Stable Plants Research Articles

Metagenomic analyses of plant virus sequences in sewage water for plant viruses monitoring.

The online version contains supplementary material available at 10.1007/s40858-023-00575-8.

Constrained linear output regulation via measurement output feedback model predictive control

Recently, the output regulation problem with input constraints for incrementally stable plants was solved by an error feedback model predictive control (MPC) algorithm. This paper further considers the case with both state and input constraints for linear systems and uses a measurement output feedback MPC algorithm to solve the problem. By relaxing two restrictions, the result of this paper can handle a larger class of linear systems than the existing result. Due to the uncontrollability of the augmented system composed of the plant and the exosystem, a key assumption in the literature fails, we manage to overcome this difficulty by introducing novel tightened constraints and the terminal constraint. As a result, the proposed algorithm solves the linear output regulation problem in the presence of both state and input constraints. The effectiveness of our design is illustrated by a numerical example.

The RLCK–VND6 module coordinates secondary cell wall formation and adaptive growth in rice

The orderly deposition of secondary cell wall (SCW) in plants is implicated in various biological programs and is precisely controlled. Although many positive and negative regulators of SCW have been documented, the molecular mechanisms underlying SCW formation coordinated with distinct cellular physiological processes during plant adaptive growth remain largely unclear. Here, we report the identification of Cellulose Synthase co-expressed Kinase1 (CSK1), which encodes a receptor-like cytoplasmic kinase, as a negative regulator of SCW formation and its signaling cascade in rice. Transcriptome deep sequencing of developing internodes and genome-wide co-expression assays revealed that CSK1 is co-expressed with cellulose synthase genes and is responsive to various stress stimuli. The increased SCW thickness and vigorous vessel transport in csk1 indicate that CSK1 functions as a negative regulator of SCW biosynthesis. Through observation of green fluorescent protein-tagged CSK1 in rice protoplasts and stable transgenic plants, we found that CSK1 is localized in the nucleus and cytoplasm adjacent to the plasma membrane. Biochemical and molecular assays demonstrated that CSK1 phosphorylates VASCULAR-RELATED NAC-DOMAIN 6 (VND6), a master SCW-associated transcription factor, in the nucleus, which reduces the transcription of a suite of SCW-related genes, thereby attenuating SCW accumulation. Consistently, genetic analyses show that CSK1 functions upstream of VND6 in regulating SCW formation. Interestingly, our physiological analyses revealed that CSK1 and VND6 are involved in abscisic acid-mediated regulation of cell growth and SCW deposition. Taken together, these results indicate that the CSK1–VND6 module is an important component of the SCW biosynthesis machinery, which coordinates SCW accumulation and adaptive growth in rice. Our study not only identifies a new regulator of SCW biosynthesis but also reveals a fine-tuned mechanism for precise control of SCW deposition, offering tools for rationally tailoring agronomic traits.

Tunable control of insect pheromone biosynthesis in Nicotiana benthamiana.

Previous work has demonstrated that plants can be used as production platforms for molecules used in health, medicine, and agriculture. Production has been exemplified in both stable transgenic plants and using transient expression strategies. In particular, species of Nicotiana have been engineered to produce a range of useful molecules, including insect sex pheromones, which are valued for species-specific control of agricultural pests. To date, most studies have relied on strong constitutive expression of all pathway genes. However, work in microbes has demonstrated that yields can be improved by controlling and balancing gene expression. Synthetic regulatory elements that provide control over the timing and levels of gene expression are therefore useful for maximizing yields from heterologous biosynthetic pathways. In this study, we demonstrate the use of pathway engineering and synthetic genetic elements for controlling the timing and levels of production of Lepidopteran sex pheromones in Nicotiana benthamiana. We demonstrate that copper can be used as a low-cost molecule for tightly regulated inducible expression. Further, we show how construct architecture influences relative gene expression and, consequently, product yields in multigene constructs. We compare a number of synthetic orthogonal regulatory elements and demonstrate maximal yields from constructs in which expression is mediated by dCas9-based synthetic transcriptional activators. The approaches demonstrated here provide new insights into the heterologous reconstruction of metabolic pathways in plants.

