# The Effect ofTrichoderma spp. on the Composition of Volatile Secondary Metabolites and Biometric Parameters of Coriander (Coriandrum sativum L.)

Authors: E. Gębarowska; M. Pytlarz-Kozicka; J. Nöfer; J. Łyczko; M. Adamski; A. Szumny
Journal: Journal of Food Quality
 (2019)

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## Abstract

The aim of this work was to investigate the influence ofTrichoderma spp. on volatile secondary metabolites and biometric parameters obtained from coriander (Coriandrum sativum L.). The fruits of coriander treated with liquid suspension spores of T. harzianum strain T22 and of T. asperellum strain B35 increased the yield of essential oil (by ∼36%); however, it was unaffected in its composition. Moreover, Trichoderma spp. influenced the yield and increased the number of seeds of coriander by ∼60%. Inoculation seeds with T. asperelleum strain B35 caused about 2-fold increase in the biomass of the aerial parts of coriander. There was also an increased root colonization by the fungus Trichoderma spp., limiting the number of phytopathogenic fungi from genus Fusarium observed.


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## Body

## 1. Introduction

In recent years, there has been a continuous increase in sales on the pharmaceutical market, or more broadly understood as the market of food, dietary supplements, natural substances, and herbs. Their usefulness in most cases is determined by pharmacopoeial standards (Polish or European Pharmacopoeia). Research shows that the quality of pharmacopoeial raw materials is influenced by many factors, such as (1) the cultivar used in cultivation, (2) type of substrate, (3) time and period of harvesting, or (4) the drying method [1]. The influence of the plant’s condition on the metabolites of secondary herbal raw materials is also known, which can be positively influenced by the use of living microorganisms. Microorganisms that support growth and protect plants from pathogens are a subject of growing interest [2–4]. The possibility of using living microorganisms to increase soil fertility and plant production efficiency has already been used in the middle of the last century [4, 5]. Among soil microorganisms, antagonistic fungi from genus Trichoderma are the most commonly studied and used in the biological plant protection and/or integrated pest management (IPM) programs [6]. These fungi are capable not only for stimulation of plant growth but also for induction of its defense mechanisms (i.e., ISR, induced systemic resistance; SAR, systemic acquired resistance) [7–9]. Trichoderma fungi produce a number of substances with antibiotic properties and hydrolytic enzymes (cellulases, chitinases, xylanases, pectinases, β-1,3-glucanases, and proteases among others), thanks to which they can quickly colonize plant roots and compete with phytopathogens for the place of infection or nutrients. Numerous reports show that several strains of Trichoderma had a significant reducing effect on plant diseases caused by soilborne and foliar pathogens (such as Rhizoctonia solani, Phytophthora spp., Pythium ultimum, Fusarium spp., Alternaria alternata, Sclerotinia spp., Gaeumannomyces graminis, Thielaviopsis basicola, Verticillium dahliae, and Botrytis cinerea) under greenhouse and field conditions [6, 9–13]. The use of Trichoderma fungi to protect herbal plants or organic crops can be an alternative to synthetic pesticides. The mechanisms of activity of the Trichoderma genus towards pathogens (e.g., antibiosis, micoparasitism, and competition) or stimulation of plant growth or induction of defense mechanisms is indicated widely in the literature [6–11]. However, there are very few papers describing the use of these antagonists in the cultivation of herbs or their impact on the content of aromatic compounds [14, 15].One of the world’s oldest herbs and species plants is coriander (Coriandrum sativum L.). Coriander is an annual, herbal, spicy, and melliferous plant, belonging to the celery family (Apiaceae). The main herbal raw materials are fruits containing large amounts of essential oil (0.2–2.6%) and polyphenolic acids (e.g., linoleic, oleic, palmitic, and stearic acids) as well as protein compounds, cellulose, pectins, mineral salts, and vitamins [14, 16]. Moreover, the coriander essential oils also have antibacterial properties against Gram-positive and Gram-negative bacteria (Staphylococcus aureus, Streptococcus haemolyticus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, and Proteus vulgaris) [14, 16].The data presented in the FEMA report (Flavor and Extract Manufacturers Association) indicate that the global annual production of coriander fruits is about 600,000 tonnes per year (http://www.femaflavor.org). Fruits of coriander are used mainly as a flavoring and aromatic additive in the production of food products (meat, snack, alcohol, and confectionery products). Moreover, coriander oils are included in the group of so-called safe (nontoxic) oils that can be used as a fragrance and supplement (in alcoholic beverages, cosmetics, soaps, tobacco, and medicines) [17–19].The aim of the present research was to determine the effect of coriander fruits treatment withTrichoderma fungi on the composition and content of volatile secondary metabolites. This study also investigated the influence of Trichoderma fungi on the biometric features of coriander and the degree of root colonization with these fungi. The degree of colonization of roots by toxicogenic fungi of the Fusarium genus was also examined.


## 2. Materials and Methods

In the study, coriander fruits (Coriandrum sativum var. microcarpum) of the Pallas variety belonging to the plants from the oil group (Legutko, Poland) were used.The study examined two strains of antagonistic fungi:T. harzianum strain T22 (commercial biological preparation Trianum, Koppert B.V., the Netherlands) and the T. asperellum strain B35.TheT. asperellum B35 strain comes from the collection of the Agricultural Microbiology Lab, Department of Plant Protection, Wrocław University of Environmental and Life Sciences, Poland. This strain stimulates the growth of plants and effectively limits a number of diseases of the root system of onions, cabbage, cucumbers, peppers, tomatoes, leek, and celery [20–22].


### 2.1. Field Experiment and Biometric Analyses

Single-factor field experiment was carried out from 8th of May up to 27th August 2015 in Pawłowice Research Station (51°09′N, 17°06′E) belonging to Wrocław University of Environmental and Life Sciences. The experiment was arranged with four replicates, on 10 m2 elementary plots. The forecrop of experiment in 2014 was oats. In 2015, prior to sowing, mineral fertilization was applied uniformly throughout the field at a dose of 40.0 kg N : 30.0 kg P : 50.0 kg K per hectare.The following treatments were applied: (1) control (dry fruit); (2) soaking fruits in sterile water for 10 min; and soaking fruits for 10 min in a conidia spores suspension of (3)T. harzianum T22 and (4) T. asperellum B35. The suspension of Trichoderma fungi spores was prepared by mixing the preparation with distilled water in an amount to obtain 1 × 107 spores 1 mL−1. Conidial densities in the suspension were determined by use of a hemocytometer under a light microscope. The coriander fruits were poured into a suspension and soaked for 10 min, mixing it constantly; then, the seeds were dried at room temperature and sown.Germination energy and capacity were determined before sowing. According to ISTA [23] regulations, germination energy was determined as the percentage of seeds that have been produced by seedlings classified as normal after 7 days of germination under optimal conditions. The germination capacity was expressed as a percentage of seeds that have produced normal seedlings after 14 days of germination under optimal conditions. The tests were performed on 100 fruits from each combination.After the growing season, 25 randomly selected plants were collected from each plot and biometric analyses were carried out. The number of fruits, the mass of about 1000 pieces (MTS), and the weight of fruits were calculated. The biomass increase of the aboveground parts and roots was also measured.


### 2.2. Root Colonization

Root colonization withTrichoderma fungi was examined in the period of full vegetation—before the harvest of the raw material for analyses on the content of active compounds. Roots were harvested from representative plants. The roots after mechanical soil removal were thoroughly washed with running tap water to remove adhering soil particles and then were rinsed with a sterile solution of 0.1 M MgSO4 × 7H2O. The roots were aseptically cut into ∼5-mm fragments and transferred on a PDA medium (Potato Dextrose Agar, Emapol, Poland) with 30 μg mL−1 streptomycin (to inhibit growth of bacteria colony). Eight fragments were placed per plate and incubated at 28°C. Growing colonies were observed daily. After incubation, the percent degree of roots colonization with Trichoderma and Fusarium fungi was determined [21, 24].


### 2.3. Analysis of the Content of Essential Oils

Determination of the composition and content of volatile metabolites was carried out by the method of hydrodistillation in the Deryng apparatus [25]. In a 250 mL round-bottom flask, 30 g of fresh coriander fruits was placed, 100 mL of redistilled water was added, and hydrodistillation was carried out for 3 hours. The collected distillate (extracted into 1 mL of cyclohexane and dried with Na2SO4) was stored at a temperature of −15°C until the GC-MS analysis was performed. The determination of the composition of volatile fractions was performed according to the chromatographic method developed by Szumny et al. [26]. About 50 μL of the previously prepared solution of the essential oils was diluted ten times in cyclohexane and subjected to the GC-MS analysis (injection of 1 μL).A gas chromatograph of the PerkinElmer Clarus 680 coupled to a mass spectrometer was used to identify volatile compounds. The separation was performed using an Elite-5MS column (30 m × 0.25 mm × 0.25μm film thickness). The identification of the compounds was carried out by (1) comparing the EI-MS spectrum of the compound with the spectrum in the NIST14 data library; (2) comparison of Kovats retention index of the identified molecules with data included in the NIST14, NIST Webbook database, and (3) comparison of retention times with commercially available standards isolated from plants.In addition, chromatographic analysis (GC-MS) of the main components of the obtained essential oils was confirmed by performing nuclear magnetic resonance spectra. The structures of compounds were confirmed on the basis of the comparison of signals obtained from NMR analysis with literature data. The spectra1H NMR, 13C NMR, and HSQC were performed on the Bruker Avance DRX-500 apparatus, using solutions in CDCl3.Specific rotation was measured on the Autopol IV Automatic Polarimeter (Rudolph) equipped with a thermostatic system. The analysis was performed in ethanol at a concentration expressed ing 100 mL−1.The enantiomeric excess of linalool was determined based on the GC analysis using a chiral column, separation on cyclodextrin associated with dimethyl polysiloxane (CP-Chirasil-DEX CB), with the 25 m, 0.25 mm, 0.25μm film thickness.


### 2.4. Statistical Analyses

The data from field experiment, biometric analyses, and root colonization were subjected to the analysis of variance using Tukey’s test (p<0.05) using STATISTICA 13.3 software for Windows.


## 2.1. Field Experiment and Biometric Analyses

Single-factor field experiment was carried out from 8th of May up to 27th August 2015 in Pawłowice Research Station (51°09′N, 17°06′E) belonging to Wrocław University of Environmental and Life Sciences. The experiment was arranged with four replicates, on 10 m2 elementary plots. The forecrop of experiment in 2014 was oats. In 2015, prior to sowing, mineral fertilization was applied uniformly throughout the field at a dose of 40.0 kg N : 30.0 kg P : 50.0 kg K per hectare.The following treatments were applied: (1) control (dry fruit); (2) soaking fruits in sterile water for 10 min; and soaking fruits for 10 min in a conidia spores suspension of (3)T. harzianum T22 and (4) T. asperellum B35. The suspension of Trichoderma fungi spores was prepared by mixing the preparation with distilled water in an amount to obtain 1 × 107 spores 1 mL−1. Conidial densities in the suspension were determined by use of a hemocytometer under a light microscope. The coriander fruits were poured into a suspension and soaked for 10 min, mixing it constantly; then, the seeds were dried at room temperature and sown.Germination energy and capacity were determined before sowing. According to ISTA [23] regulations, germination energy was determined as the percentage of seeds that have been produced by seedlings classified as normal after 7 days of germination under optimal conditions. The germination capacity was expressed as a percentage of seeds that have produced normal seedlings after 14 days of germination under optimal conditions. The tests were performed on 100 fruits from each combination.After the growing season, 25 randomly selected plants were collected from each plot and biometric analyses were carried out. The number of fruits, the mass of about 1000 pieces (MTS), and the weight of fruits were calculated. The biomass increase of the aboveground parts and roots was also measured.


## 2.2. Root Colonization

Root colonization withTrichoderma fungi was examined in the period of full vegetation—before the harvest of the raw material for analyses on the content of active compounds. Roots were harvested from representative plants. The roots after mechanical soil removal were thoroughly washed with running tap water to remove adhering soil particles and then were rinsed with a sterile solution of 0.1 M MgSO4 × 7H2O. The roots were aseptically cut into ∼5-mm fragments and transferred on a PDA medium (Potato Dextrose Agar, Emapol, Poland) with 30 μg mL−1 streptomycin (to inhibit growth of bacteria colony). Eight fragments were placed per plate and incubated at 28°C. Growing colonies were observed daily. After incubation, the percent degree of roots colonization with Trichoderma and Fusarium fungi was determined [21, 24].


## 2.3. Analysis of the Content of Essential Oils

Determination of the composition and content of volatile metabolites was carried out by the method of hydrodistillation in the Deryng apparatus [25]. In a 250 mL round-bottom flask, 30 g of fresh coriander fruits was placed, 100 mL of redistilled water was added, and hydrodistillation was carried out for 3 hours. The collected distillate (extracted into 1 mL of cyclohexane and dried with Na2SO4) was stored at a temperature of −15°C until the GC-MS analysis was performed. The determination of the composition of volatile fractions was performed according to the chromatographic method developed by Szumny et al. [26]. About 50 μL of the previously prepared solution of the essential oils was diluted ten times in cyclohexane and subjected to the GC-MS analysis (injection of 1 μL).A gas chromatograph of the PerkinElmer Clarus 680 coupled to a mass spectrometer was used to identify volatile compounds. The separation was performed using an Elite-5MS column (30 m × 0.25 mm × 0.25μm film thickness). The identification of the compounds was carried out by (1) comparing the EI-MS spectrum of the compound with the spectrum in the NIST14 data library; (2) comparison of Kovats retention index of the identified molecules with data included in the NIST14, NIST Webbook database, and (3) comparison of retention times with commercially available standards isolated from plants.In addition, chromatographic analysis (GC-MS) of the main components of the obtained essential oils was confirmed by performing nuclear magnetic resonance spectra. The structures of compounds were confirmed on the basis of the comparison of signals obtained from NMR analysis with literature data. The spectra1H NMR, 13C NMR, and HSQC were performed on the Bruker Avance DRX-500 apparatus, using solutions in CDCl3.Specific rotation was measured on the Autopol IV Automatic Polarimeter (Rudolph) equipped with a thermostatic system. The analysis was performed in ethanol at a concentration expressed ing 100 mL−1.The enantiomeric excess of linalool was determined based on the GC analysis using a chiral column, separation on cyclodextrin associated with dimethyl polysiloxane (CP-Chirasil-DEX CB), with the 25 m, 0.25 mm, 0.25μm film thickness.


## 2.4. Statistical Analyses

The data from field experiment, biometric analyses, and root colonization were subjected to the analysis of variance using Tukey’s test (p<0.05) using STATISTICA 13.3 software for Windows.


## 3. Results and Discussion

In the first stage of the study, the influence ofTrichoderma fungi on the germination energy and capacity of the coriander seeds was evaluated (Table 1). The germination energy of fruits depended on the used treatment. Soaking coriander fruits in water and spore suspension of Trichoderma spp. increased the germination energy in the range from 18% to 47%. Treatment of fruits with T. harzianum strain B35 increased the germination energy by 20% in comparison with water and Trianum treatment. The experimental combinations used did not affect the germination capacity of the coriander fruits.Table 1
Germination energy and capacity of the coriander fruits (%).
No.
Experimental combinations
Germination energy (%)
Germination capacity (%)
1.
Control
38a
80a
2.
Fruits soaked in water
48b
95a
3.
Fruits treated withT. harzianum T22
45b
98a
4.
Fruits treated withT. harzianum B35
56c
98a
The values in individual columns marked with the same letters differ significantly from the control objects (p=0.05).The development of coriander during the growing season depended on the method of seed treatment before sowing (Table2). Coriander growing on control objects had a slower development during the entire growing season, but it was fruiting earlier and had a slightly shorter growing season. A similar rate of coriander development was observed on objects with fruits soaked in water before sowing. In turn, biological treatment of coriander fruit with T22 and B35 strains accelerated the development of plants from 4 days during emergence to 6 days during flowering. It also extended the flowering period, fruit settling, and the growing season. Trichoderma species also had a positive influence on the increase in yield and fruit number in the range from 55% to 64% (Table 3). Newman et al. [13] had obtained similar effects in their study, while applying T. harzianum spores during the tomato cultivation. In our research, after inoculation fruits with T. asperellum strain B35, an ∼2-fold increase in plant biomass was obtained compared with the remaining experimental combinations. At the same time, an increased colonization of the roots by Trichoderma species was found in the treatment of fruits with T22 and B35 strains in comparison with objects a and b. Sobolewski et al. [27] investigating carrots had also observed significant increase in height and mass of the plants after applying Trichoderma spp. preparation instead of standard T 75 DS/WS (2012) preparation.Table 2
Length of phenological phases of coriander in the growing season.
Phenological phase
Control∗
Fruits soaked in water∗
Fruits treated withT. harzianum T22∗
Fruits treated withT. harzianum B35∗
Sowing
08.05
Beginning of emergence
15
12
12
12
Full emergence
4
4
3
3
Leafing
7
7
7
7
Formation of shoots
15
14
13
13
Beginning of budding
16
21
17
17
Beginning of flowering
11
11
9
9
Full flowering
6
6
4
4
Formation of fruit
10
14
16
16
Plant drying
20
18
23
23
Harvest
27.08
∗Number of days of individual development phases.Table 3
Coriander’s biometrics and roots colonization.
Specification
Control
Fruits soaked in water
Fruits treated withT. harzianum T22
Fruits treated withT. harzianum B35
Plant height (cm)
40a
39a
51b
52b
Mass of the superpowers from 1 plant (g)
29.4a
32.0a
37.5b
64.0c
The weight of roots from 1 plant (g)
3.4a
4.0a
4.3a
5.6b
Number of fruits from one plant (pcs)
112a
157b
174b
185b
Weight of fruit from 1 plant (g)
2.0a
2.6b
3.1b
3.2b
Mass of 1000 seeds (g)
9.3a
9.2a
10.7b
11.1b
Colonization of roots ofTrichoderma spp. (%)
29a
34a
71b
68b
Colonization of roots ofFusarium (%)
50a
51a
23b
26b
Colonization of roots ofF. oxysporum (%)
32a
34a
16b
18b
Colonization of roots ofF. culmorum (%)
9ab
13a
6b
6b
Colonization of roots ofF. avenaceum (%)
9a
4b
—
2b
Values in individual lines marked with the same letters differ significantly from the control objects according to Tukey’s test (p = 0.05).Moreover, from the coriander roots, phytopathogenic fungi from theFusarium genus (F. culmorum, F. oxysporum, and F. avenaceum) were isolated. The roots were colonized by these fungi most numerously, whose fruits before sowing was not biologically treated. This may be related to the faster colonization of these specific microniches by antagonistic fungi Trichoderma and their parasitic activity against these phytopathogens [12, 20–22].The obtained results unambiguously indicate the effect of coriander fruits treatment withTrichoderma strains tested on the content of essential oils (Table 4). The amount of volatile fraction for fruits treated with the Trianum preparation increased by ∼36%. A similar effect was obtained by Ratnakumari et al. (2014) [15], observing the increase in the content of the mint essential oil (Mentha arvensis) grown on the substrate inoculated with the T. harzianum strain NFCCI 2241 and T. ovosporum strain NFCCI 2689. The authors isolated 40 to 50% more volatile fractions from plants compared with controls. Singh et al. (2002) [14] observed an increase in the content of a dozen or so percent of the essential oil contained in Pogostemon cablin, in the soil inoculated with T. harzianum strain ATCC PTA-3701.Table 4
Quality and quantity composition of coriander oil and its efficiency.
No.
Name
KI exp.a
KI lit.a
Identification methodb
Control∗
Soaked fruits∗
Fruits treated withT. harzianum T22∗
Fruits treated withT. asperellum B35∗
Share %c
Share %c
Share %c
Share %c
1
Hexanal
801
800
KI, MS, S
0.04
0.02
0.03
0.02
2
Heptanal
902
894
KI, MS, S
0.03
0.01
0.03
0.06
3
Tricyclene
929
931
KI, MS
0.01
0.03
0.01
0.01
4
α-Thujen
932
932
KI, MS, S
0.03
0.04
0.02
0.02
5
α-Pinene
940
940
KI, MS, S, NMR
2.16 ± 0.05
2.19 ± 0.03
1.31 ± 0.05
1.67 ± 0.01
6
Camphene
957
954
KI, MS, S
0.38
1.11
0.42
0.39
7
Sabinene
978
977
KI, MS, S
0.21
0.2
0.14
0.17
8
β-Pinene
983
980
KI, MS, S
0.35
0.59
0.2
0.58
9
Myrcene
991
992
KI, MS, S
0.45
0.42
0.29
0.31
10
α-Terpinene
1021
1019
KI, MS, S
0.04
0.05
0.02
0.03
11
p-Cymene
1029
1026
KI, MS, S
0.54
0.02
0.4
0.62
12
Limonene
1035
1035
KI, MS, S
1.18 ± 0.12
0.45 ± 0.05
0.96 ± 0.04
0.97 ± 0.10
13
1,8-Cineol
1039
1037
KI, MS
0.05
3.33
0.23
0.12
14
γ-Terpinene
1064
1062
KI, MS, S, NMR
3.72 ± 0.09
3.47 ± 0.13
2.75 ± 0.11
3.28 ± 0.13
15
trans-Linalool oxide
1076
1076
KI, MS
0.20
0.15
0.14
0.09
16
Terpinolene
1089
1093
KI, MS, S
0.26
0.25
0.21
0.27
17
Linalool
1106
1106
KI, MS, S, NMR
77.55 ± 1.10
77 ± 0.96
82.43 ± 0.91
81.15 ± 1.17
18
Camphor
1156
1150
KI, MS, S, NMR
6.33 ± 0.26
7.07 ± 0.18
6.4 ± 0.22
4.8 ± 0.20
19
Borneol
1179
1173
KI, MS, S
0.21
0.77
0.21
0.04
20
Terpinen-4-ol
1186
1180
KI, MS, S
0.22
0.08
0.16
0.12
21
α-Terpineol
1198
1200
KI, MS, S
0.60
0.17
0.39
0.31
22
Decanal
1206
1207
KI, MS, S
0.19
0.04
0.37
2.55
23
Citronellol
1227
1232
KI, MS, S
0.10
0.04
0.05
0.04
24
Geraniol
1253
1258
KI, MS, S, NMR
4.43 ± 0.14
1.75 ± 0.15
2.31 ± 0.19
1.92 ± 0.15
25
Thymol
1296
1297
KI, MS, S
0.03
0.3
0.02
0.08
26
Geranyl acetate
1377
1383
KI, MS, S
0.69
0.44
0.5
0.38
27
Amount of oil (%)∗∗
0.55
0.64
0.75
0.68
aExp. = experimental; Lit. = literature. bKI = Kovac retention index; MS = comparison of the EI-MS spectrum obtained with the NIST11 library; S = comparison of retention time with a commercial or isolated pattern (chromatographic standard); NMR = comparison with literature shifts 13C and 1H NMR. cAverage of the three replicates; ∗percentage share acc. to the GC-MS analysis; ∗∗mean ± SD (standard deviation); ∗∗∗weight-volume percentage, determined according to the Deryng apparatus scale.Studies carried out with the GC-MS technique allowed to identify 26 compounds (Table4). In plants from the control objects a and b, monoterpenoid alcohol, linalool (77%), was the component with the highest content. In contrast, plants from the objects c and d, received 81.8% on average. The next components of the obtained oil were the monoterpenoids: camphor (∼5% to 7%) and geraniol (∼4%) as well as monoterpene α-pinene (∼2%) and γ-terpinene (∼4%). Monoterpenoids and monoterpenes accounted for 85.11% and 14.63% of the total amount of oil, respectively. The aliphatic aldehydes were found in trace amounts (less than 0.3%). With regard to the scientific literature, a number of studies on the composition and content of coriander oil were carried out. The literature data indicate that the composition and content of the oil differ depending on the climatic conditions of the region, growth conditions, and plant species. Msaada et al. [28] proved the presence of linalool (87.54%) and cis-dihydrocarvone (2.36%) as two main components of the fruits of the Tunisian coriander. Telci et al. [29] examined the chemical composition of two fruits varieties and observed differences in the content of the main component, linalool between the two varieties. According to literature data, coriander oil, isolated by hydrodistillation, contains from 58 to 81% of linalool [29]. In studies of coriander oils carried out by Orav et al. [30], the content of linalool also ranged from 58 to 80%. Other characteristic components were γ-terpinene, α-pinene, p-cymene, camphor, geranyl acetate, and geraniol. The highest amount of linalool was found in the oil isolated from Jantar coriander growing in Estonia. Furthermore, in obtained oil, higher contents of linalool and geranyl aceate were observed, than in oils obtained from coriander fruits in Netherlands, Russia, and France. In general, the fruits of European coriander in comparison with Asian ones are characterized by a higher content of linalool (∼30%) [30]. Results similar to ours were received by Sriti et al. [31] and Figueiredo et al. [32], who identified ∼40 compounds, of which the main components were also linalool, α-pinene, γ-terpinene, camphor, and geraniol. The quantitative analysis carried out by Msaada et al. [28] and Ravi et al. [33] showed the presence of 0.35 and 0.39% of the essential oils in coriander fruits.In previous studies, the presence of (S)-(+)-linalool (also called coriandrol) was found in the coriander essential oils. In fact, linalool is not a pure (S) isomer and contains from 12 to 15% of the (R) isomer [34, 35]. Oliver [36] in his work studied the samples of coriander essential oils for the presence of isomeric (S)-(+)-linalool. By means of gas chromatography (GC) and the use of a chiral column, he found the presence of 88% of (S)-(+)-linalool and 12% of (R)-(-)-linalool. The GC analysis we conducted on a column with a chiral filling and a proper rotation test confirmed the presence of (S)-(+)-linalool in the tested material. The presence of (S)-(+)-linalool in the essential oils was also found by Gaydou et al. [37] and Sadowski et al. [38]. Our research yielded very similar contents of (S)-(+)-linalool −82%.According to the European Pharmacopoeia, the fruits of coriander should contain no less than 0.3% of essential oils. The percentage content of components was also characterized: limonene (1.5–5%), geraniol (0.5–3%), geranyl acetate (0.5–4%), camphor (3–6%), linalool (65–78%),p-cymene (0.5–4%), γ-terpinene (1.5–8%), α-terpineol (0.1–1.5 %), α-pinene (3–7%). Comparing the data in the Pharmacopoeia with the data obtained in our work, it was found that the contents of camphor, α-pinene, and limonene slightly deviate from the acceptable standards.The conducted research is becoming particularly interesting in the context of recent reports that indicate the use of coriander in animal nutrition, which improves their well-being. These results were demonstrated by Abou-Elkhair et al. [39] in the broiler studies and Mohammed et al. [40] in research on the use of Awassi sheep and rams. In addition, as emphasized by Bahat et al. [41], Sriti et al. [32], Rezaei et al., [42] and Prachayasittkul et al. [43], the interest in coriander due to its antibacterial, anti-inflammatory, anticancerous, or antifungal activity and its richness of bioactive components (phytosterols, monoterpenes, monoterpenoids, etc.) is constantly growing. Therefore, it is a good premise for efforts such as those undertaken in our research to intensify its growth and improve the quantitative chemical composition.


## 4. Summary

The obtained results indicate the purposefulness of using biological control agents based onTrichoderma species in the cultivation of coriander as a source of essential oils. The comparison of efficiency of the distillates obtained with steam proves the positive effect of T. asperellum B35 on coriander. And most importantly, increasing the yield of essential oils obtained does not mean a change in composition. This oil is in the upper ranges of pharmacopoeial standards. At the same time, the use of antagonistic fungi affects the improvement of biometric parameters of the plant. An increased yield and the number of coriander fruits were found to be at the level of ∼60%. In the experimental combination with T. asperellum B35, the biomass of the aerial parts of the plant was twice as large.



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*Source: 5687032-2019-01-09.xml*