Explore the vast range of titles available.

Key message We developed an in-house Python-based image analysis pipeline to investigate the movement patterns of Cuscuta. Our analysis unveiled that the coiling and circumnutation movements of Cuscuta are regulated by its intrinsic circadian rhythm. Cuscuta spp., commonly known as dodders, are rootless and leafless stem parasitic plants. Upon germination, Cuscuta starts rotating immediately in a counterclockwise direction (circumnutation) to locate a host plant, creating a seamless vascular connection to steal water and nutrients from its host. In this study, our aim was to elucidate the dynamics of the coiling patterns of Cuscuta , which is an essential step for successful parasitism. Using time-lapse photography, we recorded the circumnutation and coiling movements of C. campestris at different inoculation times on non-living hosts. Subsequent image analyses were facilitated through an in-house Python-based image processing pipeline to detect coiling locations, angles, initiation and completion times, and duration of coiling stages in between. The study revealed that the coiling efficacy of C. campestris varied with the inoculation time of day, showing higher success and faster initiation in morning than in evening. These observations suggest that Cuscuta , despite lacking leaves and a developed chloroplast, can discern photoperiod changes, significantly determining its parasitic efficiency. The automated image analysis results confirmed the reliability of our Python pipeline by aligning closely with manual annotations. This study provides significant insights into the parasitic strategies of C. campestris and demonstrates the potential of integrating computational image analysis in plant biology for exploring complex plant behaviors. Furthermore, this method provides an efficient tool for investigating plant movement dynamics, laying the foundation for future studies on mitigating the economic impacts of parasitic plants.

We present the first chromosome-level genome assembly and annotation for the genus Cuscuta, a twining and leafless parasitic plant of the morning glory family (Convolvulaceae). C. campestris, the study species, is a widely studied model parasite, due in part to its worldwide occurrence as a weed of agricultural and natural plant communities. The species has served as a model parasite for studies of parasite biology, haustorium development, growth responses to chemical and light stimuli, gene content and expression, horizontal gene transfer, and interspecies RNA movement and has a recently developed transformation system. The 505 Mb (1C) genome is assembled into 31 chromosomes and supports annotation of 47,199 protein-coding genes, 214 small RNA loci (including 146 haustoria-specific miRNAs), and 3,238 interspecies mobile mRNA loci. C. campestris is a recent tetraploid with a high retention of duplicated genes and chromosomes, with less than 8% nucleotide divergence between homoeologous chromosomes. We also show that transformation of C. campestris with the RUBY marker system allows visualization of transformed Cuscuta-derived fluorescent mobile molecules that have entered the host stem. This genome, with an associated genome browser and BLAST server, will be of value for scientists performing fundamental research in a wide range of molecular, developmental, population, and evolutionary biology, as well as serve as a research tool for studying interspecies mobile molecules, generating genetic markers for species and genotype identification, and developing highly specific herbicides.

Cuscuta spp., (dodder) is a rootless and leafless parasitic plant posing significant agricultural challenges. In this study, we aimed to elucidate the dynamics of the coiling patterns in Cuscuta campestris and examine the role of circadian rhythms in its host-seeking ability. Using time-lapse photography, we recorded the circumnutation and coiling movements of C. campestris at different inoculation times on non-living hosts. Subsequent image analyses were facilitated through an in-house Python-based image analysis pipeline. We observed that the coiling efficacy of C. campestris varied with the inoculation time of day, showing higher success and faster initiation in morning than in evening. These observations suggest that Cuscuta, despite lacking leaves and a developed chloroplast, can discern photoperiod changes, which significantly determine its parasitic efficiency. The automated image analysis confirmed the reliability of our Python pipeline, aligning closely with manual annotations. This study provides significant insights into the parasitic strategies of C. campestris and demonstrates the potential of integrating computational image analysis in plant biology for exploring complex plant behaviors. Furthermore, this method provides an efficient tool for investigating plant movement dynamics, laying the foundation for future studies on mitigating the economic impacts of parasitic plants.Competing Interest StatementThe authors have declared no competing interest.

Cuscuta campestris, a stem parasitic plant, has served as a valuable model plant for the exploration of plant-plant interactions and molecular trafficking. However, a major barrier to C. campestris research is that a method to generate stable transgenic plants has not yet been developed. Here, we describe the development of a Cuscuta transformation protocol using various reporter genes (GFP, GUS, or RUBY) and morphogenic genes (CcWUS2 and CcGRF/GIF), ultimately leading to a robust protocol for Agrobacterium-mediated C. campestris transformation. The stably transformed and regenerated RUBY C. campestris plants produced haustoria, the signature organ of parasitic plants, and these were functional in forming host attachments. Transformed C. campestris also produced flowers and transgenic seeds exhibiting betalain pigment, providing proof of germline transmission of the RUBY transgene. The integration locations of T-DNAs were confirmed through TAIL-PCR. Furthermore, the RUBY reporter is not only a useful selectable marker for the Agrobacterium-mediated transformation but also provides insight into the movement of molecules from C. campestris to the host during parasitism. Thus, the protocol to generate transgenic C. campestris reported here overcomes a major obstacle to Cuscuta research and opens up new possibilities for studying parasitic plant interactions.Competing Interest StatementThe authors have declared no competing interest.