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At least 91 records · Page 5

Leaf spectral reflectance and transmittance, San Lorenzo, Panama, 2020

This data package contains full-range (350 – 2500 nm) leaf reflectance and transmittance spectra of leaves collected at the San Lorenzo forest canopy crane site in Panama (PA-SLZ) from January to March 2020. Around 1000 leaves were measured from 50 species and various leaf phenological stages. Leaves were sampled from both the top of the canopy and multiple heights within the canopy from 10 vertical profiles and for six core species, from the top of the canopy. This data package includes processed leaf spectra (*.csv), and raw data from the spectroradiometer (compressed as .zip). Metadata files include data descriptions (_dd.csv) for tabular data and sample information and a description of the field campaign protocol (PA-SLZ_2020_Protocol.pdf). In addition to the leaf spectral measurements included here, these samples were also used for measurement of leaf gas exchange, carbon and nitrogen content and leaf mass per unit leaf area (LMA) contained in related data packages.

54 ENVIRONMENTAL SCIENCES↗

G-LiHT Campaign Leaf Sample details & photos, March 2017: Puerto Rico

This data package includes details of leaves sampled for leaf spectra and chemistry from 5 sites in Puerto Rico, in March of 2017. Sunlit canopy and shaded leaves of 66 species were collected. Data for each sample includes species, leaf age, type of analysis (spectroscopy, gas exchange, chemistry), sample number and sample photographs. The data package includes a spreadsheet with sample information and a zip file of photographs (1.6 GB). This data was collected as part of the 2017 BNL–G-LiHT leaf spectra campaign. See related datasets for leaf spectral reflectance and transmittance, leaf mass area (LMA), and leaf chemistry. Note that leaf sample details are also included in related datasets.

54 ENVIRONMENTAL SCIENCES↗

Foliar isotopic and elemental biochemistry and leaf mass per area (LMA), Parque Natural Metropolitano, Panama, 2022

Foliar N and C isotope composition, bulk elemental N and C composition, and leaf mass per area (LMA) for leaves sampled from the Parque Natural Metropolitano (PNM), Panama, from January to April 2022 are presented. Measurements were made on leaves from 17 different tree, shrub and liana species, from sunlit canopy and understory locations on 8 vertical profiles. Full sample details and leaf area index (LAI) from the vertical profiles at PNM are also included in the data package. The aim of this measurement campaign was two-fold: to improve our understanding of the vertical variation in leaf-level water use efficiency, and to improve models which can predict leaf traits from leaf contact spectral measurements. Biochemistry data and sample metadata are presented in .csv files. The original isotopic data report, which includes details about standards and data accuracy, is provided in .xlsx format. Data and metadata meet the ESS-DIVE reporting format requirements for file level metadata (FLMD) and comma separated values (csv). The protocol details are provided as pdf documents. In addition to foliar biochemistry data reported here these samples were also used for measurement of leaf gas exchange, and leaf optical properties. These data can be linked using the unique sample ID and are provided in separate data packages (NGT0192 and NGT0193).

54 ENVIRONMENTAL SCIENCES↗

Vertical canopy gradients of respiration drive plant carbon budgets and leaf area index

Despite its importance for determining global carbon fluxes, leaf respiration remains poorly constrained in land surface models (LSMs). We tested the sensitivity of the Energy Exascale Earth System Model Land Model – Functionally Assembled Terrestrial Ecosystem Simulator (ELM-FATES) to variation in the canopy gradients of leaf maintenance respiration (R dark ). We ran global and point simulations varying the canopy gradient of R dark to explore the impacts on forest structure, composition, and carbon cycling. In global simulations, steeper canopy gradients of R dark lead to increased understory survival and leaf biomass. Leaf area index (LAI) increased up to 77% in tropical regions compared with the default parameterization, improving alignment with remotely sensed benchmarks. Global vegetation carbon varied from 308 Pg C to 449 Pg C across the ensemble. In tropical forest simulations, steeper gradients of R dark had a large impact on successional dynamics. Results show the importance of canopy gradients in leaf traits and fluxes for determining plant carbon budgets and emergent ecosystem properties such as competitive dynamics, LAI, and vegetation carbon. The high-model sensitivity to canopy gradients in R dark highlights the need for more observations of how leaf traits and fluxes vary along light micro-environments to inform critical dynamics in LSMs.

59 BASIC BIOLOGICAL SCIENCES↗

ORNL FACE Final Harvest, 2009, with data on leaf characteristics and tree mass

During the final year of the Oak Ridge National Laboratory (ORNL) free-air CO2 enrichment (FACE) experiment, 16 trees were felled and processed during July 2009. The trees had been grown for 12 years in ambient or elevated CO2. Two data sets are reported. Leaf characteristics (leaf mass per unit area, leaf thickness, average leaf size, carbon and nitrogen concentrations) are reported for each 1-meter section of the tree crowns in data set "Canopy_subsamples.csv". The measurements were made on approximately 20 subsamples collected from each 1-meter section of the canopy. Wood dry mass, total tree height, basal area, total leaf mass and leaf area are reported in "Tree_harvest.csv", based on field measurements of fresh mass and dry mass to fresh mass ratio of subsamples. The ORNL FACE experiment, including photos of the harvesting activity, is described at: https://richnorby.org/home/face-home/.

54 ENVIRONMENTAL SCIENCES↗

Salinity exposure affects lower-canopy specific leaf area of upland trees in a coastal deciduous forest

Sea level rise and increasing storm surges are likely to affect the canopy physiology, ecology, and structure of coastal forests, even well in advance of tree mortality. Laboratory and greenhouse studies have documented that saltwater exposure can trigger changes in leaf-level physiology and morphology, but few in situ studies have examined how tree-specific leaf area (SLA), the ratio of leaf area to mass and a crucial trait and model parameter, is affected by saline soils. We conducted an observational study of SLA in a mid-Atlantic (USA) coastal deciduous forest, taking advantage of a natural gradient in salinity along a tidal creek. Measured SLA of the 239 trees and seven species sampled ranged from Carya glabra (N = 6 trees, mean SLA = 277.9±36.3cm 2 /g) to Fagus grandifolia (N=60, 321.9±62.9cm 2 g); as expected, trees species and canopy position (sun versus shade) significantly affected SLA. For trees (N=100) directly exposed to the tidal creek, salinity was highly significant after accounting for species (P<0.001), with trees in the lower reaches of the creek having lower SLA. Leaf area index (LAI), computed from SLA and litter traps, ranged from 4.8 to 15.8 and was inversely related to salinity exposure; the spatial variability in leaf litter production contributed much more to LAI uncertainty than did SLA variability. These in situ results are correlative but consistent with the hypothesis, based on previous greenhouse studies, that the stress of chronic salinity exposure changes species’ leaf morphology. Our findings are useful for understanding the growing effects of saltwater intrusion into upland forests, as well as parameterizing and testing ecosystem-scale models simulating forest stressors and disturbances at the terrestrial-aquatic interface.

54 ENVIRONMENTAL SCIENCES↗

Leaf morphogenesis: The multifaceted roles of mechanics

Plants produce a rich diversity of biological forms, and the diversity of leaves is especially notable. Mechanisms of leaf morphogenesis have been studied in the past two decades, with a growing focus on the interactive roles of mechanics in recent years. Growth of plant organs involves feedback by mechanical stress: growth induces stress, and stress affects growth and morphogenesis. Although much attention has been given to potential stress-sensing mechanisms and cellular responses, the mechanical principles guiding morphogenesis have not been well understood. Here we synthesize the overarching roles of mechanics and mechanical stress in multilevel and multiple stages of leaf morphogenesis, encompassing leaf primordium initiation, phyllotaxis and venation patterning, and the establishment of complex mature leaf shapes. Moreover, the roles of mechanics at multiscale levels, from subcellular cytoskeletal molecules to single cells to tissues at the organ scale, are articulated. In conclusion, by highlighting the role of mechanical buckling in the formation of three-dimensional leaf shapes, this review integrates the perspectives of mechanics and biology to provide broader insights into the mechanobiology of leaf morphogenesis.

59 BASIC BIOLOGICAL SCIENCES↗

Spatio-temporal differences in leaf physiology are associated with fire, not drought, in a clonally integrated shrub