Backfitting of Dukovany NPP for design extension conditions

The contribution deals with the backfitting of Dukovany NPP for design extension conditions with emphasis on severe accidents. This NPP, located in Czech Republic, is equipped with 4 units of VVER-440/V213 reactors. During the last decades significant attention was paid to backfitting of these units. New safety systems aimed at mitigating the consequences of severe accidents have been designed and have been (are being) implemented. At first, special features of VVER-440/V213 reactor and containment equipped with bubbler condenser pressure suppression system are briefly described. The systematic process of plant backfitting for design extension condition is summarized further. Attention is paid to description of crucial severe accident management strategies that have been adopted and design and implementation of new safety systems that are necessary to fulfil these strategies. Than the requirements of Czech regulatory body (SUJB) set for practical elimination of early or large radioactive releases are discussed. Finally, the performances of severe accident management strategies are illustrated using the specific case of blackout scenario. For this purpose, an analysis was performed using ASTEC code developed by the French IRSN. The results of analysis are briefly described and discussed further. From the obtained results it follows that the high-pressure melting scenario is reliably avoided, corium is localised and stabilised inside the reactor vessel and the hydrogen in containment atmosphere is safely controlled by passive autocatalytic recombiners. Thus, stable plant state with preserved containment integrity is reached. Source term that has been estimated under these conditions indicates that the predicted releases are well below the criteria set by SUJB for early or large radioactive releases, and therefore long-term contamination of the environment is avoided.

Zinc-finger protein GmZF351 improves both salt and drought stress tolerance in soybean.

Abiotic stress is one of the most important factors reducing soybean yield. It is essential to identify regulatory factors contributing to stress responses. A previous study found that the tandem CCCH zinc-finger protein GmZF351 is an oil level regulator. In this study, we discovered that the GmZF351 gene is induced by stress and that the overexpression of GmZF351 confers stress tolerance to transgenic soybean. GmZF351 directly regulates the expression of GmCIPK9 and GmSnRK, leading to stomata closing, by binding to their promoter regions, which carry two CT(G/C)(T/A)AA elements. Stress induction of GmZF351 is mediated through reduction in the H3K27me3 level at the GmZF351 locus. Two JMJ30-demethylase-like genes, GmJMJ30-1 and GmJMJ30-2, are involved in this demethylation process. Overexpression of GmJMJ30-1/2 in transgenic hairy roots enhances GmZF351 expression mediated by histone demethylation and confers stress tolerance to soybean. Yield-related agronomic traits were evaluated in stable GmZF351-transgenic plants under mild drought stress conditions. Our study reveals a new mode of GmJMJ30-GmZF351 action in stress tolerance, in addition to that of GmZF351 in oil accumulation. Manipulation of the components in this pathway is expected to improve soybean traits and adaptation under unfavorable environments.

Plant-made vaccines against viral diseases in humans and farm animals.

Plants provide not only food and feed, but also herbal medicines and various raw materials for industry. Moreover, plants can be green factories producing high value bioproducts such as biopharmaceuticals and vaccines. Advantages of plant-based production platforms include easy scale-up, cost effectiveness, and high safety as plants are not hosts for human and animal pathogens. Plant cells perform many post-translational modifications that are present in humans and animals and can be essential for biological activity of produced recombinant proteins. Stimulated by progress in plant transformation technologies, substantial efforts have been made in both the public and the private sectors to develop plant-based vaccine production platforms. Recent promising examples include plant-made vaccines against COVID-19 and Ebola. The COVIFENZ® COVID-19 vaccine produced in Nicotiana benthamiana has been approved in Canada, and several plant-made influenza vaccines have undergone clinical trials. In this review, we discuss the status of vaccine production in plants and the state of the art in downstream processing according to good manufacturing practice (GMP). We discuss different production approaches, including stable transgenic plants and transient expression technologies, and review selected applications in the area of human and veterinary vaccines. We also highlight specific challenges associated with viral vaccine production for different target organisms, including lower vertebrates (e.g., farmed fish), and discuss future perspectives for the field.

Discovery of aphid-transmitted Rice tiller inhibition virus from native plants through metagenomic sequencing.

