Remediation of Mercury Induced Stress on Vigna mungo (L.)Hepper Using Martynia annua L. Leaf Powder as Biosorbent
Received: 04-Aug-2021 / Accepted Date: 18-Aug-2021 / Published Date: 25-Aug-2021 DOI: 10.4172/2155-6199.1000009
Abstract
Heavy metals are a threat to human health and ecosystem. These days, great deal of attention is being given to green technologies for remediation of metal contaminated soil. Biosorption is one among such emerging technologies, which utilizes naturally occurring waste materials to sequester heavy metals from contaminated soil. In this present study the impact of mercury chloride was analysed. Seedlings of Vigna mungo (L.) Hepper were treated with various concentration of mercury chloride such as 5 mM, 10 mM, 15 mM, 20 mM and 25 mM. After 10 days of treatment various biochemical and enzyme characteristics were analysed. Apart from the biochemical such as glucose, protein, amino acid, the activity of nitrate reductase was gradually decreased with increasing concentration of mercury chloride. But the content of proline, leaf nitrate, catalase and peroxidise activity was in reverse. When optimal concentration 15 mM of mercury chloride was treated with various amounts of the leaf powder of a weed plant namely Martynia annua L. viz., 2 gm, 4 gm and 6 gm, and the filtrate was applied on the same plant. The reduced biochemical and enzyme characteristics due to metal toxicity were found improved considerably. From this study, it was inferred that the biosorbent.
Keywords: Biosorption; Vigna mungo; Mercury; Proline; Leaf nitrate; Catalase; Peroxidise
Introduction
Heavy metal pollution of soils is of serious concern as they are persistent, non-biodegradable and become toxic to living organisms [1]. They have a tendency to accumulate in soft tissues of living organisms. The deposition may then show biochemical or physiological changes in them [2]. Extreme values may cause growth inhibition and loss in net production, prominently seen in plants [3]. Growth reduction as a result of changes in physiological and biochemical processes in plants growing on heavy metal polluted soils has been recorded [4]. Heavy metal salts are water-soluble and get dissolved in wastewater, which means they cannot be separated by physical separation methods [5]. Additionally, physico-chemical methods are ineffective or expensive when the concentration of heavy metals is very low. Alternately, biological methods like biosorption and/or bioaccumulation for removal of heavy metals may be an attractive alternative to physico-chemical methods [6]. Use of plants and plant by-product for remediation purposes is thus a possible solution for heavy metal pollution since it includes sustainable remediation technologies to rectify and re-establish the natural condition of soil.
Materials and Methods
The seeds were procured from Tamil Nadu Agricultural University, Coimbatore. The plant Martynia annua L dried leaf powder used as a biadsorbent. The various concentration of mercury chloride (5 mM, 10 mM, 15 mM, 20 mM and 25 mM) were prepared. Both control and experimental plants were allowed to grow in soil mixture (red: black: garden soil) in the ratio of 1:1:1. After 10 days, the seedlings of Vigna mungo (L.) Hepper were treated with heavy metal solution aforesaid. Various biochemical and enzymatic characteristics were analysed on the treated plants. The optimal concentration mercury (15 mm) was mixed with various amounts of Martynia annua L dried leaf powder (2 g/L, 4 g/L and 6 g/L w/v) and kept in shaker for 24 hours. The filtrate was used to treat plants. After 10 days of treatment, the same biochemical and enzymatic characteristics were analyzed as follows: protein [7], glucose [8], amino acid [8], proline [9], in vivo nitrate reductase [10], peroxidise and catalase activity in Tables 1-6 [11].
| Growth Parameters | Control | 2 mm | 4 mm | 6 mm | 8 mm | 10 mm |
|---|---|---|---|---|---|---|
| Root length (cm) | 9.66 ± 0.066 (100) | 9.13 ± 0.088 (91) |
8.7 ± 0.11 (85) |
8.2 ± 0.057 (73) |
7.26 ± 0.073 (66) |
6.46 ± 0.056 (58) |
| Shoot length (cm) | 23.91 ± 0.145 (100) |
19.8 ± 0.152 (92) |
19.16 ± 0.043 (89) |
18.13 ± 0.088 (74) |
17.23 ± 0.145 (62) |
15.16 ± 0.088 (51) |
| Leaf area (cm2) | 7.4 ± 0.152 (100) |
6.32 ± 0.115 (92) |
5.3 ± 0.057 (78) |
4.6 ± 0.152 (57) |
3.16 ± 0.033 (41) |
2.53 ± 0.176 (38) |
| Fresh weight (gm) | 0.98 ± 0.041 (100) |
0.91 ± 0.005 (89) |
0.78 ± 0.017 (71) |
0.69 ± 0.005 (54) |
0.55 ± 0.008 (36) |
0.45 ± 0.009 (21) |
| Dry weight (gm) | 0.73 ± 0.053 (100) |
0.63 ± 0.005 (88) |
0.42 ± 0.057 (65) |
0.26 ± 0.034 (49) |
0.14 ± 0.034 (31) |
0.11 ± 0.066 (28) |
Table 1: Impact of various concentrations of mercuric chloride on the morphometric characteristics of Vigna mungo (L.) Hepper.
| Growth Parameters | Control | 2 mm | 4 mm | 6 mm | 8 mm | 10 mm |
| Chlorophyll .a (mg/gLFW) |
3.34 ± 0.094 (100) |
3.02 ± 0.078 (85) | 2.78 ± 0.092 (71) |
2.57 ± 0.019 (57) |
1.84 ± 0.066 (37) |
1.24 ± 0.026 (25) |
| Chlorophyll .b (mg/gLFW) |
4.84 ± 0.036 (100) |
4.62 ± 0.140 (81) |
3.21 ± 0.208 (73) |
2.94 ± 0.138 (53) |
2.15 ± 0.105 (32) |
2.02 ± 0.089 (21) |
| Total.Chlorophyll (mg/gLFW) |
8.18 ± 0.039 (100) |
7.64 ± 0.137 (85) |
5.89 ± 0.029 (76) |
5.51 ± 0.124 (51) |
3.99 ± 0.022 (36) |
3.26 ± 0.136 (23) |
| Carotenoids (mg/gLFW) |
3.98 ± 0.062 (100) |
3.13 ± 0.227 (88) |
2.76 ± 0.224 (63) |
2.11 ± 0.164 (58) |
1.98 ± 0.028 (43) |
0.98 ± 0.114 (31) |
| Anthocyanin (µg/gLFW) |
3.02 ± 0.129 (100) |
3.73 ± 0.066 (131) |
4.05 ± 0.137 (153) |
5.89 ± 0.131 (182) |
6.47 ± 0.165 (218) |
6.98 ± 0.036 (234) |
Table 2: Impact of various concentrations of mercuric chloride on the pigment contents of Vigna mungo (L) Hepper.
| Parameters | Control | 2 mm | 4 mm | 6 mm | 8 mm | 10 mm |
|---|---|---|---|---|---|---|
| Total soluble sugar (mg/gLFW) |
9.46 ± 0.129 (100) |
8.56 ± 0.081 (88) |
7.62 ± 0.153 (78) |
7.12 ± 0.060 (73) |
4.22 ± 0.164 (43) |
3.32 ± 0.136 (34) |
| Total soluble protein (mg/gLFW) |
5.83 ± 0.043 (100) |
4.64 ± 0.013 (83) |
3.70 ± 0.021 (68) |
3.12 ± 0.057 (53) | 2.32 ± 0.086 (39) |
1.95 ± 0.049 (25) |
| Amino acid (µ mole/gLFW) |
3.32 ± 0.120 (100) |
4.82 ± 0.136 (121) | 5.42 ± 0.46 (146) |
6.94 ± 0.158 (163) |
7.25 ± 0.172 (188) |
8.23 ± 0.123 (204) |
| Proline (µ mole/g LFW) |
3.03 ± 0.064 (100) |
4.57 ± 0.118 (118) |
4.42 ± 0.036 (146) |
6.19 ± 0.037 (168) |
6.92 ± 0.037 (188) |
7.86 ± 0.173 (205) |
| Leaf nitrate (µg/gLFW) |
4.29 ± 0.120 (100) |
5.12 ± 0.138 (114) |
5.71 ± 0.184 (133) |
6.98 ± 0.114 (154) |
7.08 ± 0.228 (178) |
8.30 ± 0.177 (195) |
Table 3: Impact of various concentrations of mercuric chloride on the biochemical characteristics of Vigna mungo (L.) Hepper
| Parameters | Control | 2 mm | 4 mm | 6 mm | 8 mm | 10 mm |
|---|---|---|---|---|---|---|
| Nitrate Reductase (µ mole nitrite formed/g LFW)/hr |
7.68 ± 0.045 (100) |
7.03 ± 0.254 (83) |
6.54 ± 0.173 (74) |
5.46 ± 0.066 (59) |
4.23 ± 0.149 (47) |
3.23 ± 0.103 (38) |
| Catalase activity (µ mole/g LFW)/1 min. |
1.37 ± 0.022 (100) |
1.91 ± 0.024 (140) |
2.55 ± 0.021 (151) | 2.06 ± 0.038 (188) |
3.04 ± 0.058 (224) |
3.73 ± 0.015 (273) |
| Peroxidase activity (µ mole/g LFW)/3min. |
1.48 ± 0.017 (100) |
2.21 ± 0.067 (149) |
3.06 ± 0.053 (206) |
3.69 ± 0.102 (248) |
4.61 ± 0.025 (310) |
5.29 ± 0.103 (356) |
Table 4: Impact of various concentration of mercuric chloride on the enzyme activity characteristics of Vigna mungo (L) Hepper.
| Growth Parameters | Control water | + Mercury 6 Mm |
6 mM mercuric chloride + |
||
|---|---|---|---|---|---|
| 2 gm/100 ml leaf powder | 4 gm/100 ml leaf powder | 6 gm/`100 m leaf powder | |||
| Root length (cm) | 9.66 ± 0.066 (100) |
8.20 ± 0.057 (73) |
7.21 ± 0.145 (78) |
8.01 ± 0.057 (89) |
8.86 ± 0.120 (96) |
| Shoot length (cm) | 23.91 ± 0.145 (100) |
18.13 ± 0.088 (74) |
19.77 ± 0.213 (79) |
20.36 ± 0.202 (84) | 22.54 ± 0.167 (95) |
| Leaf area (cm2) |
7.40 ± 0.152 (100) |
4.6 ± 0.152 (57) |
5.21 ± 0.031 (63) | 6.63 ± 0.173 (81) |
7.14 ± 0.218 (98) |
| Fresh weight (gm) | 0.98 ± 0.041 (100) |
0.69 ± 0.005 (54) |
0.36 ± 0.009 (68) |
0.47 ± 0.085 (73) |
00.81 ± 0.017 (86) |
| Dry weight (gm) | 0.73 ± 0.053 (100) |
0.26 ± 0.034 (49) |
0.20 ± 0.060 (58) |
0.35 ± 0.006 (78) |
0.48 ± 0.028 (83) |
Table 5: Effect of Martynia annua L. leaf powder treated mercuric chloride on the morphometric characteristics of Vigna mungo (L.) Hepper
| Parameters | Control water | + Mercury 6 mM | 6 mM mercuric chloride + |
||
|---|---|---|---|---|---|
| 2 gm/100 ml leaf powder | 4 gm/100 ml leaf powder | 6 gm/`100 m leaf powder | |||
| Chlorophyll a mg/gLFW |
3.34 ± 0.094 (100) |
2.57 ± 0.099 (57) |
2.16 ± 0.012 (78) |
3.40 ± 0.810 (92) |
4.81 ± 0.073 (110) |
| Chlorophyll b mg/gLFW |
5.84 ±0.036 (100) |
2..94 ± 0.138 (53) |
3.44 ± 0.012 (76) |
4.75. ± 0.017 (84) | 5.23 ± 0.472 (108) |
| Total chlorophyll mg/gLFW |
8.18 ± 0.039 (100) |
5.51 ± 0.124 (51) |
5..60 ± 0.219 (84) | 8.15 ± 0.059 (93) |
9.04 ± 0.194 (110) |
| Carotenoids mg/gLFW |
3.98 ± 0.062 (100) |
2.11 ± 0.164 (58) |
2.24 ± 0.081 (67) |
3.23 ± 0.093 (88) |
4.95 ± 0.082 (107) |
| Anthocyanin mg/gLFW |
3.02 ± 0.129 (100) |
5.89 ± 0.131 (182) |
5.43 ± 0.073 (153) |
4.41 ± 0.131 (124) |
3.61 ± 0.163 (105) |
Table 6: Effect of Martynia annua L. leaf powder treated mercuric chloride on the pigment contents of Vigna mungo (L.) Hepper.
Results and Discussion
The results obtained that morphometric characters such as root length, shoot length, fresh weight, dry weight and leaf area were decreased with increasing the concentration of mecury. The photosynthetic pigments of chlorophyll and carotenoids were decreased but the level of anthocyanin content was increased. The total soluble sugar content was decreased with the increase in the concentration of mercury. The mercury has caused a considerable increase in the free amino acid content than the control. The Leaf nitrate, proline, catalase activity and peroxidase activity content was increased with the increasing concentration of mercury chloride.
There was a decrease in protein content and total soluble sugar with increasing concentration of mercury chloride. This result coincides with [12]. Decline in protein content under metal stress can be related to the inhibition of protein synthesis or increase in protein degradation Zn [13]. The free proline has also been shown to protect plants against free radical induced damage by quenching of singlet oxygen [14]. Similar increase in leaf nitrate content, reduction in vivo NR activities with increased in concentration of cadmium chloride on Vigna radiata has been reported earlier [15]. Similar observation was noticed by [16] under the barium treatment in Amaranthus caudatus L. Catalase is another-antioxidant scavenging enzyme. It is also analysed in the present study and was found to increase with the increasing concentration of mercury. Catalase is a special type of peroxidase enzymes, which catalyses the degradation of H2O2, which is a natural metabolite and also toxic to plants [17]. The removal of heavy metals from metal contaminated soil is carried out using biosorbents instead of conventional adsorbents. In the recent years, many low cost biosorbents such as algae, fungi, bacteria and agricultural byproducts have been investigated for their biosorption capacity towards heavy metal removal [18]. Plants applied with bioadsorbent treated metal solution showed increase in protein content, total soluble sugar and activity of nitrate reductase (Tables 7 and 8). In contrary amino acid content, activity of catalase and peroxidise was decreased in Tables 7 and 8.
| Biochemical parameters | Control water | + Mercury (6 mM) |
6 mM mercuric chloride | ||
|---|---|---|---|---|---|
| 2 gm/100 ml leaf powder |
4 gm/100 ml leaf powder |
6 gm/`100 m leaf powder |
|||
| Total soluble sugar (mg/g LFW) |
9.46 ± 0.129 (100) |
7.12 ± 0.060 (73) |
7.62 ± 0.057 (74) |
8.54 ± 0.031 (96) |
9.84. ± 0.026 (110) |
| Total soluble protein (mg/g LFW) |
5.83 ± 0.043 (100) |
3.12 ± 0.057 (53) |
3.93. ± 0.070 (81) |
5.24 ± 0.045 (93) | 6.30 ± 0.053 (112) |
| Amino acid (µ mole/g LFW) |
3.23 ± 0.120 (100) |
6.94 ± 0.158 (163) |
6.34. ± 0.05 (157) | 4.75 ± 0.046 (133) |
3.98 ± 0.58 (106) |
| Proline (µ mole/g LFW) |
3.32 ± 0.064 (100) |
6.19 ± 0.013 (188) |
5.14 ± 0.024 (163) |
4.63 ± 0.031 (133) |
3.68 ± 0.187 (109) |
| Leaf nitrate (µg/gLFW) |
4.29 ± 0.120 (100) |
6.98 ± 0.114 (204) |
7.34.± 0.092 (179) |
6.34 ± 0.137 (135) |
4.82 ± 0.076 (112) |
Table 7: Effect of Martynia annua L. leaf powder treated mercuric chloride on the biochemical characteristics of Vigna mungo (L.) Hepper.
| Enzymatic parameters | Control water | + Mercury (6 mM) | 6 mM mercuric chloride | ||
|---|---|---|---|---|---|
| 2 gm/100 ml leaf powder | 4 gm/100 ml leaf powder | 6 gm/`100 m leaf powder | |||
| Nitrate reductase µmole/gLFW|hr | 7.68 ± 0.045 (100) |
5.46 ± 0.066 (59) |
3.63 ± 0.041 (74) |
6.71 ± 0.063 (86) |
7.93 ± 0.114 (104) |
| Catalase activity µmole/gLFW1min. |
1.37 ± 0.115 (100) |
2.06 ± 0.038 (188) |
3.01 ± 0.093 (172) |
2.63 ± 0.081 (148) | 1.96 ± 0.023 (114) |
| Peroxidase activity µmole/gLFW|3 min |
1.48 ± 0.017 (100) |
3.69 ± 0.102 (248) |
2.73 ± 0.086 (176) | 2.06 ± 0.028 (136) |
1.74 ± 0.036 (112) |
Table 8: Effect of Martynia annua L. leaf powder treated mercuric chloride on the enzymatic characteristics of Vigna mungo (L.) Hepper.
Conclusion
From the present investigation, it is confirmed that the plant Martynia annua L dried leaf powder is nullifying the toxicity of heavy metal. Since it restores the suppressed biochemicals and enzyme activity due to metal toxicity.
Acknowledgment
Authors thank the Principal and Management of Ayya Nadar Janaki Ammal College, Sivakasi for providing facilities.
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Citation: Marisamy K, Saratha P, Ramasubramanian V (2021) Remediation of Mercuric Chloride Induced Stress on Vigna mungo (L.) Hepper using Martynia annua L. Leaf Powder as Biosorbent. J Bioremediat Biodegrad 12: 009. DOI: 10.4172/2155-6199.1000009
Copyright: © 2021 Marisamy K, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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