Search

Despite the overarching history of species divergence, phylogenetic studies often reveal distinct topologies across regions of the genome. The sources of these gene tree discordances are variable, but incomplete lineage sorting (ILS) and hybridization are among those with the most biological importance. Petunia serves as a classic system for studying hybridization in the wild. While field studies suggest that hybridization is frequent, the extent of reticulation within Petunia and its closely related genera has never been examined from a phylogenetic perspective. In this study, we used transcriptomic data from 11 Petunia, 16 Calibrachoa, and 10 Fabiana species to illuminate the relationships between these species and investigate whether hybridization played a significant role in the diversification of the clade. We inferred that gene tree discordance within genera is linked to hybridization events along with high levels of ILS due to their rapid diversification. Moreover, network analyses estimated deeper hybridization events between Petunia and Calibrachoa, genera that have different chromosome numbers. Although these genera cannot hybridize at the present time, ancestral hybridization could have played a role in their parallel radiations, as they share the same habitat and life history., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

In a production environment, delayed stress recognition can impact yield. Imaging can rapidly and effectively quantify stress symptoms using indexes such as normalized difference vegetation index (NDVI). Commercial systems are effective but cannot be easily customized for specific applications, particularly post-processing. We developed a low-cost customizable imaging system and validated the code to analyze images. Our objective was to verify the image analysis code and custom system could successfully quantify the changes in plant canopy reflectance. 'Supercascade Red', 'Wave© Purple', and 'Carpet Blue' Petunias ( Petunia × hybridia ) were transplanted individually and subjected to increasing fertilizer treatments and increasing substrate pH in a greenhouse. Treatments for the first trial were the addition of a controlled release fertilizer at six different rates (0, 0.5, 1, 2, 4, and 8 g/pot), and for the second trial, fertilizer solution with four pHs (4, 5.5, 7, and 8.5), with eight replications with one plant each. Plants were imaged twice a week using a commercial imaging system for fertilizer and thrice a week with the custom system for pH. The collected images were analyzed using an in-house program that calculated the indices for each pixel of the plant area. All cultivars showed a significant effect of fertilizer on the projected canopy size and dry weight of the above-substrate biomass and the fertilizer rate treatments ( p < 0.01). Plant tissue nitrogen concentration as a function of the applied fertilizer rate showed a significant positive response for all three cultivars ( p < 0.001). We verified that the image analysis code successfully quantified the changes in plant canopy reflectance as induced by increasing fertilizer application rate. There was no relationship between the pH and NDVI values for the cultivars tested ( p > 0.05). Manganese and phosphorus had no significance with chlorophyll fluorescence for 'Carpet Blue' and 'Wave© Purple' ( p > 0.05), though 'Supercascade Red' was found to have significance ( p < 0.01). pH did not affect plant canopy size. Chlorophyll fluorescence pixel intensity against the projected canopy size had no significance except in 'Wave© Purple' ( p = 0.005). NDVI as a function of the projected canopy size had no statistical significance. We verified the ability of the imaging system with integrated analysis to quantify nutrient deficiency-induced variability in plant canopies by increasing pH levels.

Petunia hybrida, widely grown as a bedding plant, has reduced growth and flower quality at temperatures above 30 °C (heat stress), primarily due to heat stress-induced ethylene (ET) production. The gene acdS encodes the 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD) enzyme, which is known for its role in reducing ET production by breaking down the ET precursor, ACC, in plant tissues. This study investigated the impact of heat stress on both 'Mirage Rose' WT petunia and its acdS-overexpressing transgenic lines. Heat stress-induced growth inhibition was observed in WT plants but not in transgenic plants. The increased stress tolerance of transgenic plants over WT plants was associated with lower ET production, ROS accumulation, higher SPAD values, water content, and relative water content. Furthermore, higher sensitivity of the WT to heat stress than the transgenic plants was confirmed by analysing ET signalling genes, heat shock transcription factor genes, and antioxidant- and proline-related genes, more strongly induced in WT than in transgenic plants. Overall, this study suggests the potential application of acdS overexpression in other floriculture plants as a viable strategy for developing heat stress-tolerant varieties. This approach holds promise for advancing the floricultural industry by overcoming challenges related to heat-induced growth inhibition and loss of flower quality., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Petunias are renowned ornamental species widely cultivated as pot plants for their aesthetic appeal both indoors and outdoors. The preference for pot plants depends on their compact growth habit and abundant flowering. While genome editing has gained significant popularity in many crop plants in addressing growth and development and abiotic and biotic stress factors, relatively less emphasis has been placed on its application in ornamental plant species. Genome editing in ornamental plants opens up possibilities for enhancing their aesthetic qualities, offering innovative opportunities for manipulating plant architecture and visual appeal through precise genetic modifications. In this study, we aimed to optimize the procedure for an efficient genome editing system in petunia plants using the highly efficient multiplexed CRISPR/Cas9 system. Specifically, we targeted a total of six genes in Petunia which are associated with plant architecture traits, two paralogous of FLOWERING LOCUS T (PhFT) and four TERMINAL FLOWER-LIKE1 (PhTFL1) paralogous genes separately in two constructs. We successfully induced homogeneous and heterogeneous indels in the targeted genes through precise genome editing, resulting in significant phenotypic alterations in petunia. Notably, the plants harboring edited PhTFL1 and PhFT exhibited a conspicuously early flowering time in comparison to the wild-type counterparts. Furthermore, mutants with alterations in the PhTFL1 demonstrated shorter internodes than wild-type, likely by downregulating the gibberellic acid pathway genes PhGAI, creating a more compact and aesthetically appealing phenotype. This study represents the first successful endeavor to produce compact petunia plants with increased flower abundance through genome editing. Our approach holds immense promise to improve economically important potting plants like petunia and serve as a potential foundation for further improvements in similar ornamental plant species., (© 2024. The Author(s).)

Vein-associated pigmentation (venation) is a type of floral coloration adopted by plants to attract pollinators. Several petunia (Petunia hybrida) lines generate dorsoventrally asymmetric venation patterning of the corolla tube, in which venation is only present in the dorsal tube. The molecular mechanism underlying this trait is unknown. Here, we demonstrate that miR319 is preferentially expressed in the dorsal corolla tube, leading to dorsoventrally asymmetric expression of its target genes. Transgenic lines overexpressing phy-miR319a generated uniform venation patterning of the corolla tube. Knockout of TCP genes targeted by miR319 promoted venation patterning in the ventral and dorsal tube, while overexpression of the miR319 target gene, PhTCP6, completely inhibited corolla tube venation patterning. In addition, miR319-targeted TCPs negatively regulated venation patterning, probably by repressing the regulator of venation patterning, AN4. Together, our data demonstrate that asymmetric expression of miR319 promotes venation patterning in the petunia dorsal tube alone by repressing the expression of its target TCP genes, which negatively regulate corolla tube venation patterning. These findings provide novel insights into how the dorsoventrally asymmetric distribution of venation patterning is established in zygomorphic flowers., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Physiological role of foliar application of thiamine (B1) on active chemical compounds percentages of Petunia flowers (petunia hybrida L.): From October 23, 2022, to June 4, 2023, the study was done in the Lath house of the Department of Horticulture and Landscape design at the University of Kirkuk/Agricultural Research and Experimentation Station/Al-Sayadah. The aim was to find out the Physiological role of the growth regulator thiamine was sprayed at( 0, 150, and 300 )mg. L-1 In terms of the percentages of effective chemical compounds in petunia flowers, the factorial experiment was designed according to R.C.B.D. Using organic solvents, the soxolite method was used to extract the effective chemical compounds from the flowers. A rotary evaporator was used under vacuum pressure to evaporate the solvent and obtain the essential oils. Then the gas chromatography-mass spectrometer device is used to detect the basic chemical components and estimate their percentage in plant flowers. The results for extracting and finding chemical compounds showed that the amounts of common substances in plant flower samples changed depending on the treatments. For example, 300 mg of thiamine was sprayed on the flowers. L-1 gave the best results in terms of the percentages of active chemicals in the plant flowers compared to the control treatment flowers.The study further suggests that thiamine treatments significantly impact the biosynthetic pathways of essential oils and active compounds, positively altering their abundance in petunia flowers. Consequently, the exploration of growth regulator manipulation could potentially lead to the development of novel horticultural strategies, aiding in the sustainable production of high-quality floral products.

Self-incompatibility (SI) is an intraspecific reproductive barrier widely present in angiosperms. The SI system with the broadest occurrence in angiosperms is based on an S-RNase linked to a cluster of multiple S-locus F-box (SLF) genes found in the Solanaceae, Plantaginaceae, Rosaceae, and Rutaceae. Recent studies reveal that non-self S-RNase is degraded by the Skip Cullin F-box (SCF) SLF -mediated ubiquitin-proteasome system in a collaborative manner in Petunia, but how self-RNase functions largely remains mysterious. Here, we show that S-RNases form S-RNase condensates (SRCs) in the self-pollen tube cytoplasm through phase separation and the disruption of SRC formation breaks SI in self-incompatible Petunia hybrida. We further find that the pistil SI factors of a small asparagine-rich protein HT-B and thioredoxin h together with a reduced state of the pollen tube all promote the expansion of SRCs, which then sequester several actin-binding proteins, including the actin polymerization factor PhABRACL, the actin polymerization activity of which is reduced by S-RNase in vitro. Meanwhile, we find that S-RNase variants lacking condensation ability fail to recruit PhABRACL and are unable to induce actin foci formation required for pollen tube growth inhibition. Taken together, our results demonstrate that phase separation of S-RNase promotes SI response in P. hybrida, revealing a new mode of S-RNase action., (© 2023 The Authors. Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

This experiment was carried out in the Plant Cells and Tissue Culture Laboratory of Horticulture and Landscaping Design Department / College of Agriculture / University of Kirkuk - IRAQ, to study the effect of auxin’s in different concentrations and MS media (full and ½) salt strength on rooting response of Petunia hybrida. The regenerated shoots explants from the multiplication stage were transferred to the rooting stage cultured on MS media ( full and ½ ) salts strength supplemented with different concentrations of IBA at ( 0.0, 0.25, 0.50, 0.75, and 1.0 ) mg.L-1, and NAA at ( 0.0, 0.25, 0.50, 0.75, and 1.0 ) mg.L-1. After 6 weeks of transforming, data showed that, the response percentage was ( 100% ) for MS media ( full and ½ ) salt strength supplemented with different concentrations of IBA and NAA beside control treatment. The best number of roots ( 29.70 root.part-1 ) and the length of the longest root ( 7.44 cm ) were recorded by MS ½ strength supplemented with IBA (1.0 mg.L-1) compared with other concentrations, and it was the highest results compared with MS full strength supplemented with different concentration of IBA. Moreover, results obtained that MS ½ strength supplemented with NAA ( 1.0 mg.L-1 ) recorded the best number of roots ( 18.70 root.part-1 ) and the length of the longest root ( 2.97 cm ) compared with the other concentrations, while it was the best result compared with MS full strength supplemented with different concentrations of NAA.

This study delves into the role of gibberellin (GA) in governing plant branch development, a process that remains incompletely understood. Through a combination of exogenous hormone treatment, gene expression analysis, and transgenic phenotype investigations, the impact of GA on petunia’s branch development was explored. The results showed that GA3 alone did not directly induce axillary bud germination. However, paclobutrazol (PAC), an inhibitor of GA synthesis, effectively inhibited bud growth. Interestingly, the simultaneous application of GA3 and 6-BA significantly promoted bud growth in both intact and decapitated plants compared to using 6-BA alone. Moreover, this study observed a significant downregulation of GA synthesis genes, including GA20ox1, GA20ox2, GA20ox3, GA3ox1, and CPS1, alongside an upregulation of GA degradation genes such as GA2ox2, GA2ox4, and GA2ox8. The expression of GA signal transduction gene GID1 and GA response factor RGA was found to be upregulated. Notably, the PhGID1 gene, spanning 1029 bp and encoding 342 amino acids, exhibited higher expression in buds and the lowest expression in leaves. The overexpression of PhGID1 in Arabidopsis resulted in a noteworthy rise in the number of branches. This study highlights the crucial role of GA in bud germination and growth and the positive regulatory function of GA signaling in shoot branching processes.

Floral scent emission of petunia flowers is regulated by light conditions, circadian rhythms, ambient temperature and the phytohormones GA and ethylene, but the mechanisms underlying sensitivity to these factors remain obscure. PHYTOCHROME INTERACTING FACTORs (PIFs) have been well studied as components of the regulatory machinery for numerous physiological processes. Acting redundantly, they serve as transmitters of light, circadian, metabolic, thermal and hormonal signals. Here we identified and characterized the phylogenetics of petunia PIF family members (PhPIFs). PhPIF4/5 was revealed as a positive regulator of floral scent: TRV-based transient suppression of PhPIF4/5 in petunia petals reduced emission of volatiles, whereas transient overexpression increased scent emission. The mechanism of PhPIF4/5-mediated regulation of volatile production includes activation of the expression of genes encoding biosynthetic enzymes and a key positive regulator of the pathway, EMISSION OF BENZENOIDS II (EOBII). The PIF-binding motif on the EOBII promoter (G-box) was shown to be needed for this activation. As PhPIF4/5 homologues are sensors of dawn and expression of EOBII also peaks at dawn, the prior is proposed to be part of the diurnal control of the volatile biosynthetic machinery. PhPIF4/5 was also found to transcriptionally activate PhDELLAs; a similar positive effect of PIFs on DELLA expression was further confirmed in Arabidopsis seedlings. The PhPIF4/5-PhDELLAs feedback is proposed to fine-tune GA signaling for regulation of floral scent production., (© 2024. The Author(s).)

Petunia × Calibrachoa 'Light Yellow' (× Petchoa 'Light Yellow') is a kind of perennial herbaceous flower obtained through intergeneric hybridization of Petunia and Calibrachoa with high ornamental value and wide application, facing challenges in seed acquisition. Expanding propagation through tissue culture is an economically efficient means. Hence, establishing an effective procedure for the storage of callus is essential for × Petchoa 'Light Yellow'. Cryopreservation is an effective method for the in vitro propagation and long-term preservation of × Petchoa 'Light Yellow' germplasms. For formulating the optimization of the vitrification procedure, first, an orthogonal experimental design was employed to pinpoint critical steps in the vitrification protocol (pre-culture, osmoprotection, dehydration, and dilution) for Petunia × Calibrachoa callus tissues and then five additional factors (pre-culture, osmoprotection I and II, dehydration, and dilution) were optimized to further reduce the sample water content and enhance cell viability levels. The vitrification procedure was described as follows: callus tissues were precultured in MS solid medium with 0.3 M sucrose for 5 d, incubated with osmoprotection solution I and II for 15 min at 25 °C, respectively, cryoprotected with PVS2 for 30 min at 0 °C, and rapidly immersed in liquid nitrogen. Cryopreserved callus tissues were then diluted in MS liquid medium with 1.2 M sucrose for 20 min at 25 °C and recovered on MS solid medium with 0.5 mg/L 6-BA and 0.1 mg/L NAA, and sucrose. The cell viability measured by TTC staining was approximately 16 %-18 % after 72 h-recovery. Following 45 days, the relative survival of callus reached up to 49.48 %. Furthermore, EST-SSR analysis showed no significant difference in the genetic stability of cryopreserved callus compared to the control. Based on the cryopreservation of × Petchoa 'Light Yellow' callus, we further evaluated the response of callus water contents to the osmotic stress in the optimized and original protocols (CK) for a higher cryopreservation survival. A comparative analysis of water content demonstrated that the procedure of gradual and gentle dehydration significantly improved water content and cell survival. Ultrastructural changes between cryopreserved and non-cryopreserved callus were examined and high vacuolation emerged as a key determinant, indicating its substantial impact on the low survival of cryopreserved cells, which should help us to understand the effectiveness of osmotic protectants in dehydration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

