# Field Evaluation of Twelve Clones of Oregano Grown in the Main

Authors: Lorena E. Torres; Paula C. Brunetti; Carla Baglio; Pablo G. Bauzá; Ana G. Chaves; Yamile Massuh; Sonia F. Ocaño; Marta S. Ojeda
Journal: ISRN Agronomy
 (2012)

---

## Abstract

In Argentina, oregano (Origanum spp.) is one of the most important aromatic species. Leaves and flowering tops are used as seasoning, targeting the retail consumer, industrial and less to export. Local production has low-average yields due to the variability of cultivated material, the vegetative propagation methods used, and the lack of knowledge and adaptive experimentation on advanced cultivation practices. Clones of oregano grown in the country were collected in order to characterize the germplasm used in Argentina. Twelve oregano clones, sanitized by meristems culture and micropropagated in vitro, were field evaluated in three different growing locations. Regardless of growing site, the quantitative variables with more discriminating value were essential oils yield, internode length, length of the longest branch, fresh weight, dry weight of leaf and stem, leaf/stem ratio, and leaf area. Based on the quantitative traits, oregano clones can be classified into four groups. From the observations based on botanical characteristics, it was determined that the evaluated clones belong to three different taxa: Origanum vulgare ssp. vulgare, Origanum vulgare ssp. hirtum, and Origanum x majoricum (hybrid). Within each group, the clones belong to the same taxon.


---

## Body

## 1. Introduction

The genusOriganumbelongs to the family Lamiaceae and includes more than 70 species, subspecies, varieties, and hybrids that are commonly found as wild plants in the Mediterranean areas [1]. This aromatic herb, mainly used as condiment, in liquors formulations, tomato sauces, baked goods, and salad dressing [2–10], is one of the most important aromatic spices cultivated in Argentina from the point of view of demand and acreage (80%) [11]. The main production areas are located in the provinces of Mendoza, Córdoba, and San Juan [12] although there are smaller production areas in the northwestern regions of the provinces of Salta and Jujuy [10]. The local production of oregano is mainly for domestic consumption; however, in recent years a surplus production for export was registered.Despite this surplus production, the cultivation of oregano in our country is still affected by several problems. The oregano produced often does not have the best qualities to be considered a premium product for food because it is grown regardless of the variety, health, and the harvest and postharvest practices; in this situation, it is not possible to conserve a product free of contamination. Besides there is little taxonomic identification of the plant material used for cultivation; in fact, in our country registered cultivars and/or varieties of oregano do not exist [12], and the fantasy names used to designate the different materials are local names assigned by the producers, who include in those names phenotypic characteristics of the material or its place of origin [13]; even more, those fantasy names are often given to materials that are genetically equivalent [10].Regarding to the agronomic evaluation of oregano cultivated in Argentina, in addition, the agronomic performance assessment of the growing materials [14–18], the study of different types of irrigation [19, 20], different methods of drying and storage [21–25], and the characterization of essential oils (yield and composition) [12, 26–28] was carried out. Besides, Farías et al. [10] generated a list of 41 descriptors of oregano, with the aim of facilitating the identification, and characterization of different clones.From the characterization of genotypes of oregano grown in the country, in terms of agronomic performance, morphology, botanical identification and study of essential oils, it is possible to plan the competitive development of this aromatic herb of great commercial value.According to the above, the aims of this study were (1) to evaluate and characterize twelve genotypes of oregano grown in major production areas of the country through agronomically important variables, (2) to establish traits with the highest discriminant value to distinguish the genotypes of oregano, and (3) to carry out the taxonomic identification of these clones.


## 2. Materials and Methods

### 2.1. Plant Material

During spring and summer season of 2006/2007, plants belonging to twelve oregano clones coming from different collection sites were sanitized by meristem culture and micropropagatedin vitro in order to use them as mother plants (Table 1). During autumn season of 2008, those mother plants were placed in pots containing vermiculite, irrigated with a nutrient and fungicidal solution, and maintained under the greenhouse environmental conditions. A month after transplanting, these mother plants were multiplied by binodal cuttings to get the number of plants needed to carry out the field essays. Plants grown from cuttings binodal were maintained under controlled environmental conditions until transplanting to the field in spring of 2008.Table 1
Twelve clones of oregano collected from the main production areas of Argentina.
Provenance
Germplasm
Mendoza
Criollo (Cr-Mza)
Compacto (Cm-Mza)
Mendocino (Me-Mza)
Cordobés (Cd-Mza)
Córdoba
Clon 1 (C1-FCA)
Clon 2 (C2-FCA)
Clon 3 (C3-FCA)
Clon 8 (C8-FCA)
Negrito (NB-TS)
Mendocino (Me-TS)
Verde Limón (VL-TS)
San Juan
Negrito (Ne-SJ)
Mza: Mendoza; FCA: Facultad de Ciencias Agropecuarias; TS: Traslasierra Valley; SJ: San Juan.


### 2.2. Field Essays

In spring season of 2008, three field essays were conducted in order to evaluate the agronomic performance of twelve genotypes of oregano. They were implanted in three different locations: the Agricultural Experiment Station La Consulta: INTA (33°29′ S, 69°04′ O, San Carlos, Mendoza), a private farm in Potrero de Garay (31°47′ S, 64°33′ O, Santa María, Córdoba) and the Experimental Farm of the Faculty of Agricultural Sciences at the Universidad Nacional de Córdoba, (31°29′ S, 64°00′ O, Capilla de los Remedios road, 15.5 Km, Córdoba). The weather characteristics from the three culture locations are described in Table2.Table 2
Weather characteristics from the three culture locations where the comparative essays were implanted.
Locations
Altitude (mosl)
JaMT (°C)
JuMT (°C)
MAP (mm)
LC-Mza
954
21.8
6.2
212.5
PG-Cba
810
21.3
9.9
723.3
CE-Cba
370
22.8
9.7
606.5
mosl: meters over sea level; JaMT: January mean temperature; JuMT: July mean temperature; MAP: mean annual precipitation. LC-Mza: La Consulta (Mendoza); PG-Cba: Potrero de Garay (Córdoba); CE-Cba: Campo Escuela FCA-UNC (Córdoba).In every location, the oregano plants were transplanted following a completely randomized design, where each of the 12 clones was represented by 10 plants. The plants were distributed in lines spaced at 0.70 m apart with a distance of 0.25 m between plants on the line. The essays were watered twice a week, especially during the summer, and the cultural labors required for the proper development of the culture (as weeding) were also carried out.The different genotypes were evaluated based on their morphological, phonological, and yield characteristics. Along the growing season, the longest branch length (LBL*) (cm) was measured. Three months after planting, the first harvest was carried out (all plants were harvested individually and correctly identified) and internode length (IL) (cm), fresh weight (FW*) (g plant−1), dry weight (DW*) (g plant−1), leave dry weight (LDW*) (g plant−1), and stem dry weight (SDW*) (g plant−1) were also measured. To determine the leaf area (LA) (cm2), fully expanded leaves located in the middle third of each branch were taken and scanned; for the image processing, software ImageJ (ver. X.XX, NIH, USA) was used. In order to estimate the essential oil content (EO) (%), from each one of the clones under study, three samples of dry leaves (5 g each) were taken. The oil extraction was carried out for 45 minutes, by hydrodistillation in a modified Clevenger equipment; subsequently, oil content was expressed in mL per 100 g of dry material. Besides, the ratio leave dry weight/stem dry weight (L/S*), water content (WC) (%) and fiber content (FC) (%) were estimated. One year after planting, a second harvest was carried out and the longest branch length (LBL**) (cm), number of branches (n° plant−1) fresh weight (FW**) (g plant−1), and dry weight (DW**) (g plant−1) were registered.


### 2.3. Taxonomic Identification

In order to determine the taxonomic name of the twelve evaluated clones, one flowering branch belonging to every clone was collected, herborized and conserved in the Museo Botánico del Instituto Multidisciplinario de Biología Vegetal Herbarium (IMBIV). The botanical characterization, mainly based in floral characters, was carried out using the identification keys of Ietswaart [29], Xifreda [2], and Rouquaud and Videla [3].


