Biofertilizer from vermicompost and microorganisms: effect on strawberry, blackberry, and blueberry phytochemical profile

doi:10.21203/rs.3.rs-2700804/v1

Download PDF

Research Article

Biofertilizer from vermicompost and microorganisms: effect on strawberry, blackberry, and blueberry phytochemical profile

https://doi.org/10.21203/rs.3.rs-2700804/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

The aim of this research was to compare the effect of conventional fertilization (control) and conventional fertilization supplemented with biofertilizer (‘Biovermix’) based on vermicompost and microorganisms (treatment), on the nutritional value of different berry species: strawberry (‘Senga Sengana’), blackberry (‘Čačanska Bestrna’) and blueberry (‘Aurora’). The results showed that biofertilization positively affected fruit weight (8.8 g), soluble solids content (8.9°Bx), sugars (65.6 g kg^− 1 FW) and all identified phenolic groups (phenolic acids, flavanols, flavonols and anthocyanins) in blackberry fruits. Significantly higher fruit weight (15.5 g), total sugars and organic acids content (62.4 and 14.5 g kg^− 1 FW, respectively) and flavanones content (20.06 mg 100 g^− 1 FW) in strawberries fruits were confirmed in biofertilization treatment. Biofertilizer supplementing exhibited the highest effectiveness in terms of the organic acid (21.0 g kg^− 1 FW) and anthocyanins (153.26 mg 100 g^− 1 FW) composition in blueberry fruits. Therefore, the use of biofertilizers as supplements to conventional fertilization can be considered an appropriate practice to ensure soil biological activity, which has an indirect positive effect on the sensory and nutritive value of the tested berry species.

berries

biofertilization

organic acids

sugars

phenolics

Numerous studies have shown the negative impact of chemical fertilizers on the environment through leaching and runoff of nutrients, leading to environmental degradation [1–3]. Organic (fruit) production has become very popular due to the ‘sustainable intensification’ of production with less pesticide and heavy metal residues, distinct flavour, better soil health, and higher biodiversity [4]. It is widely acknowledged that using compost and vermicomposts (VC) as amendments, rather than industrialized fertilizer and raw manure, could improve soil nutrients and promote soil health [5]. Leachates thus derived from vermicomposting are regarded as beneficial and can be used as liquid fertilizer due to a high concentration of plant nutrients [6, 7]. Also, improving of plant growth using VC may result from the impact of its nutrients and biologically active substances [8]. Many plant growth regulators of microbial origin are found in VC [9], and the application of VC significantly contributes to the activity of soil enzymes [10]. García et al. [11] stated that liquid humus obtained from VC of cattle manure has been shown to stimulate yield in crops such as cucumber, rice and broccoli. Further, due to the limited availability in the market and the difficult application of VC, there is a need for the use of liquid VC-based bioproducts.

Plant Growth-Promoting Microorganisms (PGPM), such as bacteria and fungi, are an important part of the soil microbiota known for their ability to promote biological nitrogen fixation, phosphate dissolution and indole acetic acid production, increase root surface area and improve plant growth [12, 13]. Microbial biomass also plays an important role in plant growth in the field. Therefore, fertilizers with leachates derived from VC and enriched by a microbial consortium as supplements to mineral fertilizers can help overcome the difficulties of VC application and positively affect fruit quality. There is no detail report of the effectiveness of VC leachates and like products with microorganisms on chemical composition i.e. content of primary and secondary metabolites in berries. Taking these facts into consideration, this study aimed to evaluate the effects of biofertilizer (‘Biovermix’) obtained from VC and MO as supplementation to standard fertilization, on the phytochemical profile of strawberry, blackberry and blueberry fruits. Supplementing standard fertilization with microbiological fertilizers will contribute to increasing the biological activity of the soil, which can indirectly have a positive effect on the quality of the fruit of the investigated berries species. The results obtained from this study could help growers to expand the application of biofertilizers based on VC enriched with MO in berry production in order to obtain fruits with improved nutritional properties.

2.1. Plant material and experimental design

Three berry species, strawberry (‘Senga Sengana’), blackberry (‘Čačanska Bestrna’), and highbush blueberry (‘Aurora’), were used in this study. All examined species were grown at the experimental plantings of the Fruit Research Institute, located in Čačak (43°54' N latitude, 20°22' E longitude, 225 m altitude), Western Serbia.

The plantings were located close to each other, thus they were in the same micro-climate conditions. According to records weather condition (Republic Hydrometeorological Service of Serbia) in Čačak were characterized by the mean growing season temperature of 19.1°C and total rainfall of 180.1 mm for 2018 (Supplementary Information Table S1). All plantings were established on alluvial soil with sandy-loam texture (51.95% sand and 48.1% loam), pH _KCl–5.48, humus, N_TOT, available K₂O and P₂O₅ contents were 3.95%, 0.20%, 27.00 and 22.95 mg 100 g^-1, respectively.

Strawberries were planted in single bed growing system in August 2017 at a distance 0.18 m among plants and the row distance was 0.50 m, apart. Blackberries were planted in 2010 at a spacing of 3.0 m in the row and 1.5 m within the row and trained to the three-wire trellis. Blueberries were planted in 2015 in the bush training system at spacing of 3.0 m in the row and 1.25 m within the plants.

The experiment was laid out in a randomised block design with four replications. The trial involved two fertilization treatments. The first treatment was standard fertilization (control) during the growing season for blackberry, strawberry, and blueberry applied according to commercial practice as described by Milivojević [14], while the second treatment included the addition of biofertilizer based on VC enriched with MO (‘Biovermix’). The trial was equipped with a drip irrigation system and all plants were subjected to standard pest and disease control. The standard fertilization and biofertilizer application of blackberry and blueberry involved 20 bushes (5 plants in four replications), while in strawberry involved 80 plants (20 plants in four replications).

2.2. Biofertilizer (‘Biovermix’) development and application

The biofertilizer ‘Biovermix’ was prepared at the Fruit Research Institute Čačak, Serbia and specially obtained as a liquid extract of VC of organic origin, enriched with selected strains of beneficial microorganisms. Thus, special brewing tank, with the pump for continuous aeration was filled with tap water. Then, the VC was placed in perforated cassettes, in the centre of the tank so that the VC: water ratio was 1:10 v/w. After that, molasses was added and the substrate was inoculated with a starter culture (microbial consortium). Microbiological consortium contained three soil microorganisms belonging to bacteria of the genus Azotobacter sp., Bacillus sp. and Pseudomonas sp. and fungus Trichoderma sp. During the growth phase, the pH, electrical conductivity, temperature and stirrer speed have been controlled. Upon the pre-defined time elapse, cultivation liquid was, through vents, discharged into a reception vessel. The final concentration of microbial starter has been adjusted to 10⁹ CFU mL^− 1. Biofertilizer had pH 7.67, electrical conductivity 3.95 ds m^− 1, total nitrogen content 11.5 mg kg^− 1, phosphorus 215.1 mg kg^− 1, and organic matter 17.5%.

Biofertilizer was applied three times in 2018: at the beginning of growing period (1st ), at flowering time (5–10% flowers open) (2nd ) and at beginning of ripening (3rd ) in all examined species (Supplementary Information Table S2). The watering amount of biofertilizer in each treatment was 1 L per bush for blueberry and blackberry and 0.5 L per plant for strawberry.

2.3. Fruit sampling

Fully ripe berries were harvested at the second ten-days of May for strawberry, and at the first ten-days of August for blackberry and blueberry. Fully ripe blackberries were black, glossy, and easily picked from the branches [15]. In order to obtain uniform samples fruits of blackberries and blueberries were visually chosen from the same pool of each bush at the same development stage. The strawberry harvest date was determined based on sensory properties (color, taste, and firmness), and the fruits were taken in the middle of the harvest period.

