The rapid increase in pig production has become a major contributor to environmental issues due to the mismanagement of organic waste. The sustainable and effective transformation of this waste into a fertilization resource has become an urgent topic for environmental protection, and new regulations have been imposed. The present study aimed to investigate the effects of different ratios of swine manure liquid (SML) and chemical fertilizers on soil phosphorus forms and microbial communities through field experiments cultivating spring wheat (cultivar “Jinqiang 10”) in Hebei, China. The results indicated that the application of SML in portions with traditional fertilizer can enhance soil pH and electrical conductivity (EC), as well as available phosphorus, particularly when the proportion of SML is high (SML ≥ 75%). Compared with CK, the available phosphorus content of group C3 increased by 22.3%. SML facilitated the transformation of stable phosphorus to unstable phosphorus, as well as the conversion of organic phosphorus to inorganic phosphorus. Additionally, SML increased the soil content of H₂O-P, NaHCO₃-Pi, and NaHCO₃-Po, and promoted the conversion of NaOH-Po to NaHCO₃-Po. Studies on bacterial diversity indicated that different fertilization treatments have no significant impact on the bacterial diversity in the 0–20 cm soil layer, whereas the dominant bacterial and fungal genera were positively correlated with the available phosphorus. The present study may facilitate the combined application of SML and chemical fertilizers for soil improvement and improve phosphorus availability.

S oybean vein necrosis virus (Orthotospovirus glycininecrovenae, SVNV) is an ambisense ssRNA virus in the genus Orthotospovirus first identified in Tennessee in 2008 (1). SVNV consists of three segments: S, M, and L. These encode a nucleocapsid protein (N), nonstructural proteins (NSs and NSm), glycoproteins (GN and GC), and an RNA-dependent RNA polymerase (RdRp) (2). The complete sequence of the SVNV17_Auburn_AL isolate was obtained using RNA-Seq and RACE. In 2023, soybean samples exhibiting symptoms of SVNV were collected. Total RNA was extracted from symptomatic leaves using the previous methodology (3), followed by ribosomal RNA depletion using the Illumina Ribo-Zero Plus rRNA Depletion Kit (Illumina, Cat: 20037135). Libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina and sequenced on an Illumina NovaSeq 6000 (150 bp PE, ~47 million reads). Quality control was conducted using FastQC (4), and adapter sequences were removed using BBDuk (https://sourceforge.net/projects/bbmap/). Processed reads were mapped to the SVNV-TN genome (GCA_004789395.1) using Bowtie (5). Variants (depth >80, Phred > 100) were called using BCFtools (6) and FreeBayes tool (7), and the average depth was calculated using SAMtools (6). Identified variants were visualized using IGV v2.3.57 (8). The consensus assembly was generated with BCFtools (6). For all tools, the default parameters were used. Missing terminal nucleotides were filled using RACE: 5′ ends with the Invitrogen 5′ RACE System (ThermoFisher, Cat: 18374058) using segment-specific primers (Table 1), and 3′ ends using E. coli poly(A) polymerase (NEB) followed by SuperScript III (Invitrogen) synthesis. PCR amplification used Phusion (ThermoFisher, F530S), cloned using the CloneJET PCR Cloning Kit, and 12 colonies were Sanger sequenced using an Applied Biosystems 3730xl sequencer. The genome of the SVNV17_Auburn_AL comprises 16,563 bases (2,602 bp [S], 4,948 bp [M], and 9,013 bp [L]) with a GC content of 35% and an average depth of 1,669×. The leaders are 58, 57, and 185 bases, while the trailers are 70, 91, and 30 bases for the S, M, and L segments, respectively. The first six bases (AGAGCA) at the 5′ ends are identical across all three segments and are complementary to the 3′ ends, forming a panhandle similar to other orthotospoviruses (9). Genome comparison between the SVNV17_Auburn_AL isolate and the TN strain revealed 43, 97, and 138 SNPs/indels in the S, M, and L segments (Fig. 1). To determine the impact on the protein level, the ORFs’ sequence was translated with ExPASy (10) and aligned to the TN strain with Clustal

In 2020, an evaluation of the basal level of incidence of tomato spotted wilt (TSW) in peanut was initiated in Alabama. This was done to understand the viral sequence divergence of tomato spotted wilt virus (TSWV) in a heavily managed system. From 2021 to 2023, 172 leaf samples were collected from peanut plants exhibiting symptoms of TSW from Brewton, Fairhope, and Headland, AL, to investigate genetic changes. Additionally, four thrips populations were collected in 2022 and 2023 from Fairhope and Headland, AL. The nucleocapsid protein of TSWV was sequenced from both leaf samples and thrips. A total of 175 nucleocapsids were sequenced, and their amino acid sequences were aligned to identify three conserved mutations compared to TSWV-MT2 (X61799.1) and seven conserved mutations when aligned against TSWV-BR-01 (NC_002051.1). Interestingly, only one conserved mutation was found in the thrips sequences compared to MT2, and no mutations were detected when aligned with BR-01. To identify if mutations caused a phenotype that could be measured, eight nucleocapsids with unique mutations were selected for localization in Nicotiana benthamiana. The localization patterns of these proteins were grouped into three phenotypes based on the observed protein aggregation speed. Small-plot trials assessed TSWV mutations and incidence in relation to planting date, insecticide use, and cultivar. Based on these data, although management strategies are effective at keeping levels of TSWV manageable, the virus can diversify its sequence, which causes changes in the expected phenotype of the nucleocapsid.

Soybean vein necrosis virus (SVNV) is a persistent, propagative, ambisense single-stranded RNA virus in the genus Orthotospovirus, transmitted by Nehydatothrips variabilis. To understand SVNV in the field, 33 samples exhibiting symptoms of SVNV were collected. The N, NSs, and NSm open reading frames (ORFs) were sequenced, revealing amino acid mutations in each gene. The five open reading frames of the SVNV Tennessee strain (N, NSs, NSm, GN, and GC) were fused in frame to GFP for experimentation in both plant and insect cells. N and NSs localize in plants at the cell periphery and nucleus. NSm induces cell death in plant cells, but not in insect cells, where cytoplasmic localization is observed. GN and GC glycoproteins localize to the membranes and display increased cytoplasmic localization in insect cells. The findings of this study contribute to understanding the genes of SVNV and capture sequence changes that have occurred over the past fifteen years.

Tomato spotted wilt virus (TSWV) is a major yield-limiting pathogen of peanut in the southeastern USA. To assess viral variability, field isolates were collected from symptomatic peanut plants at three locations in Alabama. Sequence analysis identified mutations in the non-structural movement protein (NSm) and the nucleocapsid protein (N). Subcellular localization studies showed that NSm fused to GFP (NSm:GFP) localized to plasmodesmata and co-localized with callose deposits by 2days postinfiltration (dpi). By 4 dpi, NSm:GFP formed cytoplasmic aggregates, and callose deposition appeared more consistent with basal plasmodesmatal patterns. The N protein localized to the nuclear periphery and cell margins at 2 dpi and later aggregated in the cytoplasm by 4 dpi. The early callose accumulation associated with NSm expression suggests a potential plant defence response, warranting further investigation.

During the screening of peanuts (Arachis hypogaea L.) for tomato spotted wilt virus (TSWV) in 2024, 36 plant samples expressing symptoms of tomato spotted wilt were collected from three locations in Alabama: Brewton Agricultural Research Unit (BARU), Brewton, AL; Wiregrass Research and Extension Center (WGREC), Headland, AL; and Gulf Coast Research and Extension Center (GCREC), Fairhope, AL. Foliar necrosis symptoms, not typically associated with TSWV, were also observed in all samples. Symptoms varied from moderate light brown to severe dark necrosis along the midveins and lateral veins. Samples were tested for four orthotospoviruses: TSWV, groundnut ringspot virus (GRSV), tomato chlorotic spot virus (TCSV), and soybean vein necrosis virus (SVNV) using ELISA (Agdia Inc., Elkhart, IN). All samples were positive for TSWV and negative for both GRSV and TCSV; however, one sample tested positive for SVNV (BARU 4). SVNV was previously reported not to infect peanuts (Zhou and Tzanetakis 2013). The 36 samples were subjected to RT-PCR using specific primers for the nucleocapsid (N) protein of SVNV (Shehata et al. 2024). Twenty-four of 36 samples were positive for SVNV (12 from BARU, two from WGREC, and 10 from GCREC). The amplified bands showed lower intensity than the positive control, possibly indicating a low titer of SVNV, which may explain the negative ELISAs. Following the manufacturer’sinstructions, these bands were cloned using the CloneJET PCR Cloning Kit (Thermo Fisher Scientific) and sent for Sanger sequencing. The resulting SVNV-N sequences were submitted to GenBank under accessions PQ821900 to PQ821905. The sequences demonstrated 98.19% nucleotide and 96.75% amino acid identities with SVNV from Tennessee (GCF_004789395.1). Nine conserved amino acid mutations were identified compared with the Tennessee strain, resembling those found in soybeans (Shehata et al. 2024). Additionally, RT-PCR was also used for TSWV-N detection (Martin et al. 2025), and 26 samples tested positive (12 from BARU, three from WGREC, and 11 from GCREC), which confirmed the presence of both SVNV and TSWV in positive samples. Further research is necessary to investigate co-infection between SVNV and TSWV, potential genome reassortment, and its mechanisms to understand the interactions between these viruses. These interactions could adversely impact legume production in Alabama, valued at $315 million in 2023 (soybean and peanut, USDA 2023). Since both TSWV and SVNV are transmitted by tobacco thrips (Frankliniella fusca) (Hameed et al. 2022; Keough et al. 2016), this is likely how SVNV was introduced to peanuts. This constitutes the first report on the detection of SVNV in peanuts in the United States, suggesting that SVNV has been adapting to new hosts since its discovery (Tzanetakis et al. 2009; Zhou et al. 2018).