Effect of Cultivars and Temperature on Synergistic Interaction Between Panicum Mosaic Virus and Satellite Panicum Mosaic Virus in Switchgrass

Panicum mosaic virus (PMV), the type species of the genus Panicovirus in the family Tombusviridae, naturally infects switchgrass (Panicum virgatum L.). PMV and its molecular partner, satellite panicum mosaic virus (SPMV), interact synergistically in co-infected millets with exacerbated disease phenotype and increased accumulation of PMV, compared to plants infected only by PMV. In this study, we examined the reaction of switchgrass cvs. Summer and Kanlow to PMV and PMV+SPMV infections at 24°C and 32°C. Switchgrass cv. Summer was susceptible to PMV at both temperatures. In contrast, cv. Kanlow was tolerant to PMV at 24°C but not at 32°C, suggesting that Kanlow harbors temperature-sensitive resistance against PMV. At 24°C, PMV was readily detected in inoculated leaves but not in upper non-inoculated leaves of Kanlow, suggesting that resistance to PMV was likely mediated by abrogation of long-distance virus transport. Co-infection by PMV and SPMV at 24°C and 32°C in cv. Summer but not in Kanlow caused increased symptomatic systemic infection and mild disease synergism with slightly increased PMV accumulation compared to plants infected only by PMV. These data suggest that the interaction between PMV and SPMV in switchgrass is cultivar dependent, manifested in Summer but not in Kanlow. However, co-inoculation of cv. Kanlow by PMV+SPMV caused an enhanced asymptomatic infection, suggesting a role for SPMV in enhancing symptomless infection in a tolerant cultivar. These data suggest that enhanced asymptomatic infections in virus-tolerant switchgrass cultivar could serve as a source for virus spread and play an important role in panicum mosaic disease epidemiology under field conditions. Our data revealed that cultivars, co-infection with SPMV, and temperature influenced the severity of symptoms elicited by PMV in switchgrass.

Determination of swithgrass (Panicum virgatum L.) seeds quality

Purpose. Developing a method for determination of the laboratory seed germination that could reduce the biological dormancy period and, accordingly, increase the intensity of germination. Methods. Laboratory, measuring and weighing, mathematical and statistical. Results. Cooling switchgrass seeds at a temperature of 10°C for 7 days on average for three years did not lead to a decrease in germination energy and germination compared to cooling for 14 days. These indexes were almost the same and amounted to 74 and 76%, 73 and 75%, respectively. There were no significant deviations in germination energy and seed germination over the years of research as affected by duration of the cool period. The production test of the developed method, carried out in the accredited control and measuring laboratory, confirmed the obtained in the laboratory results. Conclusions. Seed germination by an improved method, when pre-cooling is carried out for 7 instead of 14 days and counting of germinated seeds on 15th day instead of 20th, has reduced the time to determine germination by 13 days without reducing the quality of analysis. It is advisable to determine the 1000-seed weight in one of three ways, but the most accurate is the third way, i.e. counting the seeds in 10 repetitions.

Productivity of switchgrass (Panicum virgatum L.) agrophytocenoses for long-term use

Purpose. To determene the productivity of switchgrass (Panicum virgatum L.) agrophytocenoses under the long-term use. Methods. Field, statistical, calculation and comparative. Results. During long-term switchgrass cultivation, the lowest yield was obtained in the first vegetation year: 19.7 t/ha in the control treatment, 25.3 and 17.5 t/ha in the experimental treatments. The average raw biomass yield was as following: 30.3 t/ha in the control (variety ‘Cave-in-Rock’), 32.6 t/ha and 26.5 t/ha in the experimental treatments (varieties ‘Kanlow’ and ‘Morozko’, respectively). The yield of solid biofuel (17.3 t/ha) indicates the advantage of ‘Kanlow’ over the control (11.7 t/ha). The lowest yield (15.7 t/ha) was obtained from ‘Morozko’ variety. The energy yield over the treatments: 312.8 GJ/ha in the control, 397.5 and 367.2 GJ/ha in the experimental treatments, respectively. The cost of the grown production was as following: 416.03 UAH/t in the control (‘Cave-in-Rock’), 312.01 UAH/t in ‘Kanlow’, and 366.42 UAH/t in ‘Morozko’. The profitability of the control (‘Cave-in-Rock’) was 69.5%, ‘Kanlow’ – 101.4%, ‘Morozko’ – 66.8%. Economic evaluation of the productivity of switchgrass agrophytocenoses resulted in the following ranking: ‘Kanlow’, ‘Cave-in-Rock’, ‘Morozko’. When analyzing the energy equivalent of switchgrass biomass production and energy consumption, it was found that ‘Kanlow’ yielded 588.8 GJ/ha and the control – 468.3 GJ/ha, which is lower by 120.5 GJ/ha. Conclusions. The yield of switchgrass agrophytocenoses on average over the years of research (2014–2019) was 30.3 t/ha in ‘Cave-in-Rock’ (the control), 32.6 t/ha in ‘Kanlow’, and 26.5 t/ha in ‘Morozko’. The energy equivalent of the obtained yield was 468.3 GJ/ha in ‘Cave-in-Rock’ and 588.8 GJ/ha in ‘Kanlow’, which is by 120.5 GJ/ha higher than in the control.