Abstract In highly disturbed environments, clonality facilitates plant survival via resprouting after disturbance, resource sharing among interconnected stems and vegetative reproduction. These traits likely contribute to the encroachment of deep-rooted clonal shrubs in tallgrass prairie. Clonal shrubs have access to deep soil water and are typically thought of as relatively insensitive to environmental variability. However, how leaf physiological traits differ among stems within individual clonal shrubs (hereafter ‘intra-clonal’) in response to extreme environmental variation (i.e. drought or fire) is unclear. Accounting for intra-clonal differences among stems in response to disturbance is needed to more accurately parameterize models that predict the effects of shrub encroachment on ecosystem processes. We assessed intra-clonal leaf-level physiology of the most dominant encroaching shrub in Kansas tallgrass prairie, Cornus drummondii, in response to precipitation and fire. We compared leaf gas exchange rates from the periphery to centre within shrub clones during a wet (2015) and extremely dry (2018) year. We also compared leaf physiology between recently burned shrubs (resprouts) with unburned shrubs in 2018. Resprouts had higher gas exchange rates and leaf nitrogen content than unburned shrubs, suggesting increased rates of carbon gain can contribute to recovery after fire. In areas recently burned, resprouts had higher gas exchange rates in the centre of the shrub than the periphery. In unburned areas, leaf physiology remained constant across the growing season within clonal shrubs (2015 and 2018). Results suggest single measurements within a shrub are likely sufficient to parameterize models to understand the effects of shrub encroachment on ecosystem carbon and water cycles, but model parameterization may require additional complexity in the context of fire.

54 ENVIRONMENTAL SCIENCES↗

Canopy position influences the degree of light suppression of leaf respiration in abundant tree genera in the Amazon Forest

Leaf respiration in the dark (R dark ) and light (R day ) is poorly characterized in diverse tropical ecosystems, and little to no information exists on the degree of light suppression in common tree species within the Amazon basin, and their dependences upon plant functional traits and position within the canopy. We quantified R dark and apparent R day using the Kok method and measured key leaf traits in 26 tree individuals of different species distributed in three different canopy positions: canopy, lower canopy, and understory. To explore the relationships between the leaf traits we used the standardized major axis (SMA). We found that canopy trees had significantly higher rates of R dark and R day than trees in the understory. The difference between R dark and R day (the light suppression of respiration) was greatest in the understory (68 ± 9%, 95% CI) and lower canopy (49 ± 9%, 95% CI) when compared to the canopy (37 ± 10%, 95% CI). We also found that R day was significantly and strongly correlated with R dark (p < 0.001) for all the canopy positions. Also, leaf mass per area (LMA) and leaf Phosphorus concentration (P) had a significant relationship with R dark (p < 0.001; p = 0.003), respectively. In addition, a significant relationship was found for LMA in the canopy and lower canopy positions (p = 0.009; p = 0.048) while P was only significant in the canopy (p = 0.044). Finally, no significant relationship was found between R dark and nitrogen, sugars, and starch. Our results highlight the importance of including representation of the light suppression of leaf respiration in terrestrial biosphere models and also of accounting for vertical gradients within forest canopies and connections with functional traits.

54 ENVIRONMENTAL SCIENCES↗

Heterogeneous isotope effects decouple conifer leaf and branch sugar δ 18 O and δ 13 C

Isotope ratios of tree-ring cellulose are a prominent tool to reconstruct paleoclimate and plant responses to environmental variation. Current models for cellulose isotope ratios assume a transfer of the environmental signals recorded in bulk leaf water to carbohydrates and ultimately into stem cellulose. However, the isotopic signal of carbohydrates exported from leaf to branch may deviate from mean leaf values if spatial heterogeneity in isotope ratios exists in the leaf. We tested whether the isotopic heterogeneity previously observed along the length of a ponderosa pine (Pinus ponderosa) leaf water was preserved in photosynthetic products. We observed an increase in both sugar and bulk tissue δ 18 O values along the needle, but the increase in carbohydrate δ 18 O values was dampened relative to the trend observed in leaf water. In contrast, δ 13 C values of both sugar and bulk organic matter were invariant along the needle. Additionally, phloem-exported sugar measured in the branch below the needles did not match whole-needle values of δ 18 O or δ 13 C. Instead, there was a near-constant offset observed between the branch and needle sugar δ 13 C values, while branch δ 18 O values were most similar to δ 18 O values observed for sugar at the base of the needle. The observed offset between the branch and needle sugar δ 18 O values likely arises from partial isotope oxygen exchange between sugars and water during phloem loading and transport. An improved understanding of the conditions producing differential δ 13 C and δ 18 O isotope effects between branch phloem and needle sugars could improve tree-ring-based climate reconstructions.

