The strawberry gene Cyf1 encodes a phytocystatin with antifungal properties (original) (raw)
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New insight in the study of strawberry fungal pathogens
G3-Genes Genomes Genetics
Strawberry (Fragaria ananassa) is one of the world's most commercially important fruit crops, and is grown in many countries The commercial viability of the crop is continually subject to various risks, one of the most serious of which is the diseases caused by phytopathogenic organisms. More than 50 different genera of fungi can affect this cultivar, including Botrytis spp., Colletotrichum spp., Verticillium spp., and Phytophthora spp. The development of new molecular biology technologies, based on genomics, transcriptomics and proteomics approaches, is revealing new insights on the diverse pathogenicity factors causing fungal invasion, degradation and destruction of the fruit (in planta and during storage and transport). Researchers have focused attention on the plant's own defence mechanisms against these pathogens. In this review, advances in the study and detection of fungal plant pathogens, new biocontrol methods, and proteomic approaches are described and the natural defence mechanisms recently discovered are reported.
New Insights in the Study of Strawberry Fungal Pathogens
Strawberry (Fragaria ananassa) is one of the world's most commercially important fruit crops, and is grown in many countries The commercial viability of the crop is continually subject to various risks, one of the most serious of which is the diseases caused by phytopathogenic organisms. More than 50 different genera of fungi can affect this cultivar, including Botrytis spp., Colletotrichum spp., Verticillium spp., and Phytophthora spp. The development of new molecular biology technologies, based on genomics, transcriptomics and proteomics approaches, is revealing new insights on the diverse pathogenicity factors causing fungal invasion, degradation and destruction of the fruit (in planta and during storage and transport). Researchers have focused attention on the plant's own defence mechanisms against these pathogens. In this review, advances in the study and detection of fungal plant pathogens, new biocontrol methods, and proteomic approaches are described and the natural defence mechanisms recently discovered are reported.
Isolation and Expression Analysis of a Defensin Gene from Strawberry (Fragaria×ananassa cv. Paros)
Journal of Agricultural Science and Technology, 2018
Plant defensins are the cysteine-rich peptides that are encoded by small multi-gene families in the plant kingdom. In this study, we designed primers based on conserved regions of defensin genes to clone and identify defensin genes in strawberry (Fragaria×ananassa cv. Paros) by reverse transcription PCR technique. Sequence analysis showed that the deduced amino acid had significant similarity to other plant defensins from NCBI database and designated FaDef1. The predicted strawberry defensin protein encodes a 54 aa protein of 6.18 kDa, pI 9.22 and eight conserved cysteine residues with desired space conservation with other amino acids. Semi quantitative expressions of FaDef1 were analyzed in root, stem, leaf, flower, and fruit in three strawberry cultivars, namely, Queenelisa, Camarosa, and Paros. The results showed that the FaDef1 expression patterns were similar in different tissues of the three cultivars. The higher amount of relative expression of FaDef1 was in fruit and there w...
Journal of experimental botany, 2011
Ectopic expression of the strawberry (Fragaria×ananassa) gene FaPE1 encoding pectin methyl esterase produced in the wild species Fragaria vesca partially demethylated oligogalacturonides (OGAs), which conferred partial resistance of ripe fruits to the fungus Botrytis cinerea. Analyses of metabolic and transcriptional changes in the receptacle of the transgenic fruits revealed channelling of metabolites to aspartate and aromatic amino acids as well as phenolics, flavanones, and sesquiterpenoids, which was in parallel with the increased expression of some genes related to plant defence. The results illustrate the changes associated with resistance to B. cinerea in the transgenic F. vesca. These changes were accompanied by a significant decrease in the auxin content of the receptacle of the ripe fruits of transgenic F. vesca, and enhanced expression of some auxin-repressed genes. The role of these OGAs in fruit development was revealed by the larger size of the ripe fruits in transgeni...
Archives of Biochemistry and Biophysics, 2007
The biosynthesis of flavonoids and proanthocyanidins was studied in cultivated strawberry (Fragaria · ananassa) by combining biochemical and molecular approaches. Chemical analyses showed that ripe strawberries accumulate high amounts of pelargonidin-derived anthocyanins, and a larger pool of 3 0 ,4 0 -hydroxylated proanthocyanidins. Activities and properties of major recombinant enzymes were demonstrated by means of in vitro assays, with special emphasis on specificity for the biologically relevant 4 0 -and 3 0 ,4 0 -hydroxylated compounds. Only leucoanthocyanidin reductase showed a strict specificity for the 3 0 ,4 0 -hydroxylated leucocyanidin, while other enzymes accepted either hydroxylated substrate with different relative activity rates. The structure of late flavonoid pathway genes, leading to the synthesis of major compounds in ripe fruits, was elucidated. Complex developmental and spatial expression patterns were shown for phenylpropanoid and flavonoid genes in fruits throughout ripening as well as in leaves, petals and roots. Presented results elucidate key steps in the biosynthesis of strawberry flavonoid end products.