The phenylpropanoid pathway is one of the main carbon sinks in plants, channeling phenylalanine towards thousands of products including monolignols, stilbenes, flavonoids and volatile compounds. The enzymatic steps involved in many of these pathways are well characterized, however the physical organization of these enzymes within the plant cell remains poorly understood. Proximity-dependent labeling allows untargeted determination of both direct and indirect protein interactions in vivo, and therefore stands as an attractive alternative to targeted binary assays for determining global protein-protein interaction networks. We used TurboID-based proximity labeling to study protein interaction networks of the core phenylpropanoid and anthocyanin pathways in petunia. To do so, we coupled the endoplasmic reticulum (ER) membrane anchored cytochrome P450 cinnamic acid 4-hydroxylase (C4H, CYP73A412) from Petunia inflata to TurboID and expressed it in protoplasts derived from anthocyanin-rich petunia petals. We identified multiple soluble enzymes from the late anthocyanin pathway among enriched proteins, along with other C4H isoforms, and other ER membrane anchored CYPs. Several of these interactions were subsequently confirmed by bimolecular fluorescence complementation (BiFC). Our results suggest that C4H co-localizes with enzymes from the phenylpropanoid- and downstream anthocyanin pathways, supporting the idea that C4H may serve as ER anchoring points for downstream metabolic pathways. Moreover, this study demonstrates the feasibility of using protoplasts to perform global mapping of protein network for enzymes in their native cellular environment.

Floral homeotic MADS-box transcription factors ensure the correct morphogenesis of floral organs, which are organized in different cell layers deriving from distinct meristematic layers. How cells from these distinct layers acquire their respective identities and coordinate their growth to ensure normal floral organ morphogenesis is unresolved. Here, we studied petunia (Petunia × hybrida) petals that form a limb and tube through congenital fusion. We identified petunia mutants (periclinal chimeras) expressing the B-class MADS-box gene DEFICIENS in the petal epidermis or in the petal mesophyll, called wico and star, respectively. Strikingly, wico flowers form a strongly reduced tube while their limbs are almost normal, while star flowers form a normal tube but greatly reduced and unpigmented limbs, showing that petunia petal morphogenesis is highly modular. These mutants highlight the layer-specific roles of PhDEF during petal development. We explored the link between PhDEF and petal pigmentation, a well-characterized limb epidermal trait. The anthocyanin biosynthesis pathway was strongly downregulated in star petals, including its major regulator ANTHOCYANIN2 (AN2). We established that PhDEF directly binds to the AN2 terminator in vitro and in vivo, suggesting that PhDEF might regulate AN2 expression and therefore petal epidermis pigmentation. Altogether, we show that cell layer-specific homeotic activity in petunia petals differently impacts tube and limb development, revealing the relative importance of the different cell layers in the modular architecture of petunia petals., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

The epidermal cells of petunia (Petunia × hybrida) flowers are the main site of volatile emission. However, the mechanisms underlying the release of volatiles into the environment are still being explored. Here, using cell-layer-specific transcriptomic analysis, reverse genetics by virus-induced gene silencing and clustered regularly interspaced short palindromic repeat (CRISPR), and metabolomics, we identified EPIDERMIS VOLATILE EMISSION REGULATOR (EVER)-a petal adaxial epidermis-specific MYB activator that affects the emission of volatiles. To generate ever knockout lines, we developed a viral-based CRISPR/Cas9 system for efficient gene editing in plants. These knockout lines, together with transient-suppression assays, revealed EVER's involvement in the repression of low-vapor-pressure volatiles. Internal pools and annotated scent-related genes involved in volatile production and emission were not affected by EVER. RNA-Seq analyses of petals of ever knockout lines and EVER-overexpressing flowers revealed enrichment in wax-related biosynthesis genes. Liquid chromatography/gas chromatography-MS analyses of petal epicuticular waxes revealed substantial reductions in wax loads in ever petals, particularly of monomers of fatty acids and wax esters. These results implicate EVER in the emission of volatiles by fine-tuning the composition of petal epicuticular waxes. We reveal a petunia MYB regulator that interlinks epicuticular wax composition and volatile emission, thus unraveling a regulatory layer in the scent-emission machinery in petunia flowers., Competing Interests: Conflict of interest statement. None declared., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Drought is widely recognized as the most severe environmental danger to horticulture in almost all regions across the globe. The objective of this study was to determine if applying a foliar treatment of salicylic acid (SA) at 400 ppm could mitigate the negative effects of drought on Petunia grandiflora. Four experimental variants were implemented for this purpose, with regular watering or moderate drought conditions. These variants were observed for duration of 14 days. The level of photosynthetic pigments was markedly greater in the samples exposed to drought but treated with foliar SA, in comparison to the samples exposed to drought without any treatment (128.28±8.78 and 38.01±1.15 µg/g total chlorophyll (a+b), respectively). The total polyphenol content and antioxidant capacity of both the leaves and flowers of Petunia were assessed. The polyphenols content was significantly reduced by drought. However, following the application of SA, anthocyanin levels increased in comparison to the sample that had been subjected to drought.The results demonstrate that applying SA to the leaves mitigates the adverse impacts of drought, providing a solution to the issue of water scarcity in ornamental plants, particularly Petunia grandiflora.

Background: Shoot branching of flowering plants exhibits phenotypic plasticity and variability. This plasticity is determined by the activity of axillary meristems, which in turn is influenced by endogenous and exogenous cues such as nutrients and light. In many species, not all buds on the main shoot develop into branches despite favorable growing conditions. In petunia, basal axillary buds (buds 1-3) typically do not grow out to form branches, while more apical axillary buds (buds 6 and 7) are competent to grow., Results: The genetic regulation of buds was explored using transcriptome analyses of petunia axillary buds at different positions on the main stem. To suppress or promote bud outgrowth, we grew the plants in media with differing phosphate (P) levels. Using RNA-seq, we found many (> 5000) differentially expressed genes between bud 6 or 7, and bud 2. In addition, more genes were differentially expressed when we transferred the plants from low P to high P medium, compared with shifting from high P to low P medium. Buds 6 and 7 had increased transcript abundance of cytokinin and auxin-related genes, whereas the basal non-growing buds (bud 2 and to a lesser extent bud 3) had higher expression of strigolactone, abscisic acid, and dormancy-related genes, suggesting the outgrowth of these basal buds was actively suppressed. Consistent with this, the expression of ABA associated genes decreased significantly in apical buds after stimulating growth by switching the medium from low P to high P. Furthermore, comparisons between our data and transcriptome data from other species suggest that the suppression of outgrowth of bud 2 was correlated with a limited supply of carbon to these axillary buds. Candidate genes that might repress bud outgrowth were identified by co-expression analysis., Conclusions: Plants need to balance growth of axillary buds into branches to fit with available resources while allowing some buds to remain dormant to grow after the loss of plant parts or in response to a change in environmental conditions. Here we demonstrate that different buds on the same plant with different developmental potentials have quite different transcriptome profiles., (© 2023. BioMed Central Ltd., part of Springer Nature.)

The important feature of petunia in tissue culture is its unpredictable and genotype-dependent callogenesis, posing challenges for efficient regeneration and biotechnology applications. To address this issue, machine learning (ML) can be considered a powerful tool to analyze callogenesis data, extract key parameters, and predict optimal conditions for petunia callogenesis, facilitating more controlled and productive tissue culture processes. The study aimed to develop a predictive model for callogenesis in petunia using ML algorithms and to optimize the concentrations of phytohormones to enhance callus formation rate (CFR) and callus fresh weight (CFW). The inputs for the model were BAP, KIN, IBA, and NAA, while the outputs were CFR and CFW. Three ML algorithms, namely MLP, RBF, and GRNN, were compared, and the results revealed that GRNN (R2≥83) outperformed MLP and RBF in terms of accuracy. Furthermore, a sensitivity analysis was conducted to determine the relative importance of the four phytohormones. IBA exhibited the highest importance, followed by NAA, BAP, and KIN. Leveraging the superior performance of the GRNN model, a genetic algorithm (GA) was integrated to optimize the concentration of phytohormones for maximizing CFR and CFW. The genetic algorithm identified an optimized combination of phytohormones consisting of 1.31 mg/L BAP, 1.02 mg/L KIN, 1.44 mg/L NAA, and 1.70 mg/L IBA, resulting in 95.83% CFR. To validate the reliability of the predicted results, optimized combinations of phytohormones were tested in a laboratory experiment. The results of the validation experiment indicated no significant difference between the experimental and optimized results obtained through the GA. This study presents a novel approach combining ML, sensitivity analysis, and GA for modeling and predicting callogenesis in petunia. The findings offer valuable insights into the optimization of phytohormone concentrations, facilitating improved callus formation and potential applications in plant tissue culture and genetic engineering., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Rezaei et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Petunia stands as the top-selling bedding plant in the U.S., and improved lighting control in greenhouses holds the potential to reduce crop production time and optimize crop quality. This study investigated the impact of four distinct daily light integral (DLI) conditions with and without supplemental far-red (FR) radiation on the growth of petunia liners and subsequent development of finish plants. Two experiments were conducted in spring (9 April to 18 June 2021) and winter (28 October 2021 to 6 January 2022). Petunia cuttings were rooted in a common environment and then transferred to four greenhouse sections with different DLI treatments: 6, 9, 12, and 15 mol·m−2·d−1 for four weeks. Within each DLI condition, half of the plants were exposed to 28 μmol·m−2·s−1 supplemental FR radiation for 16 h daily (equivalent to 1.61 mol·m−2·d−1 light integral). The number of flower buds and open flowers were tracked daily. Representative liners were destructively harvested and evaluated after four weeks of lighting treatments. The remaining plants were transplanted and moved to a common DLI condition of 15 mol·m−2·d−1 for an additional three weeks before being destructively harvested and evaluated as finish plants. The primary finding reveals the promoting effect of DLI on flowering, branching, morphology, and biomass accumulation of petunia liners, with many effects persisting into the finish stage. A threshold DLI of 9 mol·m−2·d−1 was identified, as lower DLI (6 mol·m−2·d−1) resulted in extensive stem elongation, rendering the plants unmarketable. Higher DLI levels were found to be optimal in terms of flowering and morphology. Supplemental FR accelerated flowering by up to three days in the summer experiment and up to 12 days in the winter experiment. However, FR had limited impact on the number of flower buds and open flowers, branching, and shoot and root weight of the finish plants. Interactions between DLI and FR were observed on some parameters, whereby FR effects were more pronounced under lower DLI. Overall, both higher DLI and supplemental FR exhibited beneficial effects, but DLI had a more pronounced effect. Thus, DLI during petunia liner production appears more important than adding FR. This study well simulated the commercial propagation and production of petunia plants, providing practical insights for decision-making regarding lighting strategies.

Continuous efforts are required to find ways to protect crop production against the toxicity of petroleum hydrocarbons, such as diesel, and contamination of soils. There is a need for identification of candidate plants that are tolerant to diesel toxicity that might also have the potential for remediation of diesel-contaminated soils. In this study, petunia, a popular ornamental plant and a model experimental plant in research on phytoremediation of environmental pollutants, was used to evaluate a novel method for rapidly assessing diesel toxicity based on the tolerance of shoots generated through in vitro plant cell culture selection. Petunia shoot lines (L1 to L4) regenerated from diesel-treated callus were compared with those from non-diesel-treated callus (control). Significant morphological differences were observed among the tested lines and control, notably with L1 and L4 showing superior growth. In particular, L4 exhibited remarkable adaptability, with increased root development and microbial counts in a diesel-contaminated potting mix, suggesting that the shoots exhibited enhanced tolerance to diesel exposure. Here, this rapid bioassay has been shown to effectively identify plants with varying levels of tolerance to diesel toxicity and could therefore assist accelerated selection of superior plants for phytoremediation. Further research is needed to understand the genetic and physiological mechanisms underlying tolerance traits, with potential applications beyond petunias to other environmentally significant plants.

In this work, we assessed the effects of increasing ozone (O3) on four petunia varieties with different floral pigmentation (pink, red, rose-red, and white). Plants were exposed, in open-top chambers located in China, to three O3 concentrations, i.e., (i) ambient air (AA), (ii) AA + 60 ppb O3 (AA + 60), and (iii) AA + 120 ppb O3 (AA + 120), for 85 days (9 h day−1). Flower diameter and duration were assessed, together with leaf chlorophyll and flavonoid contents. White petunia showed a reduced flower diameter and longevity under AA + 60 (−7 and −6%, respectively, in comparison to AA), whereas pink and red petunias only showed this under AA + 120 (−8 and −7%, on average, respectively). Chlorophyll loss occurred in all varieties under AA + 60 (−30%, on average), and at AA + 120 in white and red petunias (−54%, on average). The total flavonoid content in the pink and white varieties increased only under AA + 120 (around +85%), while it grew at both AA + 60 and AA + 120 (+92% and two-fold higher, respectively) in the red variety. Increasing O3 concentrations did not affect particularly the red-rose variety. The white variety showed the strongest correlations among flower and leaf properties, confirming a variety-related O3 response, as well as demonstrating that it had the highest O3 sensitivity.

The structure and function of biochemical and developmental pathways determine the range of accessible phenotypes, which are the substrate for evolutionary change. Accordingly, we expect that observed phenotypic variation across species is strongly influenced by pathway structure, with different phenotypes arising due to changes in activity along pathway branches. Here, we use flower colour as a model to investigate how the structure of pigment pathways shapes the evolution of phenotypic diversity. We focus on the phenotypically diverse Petunieae clade in the nightshade family, which contains ca 180 species of Petunia and related genera, as a model to understand how flavonoid pathway gene expression maps onto pigment production. We use multivariate comparative methods to estimate co-expression relationships between pathway enzymes and transcriptional regulators, and then assess how expression of these genes relates to the major axes of variation in floral pigmentation. Our results indicate that coordinated shifts in gene expression predict transitions in both total anthocyanin levels and pigment type, which, in turn, incur trade-offs with the production of UV-absorbing flavonol compounds. These findings demonstrate that the intrinsic structure of the flavonoid pathway and its regulatory architecture underlies the accessibility of pigment phenotypes and shapes evolutionary outcomes for floral pigment production.

Multidrug and toxic compound extrusion (MATE) transporter proteins are a class of secondary transporter proteins that can transport flavonoids. Anthocyanins, a kind of flavonoid, are important secondary metabolites widely found in higher plants; they determine the flower color of most angiosperms. TT12 in Arabidopsis was the first MATE protein identified to be involved in flavonoid transport. Petunia (Petunia hybrida) is an important ornamental plant and is one of the ideal plants for studying plant flower color. However, there are few reports on anthocyanin transport in petunia. In this study, we characterized a homolog of Arabidopsis TT12 in the petunia genome, PhMATE1, that shares the highest amino acid sequence identity with Arabidopsis TT12. PhMATE1 protein contained 11 transmembrane helices. PhMATE1 showed a high transcription level in corollas. The silencing of PhMATE1 mediated by both virus-induced gene silence and RNA interference changed flower color and reduced anthocyanin content in petunia, suggesting that PhMATE1 is involved in anthocyanin transport in petunia. Furthermore, PhMATE1 silencing downregulated the expression of the structural genes of the anthocyanin synthesis pathway. The results of this study supported the hypothesis that MATEs are involved in the sequestration of anthocyanins during flower color formation., (© 2023 Scandinavian Plant Physiology Society.)

Petunia hybrida is an economically important ornamental plant species. Branching in ornamental plants is closely associated with their ornamental traits, and branching is a significant agronomic trait in petunia, which shapes plant architecture and production cost. Although there are few studies regarding the involvement of the SPEAR genes in lateral branch development, they are known to regulate the development of plant organs. The PhSPEAR1 gene in petunia, an ortholog of Arabidopsis SPEAR1, was isolated for study. According to the results of real-time quantitative PCR (qRT-PCR), PhSPEAR1 was primarily expressed in the roots. The fluorescence signal indicated that PhSPEAR1 was localized in the nucleus and cytoplasm. An increase in PhSPEAR1 expression was induced by cytokinin or decapitation. Overexpression of PhSPEAR1 increased lateral branches in Arabidopsis. Based on our findings, PhSPEAR1 participates significantly in the regulation of branch number in petunia.

Emission of scent volatiles by flowers is important for successful pollination and consequently, reproduction. Petunia (Petunia hybrida) floral scent is formed mainly by volatile products of the phenylpropanoid pathway. We identified and characterized a regulator of petunia scent production: the GRAS protein PHENYLPROPANOID EMISSION-REGULATING SCARECROW-LIKE (PES). Its expression increased in petals during bud development and was highest in open flowers. Overexpression of PES increased the production of floral volatiles, while its suppression resulted in scent reduction. We showed that PES upregulates the expression of genes encoding enzymes of the phenylpropanoid and shikimate pathways in petals, and of the core regulator of volatile biosynthesis ODORANT1 by activating its promoter. PES is an ortholog of Arabidopsis (Arabidopsis thaliana) PHYTOCHROME A SIGNAL TRANSDUCTION 1, involved in physiological responses to far-red (FR) light. Analyses of the effect of nonphotosynthetic irradiation (low-intensity FR light) on petunia floral volatiles revealed FR light as a scent-activating factor. While PHYTOCHROME A regulated scent-related gene expression and floral scent production under FR light, the influence of PES on volatile production was not limited by FR light conditions., Competing Interests: Conflict of interest statement. The authors declare no conflict of interest., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Exogenous application of chitosan has been shown to reduce plant disease severity in food crops; however, less is known about the potential use of chitosan in floriculture. The objective of this study was to investigate the effectiveness of chitosan to suppress gray mold on petunia leaves caused by Botrytis cinerea using in vitro and in planta approaches. We also aimed to determine if chitosan molecular weight influences efficacy. Medium and high molecular weight reagent grade chitosan reduced growth of B. cinerea in vitro at chitosan concentrations ranging from 1.25 to 2% (v/v), while low molecular weight reagent grade chitosan only reduced growth at 2.0% (v/v). In detached leaf assays, all reagent grade chitosan treatments reduced Botrytis lesion size on petunia leaves up to 65% compared to the water control. The commercial product Tidal Grow reduced in vitro growth of Botrytis , starting at 0.5%, and reduced disease severity at 0.75% on petunia leaves. The commercial product ARMOUR-Zen 15 reduced Botrytis growth in vitro at 3.75% and higher and reduced disease severity at 0.3 and 1.0% on petunia leaves. Under greenhouse conditions, low, medium, and high molecular weight reagent grade chitosan and ARMOUR-Zen 15 at 0.4% chitosan reduced Botrytis lesion size on petunia leaves up to 60% compared to the water control. Suppression in vitro suggests that chitosan may have direct phytotoxic effects on fungal growth, however our in planta and greenhouse trials suggest that additional modes of action may also play a role in the observed suppressive effects.

Dạ Yến Thảo là một loài cây cảnh rất được ưa chuộng trong trang trí nhà cửa, sân vườn. Nghiên cứu này được thực hiện để thiết lập một quy trình vi nhân giống hiệu quả cho cây Dạ Yến Thảo. Đế hoa Dạ Yến Thảo được khử trùng bằng ethanol 70% trong 1 phút và Javel Mỹ Hảo 25% trong 15 phút. Mẫu tái sinh chồi trên môi trường MS bổ sung sucrose 30 g/L, BA 2,5 mg/L, NAA 0,1 mg/L. Meta-topolin 1 mg/L giúp nhân nhanh chồi hiệu quả, đạt 13,7 chồi/ cụm sau 2 tuần nuôi cấy. Ở giai đoạn tạo rễ, môi trường phù hợp là môi trường MS bổ sung sucrose 30 g/L, NAA 1 mg/L và than hoạt tính 0,5 g/L, tỷ lệ tạo rễ đạt 100%, 13,7 rễ/ cây. Cây con đã ra rễ được chuyển sang chậu trồng có chứa mụn xơ dừa và phân bò (tỉ lệ 1:1) đạt tỉ lệ sống cao (80%).

Introduction Flower color is one of the most significant characteristics in ornamental plant breeding. New varieties of various plants in relation to their flower color have been obtained by monitoring the expression levels of genes involved or regulating the flavonoid and anthocyanin biosynthesis pathway. Flavonoids possess significant and diverse biological functions. They are the major pigments for flowers, fruits, seeds, and leaves. They are natural products that contain a C6-C3-C6 carbon framework and are synthesized by a branched pathway that yields both colored and colorless compounds. The gene encoding chalcone isomerase (CHI) is among the genes and enzymes identified in the flavonoid pathway. This enzyme catalyzes the isomerization of naringenin chalcone into the corresponding flavanone. CHI enzyme belongs to the family of isomerases, specifically the class of intramolecular lyases. Chalcone isomerase has a core 2-layer alpha/beta structure and has attracted much attention recently due to its role in stress response and pigment production. One of the most effective methods of genetic engineering is the reduction of flower pigments by suppression of required enzymes for their biosynthesis. RNA interference (RNAi) has provided the tool for the investigation of genes involved in the production of flower color. Silencing of any gene in the anthocyanin biosynthetic pathway can result in reduced or inhibited anthocyanin production. RNAi technology is an effective gene silencing method and a powerful tool for studying gene function and development of new traits by transformation of viral RNA or hairpin RNA (hpRNA) constructs into plants. The processing of dsRNA into 21-23-nt small interfering RNAs (siRNAs), and the mediators of RNAi, triggers cognate mRNA degradation. The hpRNAi methodology simply requires a transgene construct containing an inversely-repeated sequence of the target gene flanked with a promoter and terminator which effectively function in plants. Material and Methods In this research, with the design and construction of chiRNAi, the transformation of the RNAi construct was carried out of Petunia plants. Potted plants of P. hybrida were grown under standard greenhouse conditions (16-17°C night temperature and 21-24°C day temperature and photoperiod 16/8 (light/dark)). The RNAi construct including the 530 bp cds of the chalcone isomerase (chi) gene and 741 bp of pdk gene as intron between chi sense and antisense were used for transient RNAi-induced silencing. The pBI121-chi530 plasmids were introduced into A. tumefaciens strain LBA4404 by electroporation method. Colonies of A. tumefaciens carrying the desired plasmid were screened by PCR with specific primers for chi gene. RNAi construct co-cultured with petunia’s leave. Samples was kept in dark condition for 3 days and then transferred to branch induction media. Samples were investigated for phenotypical changes and chi gene expression by qRT-PCR. Results and Discussion Transgenic lines showed a reduced number of pigments and a faded flower color. So that, in purple petunia, was shown 5 phenotypical groups. These groups was indicated different levels of chi gene silencing. In pink petunia was seen two groups of phenotypical changes. In these plants, chi-RNAi construct was reduced pigment production and so, these plants had faded colors in petals. Also, the chi gene expression was reduced in all transgenic lines. Generally, the results of this research showed that RNAi can be used as an efficient method for gene silencing. The application of gene silencing can indicate the gene’s function in biosynthesis pathways of various components such as anthocyanins. In addition, the chalcone isomerase gene was identified as one of the effective genes in anthocyanin biosynthesis pathway in Petunia plants that could be involved in the production of color in these plants; hence, chi gene silencing resulted in clear phenotypic alterations in this plant. Conclusion In general the concentration of the target mRNA in a particular tissue could be a factor that influences silencing efficiency. At very low levels of gene expression, small amounts of the silencing target, mRNA, could be completely degraded by the RNA-induced silencing complex (RISC), whereas the presence of higher amounts of the target mRNA may result in incomplete silencing, allowing some residual functional mRNA to be translated into the corresponding protein. This research demonstrated the hpRNA construct has been successfully established for floral tissues of P. hybrida. The hpRNA construct was developed for chi-RNAi silencing of one of the key genes in the anthocyanin biosynthetic pathway in Petunia flowers. The silencing of the chi gene is a prototype for the modification of the anthocyanin biosynthetic pathway in Petunia through gene suppression. This strategy could also be useful for rapid functional analysis of other genes involved in flower development.

The process of optimizing in vitro seed sterilization and germination is a complicated task since this process is influenced by interactions of many factors (e.g., genotype, disinfectants, pH of the media, temperature, light, immersion time). This study investigated the role of various types and concentrations of disinfectants (i.e., NaOCl, Ca(ClO)2, HgCl2, H2O2, NWCN-Fe, MWCNT) as well as immersion time in successful in vitro seed sterilization and germination of petunia. Also, the utility of three artificial neural networks (ANNs) (e.g., multilayer perceptron (MLP), radial basis function (RBF), and generalized regression neural network (GRNN)) as modeling tools were evaluated to analyze the effect of disinfectants and immersion time on in vitro seed sterilization and germination. Moreover, non‑dominated sorting genetic algorithm‑II (NSGA‑II) was employed for optimizing the selected prediction model. The GRNN algorithm displayed superior predictive accuracy in comparison to MLP and RBF models. Also, the results showed that NSGA‑II can be considered as a reliable multi-objective optimization algorithm for finding the optimal level of disinfectants and immersion time to simultaneously minimize contamination rate and maximize germination percentage. Generally, GRNN-NSGA-II as an up-to-date and reliable computational tool can be applied in future plant in vitro culture studies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Rezaei et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)