This study aimed to evaluate the gene expression of HSP70 and HMA3 in the switchgrass inoculated with plant-growth-promoting-bacteria (PGPB) under cadmium (Cd) stress and to observe the benefit of PGPB in plant growth and development. Plants were grown in hydroponic culture and treated with PGPB inoculants: Pseudomonas grimontii, Pantoea vagans, Pseudomonas veronii, and Pseudomonas fluorescens with the strains Bc09, So23, E02, and Oj24, respectively. The experimental results revealed that HSP70 and HMA3 genes expressed highly in the PGPB-inoculated plants under Cd stress. In addition, the expression of HSP70 and HMA3 genes was considerably higher in the first two days after successive four-day exposure of Cd in plants compared to the last two days of exposure. Increased biomass and indole-3-acetic-acid production with reduced Cd accumulation were observed in the PGPB-inoculated plants under Cd stress compared to the Cd-control plants. These PGPB, with their beneficial mechanisms, protect plants by modifying the gene expression profile that arises during Cd-toxic conditions and increased the healthy biomass of switchgrass. This demonstrates there is a correlation among the growth parameters under Cd stress. The PGPB in this study may help to intensify agriculture by triggering mechanisms to encourage plant growth and development under heavy metal stress.

Cadmium (Cd) is a toxic heavy metal element. It is relatively easily absorbed by plants and enters the food chain, resulting in human exposure to Cd. Italian ryegrass (Lolium multiflorum Lam.), an important forage cultivated widely in temperate regions worldwide, has the potential to be used in phytoremediation. However, genes regulating Cd translocation and accumulation in this species are not fully understood. Here, we optimized PacBio ISO-seq and integrated it with RNA-seq to construct a de novo full-length transcriptomic database for an un-sequenced autotetraploid species. With the database, we identified 2367 differentially expressed genes (DEGs) and profiled the molecular regulatory pathways of Italian ryegrass with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis in response to Cd stress. Overexpression of a DEG LmAUX1 in Arabidopsis thaliana significantly enhanced plant Cd concentration. We also unveiled the complexity of alternative splicing (AS) with a genome-free strategy. We reconstructed full-length UniTransModels using the reference transcriptome, and 29.76% of full-length models had more than one isoform. Taken together, the results enhanced our understanding of the genetic diversity and complexity of Italian ryegrass under Cd stress and provided valuable genetic resources for its gene identification and molecular breeding.

Subcellular fractions and the chemical forms of cadmium (Cd) reflect its level of toxicity to plants; however, these effects of exogenous glutathione (GSH) are poorly understood. We exposed two Italian ryegrass (Lolium multiflorum) cultivars (IdyII and Harukaze) to 50 µM Cd or 200 µM GSH to investigate the effect of GSH on the Cd uptake, subcellular compartments, and chemical forms. Cd significantly inhibited the plant growth, while GSH supplementation decreased this inhibition. The application of GSH significantly improved the Cd concentration in the roots but reduced that in the shoots and decreased the Cd translocation from root to shoot. The Cd concentration of the root in the cell wall was increased while the concentration in the soluble fraction was decreased when supplied with GSH. The inorganic form (80% ethanol for Cd extraction) in the roots was significantly reduced when treated with GSH. The Cd form extracted by 2% acetic acid (HAC) with low toxicity and immobility were greatly increased. In leaves, the application GSH decreased in any form of Cd form extracted. In conclusion, exogenous GSH decreased the translocation of Cd and alleviated Italian ryegrass Cd toxicity by accumulating more Cd in the root cell wall and immobilizing more Cd in lower toxicity fractions.

Bm-SP142 is a 35 kDa protease in the silkworm, but its exact functions remain unknown. In this study, sequence alignment revealed that the His-Asp-Ser catalytic triad is embedded in the TAAHC-DIAL-GDSGGP sequence motif, establishing Bm-SP142 as a serine protease. Soluble recombinant GST-BmSP142 was expressed and purified, and serine protease activity was confirmed in vitro. RT-qPCR results indicated that Bm-SP142 was mainly expressed in the middle part of the silkworm midgut, and Bm-SP142 transcripts were significantly up-regulated at 24 hours post infection (hpi) in BmBDV-resistant strains (798) inoculated with BmBDV and BmNPV-resistant strains (NB) inoculated with BmNPV, but not in BmBDV-susceptible strains (306). Surprisingly, transcripts were significantly down-regulated at 12 hpi in BmNPV-susceptible strains (HuaBa 35) inoculated with BmNPV, compared with healthy silkworms. Recombinant BmNPV treated with purified Bm-SP142 effectively impaired its ability to infect BmN cells, and Bm-SP142 decreases the efficiency of BmNPV and BmBDV propagation in silkworms. Furthermore, overexpression of Bm-SP142 in BmN cells inhibited viral propagation.

Planting bioenergy crops with high water-use efficiency (WUE) on heavy metal-polluted land is a good practice for biomass production and phytoremediation. Switchgrass (Panicum virgatum L.), a C₄ perennial bioenergy grass, is native to the United States. The relationship between the WUE and Cd accumulation of switchgrass has seldom been studied. Here, the WUE and Cd accumulation characteristics of 14 high-biomass switchgrass cultivars were investigated under Cd stress by hydroponic culture. The main results showed that Cd inhibited the instantaneous WUE in switchgrass seedlings and that the inhibition rate was more significant in the upland types than in the lowland types of switchgrass. A positive correlation was found between relative WUE and Cd accumulation in roots and shoots. The relative expression level of stomatal control-related genes (ERECTA and EPF1) in lowland cultivars with high WUE was higher than in upland cultivars with low WUE, both in control and Cd treatment conditions. The results suggest that it would be possible to further select and cultivate switchgrass with high WUE and a high capacity for Cd accumulation for phytoremediation in Cd-contaminated land.