The results of the analysis of variance showed that the effect of selenium and nano selenium fertilizers on plant height, inflorescence length, number of inflorescences, inflorescence weight, shoot weight, root length, total biomass weight, essential oil percentage, and hypericin and hyperforin content was significant at 1% level, and on the root weight at 5% level (Table 1).
Table 1
Effect of selenium and nano-selenium on morphophysiological characteristics and growth of St. John's wort using analysis of variance
S.O.V
|
df
|
plant height
|
inflorescence length
|
number of inflorescences
|
inflorescence weight
|
shoot weight
|
root length
|
root weight
|
total biomass weight
|
essential oil percentage
|
hypericin content
|
hyperforin content
|
Replication
|
2
|
75.69*
|
26.01ns
|
9ns
|
57.76**
|
147.21**
|
87.84**
|
2.25ns
|
36.4*
|
0.00ns
|
0.75**
|
73.38**
|
Treatment
|
8
|
127.05**
|
35.45**
|
54**
|
45.20**
|
238.58**
|
39.45**
|
37.77*
|
773.08**
|
0.00**
|
3.53**
|
267.75**
|
Error
|
16
|
7.22
|
8.52
|
5.25
|
5.58
|
2.69
|
0.62
|
5.46
|
10.59
|
0.00
|
0.06
|
2.19
|
%CV.
|
|
5.52
|
13.59
|
22.91
|
5.35
|
2.90
|
3.4
|
13.03
|
3.66
|
23.06
|
10.82
|
3.60
|
Ns, * and ** indicate non-significance and significance at the level of 5 and 1%, respectively. |
Continuous of Table (1).
S.O.V
|
df
|
selenium
|
RWC
|
chlorophylls a
|
chlorophylls b
|
chlorophylls total
|
phenol
|
proline
|
CAT
|
POD
|
MDA
|
DPPH
|
Replication
|
2
|
378.56**
|
272.03**
|
0.19**
|
0.006**
|
0.26**
|
2.08**
|
44.44**
|
0.002**
|
0.40*
|
3321.6*
|
54.76**
|
Treatment
|
8
|
16804.1**
|
508.77**
|
0.09**
|
0.02**
|
0.17**
|
3.07**
|
614.33**
|
0.004**
|
0.61**
|
201147**
|
350.08**
|
Error
|
16
|
21.81
|
0.39
|
0.00
|
0.00
|
0.00
|
0.29
|
0.98
|
0.00
|
0.07
|
769.53
|
4.59
|
%CV.
|
|
3.84
|
3.88
|
4.00
|
7.04
|
3.50
|
13.06
|
3.62
|
13.48
|
15.14
|
9.28
|
3.83
|
RWC: Relative Water Content, CTA: Catalas, POD: Proxidase, MDA: Malondialdehyde |
Ns, * and ** indicate non-significance and significance at the level of 5 and 1%, respectively. |
The effect of selenium and nano-selenium on the amount of selenium accumulation in the plant, relative leaf water content, the content of chlorophylls a, b, and total, the phenol and proline content, and production and accumulation of catalase, peroxidase, malondialdehyde, and DPPH enzymes were statistically significant at the 1% level (Table 1).
Comparison of means showed that the highest plant height was obtained from control treatment (58 cm) and foliar application of 6 mg/l selenium (56.4 cm) (Table 2). Inflorescence length was the highest in control treatment and two treatment levels of 6 and 8 mg/l selenium and nano-selenium (Table 2). The number of inflorescences was the highest in the control treatment and the first treatment level (6 mg/l) of selenium and nano-selenium (Table 2).
Table 2
Comparison of means of the effect of selenium and nano-selenium on morphophysiological characteristics and growth of St. John's wort
|
plant height
|
inflorescence length
|
number of inflorescences
|
inflorescence weight
|
shoot weight
|
root length
|
root weight
|
total biomass weight
|
essential oil percentage
|
hypericin content
|
hyperforin content
|
|
cm
|
cm
|
|
g/plant
|
g/plant
|
cm
|
g/plant
|
g/plant
|
%
|
mg/g dry matter
|
mg/g dry matter
|
co
|
58a
|
25.5a
|
16a
|
21a
|
70a
|
27a
|
24a
|
115a
|
0.14a
|
2.7cd
|
44c
|
si6
|
56.4a
|
25ab
|
14ab
|
17b
|
66b
|
26ab
|
22ab
|
105b
|
0.1b
|
3.2b
|
50b
|
8
|
50bc
|
24ab
|
10cd
|
15c
|
60c
|
25bc
|
19bc
|
94cd
|
0.09bc
|
3.8a
|
44c
|
10
|
48.5bc
|
20bcd
|
8de
|
12d
|
55d
|
22d
|
17.5c
|
84.5e
|
0.07bcde
|
1.5e
|
57a
|
12
|
41de
|
18cd
|
6ef
|
10e
|
47e
|
20ef
|
16.5c
|
73.5f
|
0.05de
|
1f
|
33e
|
nanosi6
|
51.5b
|
23.8ab
|
14ab
|
17.5b
|
62c
|
25bc
|
18bc
|
97.5c
|
0.1b
|
3bc
|
46c
|
8
|
49b
|
22.5abc
|
12bc
|
15c
|
57d
|
23.8c
|
17.5c
|
89.5de
|
0.08bcd
|
2.5d
|
41d
|
10
|
45.6cd
|
18.5cd
|
6ef
|
12d
|
48e
|
21de
|
15cd
|
75f
|
0.06cde
|
1.2ef
|
34e
|
12
|
38e
|
16d
|
4f
|
9e
|
44f
|
18.8f
|
12d
|
65g
|
0.04e
|
0.8f
|
26f
|
The same letters in each column indicate that there is no significant difference between the means |
Continuous of Table (2).
|
selenium
|
RWC
|
chlorophylls a
|
chlorophylls b
|
chlorophylls total
|
phenol
|
proline
|
CAT
|
POD
|
MDA
|
DPPH
|
|
µg/g dw
|
%
|
g/100 g FM
|
g/100 g FM
|
g/100 g FM
|
Mg/l FW extract
|
µmol/g FW
|
u/g FW/min
|
U/g FW/min
|
mM/d FW
|
%
|
co
|
37h
|
84a
|
0.77e
|
0.26d
|
1.03e
|
3.5c
|
12h
|
0.05e
|
1.1d
|
68d
|
35f
|
si6
|
49.5g
|
76b
|
0.96d
|
0.3cd
|
1.26d
|
3.23c
|
15g
|
0.07de
|
1.3d
|
92d
|
47e
|
8
|
79f
|
75b
|
1.02d
|
0.32bc
|
1.34d
|
3.31c
|
25d
|
0.07de
|
2ab
|
103d
|
56c
|
10
|
120d
|
70c
|
1.13c
|
0.35b
|
1.48c
|
3.28c
|
33c
|
0.1bc
|
2.3a
|
185c
|
68a
|
12
|
137c
|
61f
|
1.23b
|
0.28cd
|
1.51c
|
4.54b
|
48b
|
0.15a
|
1.5cd
|
590b
|
61b
|
nanosi6
|
94e
|
75.5b
|
1d
|
0.32bc
|
1.32d
|
3.41c
|
17f
|
0.08cd
|
1.4cd
|
110d
|
52d
|
8
|
122.63d
|
68.4d
|
1.19bc
|
0.36b
|
1.55c
|
4.66b
|
19e
|
0.09cd
|
1.8bc
|
222c
|
71a
|
10
|
167.52b
|
66e
|
1.25ab
|
0.46a
|
1.71b
|
5.2ab
|
26d
|
0.12b
|
2.4a
|
695a
|
57c
|
12
|
287.52a
|
58g
|
1.32a
|
0.49a
|
1.81a
|
6a
|
52a
|
0.16a
|
1.5cd
|
624b
|
55cd
|
RWC: Relative Water Content, CTA: Catalas, POD: Proxidase, MDA: Malondialdehyde |
The same letters in each column indicate that there is no significant difference between the means |
The highest inflorescence weight (21 g/plant), shoot weight (70 g/plant), root weight (27 g/plant), total biomass weight (115 g/plant), and essential oil content (0.14%) was obtained from the control treatment. The highest root weight was obtained from the control treatment (24 cm) and foliar application of 6 mg/l selenium (22 cm). The highest accumulation of hypericin (3.8 mg/g dry matter) was obtained from the foliar application of 8 mg/l selenium. The highest accumulation of hyperforin (57 mg/g dry matter) was obtained from the foliar application of 10 mg/l selenium.
It was observed that the highest accumulation of selenium (287.52 µg/g dry matter), the highest content of chlorophyll a (1.32 g), chlorophyll b (0.49 g) total chlorophyll (1.81 g), phenol (6 mg), and proline (52 µmol/g) were obtained in the foliar application of 12 mg/l nano-selenium. The highest catalase content was obtained from the foliar application of 12 mg/l nano-selenium (0.16 u/g FW/min) and foliar application of 12 mg/l selenium (0.15 u/g FW/min). Peroxidase was higher in the foliar application of 8 and 10 mg/l selenium and 10 mg/l nano-selenium than other treatments. The highest malondialdehyde leakage (695 u/g FW/min) was obtained from the 10 mg/l nano-selenium treatment. The highest levels of antioxidant enzyme (71 and 68%) were obtained from the foliar application of 8 mg/l selenium and 10 mg/l nano-selenium, respectively.
The study of simple relationships between traits showed a significant positive correlation between the plant height and inflorescence length, number of inflorescences, inflorescence weight, shoot weight, root length, root weight, total biomass weight, essential oil percentage, the content of hypericin and hyperforin, relative water content, and peroxidase at the level of 1%. The plant height was also significantly correlated with the amount of selenium, chlorophyll a, b, and total chlorophyll, phenol, proline, catalase, malondialdehyde, and DPPH (Table 3). Similar to the plant height, the inflorescence length, number of inflorescences, inflorescence weight, shoot weight, root length, root weight, essential oil percentage, and amount of hypericin and hyperforin had a significant correlation with other traits (Table 3).
Table 3
Simple correlation of the effects of selenium and nano-selenium on morphophysiological characteristics and growth of St. John's wort
|
plant height
|
inflorescence length
|
number of inflorescences
|
inflorescence weight
|
shoot weight
|
root length
|
root weight
|
total biomass weight
|
essential oil percentage
|
hypericin content
|
hyperforin content
|
|
1
|
2
|
3
|
4
|
5
|
6
|
7
|
8
|
9
|
10
|
11
|
1
|
1.00
|
|
|
|
|
|
|
|
|
|
|
2
|
0.83**
|
1.00
|
|
|
|
|
|
|
|
|
|
3
|
0.84**
|
0.77**
|
1.00
|
|
|
|
|
|
|
|
|
4
|
0.55**
|
0.45*
|
0.64**
|
1.00
|
|
|
|
|
|
|
|
5
|
0.91**
|
0.85**
|
0.87**
|
0.57**
|
1.00
|
|
|
|
|
|
|
6
|
0.90**
|
0.81**
|
0.79**
|
0.35ns
|
0.93**
|
1.00
|
|
|
|
|
|
7
|
0.71**
|
0.57**
|
0.71**
|
0.61**
|
0.82**
|
0.68**
|
1.00
|
|
|
|
|
8
|
0.86**
|
0.77**
|
0.87**
|
0.76**
|
0.95**
|
0.81**
|
0.90**
|
1.00
|
|
|
|
9
|
0.80**
|
0.81**
|
0.79**
|
0.62**
|
0.89**
|
0.78**
|
0.76**
|
0.89**
|
1
|
|
|
10
|
0.77**
|
0.82
|
0.75**
|
0.52**
|
0.83**
|
0.82**
|
0.62**
|
0.79**
|
0.73**
|
1.00
|
|
11
|
0.71**
|
0.59**
|
0.54**
|
0.31ns
|
0.70**
|
0.63**
|
0.56**
|
0.64**
|
0.56**
|
0.58**
|
1.00
|
12
|
-0.78**
|
-0.68**
|
-0.75**
|
-0.74**
|
-0.79**
|
-0.66**
|
-0.81**
|
-0.87**
|
-0.74**
|
-0.73**
|
-0.68**
|
13
|
0.91**
|
0.78**
|
0.82**
|
0.60**
|
0.96**
|
0.90**
|
0.79**
|
0.93**
|
0.86**
|
0.79**
|
0.70**
|
14
|
-0.47*
|
-0.36ns
|
-0.50*
|
-0.94**
|
-0.49*
|
-0.25ns
|
-0.60**
|
-0.70**
|
-0.59**
|
-0.44*
|
-0.32ns
|
15
|
-0.48*
|
-0.39*
|
-0.55**
|
-0.65**
|
-0.50*
|
-0.34ns
|
-0.62**
|
-0.63**
|
-0.46*
|
-0.44*
|
-0.43*
|
16
|
-0.50*
|
-0.39*
|
-0.54**
|
-0.91**
|
-0.53**
|
-0.29ns
|
-0.64**
|
-0.72**
|
-0.59**
|
-0.47*
|
-0.37*
|
17
|
-0.56**
|
-0.53**
|
-0.53**
|
-0.67**
|
-0.53**
|
-0.34ns
|
-0.59**
|
-0.64**
|
-0.56**
|
-0.58**
|
-0.66**
|
18
|
-0.78**
|
-0.67**
|
-0.78**
|
-0.84**
|
-0.76**
|
-0.64**
|
-0.66**
|
-0.84**
|
-0.75**
|
-0.72**
|
-0.54*
|
19
|
-0.71**
|
-0.54**
|
-0.71**
|
-0.87**
|
-0.68**
|
-0.52*
|
-0.67**
|
-0.81**
|
-0.72**
|
-0.71**
|
-0.56*
|
20
|
-0.17ns
|
-0.24ns
|
-0.43*
|
-0.51*
|
-0.24ns
|
-0.06ns
|
-0.19ns
|
-0.33ns
|
-0.35ns
|
-0.15ns
|
0.16ns
|
21
|
-0.73**
|
-0.72**
|
-0.76**
|
-0.72**
|
-0.82**
|
-0.67**
|
-0.69**
|
-0.86**
|
-0.73**
|
-0.81**
|
-0.76**
|
22
|
-0.39*
|
-0.33ns
|
-0.42*
|
-0.63**
|
-0.45*
|
-0.28ns
|
-0.48*
|
-0.56*
|
-0.56*
|
-0.28ns
|
0.04ns
|
Ns, * and ** indicate non-significance and significance at the level of 5 and 1%, respectively. |
Continuous of Table (3).
|
selenium
|
RWC
|
chlorophylls a
|
chlorophylls b
|
chlorophylls total
|
phenol
|
proline
|
CAT
|
POD
|
MDA
|
DPPH
|
|
12
|
13
|
14
|
15
|
16
|
17
|
18
|
19
|
20
|
21
|
22
|
12
|
1.00
|
|
|
|
|
|
|
|
|
|
|
13
|
-0.80**
|
1.00
|
|
|
|
|
|
|
|
|
|
14
|
0.73**
|
-0.53**
|
1
|
|
|
|
|
|
|
|
|
15
|
0.83**
|
-0.49*
|
0.71**
|
1
|
|
|
|
|
|
|
|
16
|
0.8**
|
-0.55*
|
0.98**
|
0.84**
|
1
|
|
|
|
|
|
|
17
|
0.8**
|
-0.52*
|
0.78**
|
0.73**
|
0.81**
|
1
|
|
|
|
|
|
18
|
0.81**
|
-0.77**
|
0.69**
|
0.46*
|
0.66**
|
0.59**
|
1
|
|
|
|
|
19
|
0.83**
|
-0.71**
|
0.84**
|
0.63**
|
0.83**
|
0.78**
|
0.87**
|
1
|
|
|
|
20
|
0.26ns
|
-0.15ns
|
0.52*
|
0.51*
|
0.55**
|
0.25ns
|
0.19ns
|
0.28ns
|
1
|
|
|
21
|
0.8**
|
-0.78**
|
0.7**
|
0.64*
|
0.72**
|
0.77**
|
0.71**
|
0.82**
|
0.29ns
|
1
|
|
22
|
0.38*
|
-0.49*
|
0.68*
|
0.33ns
|
0.61**
|
0.31ns
|
0.41*
|
0.44*
|
060**
|
0.29ns
|
1
|
RWC: Relative Water Content, CTA: Catalas, POD: Proxidase, MDA: Malondialdehyde |
Ns, * and ** indicate non-significance and significance at the level of 5 and 1%, respectively. |
It was observed that the content of selenium had a significant negative correlation with morphological traits and yield of different organs and a significant positive correlation with the number of enzymes, chlorophylls, phenol, and proline. In contrast to selenium, relative water content had a significant positive correlation with agronomic traits and yields and a significant negative correlation with the number of enzymes, chlorophylls, phenol, and proline.
As can be seen in Table 3, physiological traits such as chlorophylls, enzymes, phenol, and proline had a significant positive correlation with each other.