### 2.4. Statistical Analysis

All results obtained for each of the measured variables were analyzed using the software INFOSTAT [30].The differences among clones and growing locations for each one of the evaluated variables were estimated using an ANOVA according to the following model:(1)Yijk=μ+Ci+Lj+Lj>Rk+C*Lij+εijk,
where Yijk represents the average value of each variable of the ith clone (Ci) evaluated at the jth location (Lj), Rk is the number of repetitions of each clone in every location, C*Lij is the interaction between the ith clone, and jth location and Eijk with ~N (0, S2) is the random error term associated with observation Yijk. Differences between mean values of each clone and location were determined using the posteriori test for comparison of means DGC [31].By using the principal component analysis (PCA) that was assessed the potentiality of quantitative traits measured was assessed, in order to discriminate the different clones in each growing environment. Finally, a Generalized Procrustes Analysis (GPA) was conducted in order to determine the level of consensus between the configurations obtained for all evaluated characters in each growing location, in other words to determine whether the variables have the same discriminant value in all growing environment.


## 2.1. Plant Material

During spring and summer season of 2006/2007, plants belonging to twelve oregano clones coming from different collection sites were sanitized by meristem culture and micropropagatedin vitro in order to use them as mother plants (Table 1). During autumn season of 2008, those mother plants were placed in pots containing vermiculite, irrigated with a nutrient and fungicidal solution, and maintained under the greenhouse environmental conditions. A month after transplanting, these mother plants were multiplied by binodal cuttings to get the number of plants needed to carry out the field essays. Plants grown from cuttings binodal were maintained under controlled environmental conditions until transplanting to the field in spring of 2008.Table 1
Twelve clones of oregano collected from the main production areas of Argentina.
Provenance
Germplasm
Mendoza
Criollo (Cr-Mza)
Compacto (Cm-Mza)
Mendocino (Me-Mza)
Cordobés (Cd-Mza)
Córdoba
Clon 1 (C1-FCA)
Clon 2 (C2-FCA)
Clon 3 (C3-FCA)
Clon 8 (C8-FCA)
Negrito (NB-TS)
Mendocino (Me-TS)
Verde Limón (VL-TS)
San Juan
Negrito (Ne-SJ)
Mza: Mendoza; FCA: Facultad de Ciencias Agropecuarias; TS: Traslasierra Valley; SJ: San Juan.


## 2.2. Field Essays

In spring season of 2008, three field essays were conducted in order to evaluate the agronomic performance of twelve genotypes of oregano. They were implanted in three different locations: the Agricultural Experiment Station La Consulta: INTA (33°29′ S, 69°04′ O, San Carlos, Mendoza), a private farm in Potrero de Garay (31°47′ S, 64°33′ O, Santa María, Córdoba) and the Experimental Farm of the Faculty of Agricultural Sciences at the Universidad Nacional de Córdoba, (31°29′ S, 64°00′ O, Capilla de los Remedios road, 15.5 Km, Córdoba). The weather characteristics from the three culture locations are described in Table2.Table 2
Weather characteristics from the three culture locations where the comparative essays were implanted.
Locations
Altitude (mosl)
JaMT (°C)
JuMT (°C)
MAP (mm)
LC-Mza
954
21.8
6.2
212.5
PG-Cba
810
21.3
9.9
723.3
CE-Cba
370
22.8
9.7
606.5
mosl: meters over sea level; JaMT: January mean temperature; JuMT: July mean temperature; MAP: mean annual precipitation. LC-Mza: La Consulta (Mendoza); PG-Cba: Potrero de Garay (Córdoba); CE-Cba: Campo Escuela FCA-UNC (Córdoba).In every location, the oregano plants were transplanted following a completely randomized design, where each of the 12 clones was represented by 10 plants. The plants were distributed in lines spaced at 0.70 m apart with a distance of 0.25 m between plants on the line. The essays were watered twice a week, especially during the summer, and the cultural labors required for the proper development of the culture (as weeding) were also carried out.The different genotypes were evaluated based on their morphological, phonological, and yield characteristics. Along the growing season, the longest branch length (LBL*) (cm) was measured. Three months after planting, the first harvest was carried out (all plants were harvested individually and correctly identified) and internode length (IL) (cm), fresh weight (FW*) (g plant−1), dry weight (DW*) (g plant−1), leave dry weight (LDW*) (g plant−1), and stem dry weight (SDW*) (g plant−1) were also measured. To determine the leaf area (LA) (cm2), fully expanded leaves located in the middle third of each branch were taken and scanned; for the image processing, software ImageJ (ver. X.XX, NIH, USA) was used. In order to estimate the essential oil content (EO) (%), from each one of the clones under study, three samples of dry leaves (5 g each) were taken. The oil extraction was carried out for 45 minutes, by hydrodistillation in a modified Clevenger equipment; subsequently, oil content was expressed in mL per 100 g of dry material. Besides, the ratio leave dry weight/stem dry weight (L/S*), water content (WC) (%) and fiber content (FC) (%) were estimated. One year after planting, a second harvest was carried out and the longest branch length (LBL**) (cm), number of branches (n° plant−1) fresh weight (FW**) (g plant−1), and dry weight (DW**) (g plant−1) were registered.


## 2.3. Taxonomic Identification

In order to determine the taxonomic name of the twelve evaluated clones, one flowering branch belonging to every clone was collected, herborized and conserved in the Museo Botánico del Instituto Multidisciplinario de Biología Vegetal Herbarium (IMBIV). The botanical characterization, mainly based in floral characters, was carried out using the identification keys of Ietswaart [29], Xifreda [2], and Rouquaud and Videla [3].


## 2.4. Statistical Analysis

All results obtained for each of the measured variables were analyzed using the software INFOSTAT [30].The differences among clones and growing locations for each one of the evaluated variables were estimated using an ANOVA according to the following model:(1)Yijk=μ+Ci+Lj+Lj>Rk+C*Lij+εijk,
where Yijk represents the average value of each variable of the ith clone (Ci) evaluated at the jth location (Lj), Rk is the number of repetitions of each clone in every location, C*Lij is the interaction between the ith clone, and jth location and Eijk with ~N (0, S2) is the random error term associated with observation Yijk. Differences between mean values of each clone and location were determined using the posteriori test for comparison of means DGC [31].By using the principal component analysis (PCA) that was assessed the potentiality of quantitative traits measured was assessed, in order to discriminate the different clones in each growing environment. Finally, a Generalized Procrustes Analysis (GPA) was conducted in order to determine the level of consensus between the configurations obtained for all evaluated characters in each growing location, in other words to determine whether the variables have the same discriminant value in all growing environment.


## 3. Results and Discussion

Significant differences among the twelve evaluated clones were observed in most of quantitative traits measured in all cultivation locations (Tables3, 4, and 5). The agronomic performance of 12 clones evaluated in each cultivation sites is described below.Table 3
Quantitative trait mean values for the twelve clones of oregano evaluated in La Consulta (San Carlos, Mendoza).
Clon
LBL1 (cm)
IL (cm)
LA (cm2)
FW*(g pl-1)
DW*(g pl-1)
LDW* (g pl-1)
SDW* (g pl-1)
L/S*
EO (%)
WC* (%)
FC* (%)
LBL2 (cm)
Branch (N°)
FW** (g pl-1)
DW** (g pl-1)
Me-TS
19.02a
2.27b
0.36a
60.61a
18.73a
14.34a
4.39a
3.31a
2.00d
69.15b
30.90a
47.34a
94a
272a
107.5a
VL-TS
23.42a
2.53c
0.53c
52.80a
16.79a
11.32a
5.48a
2.02a
0.72b
68.41b
31.80a
54.86c
112a
240a
106.0a
NB-TS
29.59b
2.16b
0.57c
73.72a
23.08a
17.75b
5.33a
4.07b
0.83b
68.17b
31.30a
56.01c
97a
282a
105.1a
Ne-SJ
30.93b
1.82a
0.84d
87.31b
25.51a
20.65b
4.86a
5.29b
0.35a
70.38b
29.21a
46.04a
101a
322a
108.5a
Me-Mza
28.56b
2.59c
0.49b
63.10a
19.07a
13.56a
5.51a
2.52a
2.36e
69.94b
30.22a
52.56b
96a
268a
101.0a
Cm-Mza
30.02b
2.17b
0.53c
80.41a
22.94a
18.63b
4.30a
4.48b
0.91b
71.17b
28.53a
56.77c
118a
314a
118.5a
Cd-Mza
32.86b
1.66a
0.40a
62.70a
21.08a
14.52a
6.56a
2.28a
1.42c
65.96a
33.62b
46.33a
109a
266a
110.0a
Cr-Mza
34.49b
2.13b
0.49b
50.87a
17.94a
11.81a
6.13a
1.85a
1.35c
64.42a
35.27b
57.21c
115a
353a
128.9a
C1-FCA
38.86b
1.94b
0.45b
66.97a
22.39a
15.66a
6.73a
2.33a
1.42c
66.53a
33.43b
57.28c
123a
368a
131.0a
C2-FCA
37.72b
2.04b
0.40a
74.47a
25.56a
17.73b
7.83b
2.30a
1.55c
65.76a
34.32b
57.74c
119a
334a
142.5a
C3-FCA
23.04a
1.68a
0.47b
94.12b
28.58a
20.33b
8.25b
2.78a
0.50a
69.72b
30.37a
51.68b
102a
544b
191.1b
C8-FCA
35.99b
1.75a
0.49b
98.89b
31.50b
23.66b
7.84b
3.29a
0.66b
68.24b
31.85a
51.82b
103a
544b
192.0b
C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length; IL: internode length, LA: leaf area, FW: fresh weight, DW: dry weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, WC: water content, FC: fiber content, *measured at 1st harvest, **measured at 2nd harvest, 1three month after sowing, 2one year after sowing. Different letters indicate significant differences (P≤0.05).Table 4
Quantitative trait mean values for the twelve clones of oregano evaluated in Potreto de Garay (Santa María, Córdoba).
Clon
LBL1 (cm)
IL (cm)
LA (cm2)
FW*(g pl-1)
DW* (g pl-1)
LDW* (g pl-1)
SDW* (g pl-1)
L/S*
EO (%)
WC* (%)
FC* (%)
LBL2 (cm)
Branch (N°)
FW** (g pl-1)
DW** (g pl-1)
Me-TS
28.04a
0.96a
0.68b
8.90a
5.53a
2.51a
3.02a
0.73a
3.11c
39.39a
62.13b
21.43a
30b
31.97a
13.76a
VL-TS
32.25a
1.08b
0.36a
36.50b
16.22b
10.64b
5.58a
1.68b
0.71a
49.58b
44.44a
24.16a
30b
37.79a
15.80a
NB-TS
28.75a
1.17b
1.06d
54.85c
22.47c
15.99c
6.48a
2.58c
0.43a
57.26c
40.97a
31.12b
68d
145.73c
56.30c
Ne-SJ
41.54b
1.14b
0.65b
61.70c
25.11c
18.62c
6.49a
3.30d
0.36a
58.89c
40.70a
36.41b
83d
194.51d
64.94c
Me-Mza
33.18a
0.91a
0.53b
8.55a
5.05a
1.86a
3.19a
0.52a
4.63d
38.07a
59.06b
20.76a
17a
19.42a
8.75a
Cm-Mza
26.90a
1.14b
0.92c
60.30c
25.11c
17.60c
7.51a
2.54c
0.41a
57.34c
41.64a
32.50b
76d
144.86c
57.19c
Cd-Mza
29.17a
1.09b
0.55b
21.35a
9.91b
5.90b
4.01a
1.45b
1.46b
53.94c
46.42a
31.43b
38b
66.30b
28.28b
Cr-Mza
33.60a
1.35c
0.61b
31.35b
13.21b
8.03b
5.17a
1.48b
1.19b
55.72c
42.14a
35.53b
45b
86.06b
37.23b
C1-FCA
35.42a
1.17b
0.56b
35.85b
16.80b
10.42b
6.38a
1.43b
1.49b
47.44b
46.86a
35.14b
57c
109.65b
46.49b
C2-FCA
33.27a
1.11b
0.61b
35.75b
16.81b
10.70b
6.11a
1.75b
1.56b
53.03c
47.02a
34.42b
54c
95.23b
40.07b
C3-FCA
36.91a
1.22b
0.44a
42.79b
18.28b
12.21b
7.58a
1.66b
1.00b
55.33c
42.72a
47.10d
53c
143.99c
57.38c
C8-FCA
38.16a
1.22b
0.41a
68.50c
29.54c
16.33c
13.20b
1.72b
1.01b
56.39c
43.12a
41.88c
61c
135.78c
55.02c
C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino”, Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, DW: dry weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, WC: water content, FC: fiber content, *measured at 1st harvest, **measured at 2nd harvest, 1three month after sowing, 2one year after sowing. Different letters indicate significant differences (P≤0.05).Table 5
Quantitative trait mean values for the twelve clones of oregano evaluated in Capilla de los Remedios (Río Segundo, Córdoba).
Clon
LBL1 (cm)
IL (cm)
LA (cm2)
FW*(g pl-1)
DW*(g pl-1)
LDW* (g pl-1)
SDW* (g pl-1)
L/S*
EO (%)
WC* (%)
FC* (%)
LBL2 (cm)
Branch (N°)
FW** (g pl-1)
DW** (g pl-1)
Me-TS
26.01b
1.24b
0.53a
113.55a
35.12a
25.94a
9.18a
3.22b
1.91b
69.58a
30.93b
22.91b
86a
62.04a
29.06a
VL-TS
14.03a
0.75a
0.48a
113.48a
31.85a
20.07a
11.78a
2.09a
0.52a
72.04a
28.07b
17.30a
97a
28.36a
12.55a
NB-TS
17.04a
0.90a
0.71b
227.26b
63.99b
45.92b
18.07a
3.16b
0.80a
72.03a
28.16b
25.78b
188b
134.77b
53.88b
Ne-SJ
21.22a
0.86a
0.96c
275.49b
84.81b
53.99b
30.82b
2.27a
0.41a
69.45a
30.79b
13.37a
74a
50.01a
16.75a
Me-Mza
31.86b
1.29b
0.57a
143.58a
43.60a
31.10a
12.50a
2.70a
2.38c
70.16a
30.37b
31.96b
76a
93.35b
42.15b
Cm-Mza
16.26a
0.87a
0.74b
211.80b
62.86b
43.67b
19.19a
3.60b
0.76a
70.70a
29.68b
28.51b
147a
112.26b
44.74b
Cd-Mza
26.59b
1.38b
0.62a
140.52a
41.04a
24.20a
16.84a
1.77a
1.11a
70.88a
29.21b
39.77c
125a
116.37b
55.77b
Cr-Mza
26.49b
1.34b
0.60a
145.49a
40.27a
23.91a
16.36a
1.54a
0.88a
72.24a
27.68b
40.55c
157b
162.36c
77.14c
C1-FCA
31.67b
1.22b
0.52a
125.22a
37.81a
22.60a
15.21a
1.65a
0.73a
70.03a
30.19b
29.46b
126a
109.79b
47.33b
C2-FCA
25.96b
1.24b
0.52a
132.50a
36.57a
21.85a
14.71a
1.73a
0.82a
72.43a
27.60b
34.24b
142b
119.16b
57.39b
C3-FCA
19.71a
0.87a
0.69b
240.43b
67.35b
44.27b
23.07b
2.28a
0.33a
72.17a
28.01b
22.51b
102a
86.87b
31.89a
C8-FCA
16.32a
0.82a
0.61a
188.87b
45.98a
32.18a
13.80a
2.79a
0.52a
75.12b
24.34a
24.12b
110a
80.98b
31.43a
C1-FCA: clone 1 C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, DW: dry weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, WC: water content, FC: fiber content, *measured at 1st harvest, **measured at 2nd harvest, 1three month after sowing, 2one year after sowing. Different letters indicate significant differences (P≤0.05).


### 3.1. La Consulta INTA Essay

Clones Me-TS, VL-TS, and C3-FCA showed the lower mean value for longest branch length, when this trait was measured three months after sowing. One year after sowing, the clones with the lower longest branch length mean value were Me-TS, Ne-SJ, and Cd-Mza.Regarding the fresh weight yield of the first harvest, clones C8-FCA, C3-FCA, and Ne-SJ stood out for their higher production of fresh biomass, followed by clones Cm-Mza and NB-TS. In relation to dry weight, again C8-FCA clone differed significantly from the other clones because of its greater dry biomass yields, followed by clones C3-FCA, C2-FCA, Ne-SJ, Cm-Mza and NB-TS. The production of fresh and dry biomass can be related to the length of the internodes and leaf area; clones C3-FCA, C2-FCA, Ne-SJ, Cm-Mza, and NB-TS also showed shorter internodes and greater area leaf (Table 3). In the second harvest, clones C3-FCA and C8-FCA significantly surpassed the rest of the material for both fresh and dry weight. Regardless of the clone, the fresh and dry weight yields observed in both harvests are generally similar to those reported by Droushiotis and Della [32] and Weglarz and Geszprych [33].With regard to the percentage of water that is lost during the drying process, significant differences were observed among the clones both in the first as in the second harvest. In the first harvest, the clones with the lowest content of water were Cr-Mza, C2-FCA, Cd-Mza, and C1-FCA, which in turn showed the highest percentage of fibers (66.47%, and 34.16% resp.). In the second harvest, the clones with lowest water content and higher fiber content were Cd-Mza, VL-TS, and C2-FCA, with an average percentage of 54.68% of water and 45.32% fiber. The percentage of water found in the second harvest was similar to that reported by Leto and Salamone [34], who studied 62 biotypes of oregano coming from 17 different locations. Higher fiber content (or less water loss) is equivalent to a higher dry matter yield. That is, on average, those clones with higher fiber content showed a dry matter yield 7.62% higher than the rest of the clones in the first crop and 15.85% in the second harvest. This is important when estimating production volumes of dry matter.Significant differences were also observed among the clones in terms of dry weight of leaves and stems and leaf/stem ratio (Table3), being this latter variable very important when estimating the returns as are leaves the part of the plant that is consumed [35]. Clones that showed the highest values for leaf/stem ratio were Ne-SJ, Cm-Mza, and NB-TS, these clones had short internodes, a greater number of nodes (and therefore more foliage), and greater leaf area.Based on the essential oil yield, it was possible to distinguish four groups of oregano. The first group showed an average essential oil content of 2.18% which includes the clones Me-Mza and Me-TS. The second group, including the clones C2-FCA, C1-FCA, Cd-Mza and Cr-Mza, showed an average essential oil content of 1.44%, while the third group, involving the clones Cm-Mza, NB-TS, VL-TS y C8-FCA, showed an average oil percentage of 0.78%. Finally, the group with the lowest essential oil content was constituted by clones C3-FCA and Ne-SJ, which showed an average oil yield of 0.43%. The average essential oil yield observed for the first and second group surpass in 67.7% and 10.8%, respectively, the percentages of oil reported by Weglarz and Geszprych [33], whereas the essential oil content observed in the third and fourth group was in the range reported by these authors (0.30–1.30%).Finally, the number of branches per plant ranged from 94 (Me-TS) to 123 (C1-FCA); however, no significant differences were observed among the twelve evaluated clones.Principal component analysis (PCA) showed that 73.1% of the total variability is explained by the first factorial plane (PC1 and PC2). At the level of the first axis (PC1), which explains 42.2% of the total variability, the variables with greater weight were the leave dry weight, fresh weight of the second harvest, internode length, and essential oil content, whereas the second axis (PC2) explains 30.9% of total variability, and the variables with greater inertia are the stem dry weight, leaf/stem ratio, and leaf area (Figure1).Figure 1
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in La Consulta (Mendoza). C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length; LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing); **measured at 2nd harvest (one year after sowing).


### 3.2. Potrero de Garay Essay

With regard to the length of the longest branch, measured three months after planting, clone Ne-SJ differed significantly from the other clones, showing the longest branch length mean value, while no differences were observed among the 11 remaining clones (whose branch length ranged between 26.90 and 38.16 cm) (Table4). Considering the same variable, measured one year after sowing, the clones Me-Mza, Me-TS, and VL-TS differed from the other clones by their shorter length (22.12 cm on average), and the clones C8-FCA (41.88 cm) and C3-FCA (47.10 cm) were distinguished by their greater length, while no differences were observed between the remaining clones, with an average branch length of 33.79 cm.Regarding the fresh weight yield of the first harvest, clones C8-FCA, Ne-SJ, Cm-Mza, and NB-TS were those that stood out for their higher production of fresh biomass, with an average fresh weight of 61.34 g plant−1, while the clones Me-Mza and Me-TS differed from the rest of the clones by their poor performance in fresh matter (on average 8.73 g plant−1), followed by the clone Cd-Mza with a fresh biomass yield of 21.35 g plant−1. In relation to dry weight, clones C8-FCA, Ne-SJ, Cm-Mza, and NB-TS showed dry biomass yields significantly superior from those observed in the remaining clones, followed by clones C3-FCA, C2-FCA, C1-FCA, VL-TS, Cr-Mza, and Cd-Mza, with an average dry matter yield of 15.21 g plant−1. Being the clones Me-Mza and Me-TS those showed the lowest yield in dry weight (Table 4).The higher production of fresh and dry biomass observed in clones C8-FCA, Ne-SJ, Cm-Mza and NB-TS can be associated with the largest number of branches per plant (more than 60 branches per plant), with a longer branch (C8-FCA and Ne-SJ) and greater leaf area (cm Mza and NB-TS) (Table 4). The low yields observed in the first harvest for fresh and dry weight, lower than those values reported by Droushiotis and Della [32] and Weglarz and Geszprych [33], can be attributed to the dehydration suffered by the plants during the week before the harvest, being the clones Me-Mza and Me-TS the clones that most resented the lack of water.In the second harvest, clones Ne-SJ, NS-TS, Cm-Mza, C3-FCA, and C8-FCA showed a significantly superior yield, both in fresh as in dry biomass, with an average fresh weight of 152.97 g plant−1 and an average dry weight of 58.17 g plant−1. Clones C1-FCA, C2-FCA, Cr-Mza, and Cd-Mza showed average yields in fresh and dry matter of 89.31 g plant−1, and 38.02 g plant−1 respectively; while the clones Me-Mza and Me-TS were those with the lowest yield both in fresh weight (25.70 g plant−1) as in dry weight (11.26 g plant−1), as they were in the first harvest, followed by the clone VL-TS.As regards the percentage of water that is lost during the drying process, significant differences were observed between the clones both in the first as in the second harvest. In both harvests, the clones Me-Mza and Me-TS showed the lowest water content (and the highest percentage of fibers) (Table4). At this point, it is important to note that in the week preceding the first harvest, Potrero de Garay field was exposed to water stress, so the percentage of water lost during the drying process was lower than in La Consulta and Capilla de los Remedios. For this same reason, the percentage of fibers of the different clones, measured in the first harvest, was overestimated; however, it was possible to differentiate the oregano clones based on these variables. It is reasonable to assume that a high fiber content (equivalent to less water loss) results in a higher dry weight yield. Even though clones Me-Mza and Me-TS had higher fiber content, their biomass production was so low that they fail to align their yields with those clones that have higher percentages of water.In addition, significant differences were found among the clones in terms of the dry weight of the leaves and the stems and leaf/stem ratio (Table4), being this latter variable of major economic importance when estimating amount of production since it is the leaves that are used for consumption. The clones Ne-SJ, NB-TS, and Cm-Mza were those that showed the greatest values for leaf/stem ratio, coinciding with the greatest number of branches, longest branch, and greatest leaf area (Table 4).Concerning the content of essential oils, the clones Me-Mza and Me-TS showed an essential oil yield significantly higher than the rest of the clones, with an average of 3.87% of oil, followed by the clones C2-FCA, C1-FCA, Cd-Mza, Cr-Mza, C8-FCA, and C3-FCA, which showed an average oil content of 1.29% and the clones VL-TS, NB-TS, Cm-Mza and Ne-SJ (clones with the lowest performance in oils, producing on average 0.48%). The essential oil yield observed for the clones Me-Mza and Me-TS was similar the one reported for Origanum dubium (3.30 to 5.85%) [32]. These percentages were three times higher than those reported by Weglarz & Geszprych [33] and 32.5% higher than the yield observed in Turkish oregano (Origanum onites) [36], one of the species of greater content of essential oils.The principal component analysis (PCA) showed that 82.6% of the total variability was explained by the first factorial plane (CP1 and CP2). On the level of CP1, component that by itself explains 63.8% of the total variability, the variables with higher weight were the dry weight of leaf, the fresh weight measured at the second harvest, and the content of essential oils, while, at the level of CP2, the variables with greater inertia were leaf area and length of the longest branch (measured at 400 days after planting) (Figure2).Figure 2
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in Potrero de Garay (Córdoba). C1-FCA: clone 1 C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8; Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing), **measured at 2nd harvest (one year after sowing).


### 3.3. Capilla De Los Remedios Essay

Regarding to the length of the longest branch, measured 85 days after seeding, the clones VL-TS, Cm-Mza, NB-TS, C8-FCA, C3-FCA, and Ne-SJ differed significantly from the rest, recording the lowest values for this variable (range 14.03 and 21.22 cm) (Table 5). When this variable was determined 400 days after sowing, Ne-SJ and VL-TS differed from the other clones for its smaller branch length (15.34 cm on average), while Cd-Mza and Cr-Mza were stood out for their greater length (40.16 cm). No differences were observed between the remaining genotypes, whose average branch length was 27.44 cm (Table 5).With regard to fresh weight yield of the first harvest, two significantly different groups were distinguished. The clones C8-FCA, Cm-Mza, NB-TS, C3-FCA, and Ne-SJ comprise the higher group, with an average production of fresh biomass of 228.77 g plant−1, exceeding 75.14% the group of lower values for the variable formed by the seven remaining clones, which had an average yield of fresh weight of 130.62 g plant−1. The same trend was observed in relation to dry weight, where the clones Cm-Mza, NB-TS, C8-FCA and Ne-SJ differed significantly from the rest by its higher values which averaged 69.75 g plant−1 (Table 5). As in the essays of La Consulta and Potrero de Garay, clones with higher production of fresh and dry biomass are generally characterized by the presence of a greater number of branches per plant (more than 70 branches per plant), shorter internodes, and greater leaf area. Once again, the observed values for the variables fresh weight and dry weight are within the range reported by Droushiotis and Della [32] and Weglarz and Geszprych [33].Although in the second harvest the clones differed from one to another both in fresh as dry weight (Table5), due to the strong predation caused by ants, the recorded yields did not reflect the real potential of each genotype. Although all the clones were targeted by these insects, Ne-SJ, VL-TS, Me-TS, C8-FCA, and C3-FCA were the most affected, being Ne-SJ the clone that showed the most important performance reduction for fresh and dry biomass (50.01 g plant−1 and 16.75 g plant−1, resp.). Taking into account the agronomic performance observed for the same clone (Ne-SJ) both in the essays conducted in La Consulta and Potrero de Garay as in the first harvest of this essay, it can be assumed that in normal conditions (without predation) the yield of this clone would be far higher. Furthermore, it is reasonable to assume that without ants, the second harvest would be maintained the trend observed in the first one for all the clones. Regardless of the insect damage, the clones VL-TS and Me-TS were usually the lowest performing group both in fresh as in dry biomass yield, following the same trend observed in La Consulta and Potrero de Garay. The clones NB-TS, Cm-Mza, C3-FCA, and C8-FCA (which were the top performers in the first harvest) and the clone Me-Mza (low performance in the first harvest) did not differ significantly from clones C1-FCA, C2-FCA, and Cd-Mza, with an average fresh weight yield of 106.69 g plant−1 while the clone Cr-Mza had a biomass yield significantly higher than the rest of the clones, with an average fresh weight of 162.36 g plant−1. In terms of dry weight, the clones VL-TS, Ne-SJ, Me-TS, C8-FCA, and C3-FCA had an average yield of 24.34 g plant−1; the clones Me-Mza, Cm-Mza, C1-FCA, NB-TS, Cd-Mza, and C2-FCA yielded an average of 50.21 g plant−1 while Cr-Mza was significantly higher than the rest with a fresh biomass production averaged 77.14 g plant−1 (Table 5).Respecting to the percentage of water lost during the drying process, significant differences were seen among the clones both in the first as in the second harvest. In the first harvest, C8-FCA differed significantly from the rest of the clones, with a water content exceeding 5.71% of the 11 remaining clones and being also the genotype with the lowest fiber content (Table 5). In the second harvest, the clone with highest water content was Ne-SJ (65.46%), a value that was on average 7.58% higher than that observed for clones. Regarding the fiber content in the second harvest, the clones Cd-Mza, C2-FCA, Cr-Mza, Me-TS, VL-TS, and Me-Mza had the highest value for this variable, containing average of 45.54% of fiber (Table 5). As already noted above it can be assumed that, at the same fresh weight yield, a higher fiber content is equivalent to a higher dry matter yield.Regarding the leaves and stems dry weight and leaf/stem ratio, significant differences were also observed between genotypes. The clones mainly characterized by shorter internodes, high number of branches, and the greater leaf area (clones NB-TS and Cm-Mza) and the low stems weight (clone Me-TS) were those who showed the highest values for leaf/stem ratio (Table5).Concerning the content of essential oils, Me-Mza and Me-TS differed significantly from the rest of clones, with an average essential oil content of 2.15%, while no differences were observed among the 10 remaining clones (which oil content ranged from 0.33% to 1.11%). According to Kokkini [37], based on performance of essential oils, it is possible to categorize the different genotypes of oregano into three groups: (a) those producing less than 0.05% (“poor” in oils), (b) those with essential oil content between 0.05 and 2%, and (c) those with an essential oil yield over 2% (“rich” in oil). Both in this essay as in those conducted in La Consulta and Potrero de Garay, clones of oregano belonging to the three groups were distinguished. The clones called “Mendocino” (Me-Mza and Me-TS) belong to the rich-oil group, with a performance that in the three locations exceeds 2%, while the clones called “Compacto” (Cm-Mza) and “Negritos” (NB-TS and Ne-SJ) belong to the group poor-oil group. These results are consistent with those observed by De Mastro [38]. Franz & Novak [35] propose to use differential production of oregano based on the essential oil yield, destinating for use as dried herb those materials having 1 to 3% of oil, and for essences production those clones containing levels of oil exceeding 3%. Oregano production intended for obtaining essential oils becomes more important if we consider that these essences and/or its components represent a potentially important source of pharmacologically active compounds [39] or agrochemicals (as herbicides) [40]. Particularly, it has been tested that the essential oil of oregano has a strong antioxidant activity in foods with high-fat content and fried-salted peanuts [41, 42].The principal component analysis (PCA) showed that 73% of the total variability is explained by the first factorial plane formed by the two first principal components. On the level of CP1, that by itself explains 50.1% of the total variability, the more discriminating variables were leaf dry weight, leaf area, dry weight of stems, the length of internodes and the essential oil content, while on the level of CP2 (which accounts for 22.9% of the total variability observed) were fresh weight measured in the second harvest, leaf/stem ratio, and the length of the longest branch. The PCA showed that all 12 clones of oregano evaluated tend to cluster in a manner similar to those observed in La Consulta and Potrero de Garay (Figure3).Figure 3
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in Capilla de los Remedios (Córdoba). C1-FCA: clone 1 C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8; Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing); **measured at 2nd harvest (one year after sowing).With regard to generalized procrustes analysis (GPA), combining the information obtained for quantitative traits in clones of oregano grown in the three locations, the eigenvalues indicate that the first two axes explain 100% of the variability. Figure4 shows the consensus configuration between the rankings generated by the principal components of the quantitative characters measured in La Consulta, Potrero de Garay, and Capilla de los Remedios. From the ratio between the consensus and the total sum of squares, it can be concluded that there is a 77.3% of consensus between the configurations produced by the quantitative traits measured in each one of the growing locations. From the GPA it can be said that, regardless of their agronomic performance, the 12 clones of oregano evaluated tend to be clustered in a similar way in the three locations where the essays were conducted; moreover four groups of oregano can be clearly distinguished (Figure 4).Figure 4
Arrangement the 12 clones of oregano on the plane formed by first two axes of a PGA (with minimum tree trail), considering quantitative characteristics evaluated at the three locations of cultivation. C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito.”