2.4. Determination of basic fruit quality traits

After harvest, 25 fruits of each species (n = 25) in four replicates were randomly selected (100 fruits in total) from both treatments. Fruit weight was measured using a Mettler (Toledo, Switzerland) balance (± 0.01 g accuracy) and the data were expressed in g. Total soluble solids content (SSC) was determined by a digital refractometer (Carl Zeiss, Jena, Germany) at 20°C, and data were expressed in °Brix. After fruit weight and SSC measurements, fruit samples were immediately frozen and stored at -20°C until chemical analyses.

2.5. Estimation of sugars and organic acids

Fruits of strawberry, blackberry (5.0 g), and blueberry (3.0 g) were homogenized in double-distilled water. After 30 min at 10,000 rpm (Eppendorf Centrifuge 5 810 R, Norway), they were filtered through a 0.20 µm cellulose ester filter (Macherey-Nagel, Germany) and poured into a vial before being analyzed by a high-performance liquid chromatography system (HPLC, Thermo Scientific, Finnigan Spectra System, Waltham, MA, USA). The organic acids and sugars content were analysed using the same HPLC system, as described by Smrke et al. [16]. Briefly, sugar separations were carried out using a Rezex RCM-monosaccharide column from Phenomenex (Ca + 2%) operated at 65°C (300 mm × 7.8 mm). The mobile phase was double distilled water. The duration of the analysis was 30 min. and a refractive index (RI) detector was used. The injection volume was 20 µl, and the flow rate was maintained at 0.6 ml/min. Analyses of organic acids were performed on the same HPLC system, equipped with a UV detector set at 210 nm, using a Rezex ROA-organic acid (H+ (8%)) column from Phenomenex (300 mm × 7.8 mm) coupled with a Chromsep guard column. The column temperature was set at 65°C and the elution solvent was 4 mM sulphuric acid in double distilled water. The flow rate was 0.6 ml/min.

The sugars and acids content were calculated according to a calibration curve of corresponding standard solutions. The results were expressed as grams per kilogram fresh weight for sugars and acids (g kg^− 1 FW), except for fumaric and shikimic acids that were presented as milligrams per kilogram fresh weight (mg kg^− 1 FW).

2.6. Estimation phenolic content

For the analysis of individual phenolics, berries were chopped and 5.0 g of strawberry and blueberry, and 4.0 g of blackberry were extracted with methanol with 5% formic acid in a cooled ultrasonic bath for 1 h. Supernatants were centrifuged for 10 min at 10,000 rpm, filtered through 0.2 µm polyamide filters (Macherey-Nagel, Düren, Germany), and transferred into a vial injected into the HPLC system coupled with a diode array detector (Thermo Finnigan, San Jose, CA, USA) at 280 nm, 350 nm, and 530 nm, as described by Smrke et al. [16]. The column was a Gemini C18 (150 × 4.4 mm, 3 µm; Phenomenex, Torrance, CA, USA) coupled with a Chromsep guard column operated at 25° C. The elution solvents were aqueous 0.1% formic acid in double-distilled water with 3% acetonitrile (A) and 0.1% formic acid in acetonitrile with 3% double-distilled water (B). Samples were eluted using a linear gradient of 5 to 20% B in the first 15 min, followed by a linear gradient of 20 to 30% B for 5 min, an isocratic mixture for 5 min, a linear gradient of 30 to 90% B for 5 min, and an isocratic mixture for 15 min before returning to the initial conditions.

The phenols content were identified using a mass spectrometer (LTQ XL Linear Ion Trap Mass Spectrometer, Thermo Fisher Scientific, USA) with electrospray ionization (ESI) operating in a positive (anthocyanins) or negative (all the other phenolics) ionization mode. All of the mass spectrometer conditions set were the same as reported by Smrke et al. [16]. Spectral data were elaborated using the Excalibur software (Thermo Scientific). The identification of compounds was confirmed by comparing retention times and their spectra, and by fragmentation and comparison to the literature data. The concentrations of phenols content were calculated from peak areas of samples and corresponding standards obtained using external standard compounds and expressed as milligrams per 100 g fresh weight (mg 100 g^− 1 FW).

2.7. Chemicals

The following standards were used for the determination of sugars and organic acids: fructose, glucose and sucrose; citric, malic, quinic and fumaric acid from Fluka Chemie (Buchs, Switzerland); shikimic acid from Sigma-Aldrich Chemie (Steinheim, Germany). The following standards were used for the quantification of phenolic compounds: neochlorogenic acid (5-O-caffeoylquinic acid), cryptochlorogenic acid (4-O-caffeoylquinic), caffeic acid, ellagic acid, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, quercetin-3-O-rutinoside, quercetin-3-D-xyloside and naringenin from Sigma-Aldrich Chemie (Steinheim, Germany); p-coumaric acid, quercetin-3-O-galactoside, quercetin-3-O-glucoside, kaempferol-3-O-glucoside, (−) epicatechin, apigenin-7-glucoside, procyanidin B1, peonidin-3-O-glucoside from Fluka Chemie (Buchs, Switzerland); isorhamnetin-3-O-glucoside from Extrasynthese (Genay, France); quercetin-3-arabinofuranoside, quercetin-3-arabinofuranoside and myricetin-3-rhamnoside from Apin Chemicals LTD (Compton, United Kingdom). Methanol for the extraction of phenolics was acquired from Sigma-Aldrich Chemie (Steinheim, Germany). The chemicals for the mobile phases were HPLC-MS grade acetonitrile and formic acid from Fluka Chemie (Buchs, Switzerland). Water for the mobile phase was double distilled and purified with the Milli-Q system (Millipore, Bedford, USA). For phenolic compounds of which the external standards were not available, contents are presented as equivalents of related substances.

2.8. Statistical analysis

The data were presented as mean ± standard deviation of four replicates. The results were compared by one-way analysis of variance (ANOVA) and Least Significant Difference (LSD) test were carried out to test any significant differences between means of treatments for each species (strawberry, blueberry and blackberry) using MSTAT-C statistical computer package (Michigan State University, East Lansing, MI, USA). Differences among means at 5% level (p ≤ 0.05) were considered statistically significant.

3.1. Basic fruit quality traits

In evaluating new berry cultivars special attention is paid to the large size of the fruit, which is significant from the point of view of increasing yield as well as the efficiency of manual harvesting and placement of fresh fruit. In general, the fruit weight in our study was lower in control compared to biofertilizer treatment (Table 1). The application of biofertilizer caused an increase in fruit weight by 7% (blackberry ‘Čačanska Bestrna’), 9% (blueberry ‘Aurora’), and 25% (strawberry ‘Senga Sengana’). The increasing trend in terms of fruit weight in our study corresponds to the findings reported by Hassan et al. [17] for the black cherry tomato treated with VC leachates.

Table 1

Fruit weight and SSC of berry species treated with biofertilizer (‘Biovermix’)
Species/cultivar	Treatment	Weight (g)	SSC (°Bx)
Strawberry ‘Senga Sengana’	‘Biovermix’	15.5 ± 0.3 a	9.8 ± 0.9 a
Strawberry ‘Senga Sengana’	Control	12.5 ± 0.9 b	9.4 ± 0.8 a
Blueberry ‘Aurora’	‘Biovermix’	2.2 ± 0.4 a	10.2 ± 1.3 a
Blueberry ‘Aurora’	Control	2.0 ± 0.2 a	10.1 ± 0.6 a
Blackberry ‘Čačanska Bestrna’	‘Biovermix’	8.8 ± 0.4 a	8.9 ± 0.8 a
Blackberry ‘Čačanska Bestrna’	Control	7.8 ± 0.4 b	8.4 ± 1.0 b

SSC – soluble solids content; Values within columns for each species followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

Higher SSC in strawberries (9.8°Bx), blackberries (8.9°Bx) and blueberries (10.2°Bx) has been recorded in biofertilizer application in comparison to control, despite the differences were not statistically significant, except in blackberry. Interestingly, Abud-Archila et al. [18] also observed a higher value of SSC (11°Bx) in the fruit of blackberry ‘Čačanska Bestrna’ in treatment with VC, rock phosphate and arbuscular mycorrhizae which have been frequently used in organic production. Further, Gutierez-Miceli et al. [19] found that the SSC of tomatoes was increased when the soil was amended with VC. Accordingly, Budiastuti et al. [20] emphasized that compost-tea treatments in melon fruit were superior with respect to a very important criteria, such as SSC.

3.2. Sugars and organic acids content

The sugars and acids content play important role in formation of berries flavour. Among primary metabolites, 3 sugars (glucose, fructose and sucrose) and 5 organic acids (citric, malic, quinic, shikimic and fumaric) were determined in the studied berry species. Fructose was the prevailing individual sugar in all examined species (54% in strawberry; 49% in blueberry; 53% in blackberry of total sugars content) (Table 2).

Table 2

Content of individual and total sugars of berry species treated with biofertilizer (‘Biovermix’)
Species/cultivar	Treatment	Fructose	Glucose	Sucrose	Total sugars
Species/cultivar	Treatment	(g kg^− 1 FW)
Strawberry ‘Senga Sengana’	‘Biovermix’	33.7 ± 1.6 a	26.8 ± 1.3 a	1.8 ± 0.2 b	62.4 ± 4.3 a
Strawberry ‘Senga Sengana’	Control	23.6 ± 5.5 b	18.1 ± 4.0 b	2.0 ± 0.2 a	43.8 ± 3.5 b
Blueberry ‘Aurora’	‘Biovermix’	25.4 ± 2.8 b	22.2 ± 2.6 b	6.5 ± 0.7 a	54.1 ± 2.9 a
Blueberry ‘Aurora’	Control	29.2 ± 0.9 a	24.2 ± 0.9 a	4.1 ± 0.4 b	57.6 ± 4.2 a
Blackberry ‘Čačanska Bestrna’	‘Biovermix’	34.9 ± 6.0 a	29.2 ± 4.6 a	1.5 ± 0.6 a	65.6 ± 4.4 a
Blackberry ‘Čačanska Bestrna’	Control	22.4 ± 11.0 b	18.8 ± 8.8 b	0.6 ± 0.5 b	41.8 ± 3.4 b

Values within columns for each species followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

The values of fructose, glucose and total sugars content in strawberries and blackberries were significantly higher in biofertilizer treatment, while a reverse tendency was observed in blueberries. Consistently, Singh et al. [21] reported enhancement SSC, SSC/acid ratio, and total sugars in strawberry co-inoculated with Azotobacter + Azospirillum + Pseudomonas as well as with Azotobacter + Azospirillum + Arbuscular mycorrhizal fungi over recommended dose of fertilizers. The same authors explained increasing TSS and total sugars might be due to steady supply of nutrients by bio-inoculants throughout the growing period. This increased vigour of plants and leaf area with higher synthesis of assimilates due to enhanced rate of photosynthesis. Thus, the mobility of photosynthetic products from leaves to fruits increases, which can positively affect the sugar content in fruits [22]. The highest sucrose content was found in blueberry ‘Aurora’ in both, treatment and control, followed by strawberry ‘Senga Sengana’, while blackberry ‘Čačanska Bestrna’ had lowest sucrose content. The sucrose content of blueberry ‘Aurora’ and blackberry ‘Čačanska Bestrna’ increased when treated with biofertilizer, and more importantly, these values were 1.5 and 2-fold higher, respectively, compared to the untreated berries.

The balance between the total sugars and acids content is very important for consumers to accept the taste of fruits [23]. The acids content are also useful for stabilizing ascorbic acid and anthocyanins, making them essential for fruit color and the extension the storage life of fresh and processed fruits [24]. Citric acid was the most abundant individual organic acid (46% in strawberry; 77% in blueberry; 49% in blackberry of total organic acids) followed by quinic and malic, while shikimic and fumaric acids were present in much lower amounts in the studied species (Table 3).

Table 3

Content of individual and total organic acids in berry species treated with biofertilizer (‘Biovermix’).
Species/cultivar	Treatment	Citric acid	Malic acid	Quinic acid	Shikimic acid	Fumaric acid	Total acids (g kg^− 1 FW)
Species/cultivar	Treatment	(g kg^− 1 FW)			(mg kg^− 1 FW)		Total acids (g kg^− 1 FW)
Strawberry ‘Senga Sengana’	‘Biovermix’	6.6 ± 0.5 a	3.0 ± 0.1 a	4.89 ± 0.28 a	1.2 ± 0.5 a	1.7 ± 0.5 a	14.5 ± 2.6 a
Strawberry ‘Senga Sengana’	Control	5.2 ± 1.3 b	2.4 ± 0.5 b	3.82 ± 0.90 a	0.8 ± 0.2 a	1.2 ± 0.3 b	11.5 ± 2.1 b
Blueberry ‘Aurora’	‘Biovermix’	16.2 ± 0.4 a	0.9 ± 0.1 a	3.72 ± 0.45 a	3.2 ± 0.8 a	0.6 ± 0.1 a	21.0 ± 3.7 a
Blueberry ‘Aurora’	Control	11.4 ± 0.2 b	1.0 ± 0.0 a	4.45 ± 0.18 a	2.5 ± 0.2 a	0.2 ± 0.3 b	16.9 ± 3.2 b
Blackberry ‘Čačanska Bestrna’	‘Biovermix’	8.2 ± 1.2 a	3.2 ± 0.8 a	5.06 ± 1.41 a	2.5 ± 0.7 a	0.9 ± 0.1 a	16.5 ± 2.1 a
Blackberry ‘Čačanska Bestrna’	Control	7.0 ± 0.6 b	2.8 ± 0.2 b	5.22 ± 0.27 a	3.2 ± 0.3 a	0.8 ± 0.1 b	15.1 ± 3.0 a

Values within columns for each species followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

Blueberries had the highest level of total acids content, followed by blackberries and strawberries. Variations in the metabolism of organic acids were recorded in many fruit species [25] and a large number of genetic studies showed that the accumulation of organic acids (e.g. malic acid) was controlled genes, with differences not only between species, but also among cultivars [26]. This is confirmed by the results obtained in this work where blueberry samples contained higher levels of citric acid compared to strawberry and blackberry, but these two species, on the other hand, had significantly more malic acid. Similar to our results, Skupień and Oszmianski [27] found a dominant presence of citric acid in the strawberry ‘Senga Sengana’ grown in northwestern Poland, but the value that they obtained was about two times higher compared to our results. On the other hand, fruits of blueberry ‘Aurora’ accumulated similar levels of citric acid as those obtained by Smrke et al. [16].

Considering application ‘Biovermix’, a biofertilizer based on VC + MC, the individual and total acids content significantly varied within examined treatments, except quinic and shikimic acids content. Values of citric, and fumaric acids in strawberries, blueberries, and blackberries were significantly higher in biofertilizer treatment. Significantly higher total acids content in the biofertilizer treatment was found in blueberries and strawberries, while blackberries also had higher total acids content, but no significant differences were found between treatment and control. An increase in the acidity of the fruit can be a consequence of higher synthesis of organic acids, which occurs due to the larger leaf area of plants, which shades the fruits and thus affects the lowering of the temperature, and less consumption of acid in the respiration process [28]. Mikiciuk et al. [29] failed to report any changes in fruit acidity in strawberry ‘Rumba’ inoculated with bioproducts containing arbuscular mycorrhizal fungi or plant growth promoting rhizobacteria.

3.3. Phenolic content

The public health experts are recommending increased consumption of berries because they have been linked to a reduced risk of various health maladies, including cancer, coronary heart disease, metabolic disorders, and inflammatory responses [30]. According to Paredes-López et al. [31], due to high polyphenolic concentration and diversity, berry fruits are increasingly referred to as natural functional foods. In our research, a total of 30, 36, and 35 phenolic compounds were determined in strawberry, blackberry, and blueberry, respectively. The secondary metabolism among examined species was significantly different. The main categories of phenolic compounds found in berry fruits (raspberry, blueberry, goji berry, black currant, strawberry, cranberry, and blackberry) are phenolic acids, flavonoids, tannins, and stilbenes [32]. Secondary metabolism among examined species in our study was considerable different.

Blackberry had the greatest diversity and highest content in terms of phenolic acids and was the only species that contained hydroxybenzoic acids, in addition to hydroxycinnamic acids. It is important to emphasize that the total content of phenolic acids in blackberries was ten-fold higher compared to strawberries and blueberries. Despite fact that strawberries were characterized by the greatest variety of phenols from the flavanol group (four procyanidin dimers and six procyanidin trimmers), the total flavanol amount was the highest in blackberries. The procyanidins have been reported to have several health beneficial effects by acting as antioxidant, anti-carcinogen, cardio preventive, antimicrobial, anti-viral, and neuroprotective agents; in particular, their effect is related to several mechanisms including the scavenging of free radicals, the chelation of transition metals and the mediation and inhibition of enzymes [33].

However, strawberry was the only of examined species that contained flavanones (apigenin and naringenin). Glycoside forms of quercetin were the dominant flavonols in all examined species, while quantitatively they were the most abundant in blueberry with a total content as much as twenty times higher than strawberry and four times higher compared to blackberry. In addition to high amounts of quercetin in the fruit, smaller amounts of myricetin, isorhamnetin and syringetin were found in blueberries. Anthocyanins made up a significant part of total phenols and among them glycosides of cyanidin and pelargonidin were predominantly represented in strawberries and blackberries. Canidin-3-glucoside was the prevalent form of anthocyanin pigments in blackberries and pelargonidin-3-glucoside in strawberry fruits. It is important to emphasize distinctly high canidin-3-glucoside content in blackberry related to other identified anthocyanins as well as his higher antioxidant efficiency in the prevention of oxidation of human lipoproteins of higher densities in cyanidin compared to pelargonidin [34]. Five different anthocyanins pigments were confirmed in blueberries, and in the highest amount was malvidin and delphinidin, followed by petunidin, peonidin and cyanidin. Malvidin possesses great antioxidant activity, and cytotoxicity against human monocytic leukemia cells and HT-29 colon cancer cells [35–36] and anti-hypertensive activity by inhibiting angiotensin I-converting enzyme (ACE) [37]. Generally, the highest levels of total anthocyanins were characteristic for blueberry.

Overall, regardless of the treatment, blackberry was distinguished by almost 1.5 times higher levels of total phenols compared to blueberry, which was the second richest in this group of metabolites, and three times higher content compared to strawberry, which contained the least amount of phenols.

Table 4

Effects of application biofertilizer (‘Biovermix’) on phenols content in strawberry ‘Senga Sengana’
Compounds	Strawberry ‘Senga Sengana’
	‘Biovermix’	Control
I Phenolic acids	(mg 100 g^− 1 FW)
Hydroxycinnamic acids
1. caffeic acid hexsoside	1.10 ± 0.27 a	1.46 ± 0.14 a
2. 5-caffeolyquinic acid	0.72 ± 0.12 a	0.66 ± 0.09 a
3. p-coumaric acid hexoside	0.72 ± 0.09 a	0.80 ± 0.16 a
4. 3-p-coumaroyl quinic acid	0.48 ± 0.14 a	0.24 ± 0.02 a
5. 4-p-coumaroyl quinic acid	0.25 ± 0.04 a	0.26 ± 0.04 a
6. 3-feruloylquinic acid	1.24 ± 0.12 a	1.14 ± 0.12 a
7. 5-feruloylquinic acid	0.72 ± 0.16 a	0.63 ± 0.08 a
8. cinnamoyl hexoside	2.24 ± 0.16 a	1.63 ± 0.41 a
Total phenolic acids	7.47 ± 0.11 a	6.82 ± 0.21 a
II Flavanols
9. procyanidin trimer 1	4.00 ± 0.77 a	4.27 ± 1.26 a
10. procyanidin trimer 2	3.47 ± 0.15 b	4.84 ± 0.34 a
11. procyanidin trimer 3	2.88 ± 0.22 a	5.89 ± 1.74 a
12. procyanidin trimer 4	0.08 ± 0.02 a	0.07 ± 0.01 a
13. procyanidin trimer 5	5.80 ± 0.74 a	4.43 ± 0.89 b
14. procyanidin trimer 6	0.14 ± 0.02 a	0.18 ± 0.03 a
15. procyanidin dimer 1	1.81 ± 0.08 b	2.53 ± 0.18 a
16. procyanidin dimer 2	7.23 ± 0.56 a	6.51 ± 1.33 a
17. procyanidin dimer 3	11.03 ± 0.78 a	8.33 ± 1.92 a
18. procyanidin dimer 4	1.27 ± 0.05 b	1.68 ± 0.12 a
Total flavanols	37.71 ± 0.33 a	38.73 ± 0.65 a
III Flavanones
19. apigenin acetyl hexoside	19.97 ± 1.54 a	17.31 ± 3.26 a
20. naringenin hexoside	0.09 ± 0.02 a	0.13 ± 0.02 a
Total flavanons	20.06 ± 0.96 a	17.44 ± 1.01 b
III Flavonols
21. quercetin-3-rutinoside	0.20 ± 0.03 a	0.09 ± 0.02 b
22. quercetin-3-glucoside	0.09 ± 0.01 a	0.09 ± 0.01 a
23. quercetin-3-glucuronide	0.24 ± 0.02 a	0.31 ± 0.05 a
24. kaempferol coumaroyl hexoside	0.04 ± 0.01 a	0.05 ± 0.01 a
25. kaempferol-3-glucoside	0.19 ± 0.03 a	0.15 ± 0.02 a
26. kaempferol-acetyl hexoside	0.13 ± 0.02 a	0.14 ± 0.01 a
Total flavonols	0.89 ± 0.02 a	0.83 ± 0.02 a
IV Anthocyanins
27. pelargonidin-3-glucoside	19.62 ± 3.98 a	21.83 ± 1.69 a
28. pelargonidin-3-malonyl glucoside	4.47 ± 0.73 a	5.32 ± 0.38 a
29. pelargonidin-3-rutinoside	1.14 ± 0.23 a	1.27 ± 0.10 a
30. cyanidin-3-glucoside	2.31 ± 0.35 a	2.05 ± 0.34 a
Total anthocyanins	27.54 ± 1.56 a	30.47 ± 1.11 a
TOTAL PHENOLICS	93.67 ± 1.73 a	94.29 ± 0.60 a

Values within each row followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

Namely, 8 hydroxycinnamic acids, 10 flavanols, 2 flavanones, 6 flavonols and 4 anthocyanins were detected in strawberries (Table 4). Treatment and control samples analysed in the present study contained similar concentrations of all groups of phenolics which is consistent with the report of Weber et al. [38]. Studying the benefits of biostimulant application in terms of yield and internal and external quality parameters of strawberries, these authors found that the content of flavanols, flavones and flavonols varied between the two treatments (CON-control treatment with no foliar spraying with biostimulants; OP + AN-foliar application of SiO₂ and Ascophyllum nodosum extract), but the differences were mostly not significant. The same authors state that most of these compounds are regarded as defense phenolics, which are produced in plant tissue in response to different stressors [38], so biostimulators mitigate these adverse effects and plants response in the lower synthesis of specific groups of secondary metabolites. In our study, the flavanols were present in high amounts in strawberries in both, biofertilizer (37.71 mg 100 g^− 1 FW) and control (38.73 mg 100 g^− 1 FW) and represented 40% and 41% of the total analyzed phenolic content, respectively. Procyanidin dimer 3 represented 29% of the total flavanols in biofertilizer treatment, and 22% in control.

What is more, the results obtained in our study indicate considerable share of anthocyanins (29% in biofertilizer treatment, 32% in control, respectively) and flavanones (21% in biofertilizer treatment and 19% in control, respectively) in total phenols content of strawberry ‘Senga Sengana’. However, significant differences between biofertilizer treatment and control were existed only in terms of flavanones content. The other classes of phenolics (hydroxycinnamic acids and flavonols) make a small share (approx. 8‒9%) of total phenolic content.

The results shown in Table 5 indicate that blackberry anthocyanins contributed the most in total phenols content (approx. 45%). Also, derivatives of hydroxycinnamic acids (approx. 30%) and flavanols (approx. 24%) were present in high amounts in blackberries, while flavonols contributed little overall phenols content (< to 6%). The predominant phenolic acid in blackberry was 5-caffeolyquinic acid and accounted for 95% of all phenolic acids. Caffeolyquinic acid exists in different forms with mono-, di- and tricaffeoylquinic acids being the most abundant in fruits and vegetables. Makori et al. [39] highlighted that the health benefits obtained from consuming various fruit and vegetables high in polyphenol compounds have been linked at least in part to the existence of caffeolyquinic acid esters in these food product.

Table 5

Effects of application biofertilizer (‘Biovermix’) on phenols content in blackberry ‘Čačanska Bestrna’
Compounds	Blackberry ‘Čačanska Bestrna’
	‘Biovermix’	Control
I Phenolic acids	(mg 100 g^− 1 FW)
Hydroxycinnamic acids
1. 5-caffeolyquinic acid	74.94 ± 9.72 a	43.62 ± 8.53 b
2. 4-caffeolyquinic acid	0.71 ± 0.15 a	0.91 ± 0.11 a
3. caffeic acid hexoside 1	1.87 ± 0.24 a	1.09 ± 0.21a
4. caffeic acid hexoside 2	3.01 ± 0.70 a	3.60 ± 0.73 a
5. p-coumaric acid hexoside 1	1.16 ± 0.15 a	0.68 ± 0.13 a
6. p-coumaric acid hexoside 2	0.17 ± 0.07 a	0.03 ± 0.01 a
7. 3-p-coumaroyl quinic acid	0.08 ± 0.01 a	0.05 ± 0.01 a
Hydroxybenzoic acids
8. ellagic acid pentoside 1	0.27 ± 0.06 a	0.08 ± 0.01 b
9. ellagic acid pentoside 2	0.19 ± 0.03 a	0.09 ± 0.01 b
10. methylellagic acid pentoside 1	0.21 ± 0.04 a	0.09 ± 0.01 b
11. methylellagic acid pentoside 2	0.14 ± 0.04 a	0.04 ± 0.02 a
Total phenolic acids	82.75 ± 3.21 a	50.28 ± 1.24 b
II Flavanols
12. catehin	4.87 ± 1.90 a	0.95 ± 0.21 a
13. epicatehin	4.07 ± 1.14 a	1.37 ± 0.41 a
14. procyanidin trimer 1	11.47 ± 4.47 a	2.23 ± 0.50 b
15. procyanidin trimer 2	8.34 ± 1.61 a	5.77 ± 0.89 a
16. procyanidin trimer 3	8.57 ± 1.71 a	7.30 ± 1.70 a
17. procyanidin dimer	24.12 ± 5.65 a	28.92 ± 5.88 a
Total flavanols	61.44 ± 1.58 a	46.54 ± 1.11 b
III Flavonols
18. quercetin-3-rutinoside	0.33 ± 0.20 a	0.07 ± 0.01 a
19. quercetin-3-galactoside	0.85 ± 0.22 a	0.37 ± 0.04 a
20. quercetin-3-glucoside	0.40 ± 0.10 a	0.07 ± 0.02 b
21. quercetin-3-xyloside	0.04 ± 0.01 a	0.07 ± 0.01 a
22. quercetin-3-glucuronide	0.40 ± 0.11 a	0.08 ± 0.01 b
23. quercetin-3-arabinopyranoside	0.03 ± 0.01 a	0.01 ± 0.00 b
24. quercetin-3-arabinofuranoside	0.31 ± 0.09 a	0.09 ± 0.01 a
25. quercetin-3-acetylhexoside	0.97 ± 0.31 a	0.31 ± 0.04 a
26.quercetin-3-(6″-(hydroxyl-3-methylglutaroyl)-hexoside	0.26 ± 0.07 a	0.16 ± 0.02 a
27. isorahmetin-3-glucuronide	1.13 ± 0.31 a	1.81 ± 0.22 a
28. kaempferol-hexoside	0.09 ± 0.03 a	0.02 ± 0.00 b
Total flavonols	4.81 ± 0.18 a	3.06 ± 0.03 b
IV Anthocyanins
29. cyanidin-3-glucoside	102.59 ± 12.45 a	60.45 ± 12.04 a
30. cyanidin-3-rutinoside	6.16 ± 0.75 a	3.63 ± 0.72 a
31. cyanidin-3-arabinoside	0.62 ± 0.07 a	0.36 ± 0.07 a
32. cyanidin-3-xyloside	4.14 ± 1.42 a	0.26 ± 0.06 b
33. cyanidin-3-(6″-dioxalylglucoside	7.13 ± 1.77 a	8.45 ± 1.66 a
34. cyanidin-3-(6″-malonylglucoside)	1.50 ± 0.37 a	1.78 ± 0.35 a
35. pelargonidin-3-glucoside	3.72 ± 0.93 a	2.65 ± 0.35 a
36. pelargonidin-3-rutinoside	0.26 ± 0.07 a	0.19 ± 0.02 a
Total anthocyanins	126.12 ± 2.34 a	77.77 ± 1.78 b
TOTAL PHENOLICS	275.12 ± 1.83 a	177.65 ± 1.04 b

Values within each row followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

Optimizing agrotechnical practices, primarily fertilization, can be one of the effective ways to increase phenols content [40]. Analysis of data related to the different group of phenolics in blackberry fruit showed higher levels of all groups in the biofertilizer treatment (liquid VC enriched with bacteria of the genus Azotobacter sp., Pseudomonas sp., and Pseudomonas sp.) compared to the control, indicating a positive effect of biofertilizers on the synthesis of secondary metabolites in blackberries. A similar pattern was also reported by Abud-Archila et al. [41] who studied cultivated blackberry from cuttings in soil-enriched VC, phosphate rock, Glomus mosseae and found a significant effect of VC treatment on the increase of anthocyanin content in fruits compared to control. Some species of the genus Pseudomonas produce metabolites such as antibiotics and hydrogen cyanides (HCN) [42], some produce siderophores with high affinity for Fe³⁺ absorption [43], and some auxin [44]. Bacteria of the genus Bacillus synthesize a large number of secondary metabolites that affect their environment, thus increasing the availability of nutrients to plants [45]. Additionally, numerous studies indicate that arbuscular mycorrhizal symbiosis, which also contained applied biofertilizers, affects the primary and secondary metabolism of host plants [46] causing changes in enzymatic activities and physiological mechanisms that lead to the accumulation of phenolics, such as are carotenoids and polyphenolics [47]. All these metabolites strongly influence the environment by inhibiting the growth of certain harmful microorganisms, and on the other hand they increase the availability of nutrients to plants, which most likely in this research was reflected in the content of anthocyanins and phenolic acids in blackberry fruits after use of biofertilizer ‘Biovermix’, which contains bacteria of the genus Azotobacter, Pseudomonas and Bacillus. Also, Kumar and Gupta [42] noted the VC is better than other fertilizers due to its greater nutrient availability to plants.

The health benefits of blueberries are of particular interest of lately due to their high content of phytochemicals known as (poly)phenolics [48]. In blueberry samples analized in our study, four groups of phenolic compounds were present: 9 hydroxycinnamic acids, 3 flavanols, 15 flavonols, and 8 anthocyanins (Table 6). The analysis of the total phenols content in blueberries in different nutrition conditions showed a weaker effect of biofertilization compared to blackberries.

