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PubMed2006

Characterization of LF9, an octoploid strawberry genotype selected for rapid regeneration and transformation.

Cultivated strawberry (Fragaria xananassa) is a valuable crop, yet the absence of a rapid, high-throughput transgenic system has precluded meaningful application of biotechnology and translation of information from plant models to this crop. A new octoploid strawberry genetic line Laboratory Festival #9 has been identified, selected solely for its rapid regeneration and efficient transformation. Direct organogenesis has been achieved from all tissues tested, with rapidly-growing shoot initials visible in as few as 13 days. The conditions for optimal shoot regeneration, transformant selection, root generation, and plant acclimatization are presented. The progression from explant to plant in soil can be achieved in about 60 days. The development of transformation protocols in this rapid-cycling genotype allows high-throughput studies of gene function in the octoploid strawberry genetic background.

Acclimatization↗

PubMed2006

Strawberry fruit protein with a novel indole-acyl modification.

Achenes and receptacle tissue of Fragaria vesca, L. cultivar Yellow Wonder were shown to contain conjugated indole-3-acetic acid (IAA) that was not soluble in organic solvents and yielded IAA after strong alkaline hydrolysis, suggestive of IAA attached to plant proteins. This solvent insoluble conjugated IAA accounted for between 0.4 and 4 ng of IAA per gram fresh weight of tissue in both achenes and receptacles. To investigate this strawberry conjugate class further, a polyclonal antibody was produced to IAA-glycine attached to BSA that detected neutral indole acid esters, monocarboxylic-amino acid IAA conjugates and IAA proteins. Using immunoblotting, both achenes and receptacles of strawberry were shown to have primarily an immuno-detectable band at 76 kDa. Two-dimensional polyacrylamide gel electrophoresis yielded a wide band that was analyzed by LC-MS/MS analysis following in-gel trypsin digestion. Peptides derived from the immuno-detectable band were tentatively identified by peptide fragment analysis as being from either a chaperonin related to the hsp60 class of proteins or, alternatively, an ATP synthase. This is one of the first reports of an IAA modified protein in fruit tissue.

Fragaria↗

PubMed2002

Root cooperation in a clonal plant: connected strawberries segregate roots.

The ability to selectively avoid competition with members of the same clone should be highly advantageous but has not been demonstrated in plants. We found that physical connection between plants in a clone of the wild strawberry Fragaria chiloensis induced them to segregate their roots, significantly increasing clonal performance. Such increase in performance was not found when plants were grown in containers that artificially divided their rooting zones. There was no effect of connection in a different clone of F. chiloensis with a lower degree of carbon transport between connected plants, suggesting that the mechanism for root segregation depended upon transport of a signal through the strawberry runners. We suggest that clonal integration allows some clones to coordinate below-ground resource foraging with other clone members, thus exhibiting a type of root cooperation.

Biomass↗

PubMed2004

Prey stage preference and functional response of Euseius hibisci to Tetranychus urticae (Acari: Phytoseiidae, Tetranychidae).

The aims of this study were: (a) determine the prey stage preference of female Euseius hibisci (Chant) (Phytoseiidae) at constant densities of different stages of Tetranychus urticae Koch (Tetranychidae), (b) assess the functional response of the predator females to the varying densities of eggs, larvae, or protonymphs of T. urticae, and (c) estimate the functional response of E. hibisci when pollen of Ligustrum ovalifolium was present as well. We conducted experiments on excised pieces of strawberry leaf arenas (Fragaria ananassa) under laboratory conditions of 25+/-2 degrees C, 60+/-5% RH and 12 h photophase. Our results indicated that the predator consumed significantly more prey eggs than other prey stages. Consumption of prey deutonymphs and adults was so low that they were excluded from the non-choice functional response experiments. The functional response on all food items was of type II. The two parameters of the functional response were estimated for each prey type by means of the adjusted non-linear regression model. The highest estimated value a' (instantaneous rate of discovery) and the lowest value of Th (handling time, including digestion) were found for the predator feeding on prey eggs, and a' was lowest and Th highest when fed protonymphs. Using the jack-knife method, the values for the functional response parameters were estimated. The values of a' and Th produced by the model were similar among all prey types except for the eggs, which were different. Using pollen simultaneously with prey larvae decreased the consumption of the latter over the full range of prey densities The suitability of this predator for biological control of T. urticae on strawberry is discussed.