A major threat to rice production is the disease epidemics caused by insect-borne viruses that emerge and re-emerge with undefined origins. It is well known that some human viruses have zoonotic origins from wild animals. However, it remains unknown whether native plants host uncharacterized endemic viruses with spillover potential to rice (Oryza sativa) as emerging pathogens. Here, we discovered rice tiller inhibition virus (RTIV), a novel RNA virus species, from colonies of Asian wild rice (O. rufipogon) in a genetic reserve by metagenomic sequencing. We identified the specific aphid vector that is able to transmit RTIV and found that RTIV would cause low-tillering disease in rice cultivar after transmission. We further demonstrated that an infectious molecular clone of RTIV initiated systemic infection and causes low-tillering disease in an elite rice variety after Agrobacterium-mediated inoculation or stable plant transformation, and RTIV can also be transmitted from transgenic rice plant through its aphid vector to cause disease. Finally, global transcriptome analysis indicated that RTIV may disturb defense and tillering pathway to cause low tillering disease in rice cultivar. Thus, our results show that new rice viral pathogens can emerge from native habitats, and RTIV, a rare aphid-transmitted rice viral pathogen from native wild rice, can threaten the production of rice cultivar after spillover.

Addition of the T5 exonuclease increases the prime editing efficiency in plants

Prime editing (PE) is a versatile genome editing tool without the need for double-stranded DNA breaks or donor DNA templates, but is limited by low editing efficiency. We previously fused the M-MLV reverse transcriptase to the Cas9 nickase, generating the PE2 (v1) system, but the editing efficiency of this system is still low. Here we develop different versions of PE2 by adding the 5′-to-3′ exonuclease at different positions of the nCas9-M-MLV RT fusion protein. PE2 (v2), in which the T5 exonuclease fused to the N-terminus of the nCas9-MMLV fusion protein enhances prime editing efficiency of base substitutions, deletions, and insertions at several genomic sites by 1.7- to 2.9-fold in plant cells compared to PE2 (v1). The improved editing efficiency of PE2 (v2) is further confirmed by generating increased heritable prime edits in stable transgenic plants compared to the previously established PE-P1, PE-P2, and PPE systems. Using PE2 (v2), we generate herbicide-resistant rice by simultaneously introducing mutations causing amino acid substitutions at two target sites. The PE efficiency is further improved by combining PE2 (v2) and dual-pegRNAs. Taken together, the increased genome editing efficiency of PE2 (v2) developed in this study may enhance the applications of PE in plants.

Combination of Hairy Root and Whole-Plant Transformation Protocols to Achieve Efficient CRISPR/Cas9 Genome Editing in Soybean

The new gene-editing technology CRISPR/Cas system has been widely used for genome engineering in various organisms. Since the CRISPR/Cas gene-editing system has a certain possibility of low efficiency and the whole plant transformation of soybean is time-consuming and laborious, it is important to evaluate the editing efficiency of designed CRISPR constructs before the stable whole plant transformation process starts. Here, we provide a modified protocol for generating transgenic hairy soybean roots to assess the efficiency of guide RNA (gRNA) sequences of the CRISPR/Cas constructs within 14 days. The cost- and space-effective protocol was first tested in transgenic soybean harboring the GUS reporter gene for the efficiency of different gRNA sequences. Targeted DNA mutations were detected in 71.43-97.62% of the transgenic hairy roots analyzed as evident by GUS staining and DNA sequencing of the target region. Among the four designed gene-editing sites, the highest editing efficiency occurred at the 3' terminal of the GUS gene. In addition to the reporter gene, the protocol was tested for the gene-editing of 26 soybean genes. Among the gRNAs selected for stable transformation, the editing efficiency of hairy root transformation and stable transformation ranged from 5% to 88.8% and 2.7% to 80%, respectively. The editing efficiencies of stable transformation were positively correlated with those of hairy root transformation with a Pearson correlation coefficient (r) of 0.83. Our results demonstrated that soybean hairy root transformation could rapidly assess the efficiency of designed gRNA sequences on genome editing. This method can not only be directly applied to the functional study of root-specific genes, but more importantly, it can be applied to the pre-screening of gRNA in CRISPR/Cas gene editing.

Evaluate the guide RNA effectiveness via Agrobacterium-mediated transient assays in Nicotiana benthamiana.