Table 6

Effects of application biofertilizer (‘Biovermix’) on phenols content in blueberry ‘Aurora’
Compounds	Blueberry ‘Aurora’
I Phenolic acids	‘Biovermix’	Control
Hydroxycinnamic acids
1. caffeic acid	0.01 ± 0.00 a	0.01 ± 0.00 a
2. 5-caffeoylquinic acid 1	0.75 ± 0.41 a	1.55 ± 0.54 a
3. 5-caffeolyquinic acid 2	0.75 ± 0.41a	1.55 ± 0.54 a
4. p-coumaric acid	2.75 ± 0.11 a	3.27 ± 0.20 a
5. 3-p-coumaroylquinic acid	0.04 ± 0.00 a	0.05 ± 0.00 a
6. 5-p-coumaroylquinic acid 1	0.16 ± 0.01a	0.17 ± 0.01 a
7. 5-p-coumaroylquinic acid 2	0.47 ± 0.03 a	0.50 ± 0.02 a
8. 3-feruloyquinic acid	0.33 ± 0.01 a	0.24 ± 0.01 b
9. 5-feruloylquinic acid	1.63 ± 0.08 a	1.64 ± 0.15 a
Total phenolic acids	6.89 ± 0.09 a	8.98 ± 0.11 a
II Flavanols
10. catehin	0.71 ± 0.03 a	0.53 ± 0.03 b
11. epicatehin	3.02 ± 0.08 a	3.79 ± 0.19 a
12. procyanidin trimer	5.20 ± 0.15 a	5.61 ± 0.20 a
Total flavanols	8.93 ± 0.05 a	9.93 ± 0.13 a
II Flavonols
13. quercetin-3-rutonoside	3.50 ± 0.20 a	3.28 ± 0.15 a
14. quercetin-3-galactoside	3.88 ± 0.31 a	3.74 ± 0.33 a
15. quercetin-3-glucoside	1.79 ± 0.11 a	1.47 ± 0.12 a
16. quercetin-3-xyloside	0.88 ± 0.05 a	0.95 ± 0.05 a
17. quercetin-3-arabinopyranoside	0.06 ± 0.01 a	0.04 ± 0.00 a
18. quercetin-3-arabinofuranoside	0.79 ± 0.05 a	0.82 ± 0.06 a
19. quercetin-3-rhamnoside	3.96 ± 0.23 a	3.62 ± 0.30 a
20. quercetin-3-acetyl hexoside	0.91 ± 0.01 a	0.83 ± 0.07 a
21. kaempferol-3-galactoside	0.08 ± 0.01 a	0.06 ± 0.01 a
22. kaempferol-3-glucoside	0.91 ± 0.01 a	0.94 ± 0.07 a
23. myricetin-3-hexoside 1	0.36 ± 0.05 a	0.37 ± 0.09 a
24. myricetin-3-hexoside 2	0.85 ± 0.07 a	1.07 ± 0.07 a
25. isorhamnetin hexoside 1	0.44 ± 0.13 a	0.57 ± 0.04 a
26. isorhametin hexoside 2	0.04 ± 0.00 b	0.06 ± 0.01 a
27. syringetin hexoside	0.07 ± 0.01 b	0.11 ± 0.01 a
Total flavonols	18.52 ± 0.07 a	17.93 ± 0.05 a
IV Anthocyanins
28. malvidin-3-galactoside	51.42 ± 4.47 a	56.00 ± 5.67 a
29. malvidin-3-arabinoside	25.11 ± 2.27 a	23.73 ± 2.23 a
30. delphinidin-3-galactoside	35.94 ± 3.12 a	24.93 ± 2.33 b
31. delphinidin-3-arabinoside	10.53 ± 0.70 a	7.94 ± 0.53 b
32. petunidin-3-galactoside	19.58 ± 2.02 a	5.79 ± 0.83 a
33. petunidin-3-arabinoside	1.62 ± 0.17 a	1.31 ± 0.07 a
34. cyanidin-3-galactoside	7.77 ± 0.51 a	5.86 ± 0.39 b
35. peonidin-3-glucoside	1.29 ± 0.11 a	1.40 ± 0.14 a
Total anthocyanins	153.26 ± 1.01 a	136.96 ± 0.42 b
TOTAL PHENOLICS	187.60 ± 0.31 a	176.80 ± 0.18 b

Values within each row followed by the same small letter are not significantly different at p ≤ 0.05 by LSD test.

The anthocyanins were the most abundant phenolics in both nutrition conditions (treatment and control). They were made up 81% of the total phenols content in biofertilizer treatment, and were significantly higher than control. Comparably, Schoebitz et al. [1] found that combined application of microbial consortium and humic substances improves blueberry plant performance, resulting in a synergistic effect when beneficial microorganisms and humic substances are introduced. The same authors indicate that a combined treatment with a microbial consortium and humic substances seems to be the most appropriate method to improve nutrient uptake, increase plant biomass and stimulate soil microflora. Accordingly, the optimal nutritional conditions in our study probably contributed to a higher accumulation of anthocyanins in the fruits of blueberries and blackberries treated with biofertilizer ‘Biovermix’. Phytohormones, including cytokinin, ethylene, jasmonic acid, and gibberellins, which are present in VC, are important internal factors affecting anthocyanin biosynthesis [49].

Malvidin-3-galactoside was the prevailing anthocyanin (34% of total anthocyanins in biofertilizer treatment, and 41% in control) while delphinidin-3-galactoside and petunidin-3-galactoside were represented approximately 39% (treatment) and 30% (control) of all analyzed anthocyanins in blueberry. Quantity and number of anthocyanidins and their derivatives identified in the present study correspond to data reported by Wang et al. [50] for the blueberry ‘Bluecrop’. On the other hand, no significant differences were found in the content of hydroxycinnamic acids and total flavanols between blueberry fruits harvested from bushes watered with biofertilizer and control.

There are a limited number of studies on the phytochemical profile of economically important berry species grown under conditions of supplementation with biofertilizer based on vermicomposting leachates enriched with microorganisms. Our research included comparative studies of standard fertilization and standard fertilization supplemented with ‘Biovermix’ – a biofertilizer based on VC + MC, in strawberries, blackberries and blueberries. Generally, the results of the study indicate that the application of biofertilizer contributed to an increase in the fruit weight and in the content of compounds important for fruit taste, sugars and organic acids, in all the tested species. The results underline the positive effects of biofertilizer ‘Biovermix’, emphasizing that the sugar content in blueberries and strawberries was the most affected by the application.

Despite the fact that secondary metabolites were less affected by treatment, blackberry samples had the highest contents of all groups of phenolic compound (phenolic acids, flavanols, flavonols and anthocyanins), in ‘Biovermix’ treatment compared to the control. The blueberry samples also showed high contents of anthocyanins under the biofertilization conditions, confirming their superiority over the standard fertilization. Accordingly, our research will contribute to a better understanding of the importance of the application VC leachate in combination with a microbial consortium in berry growing technology to obtain fruits with a better nutritional composition.

Vermicompost

Microorganisms

PGPM

Plant Growth-Promoting Microorganisms

Fresh weight

ANOVA

Analysis of variance

LSD

Least significant difference

SSC

Soluble solids content

Ethical Approval

Not applicable.

Consent to Participate

Not applicable.

Consent for Publication

The institutions of affiliation have approved the submission of the article for its publication.

Conflict of Interest

All authors declare that they have no competing interests.

Authors' Contributions

All authors contributed to reviewing and editing the main manuscript text. Conceptualization: Žaklina Karaklajić-Stajić, Jelena Tomić; methodology: Maja Mikulič-Petkovšek, Metka Hudina; Formal analysis and investigation: Marijana Pešaković; Writing - Review & Editing: Jerneja Jakopič, Franc Štampar; Funding acquisition: Robert Veberič; Resources: Svetlana M. Paunović. All authors read and approved the final manuscript.