Frontiers in Plant Science, 2019
Gray mold caused by Botrytis cinerea is a major cause of economic losses in strawberry fruit production, limiting fruit shelf life and commercialization. When the fungus infects Fragaria × ananassa strawberry at flowering or unripe fruit stages, symptoms develop after an extended latent phase on ripe fruits before or after harvesting. To elucidate the growth kinetics of B. cinerea on flower/fruit and the molecular responses associated with low susceptibility of unripe fruit stages, woodland strawberry Fragaria vesca flowers and fruits, at unripe white and ripe red stages, were inoculated with B. cinerea. Quantification of fungal genomic DNA within 72 h postinoculation (hpi) showed limited fungal growth on open flower and white fruit, while on red fruit, the growth was exponential starting from 24 hpi and sporulation was observed within 48 hpi. RNA sequencing applied to white and red fruit at 24 hpi showed that a total of 2,141 genes (12.5% of the total expressed genes) were differentially expressed due to B. cinerea infection. A broad transcriptional reprogramming was observed in both unripe and ripe fruits, involving in particular receptor and signaling, secondary metabolites, and defense response pathways. Membranelocalized receptor-like kinases and nucleotide-binding site leucine-rich repeat genes were predominant in the surveillance system of the fruits, most of them being downregulated in white fruits and upregulated in red fruits. In general, unripe fruits exhibited a stronger defense response than red fruits. Genes encoding for pathogenesis-related proteins and flavonoid polyphenols as well as genes involved in cell-wall strengthening were upregulated, while cell-softening genes appeared to be switched off. As a result, B. cinerea remained quiescent in white fruits, while it was able to colonize ripe red fruits.
Genomics, Transgenics, Molecular Breeding and Biotechnology of Strawberry
Strawberry is one of the most popular fruits due to its delicious flavor and attractive aspect, and the demand for strawberries, as well as its culture, is continuously increasing worldwide. This crop, however, is not absent of problems that compromise yield or fruit quality. Actually, strawberry is considered by the consumers as one of the most inconsistent commodities in the marketplace. The use of highly producer cultivars with large but poorly flavored fruits, incorrect crop management practices such as harvesting the fruits before full ripening, the short postharvest life of strawberry fruits or the extensive use of dangerous fumigants to control pest are among the main problems of this crop. Nowadays, people demand plant biology researchers to focus more efforts on safer and sustainable agriculture, avoiding as much as possible the use of chemicals, and to improve fruit quality traits. The implementation of biotechnological approaches within traditional breeding programs can be helpful to reach this goal. Besides this practical view, strawberry has been adopted as a model crop for some molecular and physiological studies, such as the ripening of non climacteric fruits, increasing their importance in the scientific world. The aim of this volume is to provide updated information about the current stage of genomics, transgenesis and biotechnology in strawberry, through the inclusion of reviews and research papers wrote by leader groups in these areas. Molecular markers have been explored in strawberry from the 80´s but in recent years the development of high throughput sequencing technologies has increased dramatically the amount of genomic data in this species. These advances in structural genomics in the genus Fragaria have been reviewed by Bonet and Monfort, and the release of the first draft of the strawberry genome is expected very soon. Most strawberry cultivars show seasonal flowering, requiring short photoperiod and/or low temperature for the induction of flowering, while the opposite environmental signals are needed for runnering. Hytönen and Elomaa have reviewed our current knowledge about candidate genes involved in flowering induction. This review is complemented by the paper of Massetani et al. who examine the effect of abiotic stresses, nutritional factors and propagation techniques on strawberry plant architecture. These studies can provide efficient tools to extend cropping season and increase berry yields. Besides attractive flavor, strawberries are a rich source of phytochemicals beneficial for human health. Hanhieneva et al. and Schmeda et al. have reviewed recent advances in metabolomics of both, cultivated strawberry and the Chilean strawberry Fragaria chiloensis, respectively. While both papers highlight the chemical composition of fruits, data about metabolomics of vegetative organs are also included. Increasing postharvest shelf life of strawberry fruit is central to strawberry research, which is very short mainly due to its fast softening rate and enhanced pathogen susceptibility usually associated with this process. Cell wall disassembly and loss of cell turgor during fruit ripening are considered the main factors responsible for fruit softening. Posé et al. have reviewed the main features of the disassembly cell wall process, and how the manipulation of some cell wall genes through transgenesis can reduce fruit softening without affecting other fruit quality parameters, while Alleva et al. examine the role of the