Animals↗

PubMed2004

A strawberry fruit-specific and ripening-related gene codes for a HyPRP protein involved in polyphenol anchoring.

A strawberry (Fragaria x ananassa cv. Chandler) fruit cDNA (Fahyprp -cDNA) and its corresponding gene (Fahyprp) showing sequence homology to higher plant hyprp genes have been isolated. The cDNA contains an open reading frame encoding a 16 kDa protein with 156 amino acids. The peptide has an amino-terminal signal sequence, a repetitive proline-rich sequence, and a cysteine-rich carboxy-terminal region homologous to other HyPRP proteins. Northern blot and QRT-PCR analysis have shown that the strawberry transcript is specifically expressed in fruit, not being detected in other plant tissues. " In situ " hybridization and immunolocalization studies have indicated that the Fahyprp gene is strongly expressed in achene sclerenchyma cells, in the vascular and receptacle cells of immature green fruit and in the vascular cells of mature red fruits. The achenes removal from unripe green fruits induced the expression of this Fahyprp gene. This induction was reverted by treatment of deachened fruit with the auxin NAA, supporting the idea that Fahyprp gene expression is regulated by auxins. Furthermore, the HyPRP protein has been localized in parenchymatic cells of immature fruits associated to structures containing condensed tannins. The results are discussed supporting a putative role of this protein in the anchoring of polymeric polyphenols in the strawberry fruit during growth and ripening.

5' Flanking Region↗

PubMed2004

Influences of vermicomposts on field strawberries: 1. effects on growth and yields.

Vermicomposts processed commercially from food wastes and paper wastes were applied, to 4.5 m(2) field plots, under high plastic hoop tunnels, at rates of 5 or 10 tha(-1) to evaluate their effects on the growth and yields of strawberries (Fragaria ananasa) var. 'Chandler'. The vermicomposts were incorporated into the top 10 cm of soil and supplemented, based on chemical analyses, with amounts of inorganic NPK fertilizers calculated to equalize the initial fertilizer rates of 85-155-125 kgha(-1) NPK applied to the inorganic fertilizer plots. All treatments were replicated four times, in a completely randomized design, at two field sites on Doles silt loam or Hoytville silty clay loam at Piketon and Fremont, Ohio, respectively. Vermicompost applications increased strawberry growth and yields significantly; including increases of up to 37% in leaf areas, 37% in plant shoot biomass, 40% in numbers of flowers, 36% in numbers of plant runners and 35% in marketable fruit weights. These responses seemed not to be dose-dependent, since strawberries at one site grew fastest and yielded most in response to the 10 tha(-1) vermicompost application rate, whereas they responded positively and similarly to both the 5 and 10 tha(-1) rates of applications at the other site. These responses could not have been mediated by availability of macronutrients, since all plots were supplemented with inorganic fertilizers, to equalize macronutrient inputs for all treatments, but based on other research in our laboratory could have been due to production of plant growth regulators by microorganisms during vermicomposting.

Agriculture↗

PubMed2005

Influences of vermicomposts on field strawberries: part 2. Effects on soil microbiological and chemical properties.

The effects of applications of food waste and paper waste vermicomposts on some soil chemical and biological properties were evaluated in field plots planted with strawberries. Six-week old strawberries (Fragaria ananasa, var. Chandler) were transplanted into 4.5 m(2) raised beds under a plastic tunnel structure measuring 9.14 x 14.6 x 3.6 m. Vermicompost were applied at rates of 5 or 10 t ha(-1) supplemented with inorganic fertilizers to balance fertilizer recommendations for strawberries of 85-155-125 kg NPK ha(-1). Effects of vermicomposts on strawberry growth and yields have been reported previously [Arancon, N.Q., Edwards C.A., Bierman P., Welch, C., Metzger, J.D., 2004. The influence of vermicompost applications to strawberries: Part 1. Effects on growth and yield. Bioresource Technology 93:145-153]. Total extractable N, NH(4)-N, NO(3)-N and orthophosphates did not differ significantly between treatments, except on the last sampling date (harvest date) in which significantly greater amounts of NH(4)-N, NO(3)-N and orthophosphates (P <or= 0.05) were recorded in vermicompost-treated soils than in the controls. Two major results of vermicompost applications to soils were increases in dehydrogenase activity and microbial biomass-N which were not dose-dependent. Increased dehydrogenase activity and microbial biomass-N was correlated positively with the increased amounts of NH(4)-N, NO(3)-N and orthophosphates in the vermicompost-treated plots than in the controls. Increases in microbial populations and activities are key factors influencing rates of nutrient cycling, production of plant growth-regulating materials, and the build-up of plant resistance or tolerance to crop pathogen and nematode attacks.

Agriculture↗

PubMed2005

In vitro antileukaemic activity of extracts from berry plant leaves against sensitive and multidrug resistant HL60 cells.

The aim of the present study was to determine in vitro antileukaemic activity of extracts obtained from selected berry plant leaves (Fragaria x ananassa Duch. cv Elsanta, raspberry Rubus ideus L. cv Polana and blueberry Vaccinium corymbosum L. cv Bluecrop) against promyelocytic HL60 cell line and its multidrug resistant sublines exhibiting two different MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/DOX (overexpressing MRP1 protein). It was found that the blueberry extract was the most efficient against sensitive HL60 cell line (about 2-fold more active than strawberry and raspberry extracts) but presented much lower activity towards resistant cells. In contrast, strawberry and raspberry extracts exhibited the high cytotoxic activity against sensitive leukaemia HL60 cell line as well as its MDR sublines. The values of resistance factor (RF) found for these extracts were very low lying in the range 0.32/2.0.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2005

Characterization of microsomal and mitochondrial phospholipase D activities and cloning of a phospholipase D alpha cDNA from strawberry fruits.

Phospholipase D alpha (PLD, EC 3.1.4.4)) is a key enzyme involved in membrane deterioration that occurs during fruit ripening and senescence. The biochemical and molecular characteristics of PLD was studied in strawberry (Fragaria ananassa Duch) fruits, which are non-climacteric fruits. PLD activity was primarily associated with the mitochondrial and microsomal fractions and showed increased activity during development. Optimal pH levels of activity were observed at 5.5 and 6.5 for mitochondrial PLD and at 5 and 7 for microsomal PLD. Calcium enhanced microsomal PLD activity at 1-40 microM levels. PLD activity followed Michaelis-Menten kinetics. Lineweaver-Burk analysis gave Km values in the range of 114 and 277 microM using dipalmitoylphosphatidylcholine (DPPC) as substrate for mitochondrial and microsomal PLD, respectively. The Vmax value for the microsomal PLD was nearly 12-fold higher than that of mitochondrial PLD. A 2874 bp full-length cDNA for PLD alpha was amplified from strawberry fruit mRNA using RT-PCR and 5'- and 3'-RACE encoding an 810 amino acid-polypeptide. The predicted strawberry PLD sequence showed the characteristic C2 domain and the phospholipase domains conferring calcium sensitivity and the enzyme activity, respectively. The strawberry PLD alpha showed a high degree of similarity to other PLD alphas from plants. The implications of PLD regulation during ripening of fruits are discussed.

Amino Acid Sequence↗

PubMedDate not supplied

Strawberry pectin methylesterase (PME): purification, characterization, thermal and high-pressure inactivation.

Pectin methylesterase (PME) was extracted from strawberries (Fragaria ananassa, cv Elsanta) and purified by affinity chromatography on a CNBr-Sepharose 4B-PME-inhibitor column. A single protein and PME activity peak was obtained. A biochemical characterization in terms of molecular mass, pI, and kinetic parameters of strawberry PME was performed. In a second step, the thermal and high-pressure stability of the enzyme was studied. Isothermal and combined isothermal-isobaric inactivation of purified strawberry PME could be described by a fractional-conversion model. Purified strawberry PME is much more stable toward high-pressure treatments in comparison to those from oranges and bananas.

Carboxylic Ester Hydrolases↗

PubMed2003

Elevated carbon dioxide increases contents of antioxidant compounds in field-grown strawberries.

The effects of elevated CO2 concentrations on the antioxidant capacity and flavonoid content in strawberry fruit (Fragaria x ananassa Duch.) were studied under field conditions. Increased CO(2) (300 and 600 micromol mol(-1) above ambient) concentrations resulted in increases in ascorbic acid (AsA), glutathione (GSH), and ratios of AsA to dehydroascorbic acid (DHAsA) and GSH to oxidized glutathione (GSSG), and a decrease in DHAsA in strawberry fruit. High anthocyanin and phenolic content were also found in fruit of CO(2) treated plants. Growing strawberry plants under CO(2) enrichment conditions significantly enhanced fruit p-coumaroylglucose, dihydroflavonol, quercetin 3-glucoside, quercetin 3-glucuronide, and kaempferol 3-glucoside contents, as well as cyanidin 3-glucoside, pelargonidin 3-glucoside, and pelargonidin 3-glucoside-succinate content. Fruit of strawberry plants grown in the CO(2) enrichment conditions also had high oxygen radical absorbance activity against ROO(*), O(2)(*-), H(2)O(2), OH(*), and (1)O(2) radicals.

Antioxidants↗

PubMed2003

Compost as a soil supplement increases the level of antioxidant compounds and oxygen radical absorbance capacity in strawberries.

Compost as a soil supplement significantly enhanced levels of ascorbic acid (AsA) and glutathione (GSH) and ratios of AsA/dehydroascorbic acid (DHAsA) and GSH/oxidized glutathione (GSSG) in fruit of two strawberry (Fragaria x ananassa Duch.) cultivars, Allstar and Honeoye. The peroxyl radical (ROO(*)) as well as the superoxide radical (O(2)(*)(-)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (OH(*)), and singlet oxygen ((1)O(2)) absorbance capacity in strawberries increased significantly with increasing fertilizer strength and compost use. The planting medium (compost) x fertilizer interaction for phenolics and flavonoids was significant. Fruit from plants grown in full-strength fertilizer with 50% soil plus 50% compost and 100% compost yielded fruit with the highest levels of phenolics, flavonol, and anthocyanin content. A positive relationship between antioxidant activities and contents of AsA and GSH and ratios of AsA/DHAsA and GSH/GSSG existed in fruit of both strawberry cultivars. Correlation coefficients for the content of antioxidant components versus antioxidant activity [against ROO(*), O(2)(*)(-), H(2)O(2), OH(*), or (1)O(2)] ranged from r( )()= 0.7706 for H(2)O(2) versus GSH/GSSH in cv. Allstar to r = 0.9832 for O(2)(*)(-) versus total flavonoids in cv. Allstar.

Anthocyanins↗

PubMed2005

Characterization of phenolic profiles of Northern European berries by capillary electrophoresis and determination of their antioxidant activity.

Berries are known to contain phenolic substances (i.e., flavonoids and phenolic acids), which comprise two large and heterogeneous groups of biologically active nonnutrients. This investigation evaluated the content and profile of the phenolic compounds present in six different berries found in Northern Europe. The latter included bilberry (Vaccinium myrtillus), cowberry (Vaccinium vitis-idaea), cranberry (Vaccinium oxycoccus), strawberry (Fragaria ananassa), black currant (Ribes nigrum), and red currant (Ribes rubrum). The study was focused on two areas. The first involved the extraction and analysis of berries for total phenolic content and determination of their antioxidant activity. The total phenolic level of berries was correlated with their antioxidant activity. Second, the berry extracts were analyzed by capillary electrophoresis to determine the content and profile of selected bioactive compounds. The analytes of interest included trans-resveratrol, cinnamic acid, ferulic acid, p-coumaric acid, quercetin, and morin.

Antioxidants↗

PubMed2006

Stable isotope and trace element profiling combined with classification models to differentiate geographic growing origin for three fruits: effects of subregion and variety.

Classifications of geographic growing origin of three fresh fruits combining elemental profiles with various modeling approaches were determined. Elemental analysis (Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, V, and Zn) of strawberry, blueberry, and pear samples was performed using inductively coupled plasma argon atomic emission spectrometer. Bulk stable carbon and nitrogen isotope analyses in pear were performed using mass spectrometry as an alternative fingerprinting technique. Each fruit, strawberry (Fragaria x ananassa), blueberry (Vaccinium caesariense/corymbosum), and pear (Pyrus communis), was analyzed from two growing regions: Oregon vs Mexico, Chile, and Argentina, respectively. Principal component analysis and canonical discriminant analysis were used for data visualization. The data were modeled using linear discriminant function, quadratic discriminant function, neural network, genetic neural network, and hierarchical tree models with successful classification ranging from 70 to 100% depending on commodity and model. Effects of Oregon subregional and variety classification were investigated with similar success rates.

Argentina↗

PubMed2006

Lipoxygenase involvement in ripening strawberry.

The enzymatic activity, subcellular localization, and immunolocalization of plant lipoxygenase (LOX) in strawberry fruits (Fragaria x ananassa, Duch) were investigated. Chemical and enzymatic properties of LOX have been characterized, and the LOX capability of oxygenating free and esterified unsaturated fatty acids into C6 volatile aldehydes has been confirmed. Fruits at unripe, turning, and ripe stages were analyzed for LOX activity and protein localization by Western blots, two-dimensional electrophoresis, and immunolocalization analyses. The ability of strawberry tissues to in vivo metabolize linolenic acid or linoleic acid into C6 volatile aldehydes and the LOX products was also analyzed. Analysis of strawberry proteins showed that a number of LOX forms, corresponding to at least two mobility groups of approximately 100 and 98 kDa and pI values ranging between 4.4 and 6.5, were present. Confocal and electron microscopy analyses support the idea that LOX proteins are associated to lipid-protein aggregates. Both exogenously supplied linoleate and linolenate were converted into hexanal and trans-2-hexenal at the three fruit-ripening stages. Our experiments suggest the presence of different LOX isoforms in strawberry fruits and that the lipoxygenase-hydroperoxide lyase pathway plays a role in converting lipids to C6 volatiles during ripening.

Aldehydes↗

PubMed2003

Engineering increased vitamin C levels in plants by overexpression of a D-galacturonic acid reductase.

L-Ascorbic acid (vitamin C) in fruits and vegetables is an essential component of human nutrition. Surprisingly, only limited information is available about the pathway(s) leading to its biosynthesis in plants. Here, we report the isolation and characterization of GalUR, a gene from strawberry that encodes an NADPH-dependent D-galacturonate reductase. We provide evidence that the biosynthesis of L-ascorbic acid in strawberry fruit occurs through D-galacturonic acid, a principal component of cell wall pectins. Expression of GalUR correlated with changing ascorbic acid content in strawberry fruit during ripening and with variations in ascorbic acid content in fruit of different species of the genus Fragaria. Reduced pectin solubilization in cell walls of transgenic strawberry fruit with decreased expression of an endogenous pectate lyase gene resulted in lower ascorbic acid content. Overexpression of GalUR in Arabidopsis thaliana enhanced vitamin C content two- to threefold, demonstrating the feasibility of engineering increased vitamin C levels in plants using this gene.

Alcohol Oxidoreductases↗

PubMed2003

Cloning and partial characterization of a beta-1,3-glucanase gene from strawberry.

Plant pathogenesis-related proteins, including beta-1,3-glucanses, are believed to be involved in plant defense mechanisms. We have cloned a beta-1,3-glucanse gene from strawberry (Fragaria x ananassa Duch). This gene, designated FaBG2-1, is a Class II glucanase gene composed of two exons and one intron. The location of a 397-base intron in the gene was confirmed by sequencing a partial cDNA clone obtained by using a rapid amplification of cDNA ends procedure. Also, based on the cDNA sequence, the transcription start site of FaBG2-1 was assigned to a -54G residue. Genomic Southern blot analysis indicated that FaBG2-1 is a member of a multi-gene family. Reverse transcriptase-polymerase chain reaction analysis revealed that this beta-1,3-glucanase gene is expressed constitutively in strawberry leaves.

Amino Acid Sequence↗

PubMed2006

Identification of a zinc transporter gene in strawberry.

We have cloned a ZRT (Zn-regulated transporter), IRT (Fe-regulated transporter)-like protein (ZIP) gene from the strawberry plant (Fragaria x ananassa Duch). This gene, designated as FaZIP1, is composed of three exons and two introns. The locations of the two introns (546 and 157 base pairs) in the gene were confirmed by sequence analysis of cDNA clones obtained by using 3' and 5' RNA ligase mediated-rapid amplification of cDNA ends (RLM-RACE) PCR. Also, based on the cDNA sequence, the transcriptional start site of FaZIP1 was assigned. FaZIP1 is predicted to code for a protein of 353 amino acids containing a signal peptide and eight potential transmembrane domains. Southern blot analysis indicated that FaZIP1 is a member of a multi-gene family. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that the FaZIP1 gene is constitutively expressed in strawberry leaves and roots.

Amino Acid Sequence↗