Characterization of Chocolate spot disease and leaf spot disease and recovery
Careful examination of figures 1-2 revealed that symptoms of chocolate spot disease appeared on leaves, stems flowers and pod tissues. Infection started as small reddish-brown circular spots on leaves, stems and flowers. The spots on leaves and stems enlarged and developed a grey, dead center with a red-brown margin. In the present study, two stages of the disease have been observed. The non-aggressive stage known by the small circular and discrete reddish spots with darker margins visible on one side of leaf; and the aggressive stage where the small spots merge and coalesce to form irregular larger dark-brown lesions involving the entire leaf surface. Under favorable conditions, the disease spread quickly causing sever defoliation, flowers drop, stem collapse, tissue necrosis and finally plant death (figures 1-2).
Careful examination of figures 3-4 showed that initially lesions were brown, water soaked, circular to irregular shape on the stem, pod, and other parts of the plant. These dark brown leaf spots often had a zoned pattern of concentric brown rings with dark margins which gave the spots a target-like appearance. The fungus showed that older leaves were usually attacked first but the disease progressed upward and affected leaves which turned yellowish, became senescent and either dried up and dropped or fell off. In the later stage of the disease, the leaves became blighted from the margin to the center and most of the diseased plants defoliated completely.
Uptake and translocation of nanomaterials
Transmission Electron Microscopy (TEM) for investigation of plant leaves (46-day-old) after applying chitosan nanoparticles, carbon nanotubes and solid lipid nanoparticles either singly or loaded with nystatin and fluconazole antifungal antibiotics was carried out in the present study to indicate the presence or absence of nanomaterials as a result of taken up through leaves of broad bean plants at late vegetative growth and developmental stage.
Careful examination of figures 1-6 derived from TEM images of broad bean leaves (46-day-old) revealed the following main points:
- Complete absence of nanomaterials either singly or loaded with nystatin and fluconazole inside epidermal cells, cortical cells, spongy tissues, or palisade tissues but they were existed in vascular system of broad bean leaves treated with chitosan nanoparticles, carbon nanotubes and solid lipid nanoparticles either singly or loaded with nystatin and fluconazole antifungal antibiotics (figures 4-9).
- Figures 4-9 showed that foliar application of the three nanoparticles either singly or in combination with nystatin and fluconazole to broad bean plants and confirm that the uptake and translocation of nanoparticles were achieved by phloem tissues, in particular, sieve tubes cells.
- It is worth to mention that the observed results concerning the uptake and translocation of nanomaterials by broad bean plants through the entire period of the experiment might lead to conclude that the main pathway for uptake and translocation of nanomaterials either singly or loaded with nystatin and fluconazole was via the cells of the phloem sieve tubes.
Changes in growth, yield and developmental parameters
Foliar application of either CSNPs, CNTs or SLNPs singly or loaded with nystatin and fluconazole antibiotics led to significant variable increases in all yield growth and developmental parameters determined as compared with control diseased values. The magnitude of increase was, in general, most pronounced with solid lipid nanoparticles loaded with nystatin and fluconazole antifungal antibiotics (tables 1 and 2).
Furthermore, harvest index, crop index and mobilization index values of broad bean plants variously treated with nanomaterials either singly or loaded with nystatin and fluconazole antifungal antibiotics were found to be variously changed corresponding to treatment type, way and levels of treatments as compared to values of diseased plants. Thus, in general, as evident from tables 1 and 2 and as compared to diseased plants, the calculated percentage ratio of increase in harvest index was as follows: strategy C > strategy A > strategy B was displayed for Botrytis fabae with respect to treatment of infected broad bean plants with nano-drug delivery strategies. In relation to control diseased broad bean plants, the calculated percentage improvement in crop index of the yielded broad bean plants was arranged as follows: SLNPs + FLZ > SLNPs + NYS > CSNPs + FLZ > CSNPs + NYS > CNTs + FLZ > CNTs + NYS > diseased plants. Furthermore, mobilization index of the yielded broad bean crop as compared with control diseased plants was as follows: SLNPs + FLZ > SLNPs + NYS > CSNPs + FLZ > CSNPs + NYS > CNTs + FLZ > CNTs + NYS > diseased plants.
As compared with control diseased plants, it is worth to mention that the growth and developmental yield parameters of broad bean plants variously treated with nanomaterials either singly or loaded with nystatin and fluconazole antifungal antibiotics showed significant improvement percentage of shoot length, number of pods per plant, pod weight per plant, number of seeds per plant, crop yield per plant and seed yield per plant. The magnitude of increased improvement in these growth and developmental yield parameters was most pronounced with solid lipid nanoparticles loaded with nystatin or fluconazole as nan-drug delivery system (tables 1 and 2).
Table 1: Effect of foliar application of different nanomaterials treatments either singly or loaded with nystatin and fluconazole on yield developmental parameters of Faba bean plants grown in clay-sandy soil (before and after infection with Botrytis fabae). In each case, the % change from the control and the % recovery from the infected plant are also shown. Each value is a mean of five replicates. *The values listed are significantly different from control at p ≥ 0.05.
Parameter
Treatment
|
Plant height
(cm plant-1)
|
%
change
|
%
Recov
|
Shoot length
(cm plant-1)
|
%
change
|
%
Recov
|
No. of pods/
plant
|
%
change
|
%
Recov
|
Pod
weight
(g pod-1)
|
%
change
|
%
Recov
|
Control
|
61.11
|
―
|
―
|
54.38
|
―
|
―
|
4.00
|
―
|
―
|
1.54
|
―
|
―
|
Healthy Plant
|
CNTs
|
60.32*
|
-1.29
|
―
|
53.21*
|
-2.15
|
―
|
4.00
|
0.00
|
―
|
1.49*
|
-3.25
|
―
|
CSNPs
|
66.71*
|
9.16
|
―
|
61.31*
|
12.74
|
―
|
5.00*
|
25.00
|
―
|
2.38*
|
54.55
|
―
|
SLNPs
|
64.12*
|
4.93
|
―
|
59.00*
|
8.50
|
―
|
4.00
|
0.00
|
―
|
1.77*
|
14.94
|
―
|
Infected Plant with Botrytis fabae
|
Infected plant
|
51.33*
|
-16.00
|
―
|
42.67*
|
-21.53
|
―
|
3.00*
|
-25.00
|
―
|
0.99*
|
-35.71
|
―
|
NYS
|
63.65*
|
4.16
|
24.00
|
55.42*
|
1.91
|
29.88
|
4.00
|
0.00
|
33.33
|
2.06*
|
33.77
|
108.08
|
FLZ
|
67.73*
|
10.83
|
31.95
|
60.91*
|
12.01
|
42.75
|
4.00
|
0.00
|
33.33
|
2.19*
|
42.21
|
121.21
|
Strategy A
|
A1: CSNPs
|
54.70*
|
-10.49
|
6.57
|
49.31*
|
-9.32
|
15.56
|
3.00*
|
-25.00
|
0.00
|
1.26*
|
-18.18
|
27.27
|
A2: CSNPs-NYS
|
65.73*
|
7.56
|
28.05
|
59.16*
|
8.79
|
38.65
|
5.00*
|
25.00
|
66.67
|
2.25*
|
46.10
|
127.27
|
A3: CSNPs-FLZ
|
64.72*
|
5.91
|
26.09
|
58.30*
|
7.21
|
36.63
|
5.00*
|
25.00
|
66.67
|
2.21*
|
43.51
|
123.23
|
Strategy B
|
B1: CNTs
|
51.78*
|
-15.27
|
0.88
|
41.94*
|
-22.88
|
-1.71
|
3.00*
|
-25.00
|
0.00
|
1.05*
|
-31.82
|
6.06
|
B2: CNTs-NYS
|
60.41*
|
-1.15
|
17.69
|
53.70*
|
-1.25
|
25.85
|
4.00
|
0.00
|
33.33
|
1.82*
|
18.18
|
83.84
|
B3: CNTs-FLZ
|
61.88*
|
1.26
|
20.55
|
51.78*
|
-4.78
|
21.35
|
4.00
|
0.00
|
33.33
|
1.71*
|
11.04
|
72.73
|
Strategy C
|
C1: SLNPs
|
54.55*
|
-10.73
|
6.27
|
46.17*
|
-15.10
|
8.20
|
3.00*
|
-25.00
|
0.00
|
1.16*
|
-24.68
|
17.17
|
C2: SLNPs-NYS
|
70.83*
|
15.91
|
37.99
|
64.49*
|
18.59
|
51.14
|
5.00*
|
25.00
|
66.67
|
2.28*
|
48.05
|
130.30
|
C3: SLNPs-FLZ
|
68.59*
|
12.24
|
33.63
|
60.25*
|
10.79
|
41.20
|
5.00*
|
25.00
|
66.67
|
2.50*
|
62.34
|
152.53
|
Cont. table 1:
Parameter
Treatment
|
No. of seeds/
pod
|
%
change
|
%
Recov
|
Seed
weight
(g seed-1)
|
%
change
|
%
Recov
|
Crop yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Seed yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Straw yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Control
|
3.00
|
―
|
―
|
0.43
|
―
|
―
|
6.59
|
―
|
―
|
5.20
|
―
|
―
|
8.84
|
―
|
―
|
Healthy Plant
|
CNTs
|
3.00
|
0.00
|
―
|
0.35
|
-18.60
|
―
|
6.37*
|
-3.34
|
―
|
4.07*
|
-21.73
|
―
|
8.14*
|
-7.92
|
―
|
CSNPs
|
3.00
|
0.00
|
―
|
0.60*
|
39.53
|
―
|
11.30*
|
71.47
|
―
|
8.97*
|
72.50
|
―
|
8.33*
|
-5.77
|
―
|
SLNPs
|
3.00
|
0.00
|
―
|
0.45
|
4.65
|
―
|
7.49*
|
13.66
|
―
|
5.48*
|
5.38
|
―
|
7.04*
|
-20.36
|
―
|
Infected Plant with Botrytis fabae
|
Infected plant
|
2.00*
|
-33.33
|
―
|
0.23*
|
-46.51
|
―
|
2.96*
|
-55.08
|
―
|
1.35*
|
-74.04
|
―
|
8.86
|
0.23
|
―
|
NYS
|
3.00
|
0.00
|
50.00
|
0.45
|
4.65
|
95.65
|
8.03*
|
21.85
|
171.28
|
5.99*
|
15.19
|
343.70
|
8.52*
|
-3.62
|
-3.84
|
FLZ
|
3.00
|
0.00
|
50.00
|
0.53*
|
23.26
|
130.43
|
9.00*
|
36.57
|
204.05
|
6.34*
|
21.92
|
369.63
|
8.40*
|
-4.98
|
-5.19
|
Strategy A
|
A1: CSNPs
|
2.00*
|
-33.33
|
0.00
|
0.38
|
-11.63
|
65.22
|
4.73*
|
-28.22
|
59.80
|
2.44*
|
-53.08
|
80.74
|
7.82*
|
-11.54
|
-11.74
|
A2: CSNPs-NYS
|
3.00
|
0.00
|
50.00
|
0.57*
|
32.56
|
147.83
|
10.93*
|
65.86
|
269.26
|
8.79*
|
69.04
|
551.11
|
7.56*
|
-14.48
|
-14.67
|
A3: CSNPs-FLZ
|
3.00
|
0.00
|
50.00
|
0.56*
|
30.23
|
143.48
|
10.85*
|
64.64
|
266.55
|
8.62*
|
65.77
|
538.52
|
7.08*
|
-19.91
|
-20.09
|
Strategy B
|
B1: CNTs
|
2.00*
|
-33.33
|
0.00
|
0.31*
|
-27.91
|
34.78
|
3.27*
|
-50.38
|
10.47
|
1.83*
|
-64.81
|
35.56
|
9.39*
|
6.22
|
5.98
|
B2: CNTs-NYS
|
3.00
|
0.00
|
50.00
|
0.47
|
9.30
|
104.35
|
7.15*
|
8.50
|
141.55
|
5.78*
|
11.15
|
328.15
|
7.12*
|
-19.46
|
-19.64
|
B3: CNTs-FLZ
|
3.00
|
0.00
|
50.00
|
0.54*
|
25.58
|
134.78
|
6.57
|
-0.30
|
121.96
|
4.03*
|
-22.50
|
198.52
|
6.87*
|
-22.29
|
-22.46
|
Strategy C
|
C1: SLNPs
|
2.00*
|
-33.33
|
0.00
|
0.31*
|
-27.91
|
34.78
|
3.25*
|
-50.68
|
9.80
|
1.84*
|
-64.62
|
36.30
|
7.04*
|
-20.36
|
-20.54
|
C2: SLNPs-NYS
|
3.00
|
0.00
|
50.00
|
0.59*
|
37.21
|
156.52
|
11.29*
|
71.32
|
281.42
|
8.72*
|
67.69
|
545.93
|
6.42*
|
-27.38
|
-27.54
|
C3: SLNPs-FLZ
|
3.00
|
0.00
|
50.00
|
0.66*
|
53.49
|
186.96
|
13.39*
|
103.19
|
352.36
|
9.30*
|
78.85
|
588.89
|
6.25*
|
-29.30
|
-29.46
|
Cont. table 1:
Parameter
Treatment
|
Harvest
index
|
%
change
|
%
Recov
|
Crop
index
|
%
change
|
%
Recov
|
Mobilization
index
|
%
change
|
%
Recov
|
Control
|
0.59
|
―
|
―
|
0.37
|
―
|
―
|
0.042
|
―
|
―
|
Healthy Plant
|
CNTs
|
0.50*
|
-15.25
|
―
|
0.33
|
-10.81
|
―
|
0.041
|
-2.38
|
―
|
CSNPs
|
1.08*
|
83.05
|
―
|
0.52*
|
40.54
|
―
|
0.062
|
47.62
|
―
|
SLNPs
|
0.78*
|
32.20
|
―
|
0.44
|
18.92
|
―
|
0.062
|
47.62
|
―
|
Infected Plant with Botrytis fabae
|
Infected plant
|
0.15*
|
-74.58
|
―
|
0.13*
|
-64.86
|
―
|
0.015
|
-64.29
|
―
|
NYS
|
0.70*
|
18.64
|
366.67
|
0.41
|
10.81
|
215.38
|
0.048
|
14.29
|
220.00
|
FLZ
|
0.75*
|
27.12
|
400.00
|
0.43
|
16.22
|
230.77
|
0.051
|
21.43
|
240.00
|
Strategy A
|
A1: CSNPs
|
0.31*
|
-47.46
|
106.67
|
0.24*
|
-35.14
|
84.62
|
0.030
|
-28.57
|
100.00
|
A2: CSNPs-NYS
|
1.16*
|
96.61
|
673.33
|
0.54*
|
45.95
|
315.38
|
0.071
|
69.05
|
373.33
|
A3: CSNPs-FLZ
|
1.22*
|
106.78
|
713.33
|
0.55*
|
48.65
|
323.08
|
0.078
|
85.71
|
420.00
|
Strategy B
|
B1: CNTs
|
0.19*
|
-67.80
|
26.67
|
0.16*
|
-56.76
|
23.08
|
0.017
|
-59.52
|
13.33
|
B2: CNTs-NYS
|
0.81*
|
37.29
|
440.00
|
0.45
|
21.62
|
246.15
|
0.063
|
50.00
|
320.00
|
B3: CNTs-FLZ
|
0.59
|
0.00
|
293.33
|
0.37
|
0.00
|
184.62
|
0.054
|
28.57
|
260.00
|
Strategy C
|
C1: SLNPs
|
0.26*
|
-55.93
|
73.33
|
0.21*
|
-43.24
|
61.54
|
0.029
|
-30.95
|
93.33
|
C2: SLNPs-NYS
|
1.36*
|
130.51
|
806.67
|
0.58*
|
56.76
|
346.15
|
0.090
|
114.29
|
500.00
|
C3: SLNPs-FLZ
|
1.49*
|
152.54
|
893.33
|
0.60*
|
62.16
|
361.54
|
0.096
|
128.57
|
540.00
|
Table 2: Effect of foliar application of different nanomaterials treatments either singly or loaded with nystatin and fluconazole on yield developmental parameters of Faba bean plants grown in clay-sandy soil (before and after infection with Alternaria alternata). In each case, the % change from the control and the % recovery from the infected plant are also shown. Each value is a mean of five replicates. *The values listed are significantly different from control at p ≥ 0.05.
Parameter
Treatment
|
Plant height
(cm plant-1)
|
%
change
|
%
Recov
|
Shoot length
(cm plant-1)
|
%
change
|
%
Recov
|
No. of pods/
plant
|
%
change
|
%
Recov
|
Pod
weight
(g pod-1)
|
%
change
|
%
Recov
|
Control
|
61.11
|
―
|
―
|
54.38
|
―
|
―
|
4.00
|
―
|
―
|
1.54
|
―
|
―
|
Healthy Plant
|
CNTs
|
60.32*
|
-1.29
|
―
|
53.21*
|
-2.15
|
―
|
4.00
|
0.00
|
―
|
1.49*
|
-3.25
|
―
|
CSNPs
|
66.71*
|
9.16
|
―
|
61.31*
|
12.74
|
―
|
5.00*
|
25.00
|
―
|
2.38*
|
54.55
|
―
|
SLNPs
|
64.12*
|
4.93
|
―
|
59.00*
|
8.50
|
―
|
4.00
|
0.00
|
―
|
1.77*
|
14.94
|
―
|
Infected Plant with Alternaria alternata
|
Infected plant
|
50.46*
|
-17.43
|
―
|
43.52*
|
-19.97
|
―
|
3.00*
|
-25.00
|
―
|
1.02*
|
-33.77
|
―
|
NYS
|
66.83*
|
9.36
|
32.44
|
58.21*
|
7.04
|
33.75
|
4.00
|
0.00
|
33.33
|
2.05*
|
33.12
|
100.98
|
FLZ
|
65.62*
|
7.38
|
30.04
|
57.40*
|
5.55
|
31.89
|
4.00
|
0.00
|
33.33
|
2.20*
|
42.86
|
115.69
|
Strategy A
|
A1: CSNPs
|
53.61*
|
-12.27
|
6.24
|
45.28*
|
-16.73
|
4.04
|
3.00*
|
-25.00
|
0.00
|
1.27*
|
-17.53
|
24.51
|
A2: CSNPs-NYS
|
63.32*
|
3.62
|
25.49
|
55.83*
|
2.67
|
28.29
|
4.00
|
0.00
|
33.33
|
1.79*
|
16.23
|
75.49
|
A3: CSNPs-FLZ
|
64.21*
|
5.07
|
27.25
|
54.33
|
-0.09
|
24.84
|
5.00*
|
25.00
|
66.67
|
2.29*
|
48.70
|
124.51
|
Strategy B
|
B1: CNTs
|
52.27*
|
-14.47
|
3.59
|
44.34*
|
-18.46
|
1.88
|
3.00*
|
-25.00
|
0.00
|
1.05*
|
-31.82
|
2.94
|
B2: CNTs-NYS
|
61.73*
|
1.01
|
22.33
|
55.59*
|
2.23
|
27.73
|
4.00
|
0.00
|
33.33
|
1.60
|
3.90
|
56.86
|
B3: CNTs-FLZ
|
60.15*
|
-1.57
|
19.20
|
54.46
|
0.15
|
25.14
|
5.00*
|
25.00
|
66.67
|
2.20*
|
42.86
|
115.69
|
Strategy C
|
C1: SLNPs
|
55.00*
|
-10.00
|
9.00
|
46.59*
|
-14.33
|
7.05
|
3.00*
|
-25.00
|
0.00
|
1.13*
|
-26.62
|
10.78
|
C2: SLNPs-NYS
|
69.63*
|
13.94
|
37.99
|
59.76*
|
9.89
|
37.32
|
5.00*
|
25.00
|
66.67
|
2.46*
|
59.74
|
141.18
|
C3: SLNPs-FLZ
|
70.63*
|
15.58
|
39.97
|
65.28*
|
20.04
|
50.00
|
5.00*
|
25.00
|
66.67
|
2.47*
|
60.39
|
142.16
|
Cont. table 2:
Parameter
Treatment
|
No. of seeds/
pod
|
%
change
|
%
Recov
|
Seed
weight
(g seed-1)
|
%
change
|
%
Recov
|
Crop yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Seed yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Straw yield
/plant
(g plant-1)
|
%
change
|
%
Recov
|
Control
|
3.00
|
―
|
―
|
0.43
|
―
|
―
|
6.59
|
―
|
―
|
5.20
|
―
|
―
|
8.84
|
―
|
―
|
Healthy Plant
|
CNTs
|
3.00
|
0.00
|
―
|
0.35
|
-18.60
|
―
|
6.37*
|
-3.34
|
―
|
4.07*
|
-21.73
|
―
|
8.14*
|
-7.92
|
―
|
CSNPs
|
3.00
|
0.00
|
―
|
0.60*
|
39.53
|
―
|
11.30*
|
71.47
|
―
|
8.97*
|
72.50
|
―
|
8.33*
|
-5.77
|
―
|
SLNPs
|
3.00
|
0.00
|
―
|
0.45
|
4.65
|
―
|
7.49*
|
13.66
|
―
|
5.48*
|
5.38
|
―
|
7.04*
|
-20.36
|
―
|
Infected Plant with Alternaria alternata
|
Infected plant
|
2.00*
|
-33.33
|
―
|
0.24*
|
-44.19
|
―
|
3.06*
|
-53.57
|
―
|
1.40*
|
-73.08
|
―
|
9.76*
|
10.41
|
―
|
NYS
|
3.00
|
0.00
|
50.00
|
0.54*
|
25.58
|
125.00
|
8.42*
|
27.77
|
175.16
|
6.52*
|
25.38
|
365.71
|
7.08*
|
-19.91
|
-27.46
|
FLZ
|
3.00
|
0.00
|
50.00
|
0.55*
|
27.91
|
129.17
|
8.66*
|
31.41
|
183.01
|
6.62*
|
27.31
|
372.86
|
7.89*
|
-10.75
|
-19.16
|
Strategy A
|
A1: CSNPs
|
2.00*
|
-33.33
|
0.00
|
0.37
|
-13.95
|
54.17
|
3.76*
|
-42.94
|
22.88
|
2.38*
|
-54.23
|
70.00
|
8.97*
|
1.47
|
-8.09
|
A2: CSNPs-NYS
|
3.00
|
0.00
|
50.00
|
0.49
|
13.95
|
104.17
|
7.15*
|
8.50
|
133.66
|
5.56*
|
6.92
|
297.14
|
7.39*
|
-16.40
|
-24.28
|
A3: CSNPs-FLZ
|
3.00
|
0.00
|
50.00
|
0.54*
|
25.58
|
125.00
|
10.12*
|
53.57
|
230.72
|
7.67*
|
47.50
|
447.86
|
7.30*
|
-17.42
|
-25.20
|
Strategy B
|
B1: CNTs
|
2.00*
|
-33.33
|
0.00
|
0.25*
|
-41.86
|
4.17
|
3.08*
|
-53.26
|
0.65
|
1.53*
|
-70.58
|
9.29
|
10.43*
|
17.99
|
6.86
|
B2: CNTs-NYS
|
3.00
|
0.00
|
50.00
|
0.44
|
2.33
|
83.33
|
6.73*
|
2.12
|
119.93
|
5.30*
|
1.92
|
278.57
|
6.94*
|
-21.49
|
-28.89
|
B3: CNTs-FLZ
|
3.00
|
0.00
|
50.00
|
0.54*
|
25.58
|
125.00
|
8.81*
|
33.69
|
187.91
|
6.47*
|
24.42
|
362.14
|
6.65*
|
-24.77
|
-31.86
|
Strategy C
|
C1: SLNPs
|
2.00*
|
-33.33
|
0.00
|
0.33*
|
-23.26
|
37.50
|
3.47*
|
-47.34
|
13.40
|
2.05*
|
-60.58
|
46.43
|
8.11*
|
-8.26
|
-16.91
|
C2: SLNPs-NYS
|
3.00
|
0.00
|
50.00
|
0.60*
|
39.53
|
150.00
|
12.18*
|
84.83
|
298.04
|
8.49*
|
63.27
|
506.43
|
7.11*
|
-19.57
|
-27.15
|
C3: SLNPs-FLZ
|
3.00
|
0.00
|
50.00
|
0.75*
|
74.42
|
212.50
|
13.21*
|
100.46
|
331.70
|
11.66*
|
124.23
|
732.86
|
6.83*
|
-22.74
|
-30.02
|
Cont. table 2:
Parameter
Treatment
|
Harvest
index
|
%
change
|
%
Recov
|
Crop
index
|
%
change
|
%
Recov
|
Mobilization
index
|
%
change
|
%
Recov
|
Control
|
0.59
|
―
|
―
|
0.37
|
―
|
―
|
0.042
|
―
|
―
|
Healthy Plant
|
CNTs
|
0.50*
|
-15.25
|
―
|
0.33
|
-10.81
|
―
|
0.041
|
-2.38
|
―
|
CSNPs
|
1.08*
|
83.05
|
―
|
0.52*
|
40.54
|
―
|
0.062
|
47.62
|
―
|
SLNPs
|
0.78*
|
32.20
|
―
|
0.44
|
18.92
|
―
|
0.062
|
47.62
|
―
|
Infected Plant with Alternaria alternata
|
Infected plant
|
0.14*
|
-76.27
|
―
|
0.13*
|
-64.86
|
―
|
0.013
|
-69.05
|
―
|
NYS
|
0.92*
|
55.93
|
557.14
|
0.48*
|
29.73
|
269.23
|
0.068
|
61.90
|
423.08
|
FLZ
|
0.84*
|
42.37
|
500.00
|
0.46*
|
24.32
|
253.85
|
0.058
|
38.10
|
346.15
|
Strategy A
|
A1: CSNPs
|
0.27*
|
-54.24
|
92.86
|
0.21*
|
-43.24
|
61.54
|
0.023
|
-45.24
|
76.92
|
A2: CSNPs-NYS
|
0.75*
|
27.12
|
435.71
|
0.43
|
16.22
|
230.77
|
0.058
|
38.10
|
346.15
|
A3: CSNPs-FLZ
|
1.05*
|
77.97
|
650.00
|
0.51*
|
37.84
|
292.31
|
0.070
|
66.67
|
438.46
|
Strategy B
|
B1: CNTs
|
0.15*
|
-74.58
|
7.14
|
0.13*
|
-64.86
|
0.00
|
0.012
|
-71.43
|
-7.69
|
B2: CNTs-NYS
|
0.76*
|
28.81
|
442.86
|
0.43
|
16.22
|
230.77
|
0.062
|
47.62
|
376.92
|
B3: CNTs-FLZ
|
0.97*
|
64.41
|
592.86
|
0.49*
|
32.43
|
276.92
|
0.074
|
76.19
|
469.23
|
Strategy C
|
C1: SLNPs
|
0.25*
|
-57.63
|
78.57
|
0.20*
|
-45.95
|
53.85
|
0.025
|
-40.48
|
92.31
|
C2: SLNPs-NYS
|
1.19*
|
101.69
|
750.00
|
0.54*
|
45.95
|
315.38
|
0.077
|
83.33
|
492.31
|
C3: SLNPs-FLZ
|
1.71*
|
189.83
|
1121.43
|
0.63*
|
70.27
|
384.62
|
0.092
|
119.05
|
607.69
|
Biosafety of yielded Vicia faba seeds
After confirming the presence of tested nanomaterials (CSNPs, CNTs and SLNPs) singly or loaded with nystatin and fluconazole inside plant tissue, in particular phloem tissue (sieve tubes), it was necessary to figure out the final fate of these nanomaterials. To this end, TEM analysis of representative yielded seeds from Vicia faba plants that had undergone various treatments was carried out.
Careful examination of figure 7 revealed the following main points:
- Yielded seeds produced from Vicia faba plants treated with different nanomaterials loaded with nystatin and fluconazole antifungal antibiotics exhibited complete absence of nanomaterials inside their tissues.
- When compared with control yielded Vicia faba seeds tissues, no change in seed tissues of the variously nanomaterials-treated plants as nano-drug delivery strategies which clearly indicates the complete absence of the different nanomaterials inside yielded Vicia faba seeds tissues.
In conclusion, comparing the three new and novel nano-drug delivery strategies under investigation, in the present study from the percentage important and recovery from illness of broad bean plants diseased with chocolate spot or leaf spot, the following sequence of treatments (nano-drug delivery strategy C > nano-drug delivery strategy A > nano-drug delivery strategy B) was, in general, displayed with respect to recovery from chocolate spot disease caused by Botrytis fabae fungal pathogen. On the other hand, except for the results obtained for mobilization index of broad bean yield, the following sequence of treatments (nano-drug delivery strategy C > nano-drug delivery strategy A > nano-drug delivery strategy B) was, in general, displayed with respect to all growth and developmental yield parameters of the percentage improvement and recovered plants from illness with leaf spot disease caused by Alternaria alternata fungal pathogens.