water channel aquaporins in the loss of cell turgor during fruit ripening. For a limited number of genes, their role on fruit ripening has been assessed by stable genetic transformation. However, the increased knowledge of genes involved in fruit ripening requires efficient and fast systems for gene function analysis. Schawb et al. show how gene down-regulation by RNA interference in agroinfiltration experiments has been successfully used to assess the role of several fruit specific genes, avoiding time-consuming and labor-intensive stable transformation systems. Genetic modification could be a successful approach in improving quality of strawberry crop, as has been demonstrated with several fruit ripening genes. The application of this technology requires robust regeneration and transformation systems for each particular genotype. Currently, Agrobacterium tumefaciens infection in combination with a leaf disk regeneration system is the most popular method to get transgenic strawberry plants. Husaini et al. analyze key factors determining the success of genetic transformation in this species, and describe a general transformation protocol that has been proven useful in several strawberry cultivars. However, the toughest impediment in the development of this technology is the poor consumer perception of transgenic crops. These concerns are examined by Schaart et al. They describe the results of a recent consumer survey indicating that transgenic crops would be better accepted if only genes from the species itself, the so called intragenic or cisgenic plants, were used. This review also describes the attempts to get intragenic strawberry plants resistant to Botrytis cinerea. Intragenic modification requires the use of native promoters to drive gene expression. In their research article Schaart et al. describe the characterization of a promoter sequence of the strawberry fruit expansin FaExp2 gene. One of the main problems of the strawberry industry is the high amount of pesticides required to avoid loss of production, mainly due to fungal infections. Natural resistance against important diseases is scarce within the genus Fragaria, and therefore, alternative approaches to get tolerant cultivars are quite desirable. Garrido et al. summarize recent advances in the methods for detection and identification of strawberry fungal pathogens as well as strategies for their biocontrol using microorganisms. In a research paper, Matsubara et al. show that arbuscular mycorrhizal colonization enhances tolerance to Fusarium wilt and this effect is related to an increased level of antioxidative ability. Finally, Shokaeva et al. describe a method for the obtainment of plants tolerant to Botrytis cinerea or Phytophtora cactorum, and also to salinity stress, through screening and selection of somaclonal variants developed after exposure to these stressors under in vitro conditions. As many people that enjoy consuming strawberries, for us too it has been a pleasure to work in the preparation of this project. We hope that readers of this special issue, biotechnologists, physiologists, breeders or simply strawberry lovers, enjoy the reviews and find them useful. We are thankful to our families for having adjusted with us, while we were busy for long hours during the preparation of this volume. We finally thank all authors for their excellent contributions, and also for their patience during the long time required to complete this hybrid book, and to the GSB Editor-in-Chief Dr. Jaime Teixeira da Silva for his generous support.
The Strawberry Plant Defense Mechanism: A Molecular Review
Plant and Cell Physiology, 2011
Strawberry, a small fruit crop of great importance throughout the world, has been considered a model plant system for Rosaceae, and is susceptible to a large variety of phytopathogenic organisms. Most components and mechanisms of the strawberry defense network remain poorly understood. However, from current knowledge, it seems clear that the ability of a strawberry plant to respond efficiently to pathogens relies first on the physiological status of injured tissue (pre-formed mechanisms of defense) and secondly on the general ability to recognize and identify the invaders by surface plant receptors, followed by a broad range of induced mechanisms, which include cell wall reinforcement, production of reactive oxygen species, phytoalexin generation and pathogenesis-related protein accumulation. Dissection of these physiological responses at a molecular level will provide valuable information to improve future breeding strategies for new strawberry varieties and to engineer strawberry plants for durable and broad-spectrum disease resistance. In turn, this will lead to a reduction in use of chemicals and in environmental risks. Advances in the understanding of the molecular interplay between plant (mainly those considered model systems) and various classes of microbial pathogens have been made in the last two decades. However, major progress in the genetics and molecular biology of strawberry is still needed to uncover fully the way in which this elaborate plant innate immune system works. These fundamental insights will provide a conceptual framework for rational human intervention through new strawberry research approaches. In this review, we will provide a comprehensive overview and discuss recent advances in molecular research on strawberry defense mechanisms against pathogens.