Subscribe Now Subscribe Today
Research Article
 

Herbicidal Effects of Marine Animal, Trochus tentorium from Gulf of Mannar, Southeastern India



C. Chellaram, T. Prem Anand, N.R. Shailaja and D. Kesavan
 
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail
ABSTRACT

The plants, animals and microorganisms of the marine environment with their wide range of chemical diversity are still an unexplored resource for the development of new agro-chemical agents. Works related to the herbicidal activity of the marine organisms are too scanty. Compared to the search for new pharmaceutical compounds, very little effort has been devoted to the exploration of agrochemical compounds from marine natural products. In the present study, the herbicidal activity of the crude extracts and partial purified fractions of Trochus tentorium was assayed using the duckweed, Lemna minor L. following a bench top bioassay. The crude acetone extract of T. tentorium decayed the fronds of Lemna plants at the concentration of 1000 mg mL-1 on the 4th day, while the results of same concentration of ethyl acetate, dichloromethane and methanol extracts showed decay of the plants on the 5th day of the experiment. This study reveals that the column-purified acetone fraction of the gastropod was able to decay L. minor to a better degree in comparison with the crude extracts. Hence, from the present study, it was noted that not only the 100% acetone fractions but also the 20:80 hexane: methanol and 80:20 acetone: methanol fractions showed a higher degree of activity against L. minor.

Services
Related Articles in ASCI
Search in Google Scholar
View Citation
Report Citation

 
  How to cite this article:

C. Chellaram, T. Prem Anand, N.R. Shailaja and D. Kesavan, 2012. Herbicidal Effects of Marine Animal, Trochus tentorium from Gulf of Mannar, Southeastern India. Asian Journal of Animal and Veterinary Advances, 7: 250-255.

DOI: 10.3923/ajava.2012.250.255

URL: https://scialert.net/abstract/?doi=ajava.2012.250.255
 
Received: October 31, 2011; Accepted: December 29, 2011; Published: January 14, 2012



INTRODUCTION

An ideal herbicide should have potent activity against weeds, minimum toxicity against living things other than plants, high selectivity between crop plants and weeds and cause no damage to the environment by residual material. The weeds often form a green blanket on the water surface and highly reduce light penetration. Among the aquatic weeds, the duckweed, Lemna minor L. is considered as one of the important weeds. Thick populations of L. minor are often found in association with other aquatic plants, particularly with those that provide protection from wind and wave action (Vernon and Vandiver, 2005). Hence, the usage of herbicides for weed control against this weed has thus become apparent. Herbicides for practical use today are mostly synthetic compounds which inhibit the photosynthetic electron transfer and affect plant hormonal action. However, sooner or later weeds acquire resistance to the existing herbicides and thus the continued development of new and potent drugs to control the weeds is always required. Hence, the development of herbicides must always take into consideration the problem of environmental pollution (Okuda, 1992).

Compared to the search for new pharmaceutical compounds, very little effort has been devoted to the exploration of agrochemical compounds from marine natural products (Fenical, 1993). One of the major impediments to successful herbicide development is obtaining compounds capable of penetrating the cell membrane of target plant (weed) (Lyewellyn and Burnell, 2000). The plant, animals and microorganisms of the marine environment with their wide range of chemical diversity are still an unexplored resource for the development of new agro-chemical agents (Crombie, 1990). In the hunt for new agro-chemical agents, the plants, animals and microorganisms of the marine environment with their wide range of chemical diversity prove to be an unexplored resource. Works related to the herbicidal activity of the marine organisms are too scanty. However, there is relatively limited number of reports on the herbicidal activity of some marine natural products (Burnell et al., 2000; Wagner et al., 2002; Hylleberg and Kilburn, 2002; Omura et al., 1990) and to date, research focused on isolating herbicidal prototype leads from marine origin has resulted in the report of about 40 active compounds. Many important herbicides are isolated from marine algae (Einhellig et al., 1985). Of the marine organisms, in particular, the molluscs have always proven to be potential sources of marine natural products with vast array of diversified bio-activities. There are massive evidences to witness marine molluscs as potential supply of potent metabolites (Ganambal et al., 2005; Hocklowski and Faulkner, 1983; Hubert et al., 1996). In the present study, the crude as well as partial purified extracts of the gastropod, Trochus tentorium associated to corals was screened for herbicidal activity against Lemna minor (duckweed).

MATERIALS AND METHODS

Extraction of mollusc: The Gastropod, Trochus tentorium were collected by hand picking using SCUBA diving from the intertidal area at a depth of 3-5 m in Tuticorin coastal waters (Lat 8°45 and Long 78°13’E) of Gulf of Mannar, southeast coast of India. They were immediately brought to the laboratory and identified up to species level using standard keys (Hubert et al., 1996; Riguera, 1997). The identified shells of Trochus tentorium were broke open by a hammer to remove the soft parts. The whole body of the samples (20 g) were cut in to small pieces and air-dried for 24 h at room temperature before extraction with solvents. Then the tissues were rinsed with sterile distilled water and the tissue samples were used for extraction using different solvents such as ethyl acetate, dichloromethane, acetone and methanol. The extracts were cold steeped overnight at -18°C and filtered with Whatman No. 1 filter paper. The filtrate was poured in previously weighed Petri plate and evaporated to dryness in rotary evaporator (Becerro et al., 1994; Riguera, 1997; Wright, 1998). The dried crude extracts were used for herbicidal activity.

Column purification of the active crude extracts: Partial purification of the crude extract of T. tentorium was carried out by the method of Chellaram et al. (2009). After initial screening, the higher activity was shown by acetone extract and it was fractionated by normal phase silica gel column chromatography by employing a step gradient solvent system from low to high polarity. Sequence of 100% hexane, 20% acetone: 80% hexane, 40% acetone: 60% hexane, 60% acetone: 40% hexane, 80% acetone: 20% hexane, 100% acetone, 80% acetone: 20% methanol, 60% acetone: 40% methanol, 40% acetone: 60% methanol, 20% acetone: 80% methanol and 100% methanol was used for elution. Each fraction thus obtained was once again evaporated, concentrated and assayed for herbicidal activity.

Features of duckweed Lemna minor (Arales: Lemnaceae): Lemna minor L. (duckweed) is a miniature aquatic monocot, Lemna plant, which consists of a central frond or mother frond with two attached daughter fronds and a filamentous root. Under normal conditions, the plant reproduces exponentially with budding of daughter fronds from pouches on the sides of the mother fronds. The sizes of the fronds generally range from 1.7 to 4.0 mm in length and from 0.80 to 4.0 mm in width. Fronds occur solitary or in groups of two or more. There is a single root extending from the base of each frond. Seen from a distance, it is difficult to distinguish individual plants in a body of water infested with duckweed because the surface of the water appears green. Earlier reports by Einhellig et al. (1985) provided the general guidelines for developing a Lemna bioassay to screen a large number of plant extracts and chemical substances for their effects on plant growth.

Herbicidal assay: The herbicidal activity of the crude extracts of T. tentorium was assayed against the duckweed, Lemna minor following a bench top bioassay described by McLaughlin et al. (1991). To assay the herbicidal activity, 20 mg of the dried crude extracts (Trochus tentorium) was dissolved in 20 mL of their respective solvents (ethyl acetate, dichloromethane, acetone and methanol). From this solution, 1000, 500, 100, 50 and 10 μL were pipetted into dram vials corresponding to 1000, 500, 100, 50 and 10 μg mL-1, respectively in triplicates. Controls were added with appropriate solvents alone without the extracts. The solvents of the control and test vials were allowed to evaporate overnight and 2 mL of E-medium (consisting of KH2PO4 -680 mg, KNO3 -1515 mg, Ca (NO3)2.4H2O-1180 mg, MgSO4.7H2O -492 mg, H3BO3 -286 mg, MnCl2.4H2O -3.62 mg, FeCl2.6H2O -5.40 mg, ZnSO4.7H2O -0.22 mg, CuSO4.5H2O -0.22 mg, Na2MoO4.2H2O -0.12 mg and EDTA -11.2 mg in one liter of distilled water) was added into each vial. A single healthy L. minor plant containing a rosette of three fronds was introduced into each vial and were placed in glass chamber with about 2-4 cm water at the bottom to maintain the moisture content of the chamber and sealed with glass plate. The fronds per vial were counted daily up to 6 days and symptoms of damage to the frond such as yellowing and decaying were also noted.

The column purified fractions were assayed for herbicidal activity and the bioassay was performed to find out which of the fractions were found to be more active in decaying the plant. The herbicidal activity of crude and column fractions of T. tentorium was tested at different concentrations 500, 300, 200, 100, 50, 25 and 10 mg and changes in the appearance of the fronds were noted.

RESULTS

The present study reveals that out of the different solvents, the acetone extract of T. tentorium decayed the fronds of Lemna plants at the concentration of 250 mg mL-1 on 6th day and ethyl acetate, dichloromethane and methanol extracts decayed the plants on 5th day of experiment at a concentration of 1000 mg mL-1 (Table 1). The acetone crude extracts of the gastropod, shown prominent herbicidal activities against L. minor. The column purified (100% acetone, 20:80 hexane: methanol and 80:20 acetone: methanol) fractions of T. tentorium shown complete decay of the Lemna plants (Table 2), however, 100% acetone fraction of the extract showed activity at a concentration of 100 mg mL-1 on the 5th day of the experiments. But the plants were found to be healthy till the 3rd day of experiment at a concentration of 10 mg mL-1. The plants were decayed by all the fractions at a concentration of 500 mg mL-1 on the 4th day of the experiments. This study reveals that the column-purified acetone fractions of gastropod were able to decay L. minor to a better degree in comparison with the crude extracts. Hence from the present study, it was noted that not only the 100% acetone fractions but also the 20:80 hexane: methanol and 80:20 acetone: methanol fractions shown a higher degree of activity against L. minor.

Table 1: Herbicidal activity of the crude extracts of T. tentorium against the Lemna minor
Image for - Herbicidal Effects of Marine Animal, Trochus tentorium from Gulf of Mannar, Southeastern India
Note: A- Acetone; EA- Ethyl acetate; DM- Dichloromethane and M- Methanol. GH- Green and healthy; G- Green; LY- Light yellow; Y- Yellow and D- Dead

Table 2: Herbicidal activity of the 100% acetone column fractions of T. tentorium against the Lemna minor
Image for - Herbicidal Effects of Marine Animal, Trochus tentorium from Gulf of Mannar, Southeastern India
Note: A- Acetone, H- Hexane and M- Methanol. GH- Green and healthy; G- Green; H- Healthy; LY- Light yellow; Y- Yellow and D- Dead

DISCUSSION

The Lemna assay data agreed with antitumour activities. McLaughlin et al. (1991) have reported that using Lemna assay, the search for biodegradable herbicides may be extended to include natural compounds and this is a simple screen for such activity. Reports are available on the herbicidal activities of 9 purified algal metabolites (Fenical, 1997). Lyewellyn and Burnell (2000) had stated that the number of free growing plants is low on coral reefs; therefore, the chemicals produced by these coral reefs can be widely developed as herbicides. Peng et al. (2003) identified 18 structurally diverse marine-derived compounds and examined for herbicidal activities and it was concluded that several new classes of compounds have been shown to be herbicidal in nature. In the present study, the acetone extracts of Trochus tentorium were found to be efficient in decaying the fronds of Lemna minor at a concentration of 1000 mg mL-1 on the 4th day of the experiment. Chellaram et al. (2004) observed that different solvents like ethyl acetate, acetone and dichloromethane were used for the extraction of the winged oyster, Pteria chinensis, however, the acetone extract was able to cause decaying of the fronds of Lemna minor at 5th day at a concentration of 1000 mg mL-1 which is same concentration to the present study and so the extracts of T. tentorium are assumed to be potent producers of novel metabolites. Anand (2002) isolated bacterial strains from the surface of sponges, seaweeds, crabs, ascidians and cephalopod eggs and reported that they were able to bring forth 90% inhibition against L. minor.

In conclusion, the results indicate that the crude and column fractions (100% acetone, 20:80 methanol: acetone and 80:20 acetone: hexane) of T. tentorium screened for herbicidal activity and extracts of 100% acetone may possess some biologically active compounds. The activity of the column purified fractions was found to be prominent in comparison to the crude counterparts. So the partially purified fraction of the 100% acetone T. tentorium undeniably provides a tool for the development of novel herbicides.

ACKNOWLEDGMENT

Authors thank sincerely the Director, Suganthi Devadason Marine Research Institute, Tuticorin, for providing laboratory facilities and diving kits for sample collection. We are also grateful to Prof. Chancellor Dr. R. Rangarajan, Vel Tech Dr. RR and Dr. SR Technical University and Director and Principal, Vel Tech Multi Tech Dr. Rangarajan Dr. Sakunthala Rangarajan Engineering College, for their unremitting encouragement and valuable advices.

REFERENCES

1:  Anand, P.T., 2002. Screening for bioactive metabolites producing marine bacteria from Tuticorin coastal waters, Southeast coast of India. Ph.D. Thesis, Manonmaniam Sundaranar University, Tirunelveli.

2:  Becerro, M.A., N.I. Lopez, X. Turon and M.J. Uniz, 1994. Antimicrobial activity and surface bacterial film in marine sponges. J. Exp. Mar. Biol. Ecol., 179: 195-205.
CrossRef  |  Direct Link  |  

3:  Burnell, J.N., J.R. Doyle, J.R. Woodward and L.E. Llewellyn, 2000. Rapid throughput biomolecular screening for marine derived C4 plant specific herbicides. Proceedings of 5th IBMC Conference, September 29-October 4, 2000, Townsville, Australia, pp: 24-

4:  Chellaram, C., K.M.E. Gnanambal and J.K.P. Edward, 2004. Antibacterial activity of the winged oyster Pteria chinensis (Pterioida: Pteridae). Indian J. Mar. Sci., 33: 369-372.
Direct Link  |  

5:  Chellaram, C., R.S. Sreenivasan, S. Jonesh, T.P. Anand and J.K.P. Edward, 2009. In vitro antibiotic bustle of coral reef associated gastropod, Drupa margariticola (Broderip, 1832) of tuticorin coastal waters, Southeastern India. Biotechnology, 8: 456-461.
CrossRef  |  Direct Link  |  

6:  Crombie. L., 1990. Natural Models for the Design of Insect Control Compounds: The Mammerins. In: Recent Advances in the Chemistry of Insect Control II, Crombie, L. (Ed.). The Royal Society of Chemistry Special Publication, Cambridge, UK., pp: 23-51

7:  Wagner, D.I., W. Beil, S. Lang, M. Meiners and H. Latch, 2002. Integrated approach to explore the potential of marine microorganisms for the production of bioactive metabolites. Adv. Biochem. Eng. Biotechnol., 74: 207-238.

8:  Einhellig, F.A., G.R. Leather and S.L. Hobbs, 1985. Use of Lemna minor L. as a bioassay in allelopathy. J. Chem. Ecol., 11: 65-65.
Direct Link  |  

9:  Fenical, W., 1993. Chemical studies of marine bacteria: Developing a new resource. Chem. Rev., 93: 1673-1683.
CrossRef  |  Direct Link  |  

10:  Fenical, W., 1997. New pharmaceuticals from marine organisms. Trend. Biotechnol., 15: 339-341.
PubMed  |  

11:  Ganambal, M.E., C. Chellaram and J. Patterson, 2005. Antibacterial activity of whole body extracts of Trochus radiatus (Mollusca: Gastropoda). SDMRI Res. Publ., 9: 182-186.

12:  Hocklowski, J.E. and D.J. Faulkner, 1983. Antibiotics from marine pulmonates, Siphonaria diemenensis. Tetrahedron Lett., 24: 1917-1920.
CrossRef  |  

13:  Hubert, F., V.D.W. Knaap, T. Noel and P. Roch, 1996. Cytonic and antibacterial properties of Mytilus galloprovincialis, Ostrea edulis and Crassostrea gigas (bivalve, molluscs) haemolymph. Aquat. Living Res., 9: 115-124.

14:  Hylleberg, J. and R.N. Kilburn, 2002. Annotated Inventory of Molluscs from the Gulf of Mannar and Vicinity. In: Zoogeography and Inventory of Marine Molluscs Encountered in Southern India, Hylleberg, J. and A. Nateewathana (Eds.). PMBC, Thailand, pp: 1970

15:  Lyewellyn, L.E. and J.N. Burnell, 2000. Marine organisms as sources of C4-Weed-Specific herbicides. Pestic. Outlook, 11: 64-67.
CrossRef  |  Direct Link  |  

16:  McLaughlin, J.L., C.J. Chang and D.L. Smith, 1991. Bench-Top Bioassays for the Discovery of Bioactive Natural Products: An Update. In: Studies in Natural Products Chemistry, Rhaman, A.U. (Ed.). Elsevier, Oxford, pp: 383-409

17:  Okuda, S., 1992. Herbicides: The Search for Bioactive Compounds from Marine Microorganisms. Springer-Verlag, New York, pp: 224-236

18:  Omura, S., Y. Tanaka, I. Kanaya, M. Shinose and Y. Takahashi, 1990. Pthoxazolin. A specific inhibitor of cellulose biosynthesis produced by a strain of Streptomyces sp. J. Antibiot., 43: 1034-1036.
PubMed  |  

19:  Peng, J., X. Shen, K.A. El-Sayed, D.C. Dunbar and T.L. Perry et al., 2003. Marine natural products as prototype agrochemical agents. J. Agri. Food. Chem., 51: 2246-2253.
CrossRef  |  PubMed  |  

20:  Riguera, R., 1997. Isolating bioactive compound from marine organisms. J. Mar. Biotechnol., 5: 187-193.
Direct Link  |  

21:  Vernon. V. and J. Vandiver, 2005. Biology and control of duckweed with herbicides. http://taylor.ifas.ufl.edu/docs/Marine_NatRes/Aquaculture/BiologyControlDuckweedWithHerbicide.pdf

22:  Wright, A.E., 1998. Isolation of Marine Natural Products. In: Methods in Biotechnology, Natural Products Isolation, Cannell, R.P.J. (Ed.). Humana Press Inc., New Jersey, ISBN: 978-0-89603-362-7, pp: 305-408

©  2021 Science Alert. All Rights Reserved