World Journal of Pharmaceutical Research Marepally et al. World Journal of Pharmaceutical Research SJIF Impact Factor 5.990 Volume 5, Issue 2, 1048-1055. Research Article ISSN 2277– 7105

STUDIES ON EFFICIENT CONCENTRATION OF DICROTOPHOS IN

CONTROLLING FLETCHERI MINOR INFESTATION IN TERMINALIA ARJUNA

Lakshmi Marepally* and G. Benarjee

Post- Doctoral Fellow-UGC, Department of Zoