Fragaria Vesca Research Articles

The effect of osmotic stress, lighting spectrum and temperature on growth and gene expression related to anthocyanin biosynthetic pathway in wild strawberry (Fragaria vesca L.) in vitro

ABSTRACT The goal of this research was to evaluate the effect of light, temperature, sucrose and PEG on the growth of Fragaria vesca in vitro and the expression of regulatory Myb10, WD40 and enzyme-coding genes CHI, CHS, DFR, EGL, F3H and UFGT, which are essential for anthocyanin biosynthesis. We observed plants’ response to osmotic stress, the decrease in growth and microshoot weight. A change in the expression of the investigated genes was evident under the suboptimal concentration of sucrose. The addition of PEG to the medium caused a decrease in microshoot weight and gene expression. Blue + red lights of the LED lighting system significantly affected microshoot growth in vitro. Red and blue + red + UV lights slightly reduced microshoot weight and caused a reddish colour of petioles, which indicate increased anthocyanin synthesis. Moreover, most of the studied genes’ expression tended to increase when shoots were exposed to blue, blue + red and blue + red + UV lights. A temperature of 15°C (vs 22°C) significantly reduced the mean fresh weight of microshoots while increasing CHI and CHS gene expression and decreasing WD40 gene expression. Exposure to a higher temperature (30°C) induced the vitrification of microshoots, although the fresh weight did not differ from that of the control. Gene expression also depended on the duration of exposure. In the case of CHS, gene expression remained the same or increased after exposure for 1 week and then decreased after exposure for 4 weeks.

Abnormal Pollen Development in the Fragaria vesca Mutant fruitless 1

Knowledge of the genes underlying a given trait is highly useful for developing molecular markers for breeding and is the foundation for future genomic crop improvements. The cultivated strawberry, F. ×ananassa, is a valuable horticultural crop. Genome sequencing revealed that of the four diploid strawberry subgenomes contributing to the F. ×ananassa octoploid genome, the woodland strawberry, F. vesca, subgenome is dominant. Thus, F. vesca is an important system for determining gene function and should be used as a source of genetic diversity for F. ×ananassa breeding. Ethyl methanesulfonate mutagenesis of H4 F7-3, an inbred line of F. vesca, resulted in one M2 line that did not produce any strawberries over a 3-year period in the greenhouse. This line was named fruitless 1. The fruitless 1 phenotype results from a single gene recessive mutation. Microscopic characterization revealed that fruitless 1 failed to produce fruit because anthers fail to develop properly before meiosis, resulting in no pollen production. This report of fruitless 1 facilitates further studies of the line.

Phased gap-free genome assembly of octoploid cultivated strawberry illustrates the genetic and epigenetic divergence among subgenomes.

The genetic and epigenetic mechanisms underlying the coexistence and coordination of the four diverged subgenomes (ABCD) in octoploid strawberries (Fragaria × ananassa) remains poorly understood. In this study, we have assembled a haplotype-phased gap-free octoploid genome for the strawberry, which allowed us to uncover the sequence, structure, and epigenetic divergences among the subgenomes. The diploid progenitors of the octoploid strawberry, apart from subgenome A (Fragaria vesca), have been a subject of public controversy. Phylogenomic analyses revealed a close relationship between diploid species Fragaria iinumae and subgenomes B, C, and D. Subgenome A, closely related to F. vesca, retains the highest number of genes, exhibits the lowest content of transposable elements (TEs), experiences the strongest purifying selection, shows the lowest DNA methylation levels, and displays the highest expression level compared to the other three subgenomes. Transcriptome and DNA methylome analyses revealed that subgenome A-biased genes were enriched in fruit development biological processes. In contrast, although subgenomes B, C, and D contain equivalent amounts of repetitive sequences, they exhibit diverged methylation levels, particularly for TEs located near genes. Taken together, our findings provide valuable insights into the evolutionary patterns of subgenome structure, divergence and epigenetic dynamics in octoploid strawberries, which could be utilized in strawberry genetics and breeding research.

If you go down to the woods today you're in for a big surprise: mapping the diverse phenotypes of the European woodland strawberry.

If you go down to the woods today you're in for a big surprise: mapping the diverse phenotypes of the European woodland strawberry.

Fungicide Resistance in Botrytis spp. and Regional Strategies for Its Management in Northern European Strawberry Production.

Grey mould, caused by Botrytis cinerea and other Botrytis spp., is a major cause of fruit rot in strawberries and other fruit crops worldwide. Repeated fungicide applications are essential in order to secure harvests. However, resistance to all currently registered single-site fungicides is widespread. The rising importance of strains with multiple resistance to most or all fungicides is of particular concern. These strains may be introduced into fields via contaminated nursery plants and/or by immigration from adjacent plots. On the basis of research conducted in northern German and Danish strawberry production, a concept to manage fungicide resistance under northern European conditions has been developed and put into regional strawberry production practice. This principally includes the testing of nursery plants for fungicide-resistant Botrytis strains prior to planting; the restricted and specific use of fungicides at flowering in the production fields, taking account of the resistance spectrum within the local Botrytis population; and crop sanitation measures such as the removal of rotting fruits at the beginning of harvest. Further options such as protected cultivation, reduced fertilisation and biological control are also discussed. The practical implementation of such a strategy in northern Germany and Denmark has been shown to reduce the occurrence of multi-resistant strains to a tolerable steady-state level.

The red/far-red light photoreceptor FvePhyB regulates tissue elongation and anthocyanin accumulation in woodland strawberry.

Light is an important environmental signal that influences plant growth and development. Among the photoreceptors, phytochromes can sense red/far-red light to coordinate various biological processes. However, their functions in strawberry are not yet known. In this study, we identified an EMS mutant, named P8, in woodland strawberry (Fragaria vesca) that showed greatly increased plant height and reduced anthocyanin content. Mapping-by-sequencing revealed that the causal mutation in FvePhyB leads to premature termination of translation. The light treatment assay revealed that FvePhyB is a bona fide red/far-red light photoreceptor, as it specifically inhibits hypocotyl length under red light. Transcriptome analysis showed that the FvePhyB mutation affects the expression levels of genes involved in hormone synthesis and signaling and anthocyanin biosynthesis in petioles and fruits. The srl mutant with a longer internode is caused by a mutation in the DELLA gene FveRGA1 (Repressor of GA1) in the gibberellin pathway. We found that the P8 srl double mutant has much longer internodes than srl, suggesting a synergistic role of FvePhyB and FveRGA1 in this process. Taken together, these results demonstrate the important role of FvePhyB in regulating plant architecture and anthocyanin content in woodland strawberry.

The dynamic arms race during the early invasion of woodland strawberry by Botrytis cinerea revealed by dual dense high-resolution RNA-seq analyses.

Necrotrophic pathogens replicate massively upon colonizing plants, causing large-scale wilting and death of plant tissues. Understanding both mechanisms of pathogen invasion and host response processes prior to symptom appearance and their key regulatory networks is therefore important for defense against pathogen attack. Here, we investigated the mechanisms of interaction between woodland strawberry (Fragaria vesca) leaves and gray mold pathogen (Botrytis cinerea) at 14 infection time points during the first 12hours of the infection period using a dense, high-resolution time series dual transcriptomic analysis, characterizing the arms race between strawberry F. vesca and B. cinerea before the appearance of localized lesions. Strawberry leaves rapidly initiated strong systemic defenses at the first sign of external stimulation and showed lower levels of transcriptomic change later in the infection process. Unlike the host plants, B. cinerea showed larger-scale transcriptomic changes that persisted throughout the infection process. Weighted gene co-expression network analysis identified highly correlated genes in 32 gene expression modules between B. cinerea and strawberry. Yeast two-hybrid and bimolecular fluorescence complementation assays revealed that the disease response protein FvRLP2 from woodland strawberry interacted with the cell death inducing proteins BcXYG1 and BcPG3 from B. cinerea. Overexpression of FvRLP2 in both strawberry and Arabidopsis inhibited B. cinerea infection, confirming these genes' respective functions. These findings shed light on the arms race process by which B. cinerea invades host plants and strawberry to defend against pathogen infection.

IN SILICO MINING AND VALIDATION OF SIMPLE SEQUENCE REPEAT (SSR) MARKERS DERIVED FROM THE FRAGARIA X ANANASSA GENOME ASSEMBLY

Strawberry (Fragaria x ananassa Duchesne ex Rozier) is an economically important fruit crop grown commercially worldwide due to its known nutritional value. The demand for strawberries in the Philippines has been increasing but varietal development is slow due to the lack of genetic information on locally available germplasm. In addition, molecular marker resources for strawberries are limited. Here, we developed Simple Sequence Repeats (SSR) markers to characterize strawberries cultivated in tropical environments, particularly in the Philippines. In silico mining generated a total of 219,239 SSR markers, from which 160,025 unique markers were identified. Polymorphism scoring and the variation on the expected alleles resulting to the synthesis of the top 160 markers were validated using representative strawberry cultivars from the core collection. All 70 markers produced successful amplicons from all the genotypes, but only 67 markers displayed polymorphism. Using UPGMA, genetic similarities among the cultivars were calculated generating a dendrogram which categorized the strawberry cultivars into 3 groups. The grouping clearly showed the separation of the diploid Fragaria vesca from the octoploid F. x ananassa cultivars, considered as group 1. Furthermore, Groups 2 and 3 contained cultivars from different geographical origins (US, Japan, and North America). The eight novel markers developed and validated here can be added to the genetic marker resource for cultivar verification, assessment of genetic diversity, and most importantly, marker-assisted breeding.

4mC-CGRU: Identification of N4-Methylcytosine (4mC) sites using convolution gated recurrent unit in Rosaceae genome

An epigenetic modification is DNA N4-methylcytosine (4mC) that affects several biological functions without altering the DNA nucleotides, including DNA conformation, cell development, replication, stability, and DNA structural changes. To prevent restriction enzyme from damaging self-DNA, 4mC performs a critical role in restriction–modification functions. Existing studies mainly focused on finding hand-crafted features to identify 4mC locations, but these methods are inefficient due to high time consuming and high costs. In our research work, we propose a 4mC-CGRU which is a deep learning-based computational model with a standard encoding method to identify the 4mC sites from DNA sequences that learned autonomous feature selection in the Rosaceae genome, particularly in Rosa chinensis (R. chinensis) and Fragaria vesca (F. vesca). The proposed model consists of a convolutional neural network (CNN) and a gated recurrent unit network (GRU)-based model for identifying 4mC sites from Fragaria vesca and Rosa chinensis in the genomes. The CNN model extracts useful features from the datasets and the GRU classifies the DNA sequences. Thus, our approach can automatically extract important features to detect relative sites from DNA sequence. The performance analysis shows that the proposed model consistently outperforms over the state-of-the-art works in detecting 4mC sites.

Comparative genomics analysis provides insights into evolution and stress responses of Lhcb genes in Rosaceae fruit crops

BackgroundLight-harvesting chlorophyll a/b binding proteins (Lhcb) play crucial roles in plant growth, development, and the response to abiotic stress in higher plants. Previous studies have reported that Lhcb genes were involved in the phytochrome regulation and responded to different light and temperature conditions in Poaceae (such as maize). However, the evolution and functions of Lhcb genes remains poorly characterized in important Rosaceae species.ResultsIn this investigation, we conducted a genome-wide analysis and identified a total of 212 Lhcb genes across nine Rosaceae species. Specifically, we found 23 Lhcb genes in Fragaria vesca, 20 in Prunus armeniaca, 33 in Malus domestica ‘Gala’, 21 in Prunus persica, 33 in Rosa chinensis, 29 in Pyrus bretschneideri, 18 in Rubus occidentalis, 20 in Prunus mume, and 15 in Prunus salicina. Phylogenetic analysis revealed that the Lhcb gene family could be classified into seven major subfamilies, with members of each subfamily sharing similar conserved motifs. And, the functions of each subfamily was predicted based on the previous reports from other species. The Lhcb proteins were highly conserved within their respective subfamilies, suggesting similar functions. Interestingly, we observed similar peaks in Ks values (0.1–0.2) for Lhcb genes in apple and pear, indicating a recent whole genome duplication event (about 30 to 45 million years ago). Additionally, a few Lhcb genes underwent tandem duplication and were located across all chromosomes of nine species of Rosaceae. Furthermore, the analysis of the cis-acting elements in the 2000 bp promoter region upstream of the pear Lhcb gene revealed four main categories: light response correlation, stress response correlation, hormone response correlation, and plant growth. Quantitative expression analysis demonstrated that Lhcb genes exhibited tissue-specific expression patterns and responded differently to low-temperature stress in Rosaceae species.ConclusionsThese findings shed light on the evolution and phylogeny of Lhcb genes in Rosaceae and highlight the critical role of Lhcb in pear’s response to low temperatures. The results obtained provide valuable insights for further investigations into the functions of Lhcb genes in Rosaceae, and these functional genes will be used for further fruit tree breeding and improvement to cope with the current climate changes.

Acclimation of circadian rhythms in woodland strawberries (Fragaria vesca L.) to Arctic and mid-latitude photoperiods

BackgroundThough many abiotic factors are constantly changing, the photoperiod is a predictable factor that enables plants to time many physiological responses. This timing is regulated by the circadian clock, yet little is known about how the clock adapts to the differences in photoperiod between mid-latitudes and high latitudes. The primary objective of this study was to compare how clock gene expression is modified in four woodland strawberry (Fragaria vesca L.) accessions originating from two different populations in Italy (IT1: Tenno, Italy, 45°N, IT4: Salorno, Italy, 46°N) and two in Northern Norway (NOR2: Alta, Norway, 69°N, NOR13: Indre Nordnes, Norway 69°N) when grown under simulated daylength conditions of an Arctic or mid-latitude photoperiod. The second objective was to investigate whether population origin or the difference in photoperiod influenced phytohormone accumulation.ResultsThe Arctic photoperiod induced lower expression in IT4 and NOR13 for six clock genes (FvLHY, FvRVE8, FvPRR9, FvPRR7, FvPRR5, and FvLUX), in IT1 for three genes (FvLHY, FvPRR9, and FvPRR5) and in NOR2 for one gene (FvPRR9). Free-running rhythms for FvLHY in IT1 and IT4 were higher after the Arctic photoperiod, while the free-running rhythm for FvLUX in IT4 was higher after the mid-latitude photoperiod. IT1 showed significantly higher expression of FvLHY and FvPRR9 than all other accessions, as well as significantly higher expression of the circadian regulated phytohormone, abscisic acid (ABA), but low levels of salicylic acid (SA). NOR13 had significantly higher expression of FvRVE8, FvTOC1, and FvLUX than all other accessions. NOR2 had extremely low levels of auxin (IAA) and high levels of the jasmonate catabolite, hydroxyjasmonic acid (OH-JA).ConclusionsOur study shows that circadian rhythms in Fragaria vesca are driven by both the experienced photoperiod and genetic factors, while phytohormone levels are primarily determined by specific accessions’ genetic factors rather than the experienced photoperiod.

DNA barcodes, ISSR, RAPD and SCAR markers as potential quality control tools for molecular authentication of black and white mulberry

Morus species, especially black mulberry (Morus nigra L.), are well known for their nutritive and economic values. Black mulberry is often a subject for deliberate and undeliberate substitution with the morphologically similar white mulberry (Morus alba L.). In the current study, the genetic diversity of Morus nigra L. and Morus alba L. was assessed using different genetic profiling and fingerprinting techniques namely namely DNA barcoding, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR). DNA barcoding was implemented using 2 plastid loci (matK, and rbcL-1) and 2 nuclear spacers (ITS and ITS2) aiming at accurate authentication of the studied species. DNA barcoding, targeting the selected loci succeeded to identify and authenticate the studies Morus species but failed to differentiate them, owing to their close genetic resemblance. Out of 52 screened RAPD primers, 27 were able to produce reproducible and clear profiles with 115 amplified fragments of which 37 were polymorphic accounting for 32.17% polymorphism. The number of amplified bands varied from 2 to 7 ranging in size from 100 to 700 bp. Moreover, 55 ISSR primers were evaluated, 108 fragments were produced, using 20 primers, of which 28 were polymorphic accounting for 25.92% polymorphism with size range of 200–800 bp. Sequence Characterized Amplified Region (SCAR) marker was developed based on reproducible banding pattern obtained by RAPD primer (OPG-03), for the purpose of accelerating the initial screening of Morus batches and enabling the preventative rejection of adulterated samples. The polymorphic band obtained was sequenced and specific primers were designed to develop a SCAR marker of approximately 558 bp exclusively for M. nigra, allowing detection of down to 1 ng, of M. nigra. The SCAR marker showed 100% success in differentiating authentic M. nigra and M. alba samples. In addition, the application of the developed SCAR marker was extended to distinguish M. nigra from its four common adulterants namely Ficus carica L., Fragaria vesca L., Ficus racemosa L. and Vitis vinifera L.. The proposed protocol was successfully implemented for the evaluation of 47 commercial M. nigra samples collected from different Egyptian locations.

Влияние экологических факторов на генетический полиморфизм в природных популяциях Fragaria vesca

Genetic polymorphism of natural cenopopulations of wild strawberry (Fragaria vesca) from the Republic of Mari El was measured by randomly amplified polymorphic DNA (RAPD). The method of molecular genome marking based on RAPD-PCR makes it possible to determine the genetic status of populations and establish interpopulation relationships. The cenopopulations that we selected for the analysis were confined to different stages of the forest phytocenosis succession: the climatic phytocenosis and the area of the forest felling. These F. vesca cenopopulations differed in the ratio of sexual and vegetative reproduction during the maintenance of the cenopopulation size. In a felling area in conditions of greater illumination the intensity of sexual and vegetative reproduction increases. However, specimens of seed origin were not found in the felling area, since in conditions of high competition with ruderal and meadow plants, slowly developing strawberry seedlings died. The germination of seeds and plants of seed origin was noted under conditions of climatic phytocenosis. For the cenopopulation confined to the climax forest, Shannon’s diversity index and Nei’s genetic distance were 0.5179 and 0.3613 respectively. In a felling area vegetative reproduction processes predominate, so Shannon’s diversity index and Nei’s genetic distance were lower, 0.4573 and 0.3132 respectively. In a felling area revealed the loss of certain genetic clusters present in the climax forest. This indicates the loss of a strawberry genetic diversity in the transition to vegetative reproduction due to anthropogenic impact. Our data confirm the idea that climax communities with their more stable conditions can act as regenerative niches of the species.

Highly efficient Agrobacterium rhizogenes-mediated transformation for functional analysis in woodland strawberry

BackgroundThe diploid woodland strawberry (Fragaria vesca) is an excellent model plant for investigating economically significant traits and several genetic resources within the Rosaceae family. Agrobacterium rhizogenes-mediated hairy root transformation is an alternative for exploring gene functions, especially the genes specifically expressed in roots. However, the hairy root transformation has not been established in strawberry.ResultsHere, we described an efficient and rapid hairy root transgenic system for strawberry using A. rhizogenes. Strain of A. rhizogenes MSU440 or C58C1 was the most suitable for hairy root transformation. The transformation efficiency was highest when tissues contained hypocotyls as explants. The optimal procedure involves A. rhizogenes at an optical density (OD600) of 0.7 for 10 min and co-cultivation duration for four days, achieving a transgenic efficiency of up to 71.43%. An auxin responsive promoter DR5ver2 carrying an enhanced green fluorescent protein (eGFP) marker was transformed by A. rhizogenes MSU440, thereby generating transgenic hairy roots capable of high eGFP expression in root tip and meristem of strawberry where auxin accumulated. Finally, this system was applied for functional analysis using jGCaMP7c, which could sense calcium signals. A significant upsurge in eGFP expression in the transgenic hairy roots was displayed after adding calcium chloride. The results suggested that this approach was feasible for studying specific promoters and could be a tool to analyze gene functions in the roots of strawberries.ConclusionWe established a rapid and efficient hairy root transformation in strawberry by optimizing parameters, which was adequate for promoter analysis and functional characterization of candidate genes in strawberry and other rosaceous plants.

Effects of anti-herbivore resistance on sensory characteristics and ripening of strawberry

Plant resistance to agricultural pests is a fundamental element of sustainable crop protection. However, concerns have been raised that strategies to increase resistance may involve phytochemicals that impact fruit ripening and the sensorial perception of the fruit. Here, we experimentally tested for these putative resistance effects by contrasting susceptible varieties of strawberry (Fragaria vesca) with varieties that were either constitutively resistant to pest insects or with resistance induced with jasmonic acid (JA). GC-MS analysis identified 11 volatile compounds, including alcohols, aldehydes, lactone, terpenoids, and esters, which showed higher concentrations in fruits from resistant/induced plants. Fruits from induced plants ripened faster in the field. In sensory analyses, using a trained analytical panel, some variation between the sensory profiles of the strawberry varieties was detected, but we found no systematic correlations between sensory attributes and the level of plant resistance/induction in the varieties. These results suggest that increased plant resistance comes with positive effects of early ripening, while not strongly affecting the overall sensory experience.