Funding

This study is part of the agreement No. 451-03-47/2023-01/200215 and financed by the Ministry of Science, and Innovation the Republic of Serbia and part of the program Horticulture P4-0013 financed by the Slovenian Research Agency (ARRS). We hereby express our sincere gratitude for the support.

Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Additional information

Authors and Affiliations

Žaklina Karaklajić-Stajić, Jelena Tomić, Marijana Pešaković, Svetlana M. Paunović

Fruit Research Institute, Department for Technology of Fruit Growing, Kralja Petra I 9, Čačak, Republic of Serbia.

Franc Štampar, Maja Mikulic-Petkovsek, Metka Hudina, Robert Veberic, Jerneja Jakopic

University of Ljubljana, Biotechnical Faculty, Department of Agronomy, Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia.

Corresponding author

Correspondence to Žaklina Karaklajić-Stajić

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Schoebitz M., López MD, Serrí H, Martínez O, Zagal E (2016) Combined application of microbial consortium and humic substances to improve the growth performance of blueberry seedlings. J Soil Sci Plant Nutr 16:1010‒1023. http://dx.doi.org/10.4067/S0718-95162016005000074
Yadav RS, Singh V, Pal S, Meena SK, Meena VS, Sarma BK, Singh HB, Rakshit A (2018) Seed bio-priming of baby corn emerged as a viable strategy for reducing mineral fertilizer use and increasing productivity. Sci Hortic 241:93‒99. https://doi.org/10.1016/j.scienta.2018.06.096
Morra L, Bilotto M, Baldantoni D, Alfani A, Baiano S (2021) A seven-year experiment in a vegetable crops sequence: Effects of replacing mineral fertilizers with Biowaste compost on crop productivity, soil organic carbon and nitrates concentrations. Sci Hortic 290: 110534. https://doi.org/10.1016/j.scienta.2021.110534
Lockie S, Halpin D, Pearson D (2006) Understanding the market for organic food. In: Kristiansen P, Taji A, Reganold J (eds) Organic Agriculture: A global perspective. Collingwood VIC, Australia, pp. 245-258.
Jack AL, Thies JE (2006) Compost and vermicompost as amendments promoting soil health. In: Uphoff N (ed.) Biological approaches to sustainable soil systems, CRS Press, New York, pp. 45-466.
Jarecki MK, Chong C, Voroney RP (2005) Evaluation of compost leachates for plant growth in hydroponic culture. J Plant Nutr 28:651‒667. https://doi.org/10.1081/PLN-200052639
Gutierrez-Miceli FA, Gracia-Gomez RC, Rincon RR, Abud-Archila M, MariaAngela OL, Gullin-Cruz MJ, Dendooven L (2008) Formulation of liquid fertilizer for sorghum (Sorghum bicolour (L.) Moench) using vermicompost leachate. Bioresour Technol 99: 6174-6180. https://doi.org/10.1016/j.biortech.2007.12.043
Warman PR, AngLopez MJ (2010) Vermicompost derived from different feedstocks as a plant growth medium. Bioresour Technol 101:4479-4483. https://doi.org/10.1016/j.biortech.2010.01.098
Atiyeh RM, Lee SS, Edwards CA, Arancon NQ, Metzger J (2002) The influence of humic acid derived from earthworm-processed organic waste on plant growth. Bioresour Technol 84:7-14. https://doi.org/10.1016/s0960-8524(02)00017-2
Albiach R, Canet R, Pomares F, Ingelmo F (2000) Microbial biomass content and enzymatic activities after the application of organic amendments to a horticultural soil. Bioresour Technol 75:43-48. https://doi.org/10.1016/S0960-8524(00)00030-4
García AC, Izquierdo FG, González OLH, De Armas MMD, López RH, Rebato SM, Balmori DM, Berbara RLL (2013) Biotechnology of humified materials obtained from vermicomposts for sustainable agroecological purposes. Afr J Biotechnol 7:625-634. https://doi.org/10.5897/AJBX12.014
Bashan Y, De-Bashan LE, Prabhu SR, Hernandez JP (2014) Advances in plant growth-promoting bacterial inoculant technology: formulations and practical perspectives (1998–2013). Plant Soil 378:1-33. https://doi.org/10.1007/s11104-013-1956-x
Viscardi S, Ventorino S, Duran P, Maggio A, De Pascale S, Mora ML, Pepe O (2016) Assessment of plant growth promoting activities and abiotic stress tolerance of Azotobacter chroococcum strains for a potential use in sustainable agriculture. J Soil Sci Plant Nutr 16:848-863. http://dx.doi.org/10.4067/S0718-95162016005000060
Milivojević J (2022) Small fruits. University of Belgrade, Faculty of Agriculture, Center for Education and Research (AgroNET), Belgrade, Republic of Serbia.
Karaklajić-Stajić Ž, Nikolić M, Tomić J, Miletić R, Pešaković M (2017) Influence of ripeness stage on the rate of pigment degradation in blackberry fruits during frozen storage. Eur J Hortic Sci 82:198-203. https://doi.10.17660/eJHS.2017/82.4.5
Smrke T, Veberic R, Hudina M, Stamic D, Jakopič J (2021) Comparison of highbush blueberry (Vaccinium corymbosum L.) under ridge and pot production. Agriculture 11: 929. https://doi.org/10.3390/agriculture11100929
Hassan MR, Julie SN, Rahul SK, Rahman M, Uddin J (2018) Influence of organic leachates on growth and yield of black cherry tomato. IJBSSR 7: 2309-7892.
Abud-Archila M, Luján-Hidalgo MC, López-Pérez JM, Ordaz-Rivera J, Ruiz-Valdiviezo VM, Oliva-Llaven M.Á, Gutiérrez-Miceli FA (2018) Growth and fruit chemical characteristics of blackberry (Rubus fruticosus) cultivated with vermicompost, Glomus mosseae and phosphate rock. Compost Sci Util 26: 225‒231. https://doi.org/10.1080/1065657X.2018.1471367
Gutierez-Miceli FA, Santiago-Borraz J, Montes Molina JA, Carlos Nafate C, Abud-Archila M,Oliva Laven MA, Rincon-Rosales R, Dendooven L (2007) Vermicompost as a soilsupplement to improve growth, yield and fruit quality of tomato (Lycopersicum esculentum). Biores Techn 98:2781-2786. https://doi.org/10.1016/j.biortech.2006.02.032
Budiastuti S, Purnomo D, Sulistyo TD, Rahardjo SP, Darsono L, Pardjo YV (2012) The enhancement of melon fruit quality by applicatin of the fertilizer and gibberellin. J Agric Sci Technol B 2:455-460. https://doi.org/10.28940/terra.v38i1.527
Singh SR, Zargar MY, Singh U, Ishaq M (2010) Influence of bio-inoculants and inorganic fertilizers on yield, nutrient balance, microbial dynamics and quality of strawberry (Fragaria x ananassa) under rainfed conditions of Kashmir valley. Indian J Agric Sci 80:275.
Singh A, Singh JN (2006) Studies on influence of biofertilizers and bioregulators on flowering, yield and fruit quality of strawberry cv. Sweet Charlie. Ann Agric Res 27:261-264.
Hudina M, Štampar F (2000) Sugars and organic acids contents of European Pyrus comminus L. and Asian Pyrus serotina Rehd. pear cultivars. Acta Aliment 29:217-230. https://doi.org/10.1556/aalim.29.2000.3.2
Talcott ST (2007) Chemical components of berry fruits. In: Zhao Y (ed.) Berry fruit, value-added products for health promotion. Taylor & Francis Group, Abingdon, pp 51-73.
Zheng YJ, Duan YT, Zhang YF, Pan QH, Li JM, Huang WD (2009) Determination of organic acids in red wine and must on only one RP-LC-column directly after sample dilution and filtration. Chromatographia 69:1391-1395. https://doi.org/10.1365/s10337-009-1085-0
Saradhuldhat P, Paull RE (2007) Pineapple organic acid metabolism and accumulation during fruit development. Sci Hortic 112:297-303. https://doi.org/10.1016/j.scienta.2006.12.031
Skupień K, Oszmiański J (2004) Comparison of six cultivars of strawberries (Fragaria × ananassa Duch.) grown in northwest Poland. Eur Food Res Technol 219:66-70. https://doi.org/10.1007/s00217-004-0918-1
Umar I, Wali VK, Kher R, Jamwal M (2009) Effect of Fym, urea and Azotobacter on growth, yield and quality of strawberry cv. Chandler. Not Bot Horti Agrobot Cluj Napoca 37:139-143.
Mikiciuk G, Sas-Paszt L, Mikiciuk M, Derkowska E, Trzcinski P, Gluzcek S, Lisek A, Wera-Bril S, Rudnicka J (2019) Mycorrhizal frequency, physiological parameters, and yield of strawberry plants inoculated with endomycorrhizal fungi and rhizosphere bacteria. Mycorrhiza 29:489-501. https://doi.org/10.1007/s00572-019-00905-2
Obrenovich ME, Li Y, Parvathaneni K, Yendluri BB, Palacios HH, Leszek Gjumrakch J (2011) Antioxidants in health, disease and aging. CNS & Neurol Disord Drug Targets 10:192-207. https://doi.org/10.2174/187152711794480375
Paredes-López O, Cervantes-Ceja ML, Vigna-Pérez M, HernándezPérez T 2010. Berries: improving human health and healthy aging, and promoting quality life‒a review. Plant Foods Hum Nut 65:299-308. https://doi.org/10.1007/s11130-010-0177-1
Pap N, Fidelis M, Azevedo L, Do Carmo MAV, Wang D, Mocan A, Penha E, Pereira R, Xavier-Santos D, Sant SA, Yang B, Granato D (2021) Berry polyphenols and human health: Evidence of antioxidant, anti-inflammatory, microbiota modulation, and cell-protecting effects. Curr Opin Food Sci 42:167-186. https://doi.org/10.1016/j.cofs.2021.06.003
Aron PM, Kennedy JA (2008) Flavan‐3‐ols: Nature, occurrence and biological activity. Mol Nutr Food Res 52:79-104. https://doi.org/10.1002/mnfr.200700137
Satue-Garcia T, Heinonen M, Frankel EN (1997) Anthocyanins as antioxidants on human low-density lipoprotein and lecithin–liposome systems. J Agric Food Chem 45:3362-3367. https://doi.org/10.1021/jf970234a
Hyun JW, Chung HS (2004) Cyanidin and malvidin from Oryza sativa cv. Heugjinjubyeo mediate cytotoxicity against human monocytic leukemia cells by arrest of G2/M phase and induction of apoptosis. J Agric Food Chem 52:2213-2217. https://doi.org/10.1021/jf030370h
Patterson SJ, Fischer JG, Dulebohn RV (2008) DNA damage in HT‐29 colon cancer cells is enhanced by high concentrations of the anthocyanin malvidin. FASEB J https://doi.org/10.1096/fasebj.22.1_supplement.890.10
Lee C, Han D, Kim B, Baek N, Baik BK (2013) Antioxidant and anti‐hypertensive activity of anthocyanin‐rich extracts from hulless pigmented barley cultivars. I J Food Sci Techn 48:984-991. https://doi.org/10.1111/ijfs.12050
Weber N, Schmitzer V, Jakopic J, Stampar F (2018) First fruit in season: Seaweed extract and silicon advance organic strawberry (Fragaria × ananassa Duch.) fruit formation and yield. Sci Hortic 242:103-109. https://doi.org/10.1016/j.scienta.2018.07.038
Makori SI, Mu TH, Sun HN (2021) Physicochemical properties, antioxidant activities, and binding behavior of 3, 5-di-O-caffeoylquinic acid with beta-lactoglobulin colloidal particles. Food Chem 347:129084. https://doi.org/10.1016/j.foodchem.2021.129084
Anttonen MJ, Karjalainen RO (2006) High-performance liquid chromatography analysis of black currant (Ribes nigrum L.) fruit phenolics grown either conventionally or organically. J Agric Food Chem 54:7530-7538. https://doi.org/10.1021/jf0615350
Abud-Archila M, Luján-Hidalgo MC, López-Pérez JM, Ordaz-Rivera J, Ruiz-Valdiviezo VM, Oliva-Llaven MÁ, Gutiérrez-Miceli FA (2018) Growth and fruit chemical characteristics of blackberry (Rubus fruticosus) cultivated with vermicompost, Glomus mosseae and phosphate rock. Compost Sci Util 26:225-231. https://doi.org/10.1080/1065657X.2018.1471367
Kumar A, Gupta RK (2018) The effects of vermicompost on growth and yield parameters of vegetable crop radish (Raphanus sativus). J Pharmacogn Phytochem 7:589-592.
Weller DM, Thomashow LS (1993) Use of rhizobacteria for biocontrol. Curr Opin Biotechnol 4:306-311. https://doi.org/10.1016/0958-1669(93)90100-B
Kloepper JW, Leong J, Teintze M, Schroth MN (1980) Pseudomonas siderophores: a mechanism explaining disease suppressive soils. Curr Microbiol 4:317-320. https://doi.org/10.1007/BF02602840
Khakipour N, Khavazi K, Mojallal H, Pazira E, Asadirahmani H (2008). Production of auxin hormone by fluorescent pseudomonads. Am Eurasian J Agric Environ Sci 4:687-692.
Barriuso J, Ramos Solano B, Lucas JA, Lobo AP, García‐Villaraco A, Gutiérrez Mañero FJ (2008) Ecology, genetic diversity and screening strategies of plant growth promoting rhizobacteria (PGPR). In: Ahmad I, Pichtel J, Hayat S. (eds.), Plant‐Bacteria Interactions: Strategies and Techniques to Promote Plant Growth. Wiley‐VCH Verlag GmbH & Co, New Jersey, pp 1-17. https://doi.org/10.1002/9783527621989
Schliemann W, Ammer C, Strack D (2008) Metabolite profiling of mycorrhizal roots of Medicago truncatula. Phytochem 69:112-146. https://doi.org/10.1016/j.phytochem.2007.06.032
Toussaint JP, Smith FA, Smith SE (2007) Arbuscular mycorrhizal fungi can induce the production of phytochemicals in sweet basil irrespective of phosphorus nutrition. Mycorrhiza 17:291-297. https://doi.org/10.1007/s00572-006-0104-3
Qi T, Song S, Ren Q, Wu D, Huang H, Chen Y, Xie D (2011) The Jasmonate-ZIM-domain proteins interact with the WD-Repeat/bHLH/MYB complexes to regulate Jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana. The Plant Cell 23:1795-1814. https://doi.org/10.1105/tpc.111.083261
Wang SY, Chen CT, Sciarappa W, Wang CY, Camp MJ (2008) Fruit quality, antioxidant capacity, and flavonoid content of organically and conventionally grown blueberries. J Agric Food Chem 56:5788-5794. https://doi.org/10.1021/jf703775r

No competing interests reported.

Supplementaryinformation.docx

Download PDF

Version 1

posted

You are reading this latest preprint version

Biofertilizer from vermicompost and microorganisms: effect on strawberry, blackberry, and blueberry phytochemical profile

Status:

Version 1

Abstract

1. Introduction

2. Material And Methods

2.1. Plant material and experimental design

2.2. Biofertilizer (‘Biovermix’) development and application

2.3. Fruit sampling

2.4. Determination of basic fruit quality traits

2.5. Estimation of sugars and organic acids

2.6. Estimation phenolic content

2.7. Chemicals

2.8. Statistical analysis

3. Results And Discussion

3.1. Basic fruit quality traits

3.2. Sugars and organic acids content

3.3. Phenolic content

4. Conclusion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

Status:

Version 1