Impact of Harvest on Switchgrass Leaf Microbial Communities

Switchgrass is a promising feedstock for biofuel production, with potential for leveraging its native microbial community to increase productivity and resilience to environmental stress. Here, we characterized the bacterial, archaeal and fungal diversity of the leaf microbial community associated with four switchgrass (Panicum virgatum) genotypes, subjected to two harvest treatments (annual harvest and unharvested control), and two fertilization levels (fertilized and unfertilized control), based on 16S rRNA gene and internal transcribed spacer (ITS) region amplicon sequencing. Leaf surface and leaf endosphere bacterial communities were significantly different with Alphaproteobacteria enriched in the leaf surface and Gammaproteobacteria and Bacilli enriched in the leaf endosphere. Harvest treatment significantly shifted presence/absence and abundances of bacterial and fungal leaf surface community members: Gammaproteobacteria were significantly enriched in harvested and Alphaproteobacteria were significantly enriched in unharvested leaf surface communities. These shifts were most prominent in the upland genotype DAC where the leaf surface showed the highest enrichment of Gammaproteobacteria, including taxa with 100% identity to those previously shown to have phytopathogenic function. Fertilization did not have any significant impact on bacterial or fungal communities. We also identified bacterial and fungal taxa present in both the leaf surface and leaf endosphere across all genotypes and treatments. These core taxa were dominated by Methylobacterium, Enterobacteriaceae, and Curtobacterium, in addition to Aureobasidium, Cladosporium, Alternaria and Dothideales. Local core leaf bacterial and fungal taxa represent promising targets for plant microbe engineering and manipulation across various genotypes and harvest treatments. Our study showcases, for the first time, the significant impact that harvest treatment can have on bacterial and fungal taxa inhabiting switchgrass leaves and the need to include this factor in future plant microbial community studies.

Ammonia-oxidizing bacterial communities are affected by nitrogen fertilization and grass species in native C4 grassland soils

Background Fertilizer addition can contribute to nitrogen (N) losses from soil by affecting microbial populations responsible for nitrification. However, the effects of N fertilization on ammonia oxidizing bacteria under C4 perennial grasses in nutrient-poor grasslands are not well studied. Methods In this study, a field experiment was used to assess the effects of N fertilization rate (0, 67, and 202 kg N ha−1) and grass species (switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii)) on ammonia-oxidizing bacterial (AOB) communities in C4 grassland soils using quantitative PCR, quantitative reverse transcription-PCR, and high-throughput amplicon sequencing of amoA genes. Results Nitrosospira were dominant AOB in the C4 grassland soil throughout the growing season. N fertilization rate had a stronger influence on AOB community composition than C4 grass species. Elevated N fertilizer application increased the abundance, activity, and alpha-diversity of AOB communities as well as nitrification potential, nitrous oxide (N2O) emission and soil acidity. The abundance and species richness of AOB were higher under switchgrass compared to big bluestem. Soil pH, nitrate, nitrification potential, and N2O emission were significantly related to the variability in AOB community structures (p < 0.05).

