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1. Evidence of a novel silencing effect on transgenes in the Arabidopsis thaliana sperm cell

   https://doi.org/10.1093/plcell/koad219  

   Ohnishi, Yukinosuke; Kawashima, Tomokazu (August 2023, The Plant Cell)

   Abstract We encountered unexpected transgene silencing in Arabidopsis thaliana sperm cells; transgenes encoding proteins with no specific intracellular localization (cytoplasmic proteins) were silenced transcriptionally or posttranscriptionally. The mRNA of cytoplasmic protein transgenes tagged with a fluorescent protein gene was significantly reduced, resulting in undetectable fluorescent protein signals in the sperm cell. Silencing of the cytoplasmic protein transgenes in the sperm cell did not affect the expression of either its endogenous homologous genes or cotransformed transgenes encoding a protein with targeted intracellular localization. This transgene silencing in the sperm cell persisted in mutants of the major gene silencing machinery including DNA methylation. The incomprehensible, yet real, transgene silencing phenotypes occurring in the sperm cell could mislead the interpretation of experimental results in plant reproduction, and this Commentary calls attention to that risk and highlights details of this novel cytoplasmic protein transgene silencing. 

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2. Enabling the study of gene function in gymnosperms: Virus‐induced gene silencing in Ephedra tweedieana

   https://doi.org/10.1002/aps3.70027  

   Garcia, Anthony_G K; Bùi, Jo Trang; Michael, Todd P; Ickert‐Bond, Stefanie M; Di_Stilio, Verónica S (December 2025, Applications in Plant Sciences)

   Abstract PremiseAs the sister clade to angiosperms, extant gymnosperms are crucial for reconstructing ancestral gene regulatory networks in seed plants. This highlights the need for model systems representing each of their distinct lineages. However, tools to quickly and effectively investigate gene function in gymnosperms are still limited due to the challenges of long life cycles and large genome sizes. Species within the xerophytic genusEphedra(Gnetales) have comparatively smaller genomes and shrubby growth habits with shorter life spans, making them better suited for greenhouse cultivation and laboratory experiments. MethodsWe implement virus‐induced gene silencing (VIGS) to manipulate gene expression inEphedra tweedieanaviaAgrobacterium‐mediated vacuum infiltration of tobacco rattle virus (TRV1 and TRV2) into seedlings. ResultsTreatment resulted in highly efficient gene silencing of theE. tweedieana PHYTOENE DESATURASE(PDS) orthologEtwPDS. The expected photobleaching phenotype was observed as early as two weeks, and lasted at least five months in stems, shoot tips, leaves, axillary meristems, and lateral branches of treated plants. DiscussionWe report on virus‐induced targeted gene silencing ofPDSin a Gnetales representative to further enable functional studies of the genetic mechanisms underpinning adaptations in gymnosperms, an important and underrepresented lineage of seed plants. 

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3. Exploiting somatic piRNAs in Bemisia tabac i enables novel gene silencing through RNA feeding

   https://doi.org/10.26508/lsa.202000731  

   Mondal, Mosharrof; Brown, Judith K; Flynt, Alex (August 2020, Life Science Alliance)

   null (Ed.)

   RNAi promises to reshape pest control by being nontoxic, biodegradable, and species specific. However, due to the plastic nature of RNAi, there is a significant variability in responses. In this study, we investigate small RNA pathways and processing of ingested RNAi trigger molecules in a hemipteran plant pest, the whitefly Bemisia tabaci . Unlike Drosophila , where the paradigm for insect RNAi technology was established, whitefly has abundant somatic piwi-associated RNAs (piRNAs). Long regarded as germline restricted, piRNAs are common in the soma of many invertebrates. We sought to exploit this for a novel gene silencing approach. The main principle of piRNA biogenesis is the recruitment of target RNA fragments into the pathway. As such, we designed synthetic RNAs to possess complementarity to the loci we annotated. Following feeding of these exogenous piRNA triggers knockdown as effective as conventional siRNA-only approaches was observed. These results demonstrate a new approach for RNAi technology that could be applicable to dsRNA-recalcitrant pest species and could be fundamental to realizing insecticidal RNAi against pests. 

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4. The epigenetic factor FVE orchestrates cytoplasmic SGS3-DRB4-DCL4 activities to promote transgene silencing in Arabidopsis

   https://doi.org/10.1126/sciadv.abf3898  

   Sun, Di; Li, Yanjun; Ma, Zeyang; Yan, Xingxing; Li, Niankui; Shang, Baoshuan; Hu, Xiaomei; Cui, Kai; Koiwa, Hisashi; Zhang, Xiuren (August 2021, Science Advances)

   null (Ed.)
5. Silencing the genome with linker histones

   https://doi.org/10.1073/pnas.2009513117  

   Hansen, Jeffrey C. (June 2020, Proceedings of the National Academy of Sciences)