ALLELOPATHIC POTENTIAL OF LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON MUNG BEAN (VIGNA RADIATA L. CV. K-851) SEEDS

ALLELOPATHIC POTENTIAL OF LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON MUNG BEAN (VIGNA RADIATA L. CV. K-851) SEEDS

Uttam Kumar Kanp ¹ and Sourav Kundu ^{* 2}

Department of Botany ¹, Narajole Raj College, Narajole, Paschim Medinipur - 721211, West Bengal, India.

Department of Botany and Forestry ², Vidyasagar University, Midnapore, Paschim Medinipur - 721102, West Bengal, India.

ABSTRACT: Chromolaena and Lantana plants were analyzed to evaluate the existence of allelopathic effect using fully viable seeds of mung bean as the bioassay material. The study shows that pretreatment of mung bean seeds with various concentrations [1:1 and 1:2 (w/v)] of Chromolaena and Lantana leaf leachate for 24 h interval reduced the percentage germination. DNA and RNA contents were significantly reduced with a remarkable increase of amylase activity in seed samples pretreated with leaf leachate of Chromolaena and Lantana. The seedling performance was found to be much poor when the seedlings were raised from seeds which underwent pretreatment with the plant leachates. Evidenced from the lower levels of chlorophyll, DNA and RNA as well as a higher level of amylase activity in leaves of seedlings. Leaf leachate causes the reduction of germinability and influenced activity of amylase enzyme being the important allelopathic indices; it can be concluded that Chromolaena and Lantana can potentially render allelopathic action on mung bean. It further reemphasizes the fact that fast-growing exotic invasive weeds like Chromolaena and Lantana having inhibiting property should be treated as a potential threat to plant diversity in a natural ecosystem.

Allelopathy, Chromolaena, Lantana, Leaf leachate, Mung bean, Nucleic acid

INTRODUCTION: The growth differentiation of plants by other plants could be due to external environmental factors like height, temperature, air water, habitat as same times due to the secretion of chemicals that may act as a growth inhibitor of plants. Here the donor is benefitted, and the receiver is affected. In a recent system of agricultural land and crop management, not only competition but also allelopathy plays as important role in natural and managed maintained ecosystem.

The term allelopathy signifies that interacting or inhibition of growth both crop and weed species, by the release of chemicals from plant parts by leaching, root exudation, volatilization residue decomposition and other processes ¹. These type of chemical signaling or interactions are widely known in different groups of plants ranging from algae, lichens, and crops as well as annual and perennial weeds ^{2, 3, 4}.

There is a huge number of examples that allelochemicals liberated from certain weeds into the soil reduce crop growth and development cause economic loss ⁵. Approximately 6700 species, out of about 3,000,000 species of the flowering plants are classified as weeds in agroecosystems of the whole world ⁶ of these, 76 weed species are specially categorized as “the World’s Worst Weeds” ⁷.

Only 15 species of the crops which supply 90% of the world’s food occupy 75% of the world’s agricultural land ⁸. Most of the food species belong to five families viz., Poaceae, Solanaceae, Convolvulaceae, Euphorbiaceae and Fabaceae, and all of these families also include most of the common weeds. In crop subsystems of the agroecosystem, crop species (often exotic) are selectively cultivated, but weeds grow themselves in crop fields without any nourishment and interact with the economically beneficial crop species in various ways, including a sharp reduction in crop yields. Thus, for economic gain, weed control measures become inevitable, which increase labor requirements and cost for weed control.

Hence, various multiple methods of weed controls, such as cultural, mechanical, and chemical have been developed and employed rapidly. Most of the synthetic herbicides are non-biodegradable, and hence cause, soil and water pollution remains in the environment for a long time by biomagnifications or other means. The development of biodegradable and environment freely herbicides and biological control method need a better understanding of crop-weed interactions in both in-vivo and in-vitro conditions especially concerning chemical interaction (allelopathy), which may help to keep the interference under reasonable control. Complete weed eradication is not possible. Therefore, a sound understanding is needed of the behavior of crop and weed plants growing together in association and sharing the common resources (nutrients and water) in agro ecosystems. These aspects of the crop weed interference viz., allelospoly and allelopathy are considered here with reference restricted mostly for Indian crops and weeds.

Allelopathy is an ecological phenomenon which are normal constituents of the terrestrial plants ^{9, 10} and is very common and frequent. These include, among others, germination behavior and other physio-biochemical responses of test species ^{11, 12, 13, 14, 15, 16}. With considerable evidences adduced during the past few decades demonstrating the presence of inhibitory compounds in a wide variety of plant types and plant parts, the recent upsurge of interest in allelopathy, with major volumes of collected papers, books, journals regularly published ^{17, 18, 19} has established the topic as one of biological importance. In recent times it is focused on establishing research procedures which may improve the importance of evaluations of the allelopathic potential of exotic nonnative weeds Chromolaena odorata and Lantana camara which have become invasive and forms monospecific stands in different ecosystems in West Bengal and surroundings ²⁰. Chromolaena and Lantana are known to be serious weeds in 47 countries owing to its wide adaptability to different environmental conditions and habitats ⁷.

A very little pioneer research has been done on the allelopathic effect of phytotoxicity of Chromolaena and Lantana to other plants. There is a general motion nowadays that invasive plants displace the local and natural endemic biodiversity through their detrimental effects, including allelopathy ^{21, 22}. Allelopathic action of any plants and parts affects germination behavior, seed metabolism and growth performance of target species which in turn may disturb a species from thriving, thus affecting the complete structure in a long time ²³.

Allelopathic effects may be due to the presence of allelochemicals in Chromolaena odorata, like different types of phenolic compounds, alkaloids eupatorine, eupatoredene, odoratamine, etc. In Lantana camara phenolic compounds (protocatechuic acid, gentisic acid, p-hydroxy benzoic acid, vanillic acid, caffeic acid, syringic acids, vanillin, p-coumaric acid, m-coumaric acid, ferulic acid, salicylic acid, o-coumaric acid, t-cinnamic acid, methyl coumarin), triterpenoids (lantadene A, lantadene B, icterogenin reduced lantadene A, reduce lantadene B, lantadene C, oleanolic acid, oleanonic acid, ursolic acid, ursonic acid, 4-epihederagonic acidlantadene D, lancamarome, lantanolic acid, lantic acid, lantanalic acid, betulonioc acid, betulic acid, lantabutulic acid) essential oils (phellandral, β-cymene, α-phell-andrene, dipentene-1- tarpinene, caryophyllene, candinene, cineol, linallool, geraniol, A-terpimeol, citral) and flavonoids (umuhengerin, 5-hydroxy-6, 6, 3, 4, 5-pentametho-xyflavone), biocides (active principles have not been characterized), Juvenile hormones (active primitives have not been characterized), growth hormones (gibberelline GA₃ like substances) are present that may acts as allelochemicals.

They may be interacting with various physiological processes and may hamper several biochemical processes. Therefore, studies were conducted to test this hypothesis, and laboratory experiments have confirmed this. Finally, the experiment concludes and summarises about the allelopathic potency of Chromolaena and Lantana leaf leachates on mung bean seeds that can elucidate a proposal regarding establishment of the standard protocol for biological assessment of the allelopathic potential of some exotic and invasive species which are generally weeds and crops.

MATERIALS AND METHODS: Fresh, mature leaves of Chromolaena odorata (L.) R. M. King & H. Rob. (Asteraceae) and Lantana camara L. (Verbenaceae) [Authentication number, SK-05, and SK-02, CNH/Tech.II/2019/14, Dated: 28/03/2019] were collected from Vidyasagar University campus, Paschim Medinipur, West Bengal, India; were thoroughly sundried for 15 days. The dry leaves (500 g each) samples of the Chromolaena and Lantana were kept separately in 300 ml double distilled water in 1000 ml beaker for 48 h, and the leachate was decanted in a separate beaker. The total volume of the leachate was then made up to 500 ml volume using double distilled water, and this was taken as the 1:1 (w/v) proportion of leaf leachate. From this stock solution, another concentration grade in the proportion of 1:2 (w/v) was prepared using double distilled water. And this was taken as the two gradation leaf leachate solutions.

Five lots of viable mung bean (Vigna radiata L. cv. K-851) seeds of 25g were surface sterilized with 0.1% HgCl₂ solution for 90 seconds. The seed lots were then separately presoaked in the two concentration grade leaf leachates and for control. Data on seed germination percentage, DNA and RNA levels, and activity of amylase enzyme in seeds were tested. Chlorophyll, DNA and RNA contents, as well as amylase activity, were recorded from 10 uniformly growing 30 days old plants raised from each leaf leachate treated seeds. The plants were grown in Vidyasagar University research field for these analyses. The percentage of seed germination can be analyzed from continuous treatment sets. Three groups of 100 fresh seeds (i.e., 300 fresh seeds) were transferred to separate Petri dishes containing filter paper moistened with 10 ml each of leaf leachates and distilled water for control. Germination data were recorded after 120 h of seed soaking following the International Rules of Seed Testing ²⁴.

