Allelopathic effect of Tectona grandis leaves on protein content changes of black gram and green gram

Transcription

1 INT J CURR SCI 2012, 4: RESEARCH ARTICLE ISSN Allelopathic effect of Tectona grandis leaves on protein content changes of black gram and green gram Manimegalai A Department of Plant Biology and Biotechnology, Arignar Anna Government Arts and Science College for Women Walajapet , Tamil Nadu, India *Corresponding author: amegala2007@gmail.com; Mobile: Abstract Allelopathic effect of leaves of Tectona grandis on protein content were investigated in the seedlings of Vigna mungo L. (Black gram) and Vigna radiata L. (Green gram). The fallen dried leaves of Tectona grandis were collected from coastal area of Cuddalore District. Leaves were again dried and grind with the help of powder wiley machine.the pulses seeds of Vigna mungo L. and Vigna radiata L. were procured from Tamil Nadu Rice Research Station, Aduthurai, Tamil Nadu. Seeds with uniform size,colour and weight were selected.the fifty seeds were sterilized and placed in petridishes containing different concentrations of teak leaf extract (0, 5, 10, 25, 50, 75 and 100%) and petridish containing seeds were irrigated with distilled water and maintained as control were kept under diffused light at room temperature (28±2 C). From the results, the protein content increased in 5% leaf extract treatment over control and a very low protein content was observed in 100% treated seedlings. Among the different concentrations 5% extract alone showed a promotory effect on protein than control while the other concentrations exhibited a inhibitory effect both in black gram and green gram seedlings. Keywords: allelochemicals, allelopathic effect, protein content, Tectona grandis, vignamungo, Vigna radiate Received: 19 th March; Revised: 16 th April; Accepted: 10 th May; IJCS New Liberty Group 2012 Introduction Allelopathy refers to the determental effects of higher plants of one species (donor) on the germination, growth or development of plants of germination and growth inhibition have been identified from plant tissues and soil, which are termed allelochemics or allelochemicals (Whittaker and Feney, 1971). Generally allelochemicals caused another species (recipient) (Narwal, 1994). a decreased in the content of total proteins on test Allelopathy is a phenomenon in which the plant through leaching, volatility, the remnant body decomposition and the root system secretion releases the chemical substance to the environment that has harmful or advantageous function to own or periphery plants including microorganism indirectly (Peng and Shao, 2001; Chai and Hang, 2003; Wang and Tang, 2003; Lv and Duan, 2005). Allelopathy includes both positive and negative effects of one plant on the other, most of the studies seem to focus only on it s deleterious impacts alone and place a key role both in natural and managed ecosystem. Allelopathy has been shown to play a crucial role in crops (Kolesnichenko and Aleikina, 1976; Bansal, 1997). Tectona grandis is a large, deciduous tree up to 40 m (131 ft) tall with gray to grayish brown branchlets. Leaves are ovate-elliptic to ovate, cm ( in) long by 8-23 cm ( in) wide, and are held on robust petioles that are 2-4 cm ( in) long. Leaf margins are entire. During the dry season the tree is leafless; in hot localities the leaves fall in January, but in moist places the tree remains green until March. At the end of the dry season, when the first monsoon rains fall, the new foliage emerges. The allelopathic effect of extracts of teak forests, influencing the composition of the leaves has been tested on Solanaceae species such vegetation growth, and also provides an explanation for the patterns of forest regeneration. Most of the as the Tomato (Lycopersicum esculentum), egg plant (Solanum melongena) and pepper (Capsicum phytotoxic substances suspected of causing annum) (Krishna et al., 2003). The extracts

2 significantly inhibited germination and growth of these plant species. Tectona grandis has also shown high allelopathic activity on wheat (Triticum aestivum) (Krishna et al., 2003). Interest in allelopathic studies in agroforestrys. In laboratory it was aimed to systems is growing, because knowledge of these interactions could provide powerful tools for the integrated management of plagues and a better exploitation of natural resources anywhere in the world. Study the allelopathic effect of teak (Tectona grandis linn. f) leaves on the germination and seedling growth of black gram and green gram. Materials and Methods Collection of Plant materials The seeds of Vigna mungo L. (Black gram) var. ADT-3 and Vigna radiata L. (Green gram) var Co-3 were procured from Tamil Nadu Rice Research station, Aduthurai and were stored in metal tins as suggested by Rao (1976). The dried teak leaf was collected from the coastal area of cuddalore district, dried again and powered 50 g leaf powder was soaked in 500 ml of distilled water and filtered after 24 h with the help of what mann No. 1 filter paper. Using this stock solution, various concentrations of extract (5, 10, 25, 50, 75 and 100%) were prepared by using distilled water. Seeds were sterilized for two minutes in 0.2% mercuric chloride solution, then thoroughly washed under tap water. Fifty seeds were placed in petridishes containing various concentrations leaf extract, kept under diffused light at room temperature (28 ±2 º C). While seeds with distilled water were maintained as control separately. 0.5 g of fresh root, stem and leaves from fifteenth day seedling in each concentration were used for the estimation of protein using lowry et al. (1951). Preparation of extract The 50 g teak leaf powder was soaked in 500 ml of distilled water. They were filtered with the help of Whatmann No. 1 filter paper and pure extracts of leaf were obtained after 24 hours. Using this stock solutions, various concentration of extract (5, 10, 25, 50, 75 and 100%) were prepared by using distilled water. Seeds were sterilized for two minutes in 0.2% mercuric chloride solutions then thoroughly washed under tap water. Fifty seeds were placed in well sterilized petridishes containing various concentrations of leaf extract kept under diffused light at room temperature ( C). While seeds with distilled water were maintained under control separately. Fresh root, stem and leaves from fifteeth day seedling in each concentration were used for the estimation of protein by using Lowry et al. (1951). Extraction Plant material was weighted for 0.5 g and macerated with the help of pestle and mortar using 10 ml of 20% trichloro acetic acid (TCA). The homogenate was centrifuged for 15 min at 600 rpm. The supernatant was discarded. To the pellet 5 ml of 0.01 N NaOH was added and centrifuged. The extract was used for the estimation of protein. Estimation The extract taken for 0.5 ml in a 10 ml test tube and 5 ml of reagent C was added. The solution was mixed well, kept in dark for 10 mts. Later 0.5 ml of folin phenol was added and kept in dark for 30 min. The sample was read at 660 nm in a spectronic - 0. Bovine s serum was used as a standard. Preparation of reagents 1. Two per cent sodium carbonate in 0.1n NaoH. Reagent A 400 mg of sodium hydroxide was dissolved in distilled water and made upto 100 ml. To this solution 2 g of sodium carbonate was added. 2. One per cent copper sulphate in 2% sodium potassium tartarate. Reagent B 2 g of sodium potassium tartarate was dissolved in distilled water and made upto 100 ml. To this solution 1 g of copper sulphate was added. Reagent C 50 ml of reagent A and 1 ml of reagent B were mixed freshly at the time of experiment. 3. Folin phenol reagent 1 ml of folin phenol reagent + 1ml of distilled water. 4. Twenty per cent trichloro acetic acid (TCA)

3 20 g of trichloro acetic acid was dissolved in distilled water and made upto 100 ml N sodium hydroxide Four hundred mg of NaoH was dissolved in distilled water and made upto 100 ml. Results and Discussion Inhibition of protein content increment in black gram and green gram was caused by the aqueous leaf extract of Tectona grandis. Inhibitory effects of both black gram and green gram seedlings are presented in tables 1, 2 and 3. The uptake of active compounds by plants of another species may induce more or less distinct inhibitory effects during the germination and development. It depends on the compounds involved, their concentration and stability in the given environment and plant response to the compounds or their microbial byproducts. Table 1. The effect of aqueous leaf extract of Tectona grandis on protein content in leaves of black gram and green gram (µg/g) (the values are mean ± SE of 7 samples) Conc.of Black gram Green gram Extract % Leaf Cont. 291± ± ± ± ± ± ± ± ± ± ± ± ± ± 8.40 Table 2. The effect of aqueous leaf extract of Tectona grandis on protein content of stem in black gram and green gram (µg/g) (the values are mean ± SE of 7 samples) Conc. of Black gram Green gram Extract Stem % Control 278± ± ± ± ± ± ± ± ± ± ± ± ± ± 8.15 From these results, aqueous leaf extract of Tectona grandis effect the protein content of black gram and green gram. Allelochemicals decreased the amount of protein in wheat seedlings. Impairment of various metabolic activities under the influence of leachate inhibited the protein synthesis and/or stimulated the degradation (Mersie et al., 1993). Normal ways of protein synthesis is inhibited in lettuce seedlings (Lactuca sativa) when treated with cinnamic acid (Einhellig, 1996). Padhy et al. (2000) have reported that the leaf litter leachate of Eucalyptus globulus decreased the protein content in both root and shoot of finger millet. These findings strongly support the present observation. Kolesnichenko and Aleikina (1976) showed that the chemical compounds secreted from the roots of Fraxinus excelsior inhibited the protein synthesis in the root of Quercus robur. The aqueous extract of Ranunculus arvensis plant materials inhibited the germination of wheat varieties and also it caused a decrease in the protein content (Bansal, 1997). Also, present results indicated that protein content in black gram and green gram were significantly increased by aqueous extract of Tectona grandis leaf. Table 3. The effect of aqueous leaf extract of Tectona grandis on protein content in root of black gram and green gram (µg/g) (the values are mean ± SE of 7 samples) Conc. of Extract % Black gram Green gram Root Control 259± ± ± ± ± ± ± ± ± ± ± ± ± ± 6.90 Protein content was increased value in 5% concentration in both crops. The leaves of 5% treatment of black gram exhibit (±15.45) and 100% shows low protein content (±9.45) than control (±14.55). In stem 5% shows (±14.80), control (±13.90), 100% exhibit (±8.80) and 5% concentrated treatment of root contains (±13.05),100% (±7.70) and in control (±12.95) protein content. Impairment of various metabolic activities under the influence of leachate inhibited the protein synthesis and/or stimulated the