CRISPR/Cas9-based genome editing system is a powerful tool for plant genetic improvement. However, the variable efficiency of guide RNA(s) (gRNA) represents a key limiting factor that hampers the broad application of the CRISPR/Cas9 system in crop improvement. Here, we employed the Agrobacterium-mediated transient assays to evaluate the effectiveness of gRNAs for editing genes in Nicotiana benthamiana and soybean. We designed a facile screening system based on indels that can be introduced by CRISPR/Cas9-mediated gene editing. A gRNA binding sequence (23 nucleotides) was inserted into the open reading frame of yellow fluorescent protein (YFP) gene (gRNA-YFP), which disrupted the YFP reading frame and results in no fluorescent signal when it was expressed in plant cells. Transiently co-expression of Cas9 and a gRNA targeting the gRNA-YFP gene in plant cells could restore the YFP reading frame and recover the YFP signals. We evaluated five gRNAs targeting Nicotiana benthamiana and soybean genes and confirmed the reliability of the gRNA screening system. The effective gRNAs targeting NbEDS1, NbWRKY70, GmKTI1, and GmKTI3 had been used to generate transgenic plants and resulted in expected mutations on each gene. While a gRNA targeting NbNDR1 was confirmed to be ineffective in transient assays. This gRNA indeed failed to trigger target gene mutations in stable transgenic plants. Thus, this new transient assay system can be used to validate the effectiveness of gRNAs before generating stable transgenic plants.

Effect of a suitable treatment period on the genetic transformation efficiency of the plant leaf disc method

BackgroundAgrobacterium tumefaciens-mediated leaf disc genetic transformation is an important way to achieve transgenics or gene editing. Ensuring stable and efficient genetic transformation is still an important problem in modern biology. It is assumed that the difference in the development status of genetic transformation cells of receptor materials is the main reason for the difference and instability of genetic transformation efficiency; the stable and efficient genetic transformation rate can be obtained by defining the appropriate treatment period of the receptor material and applying genetic transformation in a timely manner.ResultsBased on these assumptions, we studied and established an efficient and stable Agrobacterium-mediated plant transformation system with hybrid poplar (Populus alba × Populus glandulosa, 84 K) leaves, stem segments and tobacco leaves as the research objects. There were differences in the development process of leaf bud primordial cells from different explants, and the genetic transformation efficiency was significantly related to the cell development stage of the in vitro cultured materials. Among them, the genetic transformation rate of poplar and tobacco leaves was the highest on the 3rd and 2nd day of culture, reaching 86.6% and 57.3%, respectively. The genetic transformation rate of poplar stem segments was the highest on the 4th day of culture, reaching 77.8%. The best treatment period was from the development of leaf bud primordial cells to the S phase of the cell cycle. The number of cells detected using flow cytometry and 5-ethynyl-2ʹ-deoxyuridine (EdU) staining, the expression of cell cycle-related protein CDKB1; 2, CDKD1; 1, CYCA3; 4, CYCD1; 1, CYCD3; 2, CYCD6; 1, and CYCH; 1 of explants, and morphological changes of explants can be used as indicators to determine the appropriate treatment period for genetic transformation.ConclusionsOur study provides a new and universal set of methods and characteristics to identify the S phase of the cell cycle and apply genetic transformation treatments at the appropriate time. Our results are of great significance for improving the efficiency and stability of plant leaf disc genetic transformation.

The F-box protein UFO controls flower development by redirecting the master transcription factor LEAFY to new cis-elements.

In angiosperms, flower development requires the combined action of the transcription factor LEAFY (LFY) and the ubiquitin ligase adaptor F-box protein, UNUSUAL FLORAL ORGANS (UFO), but the molecular mechanism underlying this synergy has remained unknown. Here we show in transient assays and stable transgenic plants that the connection to ubiquitination pathways suggested by the UFO F-box domain is mostly dispensable. On the basis of biochemical and genome-wide studies, we establish that UFO instead acts by forming an active transcriptional complex with LFY at newly discovered regulatory elements. Structural characterization of the LFY-UFO-DNA complex by cryo-electron microscopy further demonstrates that UFO performs this function by directly interacting with both LFY and DNA. Finally, we propose that this complex might have a deep evolutionary origin, largely predating flowering plants. This work reveals a unique mechanism of an F-box protein directly modulating the DNA binding specificity of a master transcription factor.

Saturating PI Control of Stable Nonlinear Systems Using Singular Perturbations

This article presents an antiwindup propor- tional-integral (PI) controller, using a saturating integrator, for a single-input and single-output (SISO) stable nonlinear plant, whose steady-state input–output map is increasing. We prove that, under reasonable assumptions, there exists an upper bound on the controller gain such that for any constant reference input, the corresponding equilibrium point of the closed-loop system is exponentially stable, with a “large” region of attraction. When the state of the closed-loop system converges to this equilibrium point, then the tracking error tends to zero. The closed-loop stability analysis employs Lyapunov methods in the framework of the singular perturbations theory. Finally, we show that if the plant satisfies the asymptotic gain property, then the closed-loop system is globally asymptotically stable for any sufficiently small controller gain. The effectiveness of the proposed PI controller is proved by showing how it performs as part of the control algorithm of a synchronverter (a special type of dc to ac power converter).