59 BASIC BIOLOGICAL SCIENCES↗

Genetic architecture of leaf morphological and physiological traits in a Populus deltoides ‘Danhong’ × P. simonii ‘Tongliao1’ pedigree revealed by quantitative trait locus analysis

Understanding the genetic architecture of leaf morphological and physiological traits will help plant breeders develop high biomass poplar genotypes. Quantitative trait locus (QTL) studies combining next-generation sequencing techniques can advance our understanding of the genetic basis of complex traits. In this study, we measured 13 leaf morphological and physiological traits and identified quantitative trait loci (QTLs) in a Populus deltoides ‘Danhong’ × P. simonii ‘Tongliao1’ F1 population (500 progenies) using a high-density genetic map constructed by whole genome re-sequencing. This linkage map consisted of 5796 single nucleotide polymorphism (SNP) markers assigned to 19 linkage groups (LGs), spanning 2683.80 centimorgans (cM) of genetic length, with an average marker density of 0.46 cM. We identified 109 QTLs on 18 LGs for leaf morphological traits and 55 QTLs on 14 LGs for leaf physiological traits. One-hundred eight putative candidate genes were identified within the candidate genomic region. Co-expression network and gene ontology enrichment analyses suggested that these candidate genes were involved in the photosynthetic process. The differential expression patterns of the CYCLIN (Potri.015G112200) and RED CHLOROPHYLL REDUCTASE (Potri.007G043600) genes between two parents indicated their potential roles in leaf morphological and physiological traits. These findings decipher the genetic architecture of leaf morphological and physiological traits in the P. deltoides ‘Danhong’ × P. simonii ‘Tongliao1’ pedigree and provide candidate genes for future poplar genetic improvement.

59 BASIC BIOLOGICAL SCIENCES↗

Migrating Populus with climate change: Phenology, coppice management, cold spell susceptibility, leaf dynamics, and biomass production

Understanding the phenology and productivity of Populus species is crucial for effective management and conservation strategies amid climate change. We investigated leaf budbreak timing, susceptibility to cold damage, leaf dynamics, and biomass production of 168 Populus genotypes with diverse provenances in the southeastern United States. Our study revealed significant variation in budbreak timing across different taxa and years, with genotypes inheriting traits adapted to their parents’ local climates. Temperature emerged as a key factor triggering budbreak, while leaf development depended on other environmental cues such as photoperiod. Notably, budbreak occurred approximately 20 days earlier in 2023 compared to 2022 due to higher accumulated degree days (ADDs). Short-rotation-coppice (SRC) management delayed budbreak by five to ten days. Cold damage was significant in 2023, particularly for genotypes from northern provenances and those with P. maximowiczii parentage. Severe damage was also observed in eastern cottonwood ( Populus deltoides ​× ​ Populus deltoides (D ​× ​D)) genotypes, despite most having southeastern US parentages. Leaf dynamics, including leaf duration and leaf area index (LAI), varied across taxa and sites, with earlier budbreak correlating with extended growing seasons and increased LAI. Biomass production was intricately linked to phenological events, with earlier budbreak leading to increased biomass production and greater susceptibility to cold damage. Our findings highlight the importance of genetics, environment, and coppicing management in understanding and managing Populus phenology and biomass production. These insights provide valuable guidance for developing effective breeding, conservation, and management strategies for Populus species in the context of climate change.

Accumulated degree days (ADDs)↗

High Temperature Acclimation of Leaf Gas Exchange, Photochemistry, and Metabolomic Profiles in Populus trichocarpa

High temperatures alter the thermal sensitivities of numerous physiological and biochemical processes that impact tree growth and productivity. Foliar and root applications of methanol have been implicated in plant acclimation to high temperature via the C 1 pathway. Here, we characterized temperature acclimation at 35°C of leaf gas exchange, chlorophyll fluorescence and extractable metabolites of potted Populus trichocarpa saplings and examined potential influences of mM concentrations of methanol added during soil watering over a two-month period. Relative to plants grown under the low growth temperature (LGT), high growth temperature (HGT) plants showed a suppression of leaf water use and carbon cycling including transpiration (E), net photosynthesis (P n ), an estimate of photorespiration (R p ), and dark respiration (R d ), attributed to reductions in stomatal conductance and direct negative effects on gas exchange and photosynthetic machinery. In contrast, HGT plants showed an upregulation of non-photochemical quenching (NPQ t ), the optimum temperature for ETR, and leaf isoprene emissions at 40°C. A large number of metabolites (867) were induced under HGT, many implicated in flavonoid biosynthesis highlighting a potentially protective role for these compounds. Methanol application did not significantly alter leaf gas exchange but slightly reduced the suppression of R d and R p by the high growth temperature while slightly impairing ETR, Fv’/Fm’, and q p . However, we were unable to determine if soil methanol was sufficiently taken up by the plant to have a direct effect on foliar processes. A small number of extracted leaf tissue metabolites (55 out of 10,015) showed significantly altered abundances under LGT and methanol treatments relative to water controls, and this increased in compound number (222) at the HGT. The results demonstrate the large physiological and biochemical impacts of high growth temperature on poplar seedlings and highlights the enhancement of the optimum temperature of ETR as a rapid thermal acclimation mechanism. Finally, although no large effect on leaf physiology was observed, the results are consistent with methanol both impairing photochemistry of the light reactions via formaldehyde toxicity, and stimulating photosynthesis and dark respiration through formate oxidation to CO 2 .

