Plastic pollution has emerged as a global challenge affecting ecosystem health and biodiversity conservation. Terrestrial environments exhibit significantly higher plastic concentrations compared to aquatic systems. Micro/nano plastics (MNPs) have the potential to disrupt soil biology, alter soil properties, and influence soil-borne pathogens and roundworms. However, limited research has explored the presence and impact of MNPs on aquaculture systems. MNPs have been found to inhibit plant and seedling growth and affect gene expression, leading to cytogenotoxicity through increased oxygen radical production. The article discusses the potential phytotoxicity process caused by large-scale microplastics, particularly those unable to penetrate cell pores. It also examines the available data, albeit limited, to assess the potential risks to human health through plant uptake.

Summary: Cellulose is the most abundant unique biopolymer in nature with widespread applications in bioenergy and high‐value bioproducts. The large transmembrane‐localized cellulose synthase (CESA) complexes (CSCs) play a pivotal role in the biosynthesis and orientation of the para‐crystalline cellulose microfibrils during secondary cell wall (SCW) deposition. However, the hub CESA subunit with high potential homo/heterodimerization capacity and its functional effects on cell wall architecture, cellulose crystallinity, and saccharification efficiency remains unclear. Here, we reported the highly potent binding site containing four residues of Pro435, Trp436, Pro437, and Gly438 in the plant‐conserved region (P‐CR) of PalCESA4 subunit, which are involved in the CESA4‐CESA8 heterodimerization. The CRISPR/Cas9‐knockout mutagenesis in the predicted binding site results in physiological abnormalities, stunt growth, and deficient roots. The homozygous double substitution of W436Q and P437S and heterozygous double deletions of W436 and P437 residues potentially reduced CESA4‐binding affinity resulting in normal roots, 1.5–2‐fold higher plant growth and cell wall regeneration rates, 1.7‐fold thinner cell wall, high hemicellulose content, 37%–67% decrease in cellulose content, high cellulose DP, 25%–37% decrease in cellulose crystallinity, and 50% increase in saccharification efficiency. The heterozygous deletion of W436 increases about 2‐fold CESA4 homo/heterodimerization capacity led to the 50% decrease in plant growth and increase in cell walls thickness, cellulose content (33%), cellulose DP (20%), and CrI (8%). Our findings provide a strategy for introducing commercial CRISPR/Cas9‐mediated bioengineered poplars with promising cellulose applications. We anticipate our results could create an engineering revolution in bioenergy and cellulose‐based nanomaterial technologies.

Multiple gene silencing is being required to target and tangle metabolic pathways in eukaryotes and researchers have to develop a subtle method for construction of RNA interference (RNAi) cassettes. Although, several vectors have been developed due to different screening and cloning strategies but still some potential limitations remain to be dissolved. Here, we worked out a simple cloning strategy to develop multisite small interfering RNA (siRNA) cassette from different genes by two cloning steps. In this method, effective siRNA sites in the target messenger RNAs (mRNAs) were determined using in silico analysis and consecutively arranged to reduce length of inverted repeats. Here, we used one-step (polymerase chain reaction) PCR by designed long primer sets covering the selected siRNA sites. Rapid screening, cost-effective and shorten procedure are advantages of this method compare to PCR classic cloning. Validity of constructs was confirmed by optimal centroid secondary structures with high stability in plants.