Effects of canopy density on understory plant diversity in Robinia pseudoacacia plantations on the Loess Plateau of China

To solve the problems of understory plant shortage and biodiversity reduction caused by high tree density of Robinia pseudoacacia plantations on the Loess Plateau in northwest China, we analyzed the data collected by field survey and from literatures. We used the upper boundary line method to examine the impacts of canopy density on understory plant diversity. Based on field survey at the Guanshan Forest Farm of Jingchuan County of Gansu Pro-vince, the species number of understory plants in R. pseudoacacia plantations was higher than that in natural grassland (91 vs. 78). The dominant species varied with canopy density, which was completely different from that of natural grassland. A comprehensive analysis of both literature data and field survey data showed that, when mean annual precipitation (MAP) <550 mm or >550 mm, the increases in canopy density first led to a stable understory plant coverage and then a sharp or slight decrease, and understory plant biomass showed a sharp and continuous decrease or a slight increase and then decrease. Understory plant species richness and other diversity indices (Shannon, Simpson, and Pielou indices) first increase and then decrease, with bigger variation range under lower MAP. In all, the characteristics (coverage, biomass and species diversity, etc.) of understory plant community in R. pseudoacacia plantations were strongly affected by canopy density, with higher sensitivity under lower MAP. There was a general threshold range of canopy density of 0.45-0.6. When canopy density was above or below this thre-shold range, it would lead to a rapid decrease of the most characteristics of understory plant community. Therefore, keeping canopy density within the rational range of 0.45-0.6 in the management of R. pseudoacacia plantations is the key to make all the above-mentioned understory plant characteristics at relatively high level.

Evaluation of Soil Loss Tolerance and Tree Growth Features Based on Planting Ground Methods in the Alpine Center, Degraded Forestland in the Republic of Korea

The Jeongseon Alpine Center, a degraded forestland, has a very unique soil feature as it was filled with soil cut from the nearby area and has not yet been rehabilitated since the end of the 2018 Winter Olympics. Therefore, this study attempted to identify a way to establish a stable and efficient planting groundwork to successfully restore this distinctive region. Six different planting groundworks, control, biochar, leaf-mold, mulching, tillage, and polyacrylamide treatments were constructed in September 2020. Soil-related indices have been tracked while the growth and physiological responses of planted Pinus densiflora (Pd) and Larix kaempferi (Lk) seedlings were monitored until May 2022. Mulching had 2–4 times the average SWC when compared to the control. Simultaneously, there was the least variation in soil temperature change, and total soil loss was only 0.05 ton/ha. Similarly, the leaf-mold had a relatively high SWC, and soil fertility increased, while total soil loss was 6.37 tons/ha, indicating a high trend in line with biochar. The Pd leaf-mold had the highest in Amax, E, gs, and Jmax. Furthermore, following the leaf-mold, mulching showed the second-highest photosynthetic indices in general, and the height and DRC also demonstrated favorable results in the above two treatments. Mulching had 1.6 to 2.2 times higher than the control group in PIabs and PItotal, which represent photosynthetic vitality, so the impact on environmental stress was thought to be less than that of leaf-mold. The mulching of Lk showed the greatest growth and physiological responses, Nonetheless, the photosynthetic indices were rather low when compared to Pd, with no discernible differences across treatments. As a result, the considerable effect of the planting groundwork method could not be demonstrated. Therefore, in the case of Pd, the leaf-mold showed the best growth and physiological response. Nevertheless, in terms of soil temperature maintenance and soil loss for slope stability, mulching is assessed to be the most ideal planting groundwork method.