59 BASIC BIOLOGICAL SCIENCES↗

Leaf Trait Plasticity Alters Competitive Ability and Functioning of Simulated Tropical Trees in Response to Elevated Carbon Dioxide

The response of tropical ecosystems to elevated carbon dioxide (CO 2 ) remains a critical uncertainty in projections of future climate. Here, we investigate how leaf trait plasticity in response to elevated CO 2 alters projections of tropical forest competitive dynamics and functioning. We use vegetation demographic model simulations to quantify how plasticity in leaf mass per area and leaf carbon to nitrogen ratio alter the responses of carbon uptake, evapotranspiration, and competitive ability to a doubling of CO 2 in a tropical forest. Observationally constrained leaf trait plasticity in response to CO 2 fertilization reduces the degree to which tropical tree carbon uptake is affected by a doubling of CO 2 (up to -14.7% as compared to a case with no plasticity; 95% confidence interval [CI95%] -14.4 to -15.0). It also diminishes evapotranspiration (up to -7.0%, CI95% -6.4 to -7.7), and lowers competitive ability in comparison to a tree with no plasticity. Consideration of leaf trait plasticity to elevated CO 2 lowers tropical ecosystem carbon uptake and evapotranspirative cooling in the absence of changes in plant-type abundance. However, “plastic” responses to high CO 2 which maintain higher levels of plant productivity, many of which fall outside of the observed range of response, are potentially more competitively advantageous, thus, including changes in plant type abundance may mitigate these decreases in ecosystem functioning. Models that explicitly represent competition between plants with alternative leaf trait plasticity in response to elevated CO 2 are needed to capture these influences on tropical forest functioning and large-scale climate.

54 ENVIRONMENTAL SCIENCES↗

Contrasting age-dependent leaf acclimation strategies drive vegetation greening across deciduous broadleaf forests in mid- to high latitudes

Increasing leaf area and extending vegetation growing seasons are two primary drivers of global greening, which has emerged as one of the most significant responses to climate change. However, it remains unclear how these two leaf acclimation strategies would vary across forests at a large spatial scale. Here, in this study, using multiple satellite-based datasets and field measurements, we analysed the temporal changes (Δ) in maximal leaf area index (LAI max ) and length of the growing season (LOS) from 2002 to 2021 across deciduous broadleaf forests (DBFs) in the middle to high latitudes. Contrary to the widely held assumption of coordination, our results revealed a negative correlation between ΔLAI max and ΔLOS. Notably, the trade-offs between ΔLAImax and ΔLOS were strongly explained by stand age. Younger DBFs, with lower baseline LAI max , predominantly located in eastern Asia, displayed an increase in LAI max with small changes in LOS. This acquisitive strategy facilitated younger DBFs to grow more photosynthetically efficient leaves with low leaf mass per area, enhancing their light use efficiency. Conversely, older DBFs with a higher baseline LAI max , primarily located in North America and Europe, extended their LOS by increasing leaf mass per area. This conservative strategy facilitated older DBFs to produce thicker, but less photosynthetically efficient leaves, resulting in decreased light use efficiency. Our findings offer new insights into the contrasting changes in leaf area and growing season length and highlight their divergent impacts on ecosystem functioning.

Wang, Fangyi [Sun Yat-Sen Univ., Zhuhai (China)]↗

Viral‐mediated delivery of morphogenic regulators enables leaf transformation in Sorghum bicolor (L.)