Diversity of the volatilome and the fruit size and shape in European woodland strawberry (Fragaria vesca).

Woodland strawberry (Fragaria vesca subsp. vesca) is a wild relative of cultivated strawberry (F. × ananassa) producing small and typically conical fruits with an intense flavor and aroma. The wild strawberry species, F. vesca, is a rich resource of genetic and metabolic variability, but its diversity remains largely unexplored and unexploited. In this study, we aim for an in-depth characterization of the fruit complex volatilome by GC-MS as well as the fruit size and shape using a European germplasm collection that represents the continental diversity of the species. We report characteristic volatilome footprints and fruit phenotypes of specific geographical areas. Thus, this study uncovers phenotypic variation linked to geographical distribution that will be valuable for further genetic studies to identify candidate genes or develop markers linked to volatile compounds or fruit shape and size traits.

Genome-wide identification and expression analyses of the pectate lyase (PL) gene family in Fragaria vesca

BackgroundPectate lyase (PL, EC 4.2.2.2), as an endo-acting depolymerizing enzyme, cleaves α-1,4-glycosidic linkages in esterified pectin and involves a broad range of cell wall modifications. However, the knowledge concerning the genome-wide analysis of the PL gene family in Fragaria vesca has not been thoroughly elucidated.ResultsIn this study, sixteen PLs members in F. vesca were identified based on a genome-wide investigation. Substantial divergences existed among FvePLs in gene duplication, cis-acting elements, and tissue expression patterns. Four clusters were classified according to phylogenetic analysis. FvePL6, 8 and 13 in cluster II significantly contributed to the significant expansions during evolution by comparing orthologous PL genes from Malus domestica, Solanum lycopersicum, Arabidopsis thaliana, and Fragaria×ananassa. The cis-acting elements implicated in the abscisic acid signaling pathway were abundant in the regions of FvePLs promoters. The RNA-seq data and in situ hybridization revealed that FvePL1, 4, and 7 exhibited maximum expression in fruits at twenty days after pollination, whereas FvePL8 and FvePL13 were preferentially and prominently expressed in mature anthers and pollens. Additionally, the co-expression networks displayed that FvePLs had tight correlations with transcription factors and genes implicated in plant development, abiotic/biotic stresses, ions/Ca2+, and hormones, suggesting the potential roles of FvePLs during strawberry development. Besides, histological observations suggested that FvePL1, 4 and 7 enhanced cell division and expansion of the cortex, thus negatively influencing fruit firmness. Finally, FvePL1-RNAi reduced leaf size, altered petal architectures, disrupted normal pollen development, and rendered partial male sterility.ConclusionThese results provide valuable information for characterizing the evolution, expansion, expression patterns and functional analysis, which help to understand the molecular mechanisms of the FvePLs in the development of strawberries.

Warmer temperature during asexual reproduction induce methylome, transcriptomic, and lasting phenotypic changes in Fragaria vesca ecotypes.

Plants must adapt with increasing speed to global warming to maintain their fitness. One rapid adaptation mechanism is epigenetic memory, which may provide organisms sufficient time to adapt to climate change. We studied how the perennial Fragaria vesca adapted to warmer temperatures (28°C vs. 18°C) over three asexual generations. Differences in flowering time, stolon number, and petiole length were induced by warmer temperature in one or more ecotypes after three asexual generations and persisted in a common garden environment. Induced methylome changes differed between the four ecotypes from Norway, Iceland, Italy, and Spain, but shared methylome responses were also identified. Most differentially methylated regions (DMRs) occurred in the CHG context, and most CHG and CHH DMRs were hypermethylated at the warmer temperature. In eight CHG DMR peaks, a highly similar methylation pattern could be observed between ecotypes. On average, 13% of the differentially methylated genes between ecotypes also showed a temperature-induced change in gene expression. We observed ecotype-specific methylation and expression patterns for genes related to gibberellin metabolism, flowering time, and epigenetic mechanisms. Furthermore, we observed a negative correlation with gene expression when repetitive elements were found near (±2kb) or inside genes. In conclusion, lasting phenotypic changes indicative of an epigenetic memory were induced by warmer temperature and were accompanied by changes in DNA methylation patterns. Both shared methylation patterns and transcriptome differences between F. vesca accessions were observed, indicating that DNA methylation may be involved in both general and ecotype-specific phenotypic variation.

Alternative substrates in the cultivation of ornamental and vegetable plants

Research objective: The aim of this research was to evaluate different types of innovative substrate mixtures as an alternative to traditional growing media with peat and pumice on vegetable and ornamental plants Materials and Methods: The experiments, which began in November 2022, were conducted in the CREA-OF greenhouses in Pescia (Pt), Tuscany, Italy (43°54′N 10°41′E) on seedlings of Arbutus unedo, Photinia red robin, Fragaria vesca and Crassula sarcocaulis. The plants were placed in 14 diameter pots, 6 plants per 3 replications, for a total of 18 seedlings per experimental thesis. The test was divided into two parallel trials with different substrates. The first trial on Crassula sarcocaulis involved the following theses (irrigated and fertilised): peat 70% + pumice 30%; peat 65% + pumice 30% + biochar 5%; compost 60% + biochar 5% + insect humus 5% + pumice 30%; compost 60% + biochar 5% + earthworm humus 5% + pumice 30%. The second trial on Arbutus unedo, Photinia red robin and Fragaria vesca involved the following theses (irrigated and fertilised): peat 70% + pumice 30%; alternative soil 72% + biochar 5% + 3% humus + 20% pumice. On 18 May 2023, plant height, vegetative weight, root volume and length, number of fruits (strawberry) and number of microorganisms were determined. Results and Discussion: The experiment showed that the use of alternative substrates can effectively improve the vegetative and root growth of Crassula sarcocaulis, Fragaria vesca, Photinia red robin and Arbutus unedo. In general, in the first experiment on Crassula sarcocaulis, a significant increase in plant height was observed with the biochar-only substrate, followed by the insect and earthworm humus-based theses. All the theses were significantly better than the control with peat and pumice. In the second trial on Fragaria vesca, Photinia red robin and Arbutus unedo, the thesis with altenative soil, biochar and humus was significantly better in all plant species for plant height, vegetative and root weight and root length. All alternative substrates showed a significant increase in microbial biomass compared to the control. Several materials have been evaluated as substitutes for or combination with peat in growing media formulation. They include coir dust, pine bark, and wood fiber as well as sand, tuff, and pumice processed materials (expanded clay, perlite, and vermiculite). Besides satisfying the relevant technical requirements, these alternatives have to be readily available at reasonable prices and be readily available in sufficient quantities. In addition to meeting specific plant requirements, these materials have been used to create tailored growing media that are environmentally friendly and reduce production costs. Conclusions: The results of research on growing substrates must be translated into agricultural practice through various steps, in cooperation with horticultural companies. To ensure the best growing conditions for seedlings, nurseries should conduct a thorough analysis of the specific composts they use. As organic farming practitioners argue, greater involvement of compost producers and end-users could improve the quality of growing media by adding additional valuable characteristics and reducing the use of peat, which would allow for a more sustainable regime.

FaMYB5 Interacts with FaBBX24 to Regulate Anthocyanin and Proanthocyanidin Biosynthesis in Strawberry (Fragaria × ananassa).

MYB and BBX transcription factors play important roles in flavonoid biosynthesis. Here, we obtained transgenic woodland strawberry with stable overexpression of FaMYB5, demonstrating that FaMYB5 can increase anthocyanin and proanthocyanidin content in roots, stems and leaves of woodland strawberry. In addition, bimolecular fluorescence complementation assays and yeast two-hybridization demonstrated that the N-terminal (1-99aa) of FaBBX24 interacts with FaMYB5. Transient co-expression of FaBBX24 and FaMYB5 in cultivated strawberry 'Xiaobai' showed that co-expression strongly promoted the expression of F3'H, 4CL-2, TT12, AHA10 and ANR and then increased the content of anthocyanin and proanthocyanidin in strawberry fruits. We also determined that FaBBX24 is also a positive regulator of anthocyanin and proanthocyanidin biosynthesis in strawberry. The results reveal a novel mechanism by which the FaMYB5-FaBBX24 module collaboratively regulates anthocyanin and proanthocyanidin in strawberry fruit.