Chloroplasts evolved an additional layer of translational regulation based on non-AUG start codons for proteins with different turnover rates

Chloroplasts have evolved from photosynthetic cyanobacteria-like progenitors through endosymbiosis. The chloroplasts of present-day land plants have their own transcription and translation systems that show several similarities with prokaryotic organisms. A remarkable feature of the chloroplast translation system is the use of non-AUG start codons in the protein synthesis of certain genes that are evolutionarily conserved from Algae to angiosperms. However, the biological significance of such use of non-AUG codons is not fully understood. The present study was undertaken to unravel the significance of non-AUG start codons in vivo using the chloroplast genetic engineering approach. For this purpose, stable transplastomic tobacco plants expressing a reporter gene i.e. uidA (GUS) under four different start codons (AUG/UUG/GUG/CUG) were generated and β-glucuronidase (GUS) expression was compared. To investigate further the role of promoter sequences proximal to the start codon, uidA was expressed under two different chloroplast gene promoters psbA and psbC that use AUG and a non-AUG (GUG) start codons, respectively, and also showed significant differences in the DNA sequence surrounding the start codon. Further, to delineate the role of RNA editing that creates AUG start codon by editing non-AUG codons, if any, which is another important feature of the chloroplast transcription and translation system, transcripts were sequenced. In addition, a proteomic approach was used to identify the translation initiation site(s) of GUS and the N-terminal amino acid encoded when expressed under different non-AUG start codons. The results showed that chloroplasts use non-AUG start codons in combination with the translation initiation site as an additional layer of gene regulation to over-express proteins that are required at high levels due to their high rates of turnover.

Virus Occurrence and Survival in Reusable Resources: A Minireview

This work covers important aspects of the occurrence and viability of various viruses in the two most common reusable waste resources: wastewater and biomass waste. Detection of human, bacterial and plant viruses in these wastes are summarized. Historically, human viruses have been monitored in wastewater for decades. Evidence suggests that wastewater mostly contains fecal-orally transmitted viruses, which are abundant and diverse. Recently, an increasing occurrence of SARS-CoV2 in sewage water with the spreading epidemics has been confirmed but lacking biological proof of infectivity yet. Besides human pathogens, wastewater is shown to be rich in bacteriophages and plant viruses as well, which supposedly enter the water from human guts. Viruses serving as water quality indicators are also discussed here. Lastly, we focus on biomass waste treatment, showing the presence of some common and stable plant viruses which may supposedly survive the technological process.

Machine learning based crop growth management in greenhouse environment using hydroponics farming techniques

Today more than one billion out of 6.5 billion people in the world are affected by hunger. The significant growth in human population inevitably places considerable pressure on food supplies. In traditional farming, farmers require good quality soil, natural minerals and also the production relatively consumes more time. The hydroponics smart greenhouse method has been adopted as a result of technological advancements. Hydroponics is a technique of growing plants that does not require the use of soil, and the plants in hydroponics grows in a shorter duration and have higher yields. In normal hydroponics system, a portion of the greenhouse is used for harvesting and the parameters like temperature, and humidity can have an impact on plant growth, and the plants are not properly nourished. In this proposed system, in hydroponics for germination organic coconut coir medium is used rather than rock wool, because rock wool is not bio-degradable and composed of volcanic materials. In the proposed research of hydroponics, the system is automated on a wide scale covering the entire green house with different crops produced in different climatic conditions. The implementation is done on, stable greenhouse environment and plants are grown covering the entire greenhouse, under constant optimal conditions which is favorable to have maximum yield. The prediction of absolute crop growth rate of leafy vegetables is predicted using KNN algorithm under various conditions. The several forms of hydroponics for commercial crop growth has been compared and it has been concluded that NFT along with coconut coir medium is best suitable for hydroponics and it produces an accuracy of 93%. Macro average, a performance metric for F1 score produces 33% and weighted average, which represents the number of instances connected to different class labels, produces an accuracy of 93%.

Heritable transgene-free genome editing in plants by grafting of wild-type shoots to transgenic donor rootstocks

Generation of stable gene-edited plant lines using clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated protein 9 (Cas9) requires a lengthy process of outcrossing to eliminate CRISPR–Cas9-associated sequences and produce transgene-free lines. We have addressed this issue by designing fusions of Cas9 and guide RNA transcripts to tRNA-like sequence motifs that move RNAs from transgenic rootstocks to grafted wild-type shoots (scions) and achieve heritable gene editing, as demonstrated in wild-type Arabidopsis thaliana and Brassica rapa. The graft-mobile gene editing system enables the production of transgene-free offspring in one generation without the need for transgene elimination, culture recovery and selection, or use of viral editing vectors. We anticipate that using graft-mobile editing systems for transgene-free plant production may be applied to a wide range of breeding programs and crop plants.