### 3.4. Taxonomic Identification of the Evaluated Clones

The taxonomic identification of the material evaluated at field was conducted using the keys of Iestwaart [29], Xifreda [2], and Rouquaud & Videla [3]. In accordance with those keys, the classification of the different clones of oregano was largely based on floral characteristics such as shape of the calyx, the length of the bracts, the length of the staminal filaments, and the color of bracts and flowers. From the observations made, it was determined that 10 of the 12 clones of oregano evaluated belong to the genus Origanum, section Origanum, and two of them are hybrids (Table 6).Table 6
Taxonomic identification of 12 clones of oregano from the main cultivation areas.
Clone
Common name
Origin
Herbarium number
Taxa
Me-TS
“Mendocino”
Córdoba (Valle de Traslasierras)
Torres 0901 (CORD)
Origanum x majoricumCambessedes
VL-TS
“Verde Limón”
Córdoba (Valle de Traslasierras)
Torres 0909 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
NB-TS
“Negrito”
Córdoba (Valle de Traslasierras)
Torres 0910 (CORD)
Origanum vulgaressp.vulgare
Ne-SJ
“Negrito”
San Juan (Santa Lucía)
Torres 0911 (CORD)
Origanum vulgaressp.vulgare
Me-Mza
“Mendocino”
Mendoza (La Consulta)
Torres 0902 (CORD)
Origanum x majoricumCambessedes
Cm-Mza
“Compacto”
Mendoza (La Consulta)
Torres 0903 (CORD)
Origanum vulgaressp.vulgare
Cd-Mza
“Cordobés”
Mendoza (La Consulta)
Torres 0905 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
Cr-Mza
“Criollo”
Mendoza (La Consulta)
Torres 0904 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C1-FCA
—
Córdoba (FCA-UNC)
Torres 0912 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C2-FCA
“Criollo”
Córdoba (FCA-UNC
Torres 0906 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C3-FCA
—
Córdoba (FCA-UNC)
Torres 0907 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C8-FCA
—
Córdoba (FCA-UNC)
Torres 0908 (CORD)
Origanum vulgaressp.hirtum (Link) IestwaartIn our country, Rouquaud & Videla [3] reported the existence of “oregano” belonging to Origanum vulgare ssp. vulgare and the hybrid Origanum x majoricum, while the subspecies Origanum vulgare ssp hirtum is reported for the first in this paper. In addition to the taxa mentioned, specimens belonging to the species Origanum dictamnus and Origanum vulgare ssp. viride [2], Origanum vulgare ssp. virens [3], Origanum majorana and the hybrid Origanum x applii [2, 3] were described in the country. Since the “oregano” includes a group of cross-pollinated species, the genetic variability within and between them is very high [32]. Thus the variability observed between clones belonging to the same species can be attributed to genetic variation originated by recombination processes inherent in all species whose members reproduce sexually [43].


## 3.1. La Consulta INTA Essay

Clones Me-TS, VL-TS, and C3-FCA showed the lower mean value for longest branch length, when this trait was measured three months after sowing. One year after sowing, the clones with the lower longest branch length mean value were Me-TS, Ne-SJ, and Cd-Mza.Regarding the fresh weight yield of the first harvest, clones C8-FCA, C3-FCA, and Ne-SJ stood out for their higher production of fresh biomass, followed by clones Cm-Mza and NB-TS. In relation to dry weight, again C8-FCA clone differed significantly from the other clones because of its greater dry biomass yields, followed by clones C3-FCA, C2-FCA, Ne-SJ, Cm-Mza and NB-TS. The production of fresh and dry biomass can be related to the length of the internodes and leaf area; clones C3-FCA, C2-FCA, Ne-SJ, Cm-Mza, and NB-TS also showed shorter internodes and greater area leaf (Table 3). In the second harvest, clones C3-FCA and C8-FCA significantly surpassed the rest of the material for both fresh and dry weight. Regardless of the clone, the fresh and dry weight yields observed in both harvests are generally similar to those reported by Droushiotis and Della [32] and Weglarz and Geszprych [33].With regard to the percentage of water that is lost during the drying process, significant differences were observed among the clones both in the first as in the second harvest. In the first harvest, the clones with the lowest content of water were Cr-Mza, C2-FCA, Cd-Mza, and C1-FCA, which in turn showed the highest percentage of fibers (66.47%, and 34.16% resp.). In the second harvest, the clones with lowest water content and higher fiber content were Cd-Mza, VL-TS, and C2-FCA, with an average percentage of 54.68% of water and 45.32% fiber. The percentage of water found in the second harvest was similar to that reported by Leto and Salamone [34], who studied 62 biotypes of oregano coming from 17 different locations. Higher fiber content (or less water loss) is equivalent to a higher dry matter yield. That is, on average, those clones with higher fiber content showed a dry matter yield 7.62% higher than the rest of the clones in the first crop and 15.85% in the second harvest. This is important when estimating production volumes of dry matter.Significant differences were also observed among the clones in terms of dry weight of leaves and stems and leaf/stem ratio (Table3), being this latter variable very important when estimating the returns as are leaves the part of the plant that is consumed [35]. Clones that showed the highest values for leaf/stem ratio were Ne-SJ, Cm-Mza, and NB-TS, these clones had short internodes, a greater number of nodes (and therefore more foliage), and greater leaf area.Based on the essential oil yield, it was possible to distinguish four groups of oregano. The first group showed an average essential oil content of 2.18% which includes the clones Me-Mza and Me-TS. The second group, including the clones C2-FCA, C1-FCA, Cd-Mza and Cr-Mza, showed an average essential oil content of 1.44%, while the third group, involving the clones Cm-Mza, NB-TS, VL-TS y C8-FCA, showed an average oil percentage of 0.78%. Finally, the group with the lowest essential oil content was constituted by clones C3-FCA and Ne-SJ, which showed an average oil yield of 0.43%. The average essential oil yield observed for the first and second group surpass in 67.7% and 10.8%, respectively, the percentages of oil reported by Weglarz and Geszprych [33], whereas the essential oil content observed in the third and fourth group was in the range reported by these authors (0.30–1.30%).Finally, the number of branches per plant ranged from 94 (Me-TS) to 123 (C1-FCA); however, no significant differences were observed among the twelve evaluated clones.Principal component analysis (PCA) showed that 73.1% of the total variability is explained by the first factorial plane (PC1 and PC2). At the level of the first axis (PC1), which explains 42.2% of the total variability, the variables with greater weight were the leave dry weight, fresh weight of the second harvest, internode length, and essential oil content, whereas the second axis (PC2) explains 30.9% of total variability, and the variables with greater inertia are the stem dry weight, leaf/stem ratio, and leaf area (Figure1).Figure 1
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in La Consulta (Mendoza). C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length; LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing); **measured at 2nd harvest (one year after sowing).


## 3.2. Potrero de Garay Essay

With regard to the length of the longest branch, measured three months after planting, clone Ne-SJ differed significantly from the other clones, showing the longest branch length mean value, while no differences were observed among the 11 remaining clones (whose branch length ranged between 26.90 and 38.16 cm) (Table4). Considering the same variable, measured one year after sowing, the clones Me-Mza, Me-TS, and VL-TS differed from the other clones by their shorter length (22.12 cm on average), and the clones C8-FCA (41.88 cm) and C3-FCA (47.10 cm) were distinguished by their greater length, while no differences were observed between the remaining clones, with an average branch length of 33.79 cm.Regarding the fresh weight yield of the first harvest, clones C8-FCA, Ne-SJ, Cm-Mza, and NB-TS were those that stood out for their higher production of fresh biomass, with an average fresh weight of 61.34 g plant−1, while the clones Me-Mza and Me-TS differed from the rest of the clones by their poor performance in fresh matter (on average 8.73 g plant−1), followed by the clone Cd-Mza with a fresh biomass yield of 21.35 g plant−1. In relation to dry weight, clones C8-FCA, Ne-SJ, Cm-Mza, and NB-TS showed dry biomass yields significantly superior from those observed in the remaining clones, followed by clones C3-FCA, C2-FCA, C1-FCA, VL-TS, Cr-Mza, and Cd-Mza, with an average dry matter yield of 15.21 g plant−1. Being the clones Me-Mza and Me-TS those showed the lowest yield in dry weight (Table 4).The higher production of fresh and dry biomass observed in clones C8-FCA, Ne-SJ, Cm-Mza and NB-TS can be associated with the largest number of branches per plant (more than 60 branches per plant), with a longer branch (C8-FCA and Ne-SJ) and greater leaf area (cm Mza and NB-TS) (Table 4). The low yields observed in the first harvest for fresh and dry weight, lower than those values reported by Droushiotis and Della [32] and Weglarz and Geszprych [33], can be attributed to the dehydration suffered by the plants during the week before the harvest, being the clones Me-Mza and Me-TS the clones that most resented the lack of water.In the second harvest, clones Ne-SJ, NS-TS, Cm-Mza, C3-FCA, and C8-FCA showed a significantly superior yield, both in fresh as in dry biomass, with an average fresh weight of 152.97 g plant−1 and an average dry weight of 58.17 g plant−1. Clones C1-FCA, C2-FCA, Cr-Mza, and Cd-Mza showed average yields in fresh and dry matter of 89.31 g plant−1, and 38.02 g plant−1 respectively; while the clones Me-Mza and Me-TS were those with the lowest yield both in fresh weight (25.70 g plant−1) as in dry weight (11.26 g plant−1), as they were in the first harvest, followed by the clone VL-TS.As regards the percentage of water that is lost during the drying process, significant differences were observed between the clones both in the first as in the second harvest. In both harvests, the clones Me-Mza and Me-TS showed the lowest water content (and the highest percentage of fibers) (Table4). At this point, it is important to note that in the week preceding the first harvest, Potrero de Garay field was exposed to water stress, so the percentage of water lost during the drying process was lower than in La Consulta and Capilla de los Remedios. For this same reason, the percentage of fibers of the different clones, measured in the first harvest, was overestimated; however, it was possible to differentiate the oregano clones based on these variables. It is reasonable to assume that a high fiber content (equivalent to less water loss) results in a higher dry weight yield. Even though clones Me-Mza and Me-TS had higher fiber content, their biomass production was so low that they fail to align their yields with those clones that have higher percentages of water.In addition, significant differences were found among the clones in terms of the dry weight of the leaves and the stems and leaf/stem ratio (Table4), being this latter variable of major economic importance when estimating amount of production since it is the leaves that are used for consumption. The clones Ne-SJ, NB-TS, and Cm-Mza were those that showed the greatest values for leaf/stem ratio, coinciding with the greatest number of branches, longest branch, and greatest leaf area (Table 4).Concerning the content of essential oils, the clones Me-Mza and Me-TS showed an essential oil yield significantly higher than the rest of the clones, with an average of 3.87% of oil, followed by the clones C2-FCA, C1-FCA, Cd-Mza, Cr-Mza, C8-FCA, and C3-FCA, which showed an average oil content of 1.29% and the clones VL-TS, NB-TS, Cm-Mza and Ne-SJ (clones with the lowest performance in oils, producing on average 0.48%). The essential oil yield observed for the clones Me-Mza and Me-TS was similar the one reported for Origanum dubium (3.30 to 5.85%) [32]. These percentages were three times higher than those reported by Weglarz & Geszprych [33] and 32.5% higher than the yield observed in Turkish oregano (Origanum onites) [36], one of the species of greater content of essential oils.The principal component analysis (PCA) showed that 82.6% of the total variability was explained by the first factorial plane (CP1 and CP2). On the level of CP1, component that by itself explains 63.8% of the total variability, the variables with higher weight were the dry weight of leaf, the fresh weight measured at the second harvest, and the content of essential oils, while, at the level of CP2, the variables with greater inertia were leaf area and length of the longest branch (measured at 400 days after planting) (Figure2).Figure 2
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in Potrero de Garay (Córdoba). C1-FCA: clone 1 C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8; Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing), **measured at 2nd harvest (one year after sowing).


## 3.3. Capilla De Los Remedios Essay

Regarding to the length of the longest branch, measured 85 days after seeding, the clones VL-TS, Cm-Mza, NB-TS, C8-FCA, C3-FCA, and Ne-SJ differed significantly from the rest, recording the lowest values for this variable (range 14.03 and 21.22 cm) (Table 5). When this variable was determined 400 days after sowing, Ne-SJ and VL-TS differed from the other clones for its smaller branch length (15.34 cm on average), while Cd-Mza and Cr-Mza were stood out for their greater length (40.16 cm). No differences were observed between the remaining genotypes, whose average branch length was 27.44 cm (Table 5).With regard to fresh weight yield of the first harvest, two significantly different groups were distinguished. The clones C8-FCA, Cm-Mza, NB-TS, C3-FCA, and Ne-SJ comprise the higher group, with an average production of fresh biomass of 228.77 g plant−1, exceeding 75.14% the group of lower values for the variable formed by the seven remaining clones, which had an average yield of fresh weight of 130.62 g plant−1. The same trend was observed in relation to dry weight, where the clones Cm-Mza, NB-TS, C8-FCA and Ne-SJ differed significantly from the rest by its higher values which averaged 69.75 g plant−1 (Table 5). As in the essays of La Consulta and Potrero de Garay, clones with higher production of fresh and dry biomass are generally characterized by the presence of a greater number of branches per plant (more than 70 branches per plant), shorter internodes, and greater leaf area. Once again, the observed values for the variables fresh weight and dry weight are within the range reported by Droushiotis and Della [32] and Weglarz and Geszprych [33].Although in the second harvest the clones differed from one to another both in fresh as dry weight (Table5), due to the strong predation caused by ants, the recorded yields did not reflect the real potential of each genotype. Although all the clones were targeted by these insects, Ne-SJ, VL-TS, Me-TS, C8-FCA, and C3-FCA were the most affected, being Ne-SJ the clone that showed the most important performance reduction for fresh and dry biomass (50.01 g plant−1 and 16.75 g plant−1, resp.). Taking into account the agronomic performance observed for the same clone (Ne-SJ) both in the essays conducted in La Consulta and Potrero de Garay as in the first harvest of this essay, it can be assumed that in normal conditions (without predation) the yield of this clone would be far higher. Furthermore, it is reasonable to assume that without ants, the second harvest would be maintained the trend observed in the first one for all the clones. Regardless of the insect damage, the clones VL-TS and Me-TS were usually the lowest performing group both in fresh as in dry biomass yield, following the same trend observed in La Consulta and Potrero de Garay. The clones NB-TS, Cm-Mza, C3-FCA, and C8-FCA (which were the top performers in the first harvest) and the clone Me-Mza (low performance in the first harvest) did not differ significantly from clones C1-FCA, C2-FCA, and Cd-Mza, with an average fresh weight yield of 106.69 g plant−1 while the clone Cr-Mza had a biomass yield significantly higher than the rest of the clones, with an average fresh weight of 162.36 g plant−1. In terms of dry weight, the clones VL-TS, Ne-SJ, Me-TS, C8-FCA, and C3-FCA had an average yield of 24.34 g plant−1; the clones Me-Mza, Cm-Mza, C1-FCA, NB-TS, Cd-Mza, and C2-FCA yielded an average of 50.21 g plant−1 while Cr-Mza was significantly higher than the rest with a fresh biomass production averaged 77.14 g plant−1 (Table 5).Respecting to the percentage of water lost during the drying process, significant differences were seen among the clones both in the first as in the second harvest. In the first harvest, C8-FCA differed significantly from the rest of the clones, with a water content exceeding 5.71% of the 11 remaining clones and being also the genotype with the lowest fiber content (Table 5). In the second harvest, the clone with highest water content was Ne-SJ (65.46%), a value that was on average 7.58% higher than that observed for clones. Regarding the fiber content in the second harvest, the clones Cd-Mza, C2-FCA, Cr-Mza, Me-TS, VL-TS, and Me-Mza had the highest value for this variable, containing average of 45.54% of fiber (Table 5). As already noted above it can be assumed that, at the same fresh weight yield, a higher fiber content is equivalent to a higher dry matter yield.Regarding the leaves and stems dry weight and leaf/stem ratio, significant differences were also observed between genotypes. The clones mainly characterized by shorter internodes, high number of branches, and the greater leaf area (clones NB-TS and Cm-Mza) and the low stems weight (clone Me-TS) were those who showed the highest values for leaf/stem ratio (Table5).Concerning the content of essential oils, Me-Mza and Me-TS differed significantly from the rest of clones, with an average essential oil content of 2.15%, while no differences were observed among the 10 remaining clones (which oil content ranged from 0.33% to 1.11%). According to Kokkini [37], based on performance of essential oils, it is possible to categorize the different genotypes of oregano into three groups: (a) those producing less than 0.05% (“poor” in oils), (b) those with essential oil content between 0.05 and 2%, and (c) those with an essential oil yield over 2% (“rich” in oil). Both in this essay as in those conducted in La Consulta and Potrero de Garay, clones of oregano belonging to the three groups were distinguished. The clones called “Mendocino” (Me-Mza and Me-TS) belong to the rich-oil group, with a performance that in the three locations exceeds 2%, while the clones called “Compacto” (Cm-Mza) and “Negritos” (NB-TS and Ne-SJ) belong to the group poor-oil group. These results are consistent with those observed by De Mastro [38]. Franz & Novak [35] propose to use differential production of oregano based on the essential oil yield, destinating for use as dried herb those materials having 1 to 3% of oil, and for essences production those clones containing levels of oil exceeding 3%. Oregano production intended for obtaining essential oils becomes more important if we consider that these essences and/or its components represent a potentially important source of pharmacologically active compounds [39] or agrochemicals (as herbicides) [40]. Particularly, it has been tested that the essential oil of oregano has a strong antioxidant activity in foods with high-fat content and fried-salted peanuts [41, 42].The principal component analysis (PCA) showed that 73% of the total variability is explained by the first factorial plane formed by the two first principal components. On the level of CP1, that by itself explains 50.1% of the total variability, the more discriminating variables were leaf dry weight, leaf area, dry weight of stems, the length of internodes and the essential oil content, while on the level of CP2 (which accounts for 22.9% of the total variability observed) were fresh weight measured in the second harvest, leaf/stem ratio, and the length of the longest branch. The PCA showed that all 12 clones of oregano evaluated tend to cluster in a manner similar to those observed in La Consulta and Potrero de Garay (Figure3).Figure 3
Biplot plane formed by the two first principal components (CP1 and CP2), where the dots represent the 12 clones of oregano evaluated and the vectors, quantitative characteristics measured in Capilla de los Remedios (Córdoba). C1-FCA: clone 1 C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8; Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito,” LBL: longest branch length, IL: internode length, LA: leaf area, FW: fresh weight, LDW: leave dry weight, SDW: stem dry weight, L/S: ratio LDW/SDW, EO: essential oil content, *measured at 1st harvest (three month after sowing); **measured at 2nd harvest (one year after sowing).With regard to generalized procrustes analysis (GPA), combining the information obtained for quantitative traits in clones of oregano grown in the three locations, the eigenvalues indicate that the first two axes explain 100% of the variability. Figure4 shows the consensus configuration between the rankings generated by the principal components of the quantitative characters measured in La Consulta, Potrero de Garay, and Capilla de los Remedios. From the ratio between the consensus and the total sum of squares, it can be concluded that there is a 77.3% of consensus between the configurations produced by the quantitative traits measured in each one of the growing locations. From the GPA it can be said that, regardless of their agronomic performance, the 12 clones of oregano evaluated tend to be clustered in a similar way in the three locations where the essays were conducted; moreover four groups of oregano can be clearly distinguished (Figure 4).Figure 4
Arrangement the 12 clones of oregano on the plane formed by first two axes of a PGA (with minimum tree trail), considering quantitative characteristics evaluated at the three locations of cultivation. C1-FCA: clone 1, C2-FCA: clone 2, C3-FCA: clone 3, C8-FCA: clone 8, Me-Mza: clone “Mendocino,” Cm-Mza: clone “Compacto,” Cd-Mza: clone “Cordobés,” Cr-Mza: clone “Criollo,” Me-TS: clone “Mendocino,” NB-TS: clone “Negrito B,” VL-TS: clone “Verde Limón,” Ne-SJ: clone “Negrito.”


## 3.4. Taxonomic Identification of the Evaluated Clones

The taxonomic identification of the material evaluated at field was conducted using the keys of Iestwaart [29], Xifreda [2], and Rouquaud & Videla [3]. In accordance with those keys, the classification of the different clones of oregano was largely based on floral characteristics such as shape of the calyx, the length of the bracts, the length of the staminal filaments, and the color of bracts and flowers. From the observations made, it was determined that 10 of the 12 clones of oregano evaluated belong to the genus Origanum, section Origanum, and two of them are hybrids (Table 6).Table 6
Taxonomic identification of 12 clones of oregano from the main cultivation areas.
Clone
Common name
Origin
Herbarium number
Taxa
Me-TS
“Mendocino”
Córdoba (Valle de Traslasierras)
Torres 0901 (CORD)
Origanum x majoricumCambessedes
VL-TS
“Verde Limón”
Córdoba (Valle de Traslasierras)
Torres 0909 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
NB-TS
“Negrito”
Córdoba (Valle de Traslasierras)
Torres 0910 (CORD)
Origanum vulgaressp.vulgare
Ne-SJ
“Negrito”
San Juan (Santa Lucía)
Torres 0911 (CORD)
Origanum vulgaressp.vulgare
Me-Mza
“Mendocino”
Mendoza (La Consulta)
Torres 0902 (CORD)
Origanum x majoricumCambessedes
Cm-Mza
“Compacto”
Mendoza (La Consulta)
Torres 0903 (CORD)
Origanum vulgaressp.vulgare
Cd-Mza
“Cordobés”
Mendoza (La Consulta)
Torres 0905 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
Cr-Mza
“Criollo”
Mendoza (La Consulta)
Torres 0904 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C1-FCA
—
Córdoba (FCA-UNC)
Torres 0912 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C2-FCA
“Criollo”
Córdoba (FCA-UNC
Torres 0906 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C3-FCA
—
Córdoba (FCA-UNC)
Torres 0907 (CORD)
Origanum vulgaressp.hirtum (Link) Iestwaart
C8-FCA
—
Córdoba (FCA-UNC)
Torres 0908 (CORD)
Origanum vulgaressp.hirtum (Link) IestwaartIn our country, Rouquaud & Videla [3] reported the existence of “oregano” belonging to Origanum vulgare ssp. vulgare and the hybrid Origanum x majoricum, while the subspecies Origanum vulgare ssp hirtum is reported for the first in this paper. In addition to the taxa mentioned, specimens belonging to the species Origanum dictamnus and Origanum vulgare ssp. viride [2], Origanum vulgare ssp. virens [3], Origanum majorana and the hybrid Origanum x applii [2, 3] were described in the country. Since the “oregano” includes a group of cross-pollinated species, the genetic variability within and between them is very high [32]. Thus the variability observed between clones belonging to the same species can be attributed to genetic variation originated by recombination processes inherent in all species whose members reproduce sexually [43].


## 4. Conclusions

The quantitative traits analyzed allowed the characterization of the 12 clones of oregano studied, establishing four clearly distinguishable groups. Regardless of the cultivation site, most discriminate quantitative traits were essential oil content, internodes length, longest branch length (400 days), fresh weight (2nd harvest), leave dry weight, stem dry weight, ratio H/T, and foliar area. The 12 evaluated clones can be clustered as follow: (Group 1) clones Me-TS and Me-Mza; (Group 2) clones Cd-Mza, Cr-Mza, C1-FCA, and C2-FCA; (Group 3) clones Cm-Mza, NB-TS, and Ne-SJ (Group 4) clones C3-FCA and C8-FCA. The clone VL-TS is a particular case since if it shares characteristic of different groups such as long branches and long internodes (group 2) and low essential oil content (less than 1%) (group 4).Otherwise, the12 clones were identified taxonomically, confirming that two of these clones correspond to the hybridOriganum x majoricum Cambessedes, three of the clones belong to the species Origanum vulgare ssp. vulgare and the remaining 7 clones belong to the species Origanum vulgare ssp. hirtum (Link) Iestwaart. Regardless of the growing location, clones were clustered in the same way, and these groups are consistent with their taxonomic determination. Differences observed among the clones pertaining to the same species can be attributed to the variability generated in the recombination processes.



---

*Source: 349565-2012-07-16.xml*