4 degradation (Mersie et al., 1993). Similar results have been observed in green gram. From results, effect in 5% leaf extract treatment, content of protein increased allelopathic values (±15.05) in 5% and inhibitory in 100% (±8.40) than control (±14.25) of leaves of green gram. Positive influence effect observed in 5% (± 14.05), very low negative inhibitory in 100% (±8.15) than control (±12.6) of stem and 5% (±12.45) than control (±11.90), Negative influence was observed in 100% (±6.90) treated seedlings of green gram. Allelochemicals produced by leaves of Callicarpa acuminata were tested on root growth, protein synthesis and enzyme activity of seedlings of P. vulgaris, L. esculentum and Z. mays (Cruz- Ortega et al., 2002). While protein content of test crops were gradually decreased in other concentrations like 10, 25, 50 and 75% when growing on it. Very low protein content was observed in 100% treated seedlings. Padhy et al. (2000) have reported that the leaf litter leachate of Eucalyptus globules decreased the protein content in both root and shoot of finger millet. This study was designed to confirm these observations. The results obtained from this study clearly revealed that the intensity of inhibition was directly proportional to the concentrations of extract employed. Conclusion Allelopathy is an important mechanism of plant interference by the addition of plant produced phytotoxins to the plant environment. In the present investigation an attempt has been made to study the allelopathic effect of dry leaves of Tectona grandis on protein content of black gram (var. ADT-3) and green gram (var. Co-3) which are commonly cultivated in the fields of Annamalai Nagar during the months of November, December and January. In laboratory study it was observed that protein content increased in 5% teak leaf extract treatment over control, gradually decreased in other concentration (10, 25, 50 and 75%) and very low protein content was observed in 100% treated seedlings. Prasad et al. (1999) reported that aerial and shoot biomass of Rhamnus virgatus tree significantly decreased the protein content in all test crops Triticum aestivum, eleusince coracana, lens culinaris and phaseolus mungo as compared to control. As the concentrations of teak leaf extract increased the inhibitory effects of protein also increased in black gram and green gram. References Bansal GL (1997). Allelopathic effect of buttercups on wheat variets. Allelopathy Journal, 4: Chai Q, Hang GB (2003). Review on action mechanism affecting factors. Cruz-Ortega R, Ayala-Cordero G, Anaya AL (2002). Allelochemical stress produced by the aqueous leachate of Callicarpa acuminata: effects on roots of bean, maize, and tomato. Physiol. Plantarum, 116: Einhellig FA (1996). Mechanism of action of allelochemicals in allelopathy. Agron. J., 88: Kolesnichenko MV, Aleikina MV (1976). The rate of protein synthesis and absorbance by the root of Oak and ash growimg together in the forest. Fizol. Rast., (Mosc). 23: Lowry DH, Rosenbrough NJ, Farr AL, Randall (1951). R. Protein Measurement with Folinphenol reagent J. Biol Chem., 193: Lv FT, Duan YM (2005). The allelopathy applied in agriculture. Agricultural Sci., 19(3): Mersie W, M Singh (1993). Phenolic acids affect photosynthesis and protein synthesis by isolated leaf cells of velvet-leaf. J. Chem. Ecol., 19: Narwal SS (1994). Allelopathy in Crop Production. Scientific Publishers, Jodhpur, India. Padhy B, Patinaik PK, Tripathy AK (2000). Allelopathic potential of Eucalyptus leaf litter leachates on germination and seedlings growth of finger millet. Allelopathy Journal, 7: Peng SL, Shao H (2001). Research significance and foreground of allelopathy. Chinese. J. App. Ecol., 12(5): Prasad D, Pant G, Rawat MSM (1999). phytotoxicity of Rhamuns virgatus on some field crops. Allelopathy Journal, 6:

5 Rao RSN (1976). Seed viabilities studies under different storage conditions. Patnagar, J. Res, 2: 253. Wang YP, Tang LH (2003). Study on rice allelopathy. Jiangsu Journal of Agricultural Sci., 19(3): Whittaker RH, Feney PP (1971). Allelochemics: chemical interactions between species. Science, 171: