD-Zip genes on 19 chromosomes of M. sinensis. Also, evolutionary analysis showed that M. sinensis HD-Zip gene family members had more collinearity with monocotyledons and less with dicotyledons. The gene structure analysis showed that there were 93.5% of proteins with motif 1 and motif 4, while motif 10 was only found in group IV. Based on the cis-elements analysis, it appeared that most of the genes were related to plant growth and development, various hormones, and abiotic stress. Furthermore, qRT-PCR analysis showed that Misin06G303300.1 was significantly expressed in seed germination and Misin05G030000.1 and Misin06G303300.1 were highly expressed under chromium, salt, and drought stress. Results in this study will provide a basis for further exploring the potential role of HD-Zip genes in stress responses and genetic improvement of M. sinensis seed germination.

Winter wheat (Triticum aestivum L.) silage has high feeding value and has become an important roughage resource in China. To recognize the optimal fermentation time of the silage product, this study systematically evaluated the temporal dynamics of microbial communities and metabolic profiles in whole winter wheat silage at days 7, 14, 30, 50, and 70. The dry matter (DM) content slightly fluctuated with the extension of fermentation time, with 28.14% at 70 days of ensiling. The organic matter and neutral detergent fiber content gradually decreased with the extension of fermentation time. A significant decrease in pH was observed at days 30, 50, and 70 compared to days 7 and 14 (p < 0.05), with the lowest pH value of 4.4 recorded at day 70. The contents of lactic acid, acetic acid, butyric acid, and total volatile fatty acids gradually increased with the extension of fermentation time, reaching a maximum at 70 days of ensiling. The dominant bacteria were Proteobacteria and Firmicutes at the phylum level, and the predominant bacteria were Hafnia-Obesumbacterium, Enterobacter, and Lactobacillus at the genus level. The relative abundance of Hafnia-Obesumbacterium and Lactobacillus fluctuated slightly with the duration of fermentation, reaching a minimum for the former and a maximum for Lactobacillus at 50 days of ensiling. By day 70, Sporolactobacillus emerged as a distinct silage biomarker. The dominant fungi was Ascomycota at the phylum level, and the predominant fungi were Fusarium and an unidentified fungus at the genus level. The correlation analysis revealed significant pH–organic acid–microbe interactions, with pH negatively correlating with organic acids but positively with specific bacteria, while organic acids showed complex microbial associations. Collectively, under natural fermentation conditions, the optimal fermentation period for wheat silage exceeds 70 days, and Sporolactobacillus shows potential as a microbial inoculant for whole winter wheat silage. These findings provide a theoretical foundation for optimizing whole winter wheat silage utilization and enhancing fermentation quality.

Biogas slurry with rich nutrients could be applied as fertilizer to improve nitrogen absorption and soil structure. Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms that establish mutualistic relationships with the plant roots. The purpose of this study was to study the effects of AMF and biogas slurry treatment on hybrid Pennisetum growth, soil chemical properties, and soil microorganisms. The results revealed that the biomass yield of hybrid Pennisetum was significantly increased after the application of biogas slurry, and it reached the peak values when the biogas slurry dosage was 900 t/hm², which were 13,216.67 kg/hm² and 13,733.33 kg/hm² in AMF− and AMF+ treatment groups, respectively. Moreover, biogas slurry treatment has a significant promoting effect on other agronomic traits related to biomass yield. As for soil chemical indicators, the contents of total nitrogen, nitrate nitrogen, ammonia nitrogen, and available phosphorus in the soil increased with the increase in biogas slurry application, while the soil organic matter was decreased. The addition of arbuscular mycorrhizal fungi significantly increased the species diversity of soil fungi with no biogas slurry application. Furthermore, when biogas slurry was applied, it had no significant effect on soil microbial diversity and composition, no matter the AMF+ or AMF− treatment. The research results can provide a reference for the long-term utilization of biogas slurry and it also can be used in the actual production of hybrid Pennisetum.