DNA and RNA levels were analyzed as per the method described by Cherry ²⁵ modified by Choudhuri and Chatterjee ²⁶. Extraction and estimation of the enzyme amylase were done as per the method described by Khan and Faust ²⁷. For the assay of this enzyme, the blank was taken as zero time control. The activity of this enzyme was expressed as [(∆A × Tv) / (t × v)], where ∆A is the absorbance of the sample after incubation minus the absorbance of the zero time control, Tv is the total volume of the filtrate, t is the time (minutes) of incubation with the substrate and v is the volume of the filtrate taken for incubation Fick and Qualset ²⁸. Chlorophyll level was analyzed from the leaves of 30 days old plants of each treatment following the method of Arnon ²⁹. All the data were statistically analyzed after treatment and replication levels, and least significant difference (LSD) values were calculated at 95% confidence limits Panse and Sukhatme ³⁰.

RESULTS:

Effect on Germination Percentage and Changes of Amylase Activity in Mung Bean Seed Kernels Table 1: Data indicates that percentage germination of mung bean seeds was strongly inhibited by continuous treatment with two concentration grades of leaf leachate of Chromolaena and Lantana. The allelopathic effect of Chromolaena leaf leachate was more inhibitory than Lantana, and the data shows that the more concentrated leachate was more injurious. Amylase activity was increased in seed samples irrespective of treatments with two concentration grades leaf leachate of Chromolaena and Lantana.

Effect on Changes of DNA and RNA Levels in Mung Bean Seed Kernels Table 2: Treatments of the mung bean seeds with leaf leachate of all types could alter gross DNA level of the seeds. Here, both leaf leachate of Chromolaena and Lantana significantly decreased the DNA content of seeds which established the allelopathic potentiality of Chromolaena and Lantana. Leaf leachates of Chromolaena and Lantana significantly reduced RNA level in seed kernels.

Effect on Changes of Chlorophyll Level and Amylase Activity in Leaves of Mung Bean Plants Table 3: Allelochemicals remarkably reduced the level of chlorophyll in leaves of mung bean plants raised from seeds pretreated with leaf leachate of all concentration. The activity of the enzyme amylase was found to increase in leaves of mung bean plants. Here, the allelopathic effect of Chromolaena was more than Lantana, and the more concentrated leaf leachates were the best inhibitor.

Effect on Changes of DNA and RNA Levels in Leaves of Mung Bean Plants Table 4: Data reveals that the pretreating agents significantly decreased the DNA content in leaves of the mung bean plant. The same result was in case of RNA level. Here, more concentration grade, i.e. 1:1 was showed more inhibitory effect than control. Both leaf leachate of Chromolaena strongly inhibited the RNA levels of mung bean plants.

TABLE 1: EFFECT OF SEED PRETREATMENT WITH LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON PERCENTAGE GERMINATION AND AMYLASE ACTIVITY OF MUNG BEAN SEEDS

TABLE 2: EFFECT OF SEED PRETREATMENT WITH LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON DNA AND RNA LEVEL IN KERNELS OF MUNG BEAN SEEDS

TABLE 3: EFFECT OF SEED PRETREATMENT WITH LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON CHANGES OF CHLOROPHYLL CONTENT AND AMYLASE ACTIVITY IN LEAVES OF MUNG BEAN PLANTS

TABLE 4: EFFECT OF SEED PRETREATMENT WITH LEAF LEACHATES OF CHROMOLAENA AND LANTANA ON CHANGES OF DNA AND RNA CONTENT IN LEAVES OF MUNG BEAN PLANTS

DISCUSSION: At present, the investigation shows that random treatment of mung bean seeds with leaf leachate of Chromolaena and Lantana sharply reduced seed germinability and also influenced amylase activity Table 1, decreased DNA and RNA levels Table 2.

Germination behavior of seed is considered to be a reliable index of evaluation of allelopathic action ^{31, 32}. Reduced germinability is a very important effect of allelopathic action of plants, and such action is chiefly exerted by several inhibitors of diverse chemical nature ³³.

In this investigation, the leaf leachate induced inhibition of the percentage of seed germination was noted significantly higher than control, and there is clear indications of the allelopathic action of the test materials. The relatively high allelopathic potential of Chromolaena and Lantana were recorded from its stronger germination inhibitory capacity. On the other hand, more concentrated leaf leachate was more injurious than more diluted leachate solutions. More concentrated plant leachate has more inhibitory compounds. Allelopathic action of Chromolaena and Lantana can also be correlated from the present data on the leaf leachate-induced reduction of DNA and RNA levels as well as increased the activity of amylase enzyme. Various inhibitors present in plants having allelopathic property reduced the overall metabolism of plants or plant parts, and they are reported to be strongly impaired ^{34, 35}.

Results, therefore, point out that both leaf leachate of Eupatorium and Lantana possesses some chemicals which efficiently rendered allelopathic action on mung bean seeds. Reduced plant growth and slowed rate of plants establishment are also convincing evidence of allelopathic action ^{36, 37}. The leaf leachates of Chromolaena and Lantana plants show allelopathic potency, which can reduce plant metabolism like chlorophyll Table 3 DNA and RNA Table 4 contents as well as increases the amylase Table 3 activity.

CONCLUSION: The leaves of Chromolaena and Lantana possesses allelochemicals, which efficiently impair allelopathic action on the present bioassay material. It further reemphasizes the fact that fast-growing exotic invasive weeds like Chromolaena and Lantana having growth-suppressing property should be treated as a potential threat to plant biodiversity in a natural ecosystem.

Therefore, the present study calls for the proper management of Chromolaena and Lantana or other invasive weeds showing similar activity. So, it is high time that more investigation should be done regarding the invasion of plants like weed species; Chromolaena odorata and Lantana camara and their irreversible detrimental effect on economically important plants like mung bean plants.

ACKNOWLEDGEMENT: The authors gratefully acknowledge the teacher’s and staff of Vidyasagar University for their constant support and Botanical Survey of India, Central National Herbarium, Howrah for authentication of allelopathic plants of our research work.

CONFLICT OF INTEREST: Nil

REFERENCES:

1. Molisch H: Der Einfluss einer Pflanze auf die andere-Allelopathie. Fischer, Jena 1937.
2. Yadav V, Singh NB, Singh H, Singh A and Hussain I: Allelopathic invasion of alien plant species in India and their management strategies: A Review. Tropical Plant Research 2016; 3(1): 87-01.
3. Putnam AR: Allelopathic Research in Agriculture: Past Highlights and Potential. In: Thompson, A.C., Ed., The Chemistry of Allelopathy: Biochemical Interactions among Plants, American Chemical Society, Washington DC 1985; 268: 1-8.
4. Horsley SB: Allelopathy. In: Avery, M. E., G. R. Cannel and C. K. Ong (Eds.). Biophysical research for Asian agroforestry. Winrock International, Arlington, Virginia; South Asia Books, USA 1991; 167-83.
5. Chauhan PS, Bhisht S and Choudhari N: Allelopathic effects of Lantana camara on Seed Growth behavior of Wheat (Triticum aestivum) and Mustard (Brassica campestris). International Research Journal of Biological Sciences 2016; 5(12): 43-48.
6. Holm L, Pancho JV, Herberger JP and Plucknett DL: A geographical atlas of world weeds. New York: John, Wiley & Sons, 1979.
7. Holm LG, Plucknett DL, Pancho JV and Herberger JP: The World’s Worst Weeds: Distribution and Biology. University of Hawaii, Honolulu, USA 1977; 609.
8. Harlan JR: Crops and Man. American Soc. Agron& Crop Sci. Soc. America, 1975.
9. Rice EL: Allelopathy. Academic Press, Orlando, 2^nd Edition 1984.
10. Datta SC and Singha-Roy SP: Allelopathy and inhibitors. Science and Culture 1974; 40: 47-59.
11. Ved A, Arsi T, Prakash O and Gupta A: A review on phytochemistry and pharmacological activity of Lantana camara International Journal of Pharmaceutical Sciences and Research 2018; 9(1): 37-43.
12. Veraplakorn V: Allelopathic hormesis and slow release of lantana (Lantana camara) callus extract. Agricultural and Natural Resources 2018; 52: 335-40.
13. Veraplakorn V: In-vitro micropropagation and allopathic effect of lantana (Lantana camara). Agriculture and Natural Resources 2017; 51: 478-84.
14. Patil BJ and Khade HN: Allelopathic effect of Parthenium hysterophorus on photosynthetic pigments and biochemical constituents of Vigna aconitifolia L. International Journal of Life Sciences 2017; 5(4): 677-82.
15. Bhakat RK, Bhattacharjee A, Kanp UK and Das RK: Allelopathic influence of Ipomoea pes-caprae (L.) Roxb. on Phaseolus mungo Vidyasagar University, Journal of Biological Sciences 2002; 8: 44-50.
16. Bhattacharjee A, Bhakat RK, Kanp UK and Das RK: An investigation on allelopathic action of Casuarina equsetifolia R. and Ipomoea pes-capre (L.) Roxb. Environment and Ecology 2003; 21(2): 283-89.
17. Sikolia SF and Elizabeth A: Allelopathic effects of saligna on germination growth and development of Vigna unguiculata L. Walp. Journal of Environmental Science, Toxicology and Food Technology 2018; 12(3): 15-24.
18. El-shora HM, El-Gawad AMA, El-shobanky AM and Hala ST: Influence of stramonium leaf extract on anti-oxidants and activities of metabolic enzymes of Trigonella foenum-graecum and L. sativum. International Journal of Current Research and Academic Review 2018; 6(2): 1-11.
19. Chimouriya S, Shrestha I, Piya S, Lamichhane J and Gauchan D: Allelopathic effects of Adhatoda vasica and Eupatorium adenophorum on germination and growth behavior of Capsicum annum. International Journal of Innovative Science and Res Technol 2018; 3(9): 362-68.
20. Mushtaq W and Siddiqui MB: Allelopathy Studies in Weed Science in India-A Review. International Journal of Natural Resource Ecol and Manag 2017; 2(6): 99-03.
21. Cronk QCB and Fuller JL: Plant Invaders. Chapman and Hall, London 1995.
22. Shiva V: Species invasion and the displacement of biological and cultural diversity. In: A. R. Putnam, C. S. Tang (eds.). Invasive Species and Biological Diversity management. Kluwer Academic Publishers, Dordrecht 1999; 33-45.
23. Ghosh KN and Datta SC: A glimpse into the phenomenon of allelopathy. Bullet. Bot. Soci Beng 1989; 43: 13-25.
24. International Seed Testing Association International Rules for seed testing. Seed Sci and Technol 1976; 4: 51-177.
25. Cherry JH: Nucleic acid determination in storage tissues of higher plants. Plant Physiology 1962; 37(5): 670-78.
26. Choudhuri MA and Chatterjee SK: Seasonal changes in the level of some cellular components in the abscission zone of coleus leaves of different ages. Annals of Botany 1970; 34(2): 275-87.
27. Khan AA and Faust MA: Effect of growth retardants on α–amylase production in germinating barley seeds. Physiologia Plantarum 1967; 20(3): 673-81.
28. Fick NG and Qualset CO: Genetic control of endosperm amylase activity and gibberellic acid responses in standard-height and short-statured wheats. Proceedings of National Academy of Sciences 1975; 72(3): 892-95.
29. Arnon DI: Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiology 1949; 24(1): 1.
30. Panse VG and Sukhatme PT: Statistical Methods for Agricultural Workers. 2^nd edition, Indian Council of Agricultural Research, New Delhi, India 1967; 150-157.
31. Raj S and Jha AK: Evaluation of the effect of leaf extract of hysterophorus L. on seed germination, seedling growth and fresh weight of Phaseolus mungo. American Journal of Research Communication 2016; 4(2): 86-03.
32. Nadirsha PSN and Yogamoorthi A: Allelopathic effects of aqueous extract of Lantana camara on seed germination of Black gram Vigna mungo L. Environmental Science: An Indian Journal 2016; 12(11): 122.
33. Maiti P, Bhakat KR, Jha Y and Bhattacharjee A: Allelopathic potential of Hyptis suaveolens on physio-biochemical changes of mung bean seeds. Communications in Plant Sciences 2015; 5(4): 67-75.
34. Datta SC and Chakrabarty SD: Allelopathic potential of Clerodentrum viscosum in relation to germination and seedling growth of weeds. Flora 1982; 172: 85-91.
35. Nayek A: Studies on allelopathic potential of Eucalyptus and Parthenium M.Sc. Dissertation, Vidyasagar University, India 2000.
36. Rusdy M and Alko A: Allelopathic effect of Lantana camara and Chromolaena odorata on germination and seedling growth of pubescens. International Journal of Applied Environmental Sciences 2017; 12(10): 1769-76.
37. Madane AN and Bhimrao JP: Allelopathic effect of Eupatorium odoratum on amylase activity during seed germination of Cicer arietinum L. and Cajanus cajan (L) Millsp. Bioscience Discovery 2017; 8(1): 82-86.
    

    ---

    How to cite this article:

    Kanp UK and Kundu S: Allelopathic potential of leaf leachates of chromolaena and lantana on mung bean (Vigna radiata L. Cv. K-851) seeds. Int J Pharm Sci & Res 2019; 10(9): 4245-50. doi: 10.13040/IJPSR.0975-8232.10(9).4245-50.

    ---

    All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

    ---