Proteomic variation is as large within as between strawberry varieties

In search for a strawberry (Fragaria ananassa) with low allergen content, we determined the proteomic variation within and between different varieties. Proteomics data were generated by DIGE and proteins identified with MALDI-MS/MS. The amount of the strawberry allergen Fra a 1 varied between different strawberry varieties (CV = 39%). The variation was at the same level, or even slightly larger, due to different growth conditions (CV = 43%). For 153 other proteins, the biological variation was more affected by different growth conditions than by different varieties (mean CV = 52% and 43%, respectively) due to variation in a subset of proteins. Thus, the allergen variation due to growth conditions must be taken into consideration in attempts to obtain a low-allergen strawberry. However, the allergen content was always lower in colorless (white) strawberry varieties than in the red ones. Moreover, of the spots whose expression correlated with the allergen and the color (32 and 68, respectively), only 3 were the same. This implies that these two phenotypic traits are not inseparable, and it may be possible to breed a red strawberry with low amount of allergen.     

Ability of the antagonistic yeast Cryptococcus albidus to control Botrytis cinerea in strawberry

The yeast Cryptococcus albidus, originally isolated from mature strawberry fruits, was tested for antagonistic activity against Botrytis cinerea, the causal agent of grey mould in strawberries. Conidial germination and germ tube growth of conidia of B. cinerea were inhibited by a cell suspension of the antagonist in aqueous strawberry fruit pulp suspension (1%) after 6 and 24 hours of incubation. Application of a cell suspension (1 × 10⁶ cells/ml) on detached strawberry leaf disks incubated at 10°C reduced incidence and conidiophore density of B. cinerea by 86 and 99%, respectively, but effectiveness was reduced at higher temperatures. Treatments with C. albidus during bloom of strawberries reduced incidence of grey mould on ripe strawberry fruits after harvest by 33, 28 and 21% in three years of field trials. The effectiveness of the yeast was increased when formulation substances (alginate, xanthan and cellulose) were added to the cell suspension.     

The control of red colour by a family of MYB transcription factors in octoploid strawberry (Fragaria × ananassa) fruits

Octoploid strawberry (Fragaria × ananassa Duch.) is a model plant for research and one of the most important non‐climacteric fruit crops throughout the world. The associations between regulatory networks and metabolite composition were explored for one of the most critical agricultural properties in octoploid strawberry, fruit colour. Differences in the levels of flavonoids are due to the differences in the expression of structural and regulatory genes involved in flavonoid biosynthesis. The molecular mechanisms underlying differences in fruit colour were compared between red and white octoploid strawberry varieties. FaMYB genes had combinatorial effects in determining the red colour of fruit through the regulation of flavonoid biosynthesis in response to the increase in endogenous ABA at the final stage of fruit development. Analysis of alleles of FaMYB10 and FaMYB1 in red and white strawberry varieties led to the discovery of a white‐specific variant allele of FaMYB10, FaMYB10‐2. Its coding sequence possessed an ACTTATAC insertion in the genomic region encoding the C‐terminus of the protein. This insertion introduced a predicted premature termination codon, which suggested the loss of intact FaMYB10 protein playing a critical role in the loss of red colour in white octoploid strawberry.     

Isolation of an osmotin-like protein gene from strawberry and analysis of the response of this gene to abiotic stresses

A strawberry genomic clone containing an osmotin-like protein (OLP) gene, designated FaOLP2, was isolated and sequenced. FaOLP2 is predicted to encode a precursor protein of 229 amino acid residues, and its sequence shares high degrees of homology with a number of other OLPs. Genomic DNA hybridization analysis indicated that FaOLP2 represents a multi-gene family. The expression of FaOP2 in different strawberry organs was analyzed using real-time PCR. The results showed that FaOLP2 expressed at different levels in leaves, crowns, roots, green fruits and ripe red fruits. In addition, the expression of FaOLP2 under different abiotic stresses was analyzed at different time points. All of the three tested abiotic stimuli, abscisic acid, salicylic acid and mechanical wounding, triggered a significant induction of FaOLP2 within 2-6h post-treatment. Moreover, FaOLP2 was more prominently induced by salicylic acid than by abscisic acid or mechanical wounding. The positive responses of FaOLP2 to the three abiotic stimuli suggested that strawberry FaOLP2 may help to protect against osmotic-related environmental stresses and that it may also be involved in plant defense system against pathogens.     

beta-xylosidase activity and expression of a beta-xylosidase gene during strawberry fruit ripening.

Strawberry fruit shows a marked softening during ripening and the process is associated with an increment of pectin solubility and a reduction of the molecular mass of hemicelluloses. In this work, we report the activity of beta-xylosidase and the expression of a beta-xylosidase gene in strawberry fruit. We have cloned a cDNA fragment encoding a putative beta-xylosidase (FaXyl1) from a cDNA library obtained from ripe strawberry fruit. The analysis of the deduced amino acid sequence revealed that FaXyl1 is closely related to other beta-xylosidases from higher plants. The expression of FaXyl1 was strongly associated to the receptacle tissue although a low expression level was detected in achenes and ovaries. The accumulation of FaXyl1 mRNA is ripening-related, starting in white fruit, reaching the maximum at 25-50% red fruit and decreasing thereafter. The total beta-xylosidase enzyme activity was detected in all ripening stages with the maximum in 25-50% red fruit. The low activity level detected in immature stages, where no expression of FaXyl1 was found, suggests the presence of other beta-xylosidases-like genes. Both the expression of FcaXyl1 and the total beta-xylosidase activity were down regulated by auxins, as occurs for most of the ripening-related processes in strawberry fruit. A putative role of FaXyl1 and beta-xylosidase is discussed.     

Companion planting to attract pollinators increases the yield and quality of strawberry fruit in gardens and allotments

1. Global pollinator declines have led to concern that crop yields might fall as a result of a pollination deficit. Companion planting is a traditional practice thought to increase yield of insect pollinated crops by planting a co-flowering species next to the crop. 2. Using a combination of conventional researcher-led experiments and observational citizen scientist data, we tested the effectiveness of bee-friendly borage (Borago officinalis) as a companion plant to strawberry (Fragaria x ananassa). Insect visitors to the ‘Test’ (strawberry + borage) versus ‘Control’ (strawberry only) plants were observed, and strawberry fruit collected. Strawberries collected during the researcher-led experiment were also subject to fruit measurements and assessments of market quality. 3. Companion plants were found to significantly increase both yield and market quality of strawberries, suggesting an increase in insect pollination per plant. Test strawberries companion planted with borage produced an average of 35% more fruits, and 32% increased yield by weight. Test strawberry plants produced significantly more fruit of higher aesthetic quality when assessed by Marketing Standards for Strawberries. 4. Although there was no significant difference in the overall insect visits, when broken down by broad insect group there were significantly more flies visiting the test strawberries than controls. 5. These results could have implications for both gardeners and commercial growers. As consumers prefer a cosmetically perfect fruit, the production of fruit with increased aesthetics aids food waste reduction.     

Acarofauna present in organic strawberry fields and associated weed species in southern Brazil

The presence of weeds in the margins of strawberry crops can enhance populations of predatory mites as a measure to support conservation biological control. The aims of this study were (i) to assess the composition of the acarofauna associated with strawberries and the accompanying herbaceous plants in an organic farming system, and (ii) to evaluate the possible relationships between phytophagous and predatory mites occurring in this system. Strawberry leaves and whole plants of weeds were sampled biweekly from August 2014 to February 2015 in Lapa (Paraná, Brazil). In total, 23 weed species belonging to 10 families were identified; 3768 mite individuals (from 15 families and 4 suborders) were recovered, 77% on strawberries and 23% on weeds. Abundance of predatory mites on weeds was greater than on strawberry cultivars. On strawberries, the most abundant family was Tetranychidae (84%) followed by Phytoseiidae (11.6%). In total, 16 predatory mite species from the Phytoseiidae family were identified, 13 of them occurring on strawberry leaflets. Typholodromalus aripo, Neoseiulus californicus and Typhlodromips mangleae were the most abundant mite species on strawberry leaves. On weeds, most individuals were predatory mites (59%), whereas phytophagous mites represented 17.2%. The most abundant family was Phytoseiidae (36.4%). On weeds, the phytoseiid mite T. aripo was the most abundant species, representing 34.7%. Besides being found on strawberry leaflets, T. aripo was associated with 15 weed species. Among the weeds, Bidens pilosa showed the highest values of the Shannon index (1.97), Margalef index (3.04), and Pielou’s evenness index (0.95). This study emphasizes the importance of surrounding weeds as a shelter for beneficial mitefauna in strawberry fields, likely contributing to enhance conservation biological control.

     

Decision-making tools for Frankliniella occidentalis management in strawberry: consideration of target markets

The western flower thrips (WFT), Frankliniella occidentalis Pergande (Thysanoptera: Thripidae), a cosmopolitan pest of many crops, is considered a major pest of low tunnel and greenhouse strawberries. The extent of damage to strawberry is unclear because different studies have produced contradictory results. Also, economic thresholds published for WFT in strawberry vary greatly, and most fail to incorporate economic factors. This study was aimed at developing a decision‐making tool for WFT management in strawberries in Israel. Toward this end, economic injury levels (EIL) and economic thresholds were calculated, based on target markets (export vs. domestic). Results indicate that serious infestation of ripe berries may cause a dull, rough appearance, and the fruit may be soft and have a reduced shelf life, rendering it unsuitable for export. Most fruit damage occurred at green and turning‐red stages of development. Two decision‐making tools were developed, one for winter, when WFT populations increase slowly but crop value is high (export market); and the second for spring, when the pest increases rapidly but crop value is low (local markets). Economic thresholds of 10 and 24 WFT/flower were calculated for winter and spring strawberries, respectively, based on direct thrips damage to fruit. This calculation does not take into account the recorded WFT damage to flowers, or its role in facilitating Botrytis cinerea fruit infection. Western flower thrips has proved only an occasional economic pest in Israeli strawberries, and no routine control measures are warranted. Furthermore, augmentative releases of Orius laevigatus or Neoseilus cucumeris against WFT are not justified in this system, because Orius colonizes strawberry fields spontaneously in high numbers when no broad spectrum insecticides are used.     

Brassinosteroid is involved in strawberry fruit ripening

Although brassinosteroid (BR) has been suggested to play a role in strawberry fruit ripening, the defined function of this hormone remains unclear in the fruit. Here, BR content and BR receptor gene FaBRI1 expression were analysed during ??Akihime?? strawberry fruit development. We found that BR levels increased during the later developmental stages, and the mRNA expression levels of FaBRI1 increased rapidly from white to initial red stages, suggesting that BR is associated with fruit ripening. This was further confirmed by exogenous application of BR and its inhibitor brassinazole (BZ) to big-green fruit, which significantly promoted and inhibited strawberry fruit ripening, respectively. More importantly, down-regulation of FaBRI1 expression in de-greening fruit markedly retarded strawberry red-colouring. In conclusion, we have provided physiological and molecular evidence to demonstrate that BR plays a role in strawberry fruit ripening. In addition, both BR content and FaBRI1 expression reached their peak levels in small-green fruit, suggesting that BR might also be involved in early strawberry fruit development. Further experiments are required to validate the role of BR in strawberry fruit cell division.     

草莓果实成熟期间RNA代谢的变化

现已证明,鳄梨和番茄等跃变型果实的成熟涉及基因表达的改变和新蛋白质(酶)的合成(Brady 1987)。但是,关于草莓这类非跃变型果实成熟的生化控制机理却很少报道,它们是否也如同跃变型果实那样与新的核酸和蛋白质合成有关?本实验测定了草莓果实成熟期间poly(A)+RNA含量的变化和poly(A)+RNA的体外翻译活性,目的在于探讨草莓果实的成熟机     

Identification of a strawberry gene encoding a non-specific lipid transfer protein that responds to ABA,wounding and cold stress

cDNA and genomic clones encoding a strawberry (Fragariaxananassa cv. Chandler) non-specific lipid transfer protein (Fxaltp gene) were isolated and characterized. The spatio-temporal expression pattern and structural features of this gene were studied for the first time in strawberry, a non-climacteric fruit of agricultural importance. The architecture and the encoded amino acid sequence of this non-climacteric fruit ltp gene were similar to those of other plant LTPs previously reported, and presents the eight cysteine residues and other features characteristic of plant LTPs. In addition, the deduced protein posseses an N-terminal signal peptide and lacks the K/HDEL retention signal, indicating that the strawberry LTP protein would enter the secretory pathway. In situ studies have shown that the Fxaltp gene is expressed in the epidermal cell layer of the strawberry fruit receptacle and achenes, flowers, and within the cell layer surrounding the endosperm. These results suggest that this Fxaltp gene promoter could be used as an endogenous promoter for biotechnological purposes in strawberry. Computer analysis using the PLACE database predicted the presence of several putative cis-regulatory sequences in response to abscisic acid and cold or wounding stresses within the Fxaltp 5'-flanking region. Accordingly, the strawberry gene responds to ABA and SA, but not to salt and heat stresses. It is also reported that ltp gene expression in strawberry is stimulated by wounding and repressed by cold stresses.     

Arbuscular mycorrhizae formed by Penicillium pinophilum improve the growth, nutrient uptake and photosynthesis of strawberry with two inoculum-types

Penicillium pinophilum was isolated from the soil in a commercial strawberry field. The strain readily formed arbuscular mycorrhizae (AM) with the roots of strawberry 'Zoji' (Fragaria x ananassa Duch. CV.) when plants were inoculated with either fresh cultured hyphae or root/soil mixtures. Fresh hyphae, however, resulted in higher amounts of colonization than root/soil inoculum. Compared with uninoculated strawberries, inoculation increased plant dry weight by 31%, as well as nitrogen content (47%), phosphorus content (57%), and photosynthetic rate (71%). AM inoculation also shortened the blossom and ripening date by 3 and 4 days, respectively. This is the first report of a P. pinophilum strain resulting in mycorrhiza with strawberry roots. The significant advantages of this strain are that it is easy to culture and inoculation of plants results in significant growth benefits that may be useful in strawberry production.     

Downregulation of RdDM during strawberry fruit ripening

Background

Recently, DNA methylation was proposed to regulate fleshy fruit ripening. Fleshy fruits can be distinguished by their ripening process as climacteric fruits, such as tomatoes, or non-climacteric fruits, such as strawberries. Tomatoes undergo a global decrease in DNA methylation during ripening, due to increased expression of a DNA demethylase gene. The dynamics and biological relevance of DNA methylation during the ripening of non-climacteric fruits are unknown.

Results

Here, we generate single-base resolution maps of the DNA methylome in immature and ripe strawberry. We observe an overall loss of DNA methylation during strawberry fruit ripening. Thus, ripening-induced DNA hypomethylation occurs not only in climacteric fruit, but also in non-climacteric fruit. Application of a DNA methylation inhibitor causes an early ripening phenotype, suggesting that DNA hypomethylation is important for strawberry fruit ripening. The mechanisms underlying DNA hypomethylation during the ripening of tomato and strawberry are distinct. Unlike in tomatoes, DNA demethylase genes are not upregulated during the ripening of strawberries. Instead, genes involved in RNA-directed DNA methylation are downregulated during strawberry ripening. Further, ripening-induced DNA hypomethylation is associated with decreased siRNA levels, consistent with reduced RdDM activity. Therefore, we propose that a downregulation of RdDM contributes to DNA hypomethylation during strawberry ripening.

Conclusions

Our findings provide new insight into the DNA methylation dynamics during the ripening of non-climacteric fruit and suggest a novel function of RdDM in regulating an important process in plant development.