Recent advancements in monocot transformation, using leaf tissue as explant material, have expanded the number of grass species capable of transgenesis. However, the complexity of vectors and reliance on inducible excision of essential morphogenic regulators have so far limited widespread application. Plant RNA viruses, such as Foxtail Mosaic Virus (FoMV), present a unique opportunity to express morphogenic regulator genes, such as Babyboom (Bbm), Wuschel2 (Wus2), Wuschel-like homeobox protein 2a (Wox2a) and the GROWTH-REGULATING FACTOR 4 (GRF4) GRF-INTERACTING FACTOR 1 (GIF1) fusion protein transiently in leaf explant tissues. Furthermore, altruistic delivery of conventional and viral vectors could provide opportunities to simplify vectors used for leaf transformation—facilitating vector optimization and reducing reliance on morphogenic regulator gene integration. In this study, both viral and conventional T-DNA vectors were tested for their ability to promote the formation of embryonic calli, a critical step in leaf transformation protocols, using Sorghum bicolor leaf explants. Although conventional leaf transformation vectors yielded viable embryonic calli (43.2 ± 2.9%: GRF4-GIF1, 50.2 ± 3%: Bbm/Wus2), altruistic conventional vectors employing the GRF4-GIF1 morphogenic regulator resulted in improved efficiencies (61.3 ± 4.7%). Altruistic delivery was further enhanced with the use of viral vectors employing both GRF4-GIF1 and Bbm/Wus2 regulators, resulting in 75.1 ± 2.3% and 79.2 ± 2.5% embryonic calli formation, respectively. Embryonic calli generated from both conventional and viral vectors produced shoots expressing fluorescent reporters, which were confirmed using molecular analysis. This work provides an important proof-of-concept for the use of both altruistic vectors and viral-expressed morphogenic regulators for improving plant transformation.

59 BASIC BIOLOGICAL SCIENCES↗

Few-shot leaf segmentation

In this work, we use few-shot learning to segment the body and vein architecture of Populus trichocarpa leaves from high-resolution scans obtained in the UC Davis common garden. Leaf and vein segmentation are formulated as separate tasks, in which convolutional neural networks (CNNs) are used to iteratively expand partial segmentations until reaching stopping criteria. Our leaf and vein segmentation approaches use just 50 and 8 manually traced images for training, respectively, and are applied to a set of 2,634 top and bottom leaf scans. We show that both methods achieve high segmentation accuracy, in some cases exceeding even human-level segmentation. The leaf and vein segmentations are subsequently used to extract 68 morphological traits using traditional open-source image processing tools, which are validated using real-world physical measurements. For a biological perspective, we perform a genome-wide association study using the "vein density" trait to discover novel genetic architectures associated with multiple physiological processes relating to leaf development and function. In addition to sharing all of the few-shot learning code, we are releasing all images, manual segmentations, model predictions, 68 extracted leaf phenotypes, and a new set of SNPs called against the v4 P. trichocarpa genome for 1,419 genotypes.

Lagergren, John [Oak Ridge National Lab. (ORNL), O↗

Data for "Viral-mediated Delivery of Morphogenic Regulators Enables Leaf Transformation in Sorghum bicolor (L.)"

Recent advancements in monocot transformation, using leaf tissue as explant material, have expanded the number of grass species capable of transgenesis. However, the complexity of vectors and reliance on inducible excision of essential morphogenic regulators have so far limited widespread application. Plant RNA viruses, such as Foxtail Mosaic Virus (FoMV), present a unique opportunity to express morphogenic regulator genes, such as Babyboom ( Bbm ), Wuschel2 ( Wus2 ), Wuschel-like homeobox protein 2a ( Wox2a ) and the GROWTH-REGULATING FACTOR 4 (GRF4) GRF-INTERACTING FACTOR 1 (GIF1) fusion protein transiently in leaf explant tissues. Furthermore, altruistic delivery of conventional and viral vectors could provide opportunities to simplify vectors used for leaf transformation—facilitating vector optimization and reducing reliance on morphogenic regulator gene integration. In this study, both viral and conventional T-DNA vectors were tested for their ability to promote the formation of embryonic calli, a critical step in leaf transformation protocols, using Sorghum bicolor leaf explants. Although conventional leaf transformation vectors yielded viable embryonic calli (43.2 ± 2.9%: GRF4-GIF1, 50.2 ± 3%: Bbm/Wus2), altruistic conventional vectors employing the GRF4-GIF1 morphogenic regulator resulted in improved efficiencies (61.3 ± 4.7%). Altruistic delivery was further enhanced with the use of viral vectors employing both GRF4-GIF1 and Bbm/Wus2 regulators, resulting in 75.1 ± 2.3% and 79.2 ± 2.5% embryonic calli formation, respectively. Embryonic calli generated from both conventional and viral vectors produced shoots expressing fluorescent reporters, which were confirmed using molecular analysis. This work provides an important proof-of-concept for the use of both altruistic vectors and viral-expressed morphogenic regulators for improving plant transformation.

gene editing↗