Sustainability Trait Modeling of Field-Grown Switchgrass (Panicum virgatum) Using UAV-Based Imagery

Unmanned aerial vehicles (UAVs) provide an intermediate scale of spatial and spectral data collection that yields increased accuracy and consistency in data collection for morphological and physiological traits than satellites and expanded flexibility and high-throughput compared to ground-based data collection. In this study, we used UAV-based remote sensing for automated phenotyping of field-grown switchgrass (Panicum virgatum), a leading bioenergy feedstock. Using vegetation indices calculated from a UAV-based multispectral camera, statistical models were developed for rust disease caused by Puccinia novopanici, leaf chlorophyll, nitrogen, and lignin contents. For the first time, UAV remote sensing technology was used to explore the potentials for multiple traits associated with sustainable production of switchgrass, and one statistical model was developed for each individual trait based on the statistical correlation between vegetation indices and the corresponding trait. Also, for the first time, lignin content was estimated in switchgrass shoots via UAV-based multispectral image analysis and statistical analysis. The UAV-based models were verified by ground-truthing via correlation analysis between the traits measured manually on the ground-based with UAV-based data. The normalized difference red edge (NDRE) vegetation index outperformed the normalized difference vegetation index (NDVI) for rust disease and nitrogen content, while NDVI performed better than NDRE for chlorophyll and lignin content. Overall, linear models were sufficient for rust disease and chlorophyll analysis, but for nitrogen and lignin contents, nonlinear models achieved better results. As the first comprehensive study to model switchgrass sustainability traits from UAV-based remote sensing, these results suggest that this methodology can be utilized for switchgrass high-throughput phenotyping in the field.

Overexpression of PvWOX3a in switchgrass promotes stem development and increases plant height

Switchgrass (Panicum virgatum L.) is an important perennial, noninvasive, tall ornamental grass that adds color and texture to gardens and landscapes. Moreover, switchgrass has been considered a forage and bioenergy crop because of its vigorous growth, low-input requirements, and broad geography. Here, we identified PvWOX3a from switchgrass, which encodes a WUSCHEL-related homeobox transcription factor. Transgenic overexpression of PvWOX3a in switchgrass increased stem length, internode diameter, and leaf blade length and width, all of which contributed to a 95% average increase in dry weight biomass compared with control plants. Yeast one-hybrid and transient dual-luciferase assays showed that PvWOX3a can repress the expression of gibberellin 2-oxidase and cytokinin oxidase/dehydrogenase through apparently direct interaction with their promoter sequences. These results suggested that overexpression of PvWOX3a could increase gibberellin and cytokinin levels in transgenic switchgrass plants, which promotes cell division, elongation, and vascular bundle development. We also overexpressed PvWOX3a in a transgenic miR156-overexpressing switchgrass line that characteristically exhibited more tillers, thinner internodes, and narrower leaf blades. Double transgenic switchgrass plants displayed significant increases in internode length and diameter, leaf blade width, and plant height but retained a tiller number comparable to that of plants expressing miR156 alone. Ultimately, the double transgenic switchgrass plants produced 174% more dry-weight biomass and 162% more solubilized sugars on average than control plants. These findings indicated that PvWOX3a is a viable potential genetic target for engineering improved shoot architecture and biomass yield of horticulture, fodder, and biofuel crops.

Producción forrajera y semillas de Panicum virgatum, Tripsacum dactyloides y Sporobolus airoides en Tulancigo, Hidalgo

Ante la necesidad de transferir tecnología de pastos de menor uso en agua de riego en el Valle de Tulancingo, el objetivo del estudio fue evaluar tres pastos mexicanos Panicum virgatum, Sporobolus airoides y Tripsacum dactyloides en producción de forraje, densidad de población tallos, producción de semilla y caracterización de semillas de manera física y fisiológica. El estudio se llevó a cabo de abril a agosto y de agosto a diciembre de 2020 en condiciones de riego por goteo con 10 trasplantes en 12 m2 a partir de plantas adultas. El rendimiento mayor de forraje (suma de dos ciclos) y hojas se observó en P. virgatum (3.59 t MS ha-1; P < 0.0001), seguido por T. dactyloides (1.15 t MS ha-1) y S. airoides (0.719 t MS ha-1). La población de tallos vivos acumulada en T. dactyloides fue 216, en P. virgatum 187 y en S. airoides 82 tallos por planta-1 (P < 0.0001). La producción de diásporas en suma de los dos ciclos, en P. virgatum fue 147 kg ha-1, similar (P > 0.05) a T. dactyloides, sin embargo, las cúpulas con incipiente pureza física (0.79 %). La mayor pureza física (14.08 kg ha-1; P < 0.0001) y semilla pura viable (12.3 kg ha-1; P < 0.0001) se observó en Sporobolus airoides.