Ukrainian Journal of Ecology
UkrainianJournal of Ecology, 2017, 7(4), 432-439, doi: 10.15421/2017_128
ORIGINAL ARTICLE
Do we have infraspecific taxa of Salvia multicaulisVahl. (Lamiaceae) in Iran?
Seyed Mehdi Talebi*1, Reza Rezakhanlou2, Alex Matsyura3
1 Department of Biology, Faculty of Sciences, Arak University Arak, 38156-8-8349 Iran. Phone: 098-863-4173317. E-mail: seyedmehdi [email protected] 2 Department of Agronomy, Saveh Branch, IslamicAzad University, Saveh, Iran.
3 Altai State University, Lenin St., 61, 656049, Barnaul, Russia Submitted: 30.10.2017. Accepted: 04.12.2017
Salvia multicaulis is a widespread species of Lamiaceae family in Iran. There are many discussions about its infraspecific variations. Although some varieties were definite for this species in various parts of the world, no infraspecific taxon was reported in Iran and all samples of this species were named as S. multicaulis. In this study, morphological characteristics of S. multicaulis populations, naturally growing in Iran, was examined. Twenty-two traits were examined in 94 individuals of this species to identify their phenotypic difference. Most of the investigated features were showing a high degree of variability, but it was highly pronounced for some characteristics such as basal leaf shape, basal leaf width, basal leaf length/ width ratio and basal petiole length. Significant positive/negative correlations were observed between some morphological variables. Furthermore, significant negative correlations were found between the eastern distribution of populations with basal leaf petiole length and basal leaf length/ width ratio. Based on the UPGMA cluster analysis, populations were divided into two main branches. The first branch contained four populations, while the second branch was bigger and clustered in two sub-branches. In one of them, three populations and in another one the rest populations arranged in two groups. CA joined plot confirmed that each of studied populations or group of populations had distinct morphological trait(s), which were useful in identification of them. Our findings supported population no. 13 had unique morphological traits such as the largest bracts and basal leaf petiole, highest flower number of each inflorescence cycle, widest and largest calyx. The conservation of the highly diverse populations of Iranian S. multicaulisis recommended. Key words: morphology; population; Salvia multicaulis; infraspecific
Introduction
Salvia L. is the largest genus of Lamiaceae that comprises of more than 900 taxa in the world (Davis 1982). This genus has 62 species in Iran (Jamzad, 2012). Different species of the genus have been used in folk medicine for the treatment of many disorders such as diabetes (Jimenez et al. 1986) and skin diseases like psoriasis and eczema (Topcu et al. 2007). Salvia multicaulisVahl (Lamiaceae) is an aromatic plant of the genus that widely distributed in different parts of Iran (Jamzad, 2012). This species is also used in folk medicine to treat various diseases, such as colds (^akilcioglu et al. 2010), the flu and tonsillitis (Tetik et al. 2013).
Ulubelen et al. (1997) reported seven new aromatic secondary compounds from the roots of this species. They were: norditerpenoids, multicaulin, 12-demethylmulticauline, multiorthoquinone, 12-demethylmultiorthoquinone, 12-methyl-5-dehydrohorminone, 12-methyl-5-dehydroacetylhorminone and salvipimarone. Moreover, they observed the antituberculous activity of these compounds against Mycobacterium tuberculosis strain H37Rv.
Investigations of infraspecific morphological diversity have been performed for different species of Labiatae family (e.g., Talebi, 2014, Talebi, 2015). However, despite many reports about essential oil composition of S. multicaulis, we still do not have comprehensive information about the morphological variations of this species. Therefore, the main objective of this investigation was to identify the variability in 94 accessions of 14 populations of this species growing wild in Iran, to determine the possible correlation between the characteristics, to identify the most useful features for discrimination between the populations and to detect relationships between them.
Materials and method
Plant material
Six qualitative and sixteen quantitative morphological traits of fourteen populations of S. muiticauiis were examined (Table 1). These samples were elected from natural populations of the species during spring 2016 and identified based on the descriptions provided in valuable references like Flora Iranica (Rechinger, 1982) and Flora of Iran (Jamzad, 2012). On the basis of population size, up to eight individuals were selected for each population. The mean and also standard deviation was determined for quantitative features (Table 2). The studied variables were: stem length, shapes of basal and floral leaves margin, blade and their length, width, length/wide ratio and petiole length, calyx and corolla color, length, width, their length/ width ratio and pedicle length. Statistical analysis
Analysis of variance was performed for morphological traits using SPSS software. The mean and standard deviation and simple correlation coefficient were calculated to determine the relationships between the studied morphological features using the Pearson correlation coefficient by SPSS.
Relationships among the populations were studied by principal component analysis (PCA). The morphological similarity coefficients according to Manhattan method were calculated using the MVSP ver. 3.2 program of the numerical taxonomy multivariate analysis and the dendrogram were constructed using the unweighted pair group method with arithmetic means (UPGMA). PCA and PCO scatter plots were constructed for representing morphological similarity among the studied populations. The CA-Joined plot and PCA-biplot representing morphological similarity relationships among populations, linking the morphometric traits (Podani, 2000).
Table 1. Localities address and coding of studied S. multicaulis populations.
Population cods Habitat
1 Markazi province, Zarandiyeh, Noshveh, 1814 m.
2 Markazi province, Zarandiyeh, Vidar, 1771 m.
3 Kurdistan province, Sanandaj, Husain Abad 1810 m.
4 Hamedan province, Hamehkasi, 2201 m.
5 Kurdistan province, Marivan-Saqqez, 1585 m.
6 Kurdistan province, Marivan, 1363m.
7 Hamedan province, Gol-e-e khandan, 1734 m.
8 Markazi province, Zarandiyeh, Vardeh 1810 m.
9 Tehran province, Tehran, Lashkarak , 2225 m.
10 West Azerbaijan, Tekab, 2047 m.
11 Markazi province, Saveh, Chamran, 1775m.
12 Markazi province, Arak, Sefidkhani, 2180m
13 Markazi province, Saveh-boen Zahra road 1931m.
14 West Azerbaijan, Urmia, Sero, 1680 m.
Results
ANOVA test showed significant differences (p< 0.05) for all of the quantitative studied characteristics with except basal lea f width, floral leaf length/width ratio and calyx leaf length/width ratio (Table 3). Quantitative features such as floral and basal leaf blade shape and their marginal shape differed between the populations. They were oblong, ovate, obovate, lanceolate, oblanceolate, linear and linear-lanceolate. Moreover, the color of calyx was purple or violet in all of the studied populations, with except population number 13, which had lavender calyx. It means that most of the morphological traits varied significantly among the populations. PCA analyses proved that seven characteristics: stem length, basal leaf shape, basal leaf width, basal leaf length/ width ratio and basal petiole length comprises 72.47% of variations. Therefore these traits were more variable among the examined characters (Table 4).
Significant positive/negative correlations were seen between some morphological traits. For example, corolla length had significant negative correlations (P < 0.05) with floral leaf length and width, calyx width, flower number of each inflorescence cycle. Significant positive correlations (P < 0.01) were found between calyx length and width with floral leaf length, pedicle length, and flower number per each inflorescence cycle.
Basal leaf length and width had significant positive correlations (P < 0.01) with stem length, floral leaf length and width a nd pedicle length.
The studied populations separated from each other in UPGMA dendrogam (fig 1); in addition, PCA, as well as PCO plots (figs 2, 3) created similar results. So population's arrangement in the dendrogam was discussed here: it is composed of two branches (A, B). The smallest one (A) had two sub-branches. Populations number 5, 7, and 10 were grouped in the bigger sub-branch, while population no. 13 was found in the smaller one far from others. The largest branch (B) divided into two sub-branches; one of them was small (C) and composed of three populations number 4, 8, and 14. In the bigger sub-branch (D), populations constructed two groups. Populations number 1, 2, and 3 arranged in one group (E) and the rest populations were clustered in the other group (F), so that populations number 6 and 11 placed closely and the population no. 12 were far from the others.
Table 3. Results on the ANOVA analysis to assess for differences in morphological traits of studied S. multicaulis populations. d.f.: degrees of freedom; F: F-statistic; P: probability.
Sum of Squares df Mean Square F P
Stem length Between Groups 3238.748 13 249.134 8.569 .000
Within Groups 2326.028 80 29.075
Total 5564.777 93
Basal leaf length Between Groups 18.052 13 1.389 2.817 .002
Within Groups 39.433 80 .493
Total 57.486 93
Basal leaf petiole Between Groups 38.988 13 2.999 1.885 .044
length Within Groups 127.284 80 1.591
Total 166.272 93
Basal leaf width Between Groups 2.146 13 .165 .916 .540
Within Groups 14.413 80 .180
Total 16.559 93
Basal leaf length/ Between Groups 9.584 13 .737 2.871 .002
width ratio Within Groups 20.545 80 .257
Total 30.130 93
Floral leaf length Between Groups 41.700 13 3.208 3.590 .000
Within Groups 71.488 80 .894
Total 113.188 93
Floral leaf width Between Groups 2.073 13 .159 2.353 .010
Within Groups 5.421 80 .068
Total 7.494 93
floral leaf length/ Between Groups 38.803 13 2.985 1.492 .139
width ratio Within Groups 160.033 80 2.000
Total 198.837 93
Corolla length Between Groups 3.397 13 .261 4.550 .000
Within Groups 4.595 80 .057
Total 7.992 93
Calyx length Between Groups 6.484 13 .499 14.197 .000
Within Groups 2.810 80 .035
Total 9.294 93
Pedicle length Between Groups .546 13 .042 4.148 .000
Within Groups .810 80 .010
Total 1.357 93
Calyx width Between Groups 20.063 13 1.543 15.372 .000
Within Groups 8.032 80 .100
Total 28.095 93
Calyx length/width Between Groups 2.623 13 .202 1.357 .199
ratio Within Groups 11.890 80 .149
Total 14.513 93
Flower number of Between Groups 471.645 13 36.280 4.892 .000
each cycle Within Groups 593.259 80 7.416
Total 1064.904 93
Bract length Between Groups 4.565 13 .351 3.482 .000
Within Groups 8.068 80 .101
Total 12.632 93
C.A joined plot showed that each of studied populations or identified group of populations had the distinct morphological variable(s), which were useful in identification of them (fig. 4). For example, floral leaf shape, the ratio of basal leaf blade length/ petiole length and calyx length/ pedicle length ratio were valuable traits for populations no. 8, 14 and 4, respectively. Furthermore, petal color putts population no. 14 far from populations no. 8 and 4. Stem length was a distinguishing character for branch A. Then, each of these populations had distinct features and separated from each other. Floral leaf length was a specific trait for populations no. 5 and 10. Pedicle length, calyx length and width and flower number of each inflorescence cycle separated population no. 13.
Fig. 1. UPGMA dendrogam of the studied populations based on the morphometric data. Numbers indicate population's code 1-14 as in Table 1.
PCA case scores
3.3T
2.6—
2.0—
14 ▲
1.3
0.7
9
13 ▲
10 ▲
4
-2.6
4
A -20
6
-1.3A
1-10.7
-0.7
-1.3
3
A
2 A
0.7
1
A
1.3
2.0
2.6
3.3
8
-2.0—
-2.6 Axis 1
Fig. 2. PCA Scatter plot for the studied S. muiticauiispopulations based on morphological characters. Numbers indicate population's code 1-14 as in Table 1.
Fig. 3. PCO plot of the studied populations on the bases of morphological traits. Numbers indicate population's code 1-14 as in Table 1.
CA joint plot
14
A
basallefpetio
V
flowernu
V
calywi
V
calylen
V
petal color bractlen
_I_lü
2.7—T
2.1
13
A
1.6 pe
1.1
florallea
V
0.5-
edicl
V
fwi
10
5AA
floaleaflen z-i
V *
stemlen basLel^ pet
-2.7 -2.1
floralshap
V
-1.6
b-Ssslienafi;ish-0lE5
V V
basal
bas, -0.5
enwide Tjj,len y
0.1 v 1.1 _________id A
yv A
'Tïv
1.6
2.1
2.7
4
A
calypedi
V
corola-em^ ca^widle
Wi.6-
calxc>
1'
-2.1
-2.7 Axis 1
1
A
2 A
orol
Fig. 4. CA joined plot representing morphological similarity relationships among populations of S. multicaulis, linking the morphometric traits. Numbers indicate population's code 1-14 as in Table 1. Abbrivations: stemlen: Stem length, basalelen: Basal leaf length, basal pet: Basal leaf petiole length, basallewid: Basal leaf width, basallenwide: Basal leaf length/ width ratio, floralshap: floral leaf shape, floraleaflen: Floral leaf length, floralleafwi: Floral leaf width, floral leaf length/ width ratio, corolalen: Corolla length, calylen: Calyx length, pedid: Pedicle length, calywi: Calyx width, calywidlen: Calyx length/width ratio, flowerno: Flower number of each cycle, bractlen: Bract length, calypedi: calyx/pedicle length ratio.
Table 4. PCA analyses of the studied morphological variables between the studied populations.
Extraction Sums of Squared
Initial Eigenvalues Loadings Rotation Sums of Squared Loadings
% of Cumulative % of Cumulative % of Cumulative
Component Total Variance % Total Variance % Total Variance %
Stem length 5.249 23.860 23.860 5.249 23.860 23.860 3.622 16.461 16.461
Basal leaf shape 3.128 14.220 38.080 3.128 14.220 38.080 3.339 15.177 31.638
Basal leaf length 2.139 9.724 47.804 2.139 9.724 47.804 1.931 8.779 40.417
Basal leaf petiole length 1.593 7.240 55.044 1.593 7.240 55.044 1.892 8.598 49.015
Blade length petiole length ratio 1.416 6.435 61.478 1.416 6.435 61.478 1.713 7.785 56.800
Basal leaf width 1.278 5.809 67.288 1.278 5.809 67.288 1.637 7.442 64.243
Basal leaf length/ width ratio 1.142 5.189 72.477 1.142 5.189 72.477 1.469 6.676 70.918
Basal margin shape 1.071 4.870 77.347 1.071 4.870 77.347 1.414 6.429 77.347
Floral leaf length .918 4.174 81.521
Floral leaf width .814 3.701 85.222
floral leaf length/ width ratio .708 3.220 88.442
floral leaf shape .579 2.633 91.075
Corolla length .505 2.294 93.370
Calyx length .417 1.894 95.263
Corolla/ calyx length ratio .326 1.480 96.744
Pedicle length .283 1.285 98.029
Calyx/pedicle length ratio .174 .790 98.818
Calyx width .109 .496 99.314
Calyx length/width ratio .063 .288 99.603
Flower number of each cycle .049 .221 99.823
Bract length .024 .111 99.934
Petal color
.014
.066
100.000
Discussion
S. multicaulis has several populations in various parts of Iran, which growth under different ecological conditions. Therefore, this species can definite as an ecological generalist plant species. These plants have highly plastic genotypes and phenotypes (Sultan, 1995), but taxa whose constituent individuals express limited adaptive plasticity might be restricted to narrower, 'specialist' ecological ranges. Williams et al. (1995) stated that plasticity might facilitate the rapid distribution of introduced and also native species into new habitats without the evolutionary lag time needed to adapt to new habitats through natural selection.
Our results revealed high morphological diversity between different populations of this species. ANOVA and PCA tests showed significant differences for most of the studied morphological traits between the studied populations and different shapes and amounts of qualitative as well as qualitative characteristics were observed between the populations. For this reason, high degree of phenotypic plasticity is seen in this species. Sultan (2000) stated that a single genotype can create various phenotypes in different ecological conditions. This phenomenon is definite as phenotypic plasticity. More investigations have proved that plants are plastic for a remarkable array of environmental important factors. Comparative, quantitative genetics and molecular approaches are leading to new insights into the adaptive nature of plasticity, its underlying mechanisms and its role in the ecological distribution and evolutionary diversification of plants (Sultan, 2000).
Previous studies showed high infraspecific variations in essential oil compositions of this species in Iran. Ahmadi and Mirza (1999) reported bornyl acetate (18.1%), p-caryophyllene (16.5%), a-pinene (15.6%), 1,8-cineole (8.3%) and limonene (8.3%) as
Ukrainian Journal of Ecology, 7(4), 2017
major constituents of essential oil compositions of this plant, while results of Morteza-Semnani et al (2005) studies showed that the major constituents of the essential oil of S. muiticauiis are camphor (11.0%), 1,8-cineole (10.7%), borneol (8.6%) and a-pinene (7.5%).
On the basses of mentioned essential oil researches on this species, it seems that a phenomenon beyond the phenotypic plasticity creates such morphological variations between the populations of S. multicaulis. Although, no infraspecific taxon definite for this species in Flora Iranica (Rechinger 1982), Flora of Iran (Jamzad 2012) and Flora of Turkey (Davis, 1982), in Flora of Lebanon one variety of this species, Salvia muiticauiisVahl. var. simpiicifoiia Boiss, is present (Senatore et al 2004). The population no. 13 had special morphological characteristics. When morphological traits of it were compared with morphological descriptions of S. muiticauiis in Flora Iranica (Rechinger, 1982), Flora of Iran (Jamzad, 2012) and Flora of Turkey (Davis, 1982), it was cleared that some main morphological traits of this population that were useful in identification of this species were different from S. muiticauiis. For example, the color of calyx and corolla length of this population was not similar to S. muiticauiis description. So these characteristics are used in identification key of this species, therefore we predict that this population was a new variety or subspecies of this species in Iran.
References
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Ahmadi, L., Mirza, M. (1999). Essential oil of Saivia muiticauiis Vahl from Iran. Journal of Essential Oil Research, 11, 289-290.
^akilcioglu, U., §engun, M.T., Turkoglu, I. (2010). An ethnobotanical survey of medicinal plants of Yazikonak and Yurtbafi districts
of Elazig province, Turkey. Journal of Medicinal Plants Research, 4,567-572.
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Jimenez, J, Risco, S, Ruiz, T et al. Hypoglycemic activity of Saiviaiavenduiifoiia. Planta Medica,52, 260-262, 1986.
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Podani, J. (2000). Introduction to the Exploration of Multivariate Data, Backhuyes, Leide Ltd. Rechinger, K.H. (1982). Labiatae in Flora Iranica. Akademische Druckund Verlagsanstalt, Graz, Austria.
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simpiicifoiia Boiss. growing wild in Lebanon. Journal of Chromatography A, 1052, 237-240.
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Citation:
Talebi, S.M., Rezakhanlou, A., Matsyura, A. (2017). Do we have infraspecific taxa of Salvia multicaulis Vahl (Lamiaceae) In Iran?
Ukrainian Journal of Ecology, 7{4), 432-439. I Thk work Is licensed under a Creative Commons Attribution 4.0. License
Table 2. Mean and standard deviation of some examined morphological characteristics between the studied populations (all values are in cm). Numbers indicate population's code 1-14 as in Table 1.
populations Stem length
1 Mean 28.8
N 8
Std. Dev 5.79
2 Mean 27.50
N 8
Std. Dev 3.38
3 Mean 27.60
N 5
Std. Dev. 4.39
4 Mean 18.80
N 5
Std. Dev. 9.33
5 Mean 33.00
N 8
Std. Dev. 4.27
6 Mean 22.12
N 8
Std. Dev. 3.64
7 Mean 38.00
N 8
Std. Dev. 7.91
8 Mean 18.11
N 9
Std. Dev. 3.14
9 Mean 24.00
N 8
Std. Dev. 5.31
10 Mean 31.00
N 4
Std. Dev. 3.16
11 Mean 23.87
N 8
Std. Dev. 7.01
12 Mean 26.20
N 5
Std. Dev. 5.93
13 Mean 32.42
N 7
Std. Dev. 4.23
14 Mean 17.00
N 3
Std. Dev. 3.60
Basal leaf shape
elliptic
lanceolat e
elliptic
Linear-lanceolat e
elliptic
oblanceol ate
elliptic
Basal ' leaf length Basal petiole length Basal leaf width Basal leaf margin Floral leaf length Floral leaf length/wid th Floral leaf width Floral leaf shape Corolla length Calyx length Pedicle Calyx length width Calyx length/width Flower number of each cycle Bract length Petal color
2.45 3.26 1.37 2.42 0.71 3.63 1.38 1.52 0.95 0.27 0.93 1.14 2.50 1.06
8 8 8 serralate 8 8 8 8 8 8 8 8 8 8 8 violet
1.11 1.62 0.52 0.92 0.15 1.74 0.79 0.20 0.10 0.12 0.34 0.47 0.53 0.35
2.05 3.51 1.11 1.93 0.45 4.13 0.78 1.66 0.98 0.16 0.93 1.1838 3.25 0.73
8 8 8 serralate 8 8 8 8 8 8 8 8 8 8 8 violet
0.83 1.21 0.35 0.96 0.07 1.29 0.24 0.23 0.20 0.05 0.23 0.68 0.88 0.20
2.14 3.42 1.18 1.86 0.52 1.99 1.95 2.10 1.20 0.36 1.08 1.16 4.20 0.78
5 5 5 serrate 5 5 5 5 5 5 5 5 5 5 5 purple
0.83 0.89 0.24 2.13 0.50 2.08 1.41 0.33 0.44 0.08 0.40 0.33 1.48 0.16
1.58 2.04 1.06 1.36 0.36 3.12 2.45 1.40 0.84 0.16 0.68 1.22 4.20 0.98
5 5 5 serralate 5 5 5 5 5 5 5 5 5 5 5 purple
0.25 0.28 0.33 0.78 0.21 1.88 1.44 0.14 0.15 0.05 0.04 0.18 1.92 0.39
3.18 3.55 1.23 3.80 0.86 4.56 1.12 1.46 1.01 0.32 1.07 1.08 7.62 0.66
8 8 8 serrate 8 8 8 8 8 8 8 8 8 8 8 purple
0.78 1.53 0.42 0.56 0.18 1.11 1.02 0.32 0.24 0.19 0.53 0.41 3.20 0.19
1.98 2.67 1.13 1.88 0.60 3.29 1.37 1.60 0.86 0.22 0.76 1.16 4.75 0.66
8 8 8 serrate 8 8 8 8 8 8 8 8 8 8 8 purple
0.49 0.80 0.30 0.50 0.20 0.77 1.32 0.29 0.15 0.04 0.19 0.22 1.16 0.14
1.77 3.68 1.20 2.47 0.82 2.98 1.53 1.37 0.83 0.25 0.67 1.43 8.25 1.03
8 8 8 serralate 8 8 8 8 8 8 8 8 8 8 8 purple
0.70 1.77 0.43 1.31 0.34 1.04 0.98 0.18 0.11 0.07 0.31 0.52 3.32 0.25
1.75 2.22 1.13 1.58 0.61 2.90 3.22 1.58 0.86 0.22 0.82 1.10 6.88 0.68
9 9 9 serralate 9 9 9 9 9 9 9 9 9 9 9 purple
0.42 1.07 0.30 0.26 0.21 1.08 0.66 0.25 0.12 0.13 0.16 0.32 5.06 0.20
2.00 3.75 1.25 2.48 0.70 3.90 1.59 1.63 0.91 0.31 0.83 1.15 6.87 0.63
8 8 8 serrate 8 8 8 8 8 8 8 8 8 8 8 purple
0.70 1.74 0.43 0.78 0.28 1.27 1.26 0.24 0.09 0.08 0.23 0.32 2.23 0.23
2.50 4 3.37 4 1.10 4 serrate 3.12 4 0.70 4 4.54 4 0.87 4 1.45 4 0.92 4 0.37 4 1.15 4 0.80 4 7.25 4 0.95 4 purple
0.49 1.10 0.60 1.35 0.21 1.80 0.14 0.17 0.15 0.09 0.12 0.15 1.70 0.70
1.90 3.18 1.01 1.48 0.51 3.47 1.71 1.58 0.95 0.22 0.88 1.14 4.62 0.87
8 8 8 serrate 8 8 8 8 8 8 8 8 8 8 8 purple
0.55 1.09 0.40 0.39 0.24 1.80 1.20 0.19 0.10 0.07 0.24 0.37 1.84 .26
2.14 5 2.06 5 1.18 5 serralate 2.16 5 .60 5 3.54 5 2.14 5 1.70 5 1.36 5 0.24 5 1.42 5 1.02 5 6.20 5 0.92 5 Medium purple
0.25 0.94 0.24 0.91 0.14 0.99 1.38 0.33 0.15 0.05 0.46 0.28 1.92 0.42
2.5000 3.88 1.51 2.82 0.87 3.64 2.60 1.17 1.81 0.44 2.51 0.72 10.71 1.44
7 7 7 serrate 7 7 7 7 7 7 7 7 7 7 7 lavender
0.87 0.94 0.70 0.86 0.30 1.61 1.19 0.11 0.21 0.07 0.39 0.08 3.72 0.51
1.26 1.80 0.73 1.73 0.76 2.55 2.16 1.36 1.33 0.26 1.50 0.90 8.33 1.000 0
3 3 3 serrate 3 3 3 3 3 3 3 3 3 3 3 purple
0.49 0.43 0.25 1.16 0.51 0.92 1.25 0.05 0.28 .05774 0.43 0.08 4.04 0.40