Fatty acid composition of seed oil of T. sebifera
The fatty acid composition and distribution of T. sebifera are presented in Table 1. The SO of T. sebifera gives a yield of 50% (v/w). The saturated fatty acids are present in the SO were palmitic acid (4.99%) and stearic acid (1.38%.). The unsaturated fatty acids are oleic acid (8.78%), linoleic acid (15.42%), and palmitoleic acid (2.13%). The distribution of saturated fatty acids is less (6.73%) as compared to unsaturated fatty acids (26.33%).
Table 1
Chemical composition of fatty acid methyl esters present in seed oil of Triadica sebifera; Note: RIa = Calculated Retention Indices; RIb = Retention Indices from literature
Sr. No. | Compound’s name | Area (%) | RIa | RIb | Mass fragmentation |
1 | 9-Hexadecenoic acid, methyl ester (Palmitoleic acid, C6: 1) | 2.13 | 1933 | 193141 | 268 [M+], 236, 207, 194, 152, 138, 123, 97, 74, 69, 55, 41 |
2 | Hexadecanoic acid, methyl ester (Palmitic acid, C16: 0) | 4.99 | 1986 | 198442 | 270 [M+], 239, 227, 199, 185, 171, 143, 129, 101, 87, 74, 57, 41 |
3 | 9,12-Octadecadienoic acid (Z, Z), methyl ester (Linoleic acid, C18: 2) | 15.42 | 2092 | 209443 | 294 [M+], 262, 178, 164, 150, 135, 123, 109, 95, 81, 67, 55, 41 |
4 | 9-Octadecenoic acid, methyl ester (Oleic acid, C18: 1) | 8.78 | 2100 | 210144 | 296 [M+], 264, 222, 180, 166, 152, 137, 123, 97, 83, 69, 55, 41 |
5 | Octadecanoic acid, methyl ester (Stearic acid, C18: 0) | 1.38 | 2121 | 212845 | 298 [M+], 255, 213, 199, 143, 129, 101, 87, 74, 57, 43, 41 |
Structure Elucidation Of Isolated Compounds
Compound 1 was isolated as a white amorphous powder. The ESI-QTOF-MS of the compound showing the molecular ion peak at m/z 471.08 (M+Na)+ indicated the molecular formula as C21H20O11 (Fig. S1). Aromatic signals observed in proton spectrum at δH7.98 d (2H, J=8.4 Hz), 6.81 d (2H, J=8.4 Hz), 6.30 s (1H), and 6.11 s (1H) showing the presence of two sets of equivalent protons and two singlet protons which confirms the presence of two aromatic rings in the structure. Also, the presence of a doublet signal at δH5.16 d (1H, J=7.2 Hz) which showing the HSQC correlation with δc 104.2 indicates the presence of a β-D-glucose moiety, which was also confirmed by COSY and HMBC correlations. The carbon value at δc 179.4 showed the presence of the C=O group. Hence from the above spectral data and comparison with literature data12, compound 1 was identified as Kaempferol-3-O-glucoside (Fig. S2).
Kaempferol-3-O-glucoside: White amorphous powder; 128 mg; mp. 178oC; ESI-QTOF-MS (Positive) m/z:471.08 (M+Na)+ 1H-NMR (600 MHz, CD3OD) (Fig. S3 and Table S1), 13C-NMR (150 MHz, CD3OD) (Fig. S4 and Table S1).
Compound 2 was isolated as a white amorphous powder. The molecular formula C21H20O11 was calculated after ESI-QTOF-MS, which was showing the molecular ion peak at m/z 487.08 (M+Na+) (Fig. S5). Compound 2 was identified by comparing observed data with literature-reported data13.After comparison of observed data with literature, compound 2 was identified as Quercetin-3-O-glucoside (Fig. S2).
Quercetin-3-O-glucoside: White amorphous powder; 150 mg; mp. 226oC; ESI-QTOF-MS (Positive) m/z:487.08 (M+Na+). 1H-NMR (600 MHz, CD3OD) (Fig. S6 and Table S1), 13C-NMR (150 MHz, CD3OD) (Fig. S7 and Table S1).
Compound 3 was obtained as a white powder. The molecular ion peak was observed at m/z 341.03 (2M+H)+ in ESI-QTOF-MS (Fig. S8), which indicates the formation of a non-covalent dimer in ESI-MS14.Hence the stable molecular ion peak was formed in the form of a non-covalent dimer (2M+H)+. From spectral analysis and literature reports15, compound 3 was identified as gallic acid (Fig. S2).
Gallic acid: White powder; 124 mg; mp. 260oC; ESI-QTOF-MS (Positive) m/z: 341.03 (2M+H)+; 1H-NMR (600 MHz, CD3OD) (Fig. S9 and Table S2), 13C-NMR (150 MHz, CD3OD) (Fig. S10 and Table S2).
Similarly, compound 4 was also isolated as a white powder. The molecular ion peak for compound 4 was obtained at m/z 349.32 (2M+H)+(Fig. S11), which was also the molecular ion peak of the non-covalent dimer peak of compound 4. ESI-QTOF-MS analyses of compound 4 were indicated its molecular formula C7H10O5 and after analysis and comparison of observed data with literature16, compound 4 was identified as Shikimic acid (Fig. S2).
Shikimic acid: White powder; 108 mg; mp. 184-185oC, ESI-QTOF-MS (Positive) m/z: 349.32 (2M+H)+; 1H-NMR (600 MHz, CD3OD) (Fig. S12 and Table S2), 13C-NMR (150 MHz, CD3OD) (Fig. S13 and Table S2).
Gas chromatography-mass spectrophotometry (GC-MS) analysis of n-hexane fractions
The metabolites and their mass fragmentation, identified through GC-MS in the n-hexane fractions of leaf and bark were presented in Table 2. In the n-hexane fraction of the leaf, the major metabolites were n-Hexadecanoic acid (15.61%) and were followed by octadecanoic acid, ethyl ester (9.85%) and neophytadiene (5.87%). In the case of the n-hexane fraction of the bark, the major metabolites were galaxolide (44.73%) and were followed by ethyl phthalate (28.43%) and 1-Octadecene (2.69%).
Table 2
Chemical composition of n-hexane fraction from leaf and bark of Triadica sebifera; Note:RIa = Calculated Retention Indices; RIb = Retention Indices from literature
Metabolites
|
Molecular formula
|
Area (%)
|
RIa
|
RIb
|
Mass fragmentation
|
Leaf
|
|
|
|
|
|
1,8-Cineole
|
C10H18O
|
1.61
|
1037
|
103746
|
m/z 154 (M+), 139, 125,108, 84, 81, 69, 43, 41
|
Fenchyl acetate
|
C12H20O2
|
0.70
|
1230
|
122347
|
m/z 154 (M+), 136, 121, 108, 95, 80, 69, 43, 41, 27
|
Neophytadiene
|
C20H38
|
5.87
|
1841
|
183648
|
m/z 137 (M+), 123, 109, 95, 82, 68, 43, 41
|
n-Hexadecanoic acid
|
C16H32O2
|
15.61
|
1961
|
196249
|
m/z 256 (M+), 227, 213, 199, 185, 171, 157, 143, 129, 115, 98, 85, 73, 60, 43, 41
|
cis, cis, cis-7,10,13-Hexadecatrienal
|
C16H26O
|
3.06
|
1992
|
198950
|
m/z 264 (M+), 222, 149, 135, 121, 108, 95, 79, 67, 55, 41
|
Octadecanoic acid, ethyl ester
|
C20H40O2
|
9.85
|
2189
|
218951
|
m/z 312 (M+), 269, 213, 157, 143, 129, 115, 101, 88, 70, 57, 43, 41
|
Bark
|
|
|
|
|
|
Hexanoic acid
|
C6H12O2
|
0.31
|
984
|
98952
|
m/z 87 (M+), 73, 60, 43, 41, 40
|
2-Decenal
|
C10H18O
|
0.23
|
1263
|
126053
|
m/z 121 (M+), 98, 84, 70, 57, 43, 41, 40
|
Ethyl phthalate
|
C12H17O4
|
28.43
|
1587
|
158554
|
m/z 222 (M+), 177, 149, 121, 105, 93, 76, 65, 50
|
1-Octadecene
|
C18H36
|
2.69
|
1803
|
180055
|
m/z 125 (M+), 111, 97, 83, 57, 41, 40
|
Galaxolide
|
C18H26O
|
44.73
|
1834
|
183756
|
m/z 258 (M+), 243, 213, 185, 171, 157, 143, 128
|
Residual toxicity of leaf/bark extracts and seed oil of T. sebifera against A. craccivora
The residual toxicity of LEE, LME, BEE, BME and SO of T. sebifera against aphid with respect toLC50 values are presented in Table 3. Bark extracts are most effective than leaf extracts. Among leaf extracts, LEE 80% was found most effective (LC50= 9590.49 mg L−1) against A. craccivora after 72 h and was followed by LME 100%, 80%, and 50% (LC50=9627, 10800, and 11540 mg L−1, respectively) as compared to LEE 100% (LC50=14100 mg L−1). Similarly, 96 h after treatment also, LEE 80% was found more effective (LC50=6756.42 mg L−1) and was followed by LME 80 and 100% (LC50=7120.27 and 7528.56 mg L−1, respectively) as compared to LME 50% and LEE 100% (LC50=7579.55 and 8702.07 mg L−1, respectively).
Table 3
Efficacy of ethanolic/methanolic leaf/bark aqueous extracts and seed oil of Triadica sebifera against Aphis craccivora; Note: LC50=Lethal concentration to kill 50% of test insect; Mean of three replications; n=150 insects per treatment; LC50 was calculated for fractions showing > 50% mortality using Probit analysis
Leaf extracts
|
LC50
(mg L−1)
|
Confidence limits
(mg L−1)
|
Slope ± SE
|
Chi square
|
P value
|
LEE 100% (72 h)
|
14100.0
|
11764.41 – 17527.10
|
2.31 ± 0.53
|
0.36
|
0.99
|
LEE 100% (96 h)
|
8702.07
|
6880.35 – 10137.00
|
3.08 ± 0.52
|
0.61
|
0.99
|
LEE 80% (72 h)
|
9590.49
|
7706.54 – 11128.43
|
2.91 ± 0.52
|
1.81
|
0.87
|
LEE 80% (96 h)
|
6756.42
|
5342.84 – 7885.95
|
3.97 ± 0.58
|
1.84
|
0.87
|
LEE 50% (72 h)
|
38860.0
|
24892.46 – 341196.32
|
2.07 ± 0.72
|
0.67
|
0.98
|
LEE 50% (96 h)
|
28570.0
|
21383.79 – 71689.83
|
2.51 ± 0.71
|
0.68
|
0.98
|
LME 100% (72 h)
|
9627.0
|
6881.53 – 11725.61
|
2.07 ± 0.49
|
2.02
|
0.85
|
LME 100% (96 h)
|
7528.56
|
5691.95 – 8931.12
|
3.04 ± 0.52
|
5.07
|
0.41
|
LME 80% (72 h)
|
10800.0
|
8603.89 – 12754.79
|
2.45 ± 0.50
|
2.24
|
0.81
|
LME 80% (96 h)
|
7120.27
|
5593.04 – 8326.45
|
3.63 ± 0.55
|
6.95
|
0.22
|
LME 50% (72 h)
|
11540.0
|
9689.45 – 13331.76
|
2.86 ± 0.53
|
3.76
|
0.58
|
LME 50% (96 h)
|
7579.55
|
6239.59 – 8676.68
|
4.15 ± 0.58
|
5.80
|
0.33
|
Seed oil (72 h)
|
2504.59
|
1675.92 – 3562.91
|
1.18 ± 0.21
|
0.86
|
0.97
|
Seed oil (96 h)
|
850.938
|
533.52 – 1171.05
|
1.69 ± 0.25
|
3.28
|
0.66
|
Bark extracts
|
|
|
|
|
|
BEE 100% (72 h)
|
8325.46
|
6958.41 – 9455.76
|
4.05 ± 0.57
|
7.39
|
0.19
|
BEE 100% (96 h)
|
5228.89
|
4038.43 – 6165.80
|
4.88 ± 0.78
|
3.83
|
0.57
|
BEE 80% (72 h)
|
7300.57
|
5889.44 – 8435.86
|
3.97 ± 0.57
|
6.17
|
0.29
|
BEE 80% (96 h)
|
5115.98
|
3613.44 – 6219.77
|
4.04 ± 0.75
|
1.31
|
0.73
|
BEE 50% (72 h)
|
10650.0
|
8244.02 – 12743.89
|
2.25 ± 0.50
|
3.73
|
0.59
|
BEE 50% (96 h)
|
7098.41
|
5159.65 – 8546.32
|
2.91 ± 0.52
|
7.32
|
0.20
|
BME 100% (72 h)
|
8737.64
|
6586.26 – 10377.96
|
2.61 ± 0.50
|
2.12
|
0.83
|
BME 100% (96 h)
|
5701.69
|
4147.42 – 6880.23
|
3.73 ± 0.63
|
2.34
|
0.67
|
BME 80% (72 h)
|
9490.58
|
7504.78 – 11085.89
|
2.78 ± 0.51
|
0.45
|
0.99
|
BME 80% (96 h)
|
5779.72
|
4187.11 – 7007.96
|
3.55 ± 0.58
|
1.42
|
0.92
|
BME 50% (72 h)
|
10580.0
|
8431.88 – 12452.44
|
2.51 ± 0.51
|
1.43
|
0.92
|
BME 50% (96 h)
|
5233.81
|
2941.71 – 6869.58
|
2.51 ± 0.53
|
7.29
|
0.20
|
Azadirachtin (72 h)
|
2642.32
|
2013.70 – 3816.64
|
1.53 ± 0.22
|
0.99
|
0.80
|
Azadirachtin (96 h)
|
1174.22
|
973.61 – 1416.60
|
2.28 ± 0.25
|
5.16
|
0.16
|
Among bark extracts, BEE 80% (LC50=7300.57 mg L−1) was most effective against aphid after 72 h, then BEE and BME 100% (LC50=8325.46 and 8737.64 mg L−1, respectively) as compared to BME 80%, 50% and BEE 50% (LC50=9490.58, 10580 and10650 mg L−1, respectively). Similarly, 96 h after treatment, BEE 80% (LC50=5115.98 mg L−1) was found to be more effective and was followed by BEE 100% (LC50=5228.89 mg L−1) as compared to BME 50%, 100%, 80%, and BEE 50% (LC50=5233.81, 5701.69, 5779.72 and 7098.41 mg L−1, respectively).
The SO of T. sebifera in the current study was found to be more effective after 72 h and 96 h of treatment (LC50=2504.59 and 850.94 mg L−1, respectively) than BEE and LEE (LC50=5115.98-6756.42 mg L−1). All the leaf and bark extracts were not superior to the positive control, Indo-neem (Azadirachtin 0.15% EC) after 72 h and 96 h (LC50= 2642.32 and 1174.22 mg L−1, respectively). However, the SO is more superior to leaf and bark extracts.
Residual toxicity of thecombination of seed oil with leaf/bark extract of T. sebifera and itssynergistic activity against A. craccivora under laboratory
The residual toxicity of thecombination of SO with 80% LEE/BEE and its synergistic activity of T. sebifera against A. craccivoraunderlaboratory conditions were presented in Table S3-S4. Results showed that among combinations studied, a mixture of LEE 80% + SO and BEE 80% {(1+1):2} was found more effective (LC50=239.94 mg L−1) against nymphs of A. craccivora after 72 h and was followed by a mixture of BEE + SO (1:1) (LC50=263.56 mg L−1), SO + LEE at 1:1 ratio (LC50=303.29 mg L−1) and LEE+BEE+SO at 1:1:1 ratio (LC50=337.33 mg L−1) as compared to other mixtures/blends. Based on the fractional effect index (FEI), all the combinations/blends evaluated against A. craccivora showed synergistic interaction after 72 h. Among them, LEE+BEE at 1:3, 3:1, and 1:1 ratio showed the most significant synergistic interaction (FEI= 0.088, 0.092 and 0.122, respectively), as compared to other blends/combinations. Similarly, 96 h after treatment, BEE + SO (1:1 ratio) was found to be more effective (LC50=144.26 mg L−1) againstA. craccivoraand was followed by SO+LEE at 1:1 ratio (LC50=168.9 mg L−1), LEE+SO+BEE at {(1+1):2} ratio (LC50=170.46 mg L−1), LEE +BEE at 1:3 ratio (LC50=179.31 mg L−1) as compared to other mixtures/blends. Based on FEI, all the combinations/blends evaluated against A. craccivora showed a synergistic effect. Among them, LEE+BEE at 1:3, 3:1, and 1:1 ratio showed significant synergistic interaction (FEI= 0.061, 0.070, and 0.100, respectively), as compared to other blends/mixtures.
Residual toxicity of seed oil, leaf/bark extracts and its binary mixtures of T. sebifera and itssynergistic activity against A. craccivoraunder plant growth chamber
Residual toxicity of seed oil, leaf/bark extracts, and its binary mixtures (1:1) of T. sebiferaagainst A. craccivoraunder plant growth chamber conditions are presented in Table S5.Results showed that the binary mixture of SO+LEE (1:1 ratio) was found to be more effective against A. craccivora (LC50=264.05 mg L−1) after 72 h and was followed by SO+BEE (LC50=362.76 mg L−1) as compared to SO, LEE, and BEE (LC50=1100.22, 1840.84 and 5073.99 mg L−1, respectively). Based on FEI values, both the blends (SO+LEE and SO+BEE) showed synergistic interaction (FEI=0.38 and 0.40) against A. craccivora (Table S5). Similarly, 96 h after treatment, binary mixtures of SO+LEE was were more effective against A. craccivora (LC50=223.82 mg L−1) and was followed by SO+BEE (LC50=247.54 mg L−1) as compared to LEE, SO, and BEE (LC50=685.47, 706.53 and 2328.79 mg L−1, respectively). Based on FEI values, SO+BEE showed synergistic interaction (FEI=0.46). A binary mixture of SO+LEE showed additive interaction after 96h of treatment.
Residual toxicity of leaf and bark fractions of T. sebifera against A. craccivora
The residual toxicity of leaf/bark fractions of T. sebifera against A. craccivora was presented in Table 4.Results showed that, among leaf fractions, n-hexane fraction (LC50= 425.73 mg L−1) was more effective than ethyl acetate and n-butanol (LC50=838.89 and 1527.84 mg L−1, respectively) as compared to water fraction (LC50= 2702.82 mg L−1) after 72 h. Similarly, 96 h after treatment, n-hexane fraction (LC50=196.61 mg L−1) was the most promising efficacy than ethyl acetate (LC50=367.75 mg L−1) as compared to water and n-butanol fraction (LC50= 864.68 and 1527.84 mg L−1, respectively).
Table 4
Efficacy of fractions of Triadica sebifera leaf and bark against Aphis craccivora; Note: LC50=Lethal concentration to kill 50% of test insect; Mean of three replications; n=150 insects per treatment; LC50 was calculated for fractions showing > 50% mortality using Probit analysis
Leaf fractions
|
LC50
(mg L−1)
|
Confidence limits (mg L−1)
|
Slope ± SE
|
Chi square
|
P value
|
n-hexane (72 h)
|
425.73
|
196.38 – 679.40
|
1.09 ± 0.21
|
1.55
|
0.82
|
n-hexane (96 h)
|
196.61
|
76.21 – 316.54
|
1.57 ± 0.33
|
2.45
|
0.65
|
Ethyl acetate (72 h)
|
838.89
|
558.64 – 1178.77
|
1.39 ± 0.21
|
3.96
|
0.41
|
Ethyl acetate (96 h)
|
367.75
|
230.65 – 503.57
|
1.88 ± 0.32
|
0.75
|
0.94
|
n-butanol (72 h)
|
1527.84
|
1123.04 – 2093.81
|
1.60 ± 0.22
|
1.14
|
0.89
|
n-butanol (96 h)
|
990.22
|
746.43 – 1294.96
|
1.92 ± 0.25
|
2.68
|
0.61
|
Water (72 h)
|
2702.82
|
1799.12 – 4556.10
|
1.12 ± 0.19
|
1.34
|
0.85
|
Water (96 h)
|
864.68
|
643.83 – 1133.25
|
1.89 ± 0.25
|
2.07
|
0.72
|
Bark fractions
|
|
|
|
|
|
n-hexane (72 h)
|
1659.98
|
1211.70 – 2310.56
|
1.54 ± 0.21
|
3.62
|
0.46
|
n-hexane (96 h)
|
1130.95
|
867.74 – 1467.86
|
2.02 ± 0.26
|
1.95
|
0.75
|
Ethyl acetate (72 h)
|
3629.52
|
2322.33 – 6984.03
|
1.05 ± 0.19
|
3.60
|
0.46
|
Ethyl acetate (96 h)
|
813.45
|
613.57 – 1052.33
|
2.04 ± 0.27
|
0.80
|
0.94
|
n-butanol (72 h)
|
3539.63
|
2343.17 – 6296.58
|
1.15 ± 0.20
|
1.92
|
0.75
|
n-butanol (96 h)
|
1071.81
|
762.69 – 1472.81
|
1.52 ± 0.22
|
5.26
|
0.26
|
Water 72 h)
|
3049.50
|
2087.70 – 4995.85
|
1.24 ± 0.20
|
5.94
|
0.20
|
Water (96 h)
|
915.15
|
684.74 –1198.41
|
1.90 ± 0.25
|
2.55
|
0.63
|
Azadirachtin (72 h)
|
2642.32
|
2013.70 – 3816.64
|
1.53 ± 0.22
|
0.99
|
0.80
|
Azadirachtin (96 h)
|
1174.22
|
973.61 – 1416.60
|
2.28 ± 0.25
|
5.16
|
0.16
|
Among bark fractions, n-hexane fraction (LC50=1659.98 mg L−1) was most promising against A. craccivora after 72 h than water (LC50=3049.50 mg L−1) as compared to n-butanol and ethyl acetate (LC50=3539.63 and 3629.52 mg L−1, respectively). Similarly, 96 h after treatment, ethyl acetate fraction was most effective (LC50=813.45 mg L−1) than water (LC50=915.15 mg L−1) as compared to n-butanol and n-hexane (LC50=1071.81 and 1130.95 mg L−1, respectively). All the leaf and bark fractions are more superior than the positive control, Indo-neem (Azadirachtin 0.15% EC) after 72 and 96h (LC50= 2642.32 and 1174.22 mg L−1, respectively).
Residual toxicity of isolated compounds of T. sebifera against A. craccivora
The experimental results on residual toxicity of the isolated compounds againstA. craccivora with respect to LC50 values are shown in Table 5.Among four compounds isolated from leaf fractions of T. sebifera, gallic acid was most effective (LC50=1303.68 mg L−1) against A. craccivora and was followed by shikimic acid and quercetin-3-O-glucoside (LC50=1725.09-1855.93 mg L−1) as compared to kaempferol-3-O-glucoside (LC50=3762.69 mg L−1). With respect to per cent mortality, gallic acid at 5000 mg L−1 also reported 98% mortality (F4,49=135.79; p<0.0001) and was followed by shikimic acid, quercetin-3-O-glucoside and kaempferol-3-O-glucoside (88, 80 and 58%, respectively) (F4,49=129.71, 106.32 and 61.00;(***P< 0.0001) (Fig. S14). All the tested compounds are not superior to the positive control, Indo-neem (Azadirachtin 0.15% EC) after 72h and 96h (LC50=2642.32 and 1174.22 mg L−1, respectively) except gallic acid after 72 h (LC50= 2339.69 mg L−1).
Table 5
Efficacy of compounds isolated from leaf fractions of Triadica sebifera against Aphis craccivora; Note: LC50=Lethal concentration to kill 50% of test insect; Mean of five replications; n=300 insects per treatment; LC50 was calculated for fractions showing > 50% mortality using Probit analysis
Compounds
|
LC50
(mg L−1)
|
Confidence limits (mg L−1)
|
Slope ± SE
|
Chi square
|
P value
|
Kaempferol-3-O-glucoside (72h)
|
4512.54
|
3455.14 – 6789.22
|
2.02 ± 0.31
|
0.94
|
0.82
|
Kaempferol-3-O-glucoside (96h)
|
3762.69
|
2924.95 – 5398.97
|
1.95 ± 0.28
|
1.36
|
0.72
|
Quercetin-3-Oglucoside (72h)
|
3068.62
|
2462.99 – 4093.71
|
2.09 ± 0.28
|
2.21
|
0.53
|
Quercetin-3-Oglucoside (96h)
|
1855.93
|
1550.48 – 2261.15
|
2.39 ± 0.27
|
2.60
|
0.46
|
Gallic acid (72h)
|
2339.69
|
1928.91 – 2933.01
|
2.24 ± 0.27
|
2.03
|
0.57
|
Gallic acid (96h)
|
1303.68
|
1118.34 – 1520.76
|
3.07 ± 0.32
|
1.44
|
0.70
|
Shikimic acid (72h)
|
2826.31
|
2320.31 – 3606.86
|
2.30 ± 0.29
|
1.68
|
0.64
|
Shikimic acid (96h)
|
1725.09
|
1447.64 – 2080.49
|
2.47 ± 0.27
|
0.23
|
0.97
|
Azadirachtin (72 h)
|
2642.32
|
2013.70 – 3816.64
|
1.53 ± 0.22
|
0.99
|
0.80
|
Azadirachtin (96 h)
|
1174.22
|
973.61 – 1416.60
|
2.28 ± 0.25
|
5.16
|
0.16
|
Detoxification enzyme activities of LEE, BEE and SO against A. craccivora
Detoxifying enzymes (GST and AChE) activities in A. craccivora fed with bean leaf discs treated with different concentrations of LEE, BEE and SO is presented in Fig. 1. Results showed that AChE activity of nymphs of A. craccivora indicated that all the concentrations of LEE, BEE and SO significantly inhibit the AChE activity (F4,14=631.00 to 968.50, p<0.0001) as compared to control. However, LEE, BEE and SO at 2% reported higher inhibition of AChE (0.19±0.00 to 0.75±0.30 mU/mg) and was followed by 1% (0.28±0.20 to 1.45±0.13 mU/mg). Among them, BEE 2% (0.19±0.00 mU/mg) showed higher inhibition of AChE and was followed by SO and LEE (0.42±0.03 and 0.75±0.30 mU/mg, respectively) as compared to lower concentrations (0.25-0.5%) (Fig. 1a).
Similarly for GST assay, all the concentrations of LEE, BEE and SO significantly inhibit the GST activity (F4,14=31.79 to 500.37, p<0.0001) as compared to control. Among them, BEE at 2% (5.37±0.50 mU/mg) showed higher inhibition of GST and was followed by LEE and SO (9.41±1.13 and 35.01±0.63 mU/mg, respectively) as compared to lower concentrations (0.25-0.5%). However, LEE, BEE and SO at 2% reported higher inhibition of GST (5.37±0.50 to 35.01±0.63 mU/mg) and was followed by 1% (7.77±0.11 to 41.26±1.88 mU/mg) (Fig. 1b).