The 2025 In Vitro Biology Meeting hosted an outstanding oral presentation competition for Plant Biotechnology Postdoctoral Associates, celebrating both groundbreaking research and exceptional presentation skills. This inspiring event not only brought forward novel scientific discoveries but also provided a valuable platform for early-career scientists to strengthen their communication and professional development. Presenters were evaluated on multiple criteria, including experimental design, data analysis, results interpretation, study originality, technical complexity, and presentation quality, by an expert panel of judges representing both from academia and industry. The judges included Dr. Christopher Bagley (Inari Agriculture, Inc.), Dr. Keunsub Lee (Iowa State University), Dr. Daniel Rodriguez Leal (University of Maryland), and Dr. Qingzhen Jiang (Boyce Thompson Institute). From a highly competitive pool of submissions, four outstanding postdoctoral researchers were selected to present at the annual meeting. After careful deliberation, first place was awarded to Greg S. Goralogia (Oregon State University, USA) for his work on a hairy root transgene excision system for woody plants. Second place went to Arjun Ojha Kshetry (Texas Tech University, USA) for developing an in planta transformation toolkit using developmental regulators, while Chi Nguyen (University of Florida, USA) earned third place for her research on lettuce regulatory networks. All winners received certificates and cash awards in recognition of their exceptional contributions. Here, we strongly encourage all Plant Biotechnology Postdoctoral Associates to consider participating in future meetings. This competition offers an excellent opportunity to showcase your research, receive expert feedback, and build vital presentation skills in a supportive, professional environment. We look forward to seeing your groundbreaking research at next year’s event!
Submitted by Bin Tian
First Place
RESET: A Hairy Root-to-Shoot Transgene Excision System for Improved Transformation and Clean Editing in Clonally Propagated Plants
Greg S. Goralogia
Many plant species remain recalcitrant to transformation, impeding the use of continually evolving gene editing technologies outside of established in vitro systems. This is especially true for clonally propagated perennial plants which include many forest trees and specialty crops. An additional challenge in these species is the removal of editing machinery inserted as stable transgenes into the plant genome, where conventional removal by crossing and segregation is undesirable. Hairy root culture using Agrobacterium rhizogenes is an accessible technique in many of these species, and many of the genes that encode this response on Ri plasmids are well characterized. Using the large-cargo enabling GAANTRY transgene assembly system, we designed and tested a synthetic gene circuit which leverages hairy root rol genes for hairy root culture, inducible shoot-inducing genes including WUSCHEL and ipt, and a Cre-lox excision system which acts on recognition sites near the T-DNA borders. Using this system, hairy roots can be cultured using a variety of approaches, then are segmented and exposed to a heat-shock treatment that induces shoot formation from roots. The heat treatment also triggers Cre-lox excision of the large majority of the transgenic T-DNA (leaving a small footprint). Afterwards, shoots containing edits but that lack most of the transgene can be easily identified using reporter genes. When testing the system in poplar, we achieved high rates of hairy root recovery in two genotypes (average of 42% of explants). Using a two-week heat shock pulse treatment (39 C for 4 hours per day), two-thirds of explants regenerated at least one shoot in culture, with one-third having more than 10 shoots per explant. One-third of these shoots had the transgene excised, and nearly all of the excised shoots appear to be edited in at least one allele of two CRISPR/Cas9-targeted genes. Given the broad host range for which hairy root induction is feasible, we believe this system will be an efficient method to generate CRISPR-Cas-free, edited clonally propagated plants in a variety of species and genotypes.
Greg S. Goralogia, Department of Forest Ecosystems and Society, Oregon State University, Corvallis, OR. Abstract Presentation: P-1000
Second Place
Shoot at Site: Advancing in Planta Transformation, Regeneration and Gene-editing Through a Cascade of Wounding-mediated Developmental Regulators
Arjun Ojha Kshetry
Developing transgenic and/or gene-edited plants largely depends on tedious, lengthy, and costly in vitro regeneration protocols. While plants have remarkable regeneration ability, not all species, genotypes or even explants exhibit the same transformation and regeneration potential under in vitro conditions. To tackle this bottleneck, we have developed a seamless and user-friendly system to induce transgenic and gene-edited de novo meristems via a synthetic cascade comprising a wound-induced regeneration pathway, plant developmental regulators (DRs) and gene-editing reagents. WOUND INDUCED DEDIFFERENTIATION 1 (WIND1) is used as a transcriptional regulator to control the expression of various DR genes through ENHANCER OF SHOOT REGENERATION 1 (ESR1) promoter. This cascade was strategically applied in planta to the non-meristematic internode of N. benthamiana to induce meristematic activity and regenerate de novo shoots with knock-out mutations of the phytoene desaturase (PDS) gene. This synthetic toolkit was further applied successfully to tomato and soybean. This methodology offers a transformative approach to overcome barriers in plant biotechnology, potentially accelerating the generation of transgenic and gene-edited plants without reliance on conventional tissue-culture intermediates.
Arjun Ojha Kshetry, Institute of Genomics for Crop Abiotic Stress Tolerance (IGCAST), Department of Plant and Soil Science, Texas Tech University, Lubbock, TX. Abstract Presentation: P-1003
Third Place
Unraveling Molecular Mechanisms of Seed Germination and Flowering Time in Lettuce Through Genetic and Transcriptomic Approaches
Chi Nguyen
Lettuce (Lactuca sativa) production is highly susceptible to environmental stresses, particularly high temperatures which induce seed thermoinhibition and premature bolting. To elucidate the molecular mechanisms governing seed dormancy and flowering time, we employed CRISPR/Cas9 gene editing, RNA interference (RNAi), and overexpression approaches to manipulate key regulators, including ABA-Hypersensitive Germination 1 (AHG1), ABA-Hypersensitive Germination 3 (AHG3), DELAY OF GERMINATION 1 (DOG1), miR156, and miR172. Phenotypic analyses revealed that lsahg1 and lsahg3 mutants exhibited significantly delayed germination, increased ABA sensitivity, and impaired developmental transitions, consistent with their role as negative regulators in the ABA signaling pathway. DOG1 silencing led to early flowering, while miR156 overexpression and miR172 silencing strongly delayed flowering. RNA sequencing of lsahg3 mutants compared to wild-type lettuce identified differentially expressed genes involved in ABA signaling and seed dormancy. Principal component analysis (PCA) revealed distinct clustering of lsahg3 and wild-type samples, confirming widespread transcriptional reprogramming. Furthermore, lsahg3 mutants exhibited differential expressions of SnRK2 and PYL genes, indicating a disruption in ABA perception and signal transduction. These findings provide novel insights into the genetic and molecular pathways controlling seed dormancy and flowering time in lettuce. Understanding these regulatory networks is crucial for developing climate-resilient lettuce cultivars with improved germination and delayed bolting under high-temperatures.
Chi Nguyen, Crop Transformation Center, Horticultural Sciences Department, University of Florida, Gainesville, FL and Mid-Florida Research & Education Center, University of Florida, Apopka, FL. Abstract Presentation: P-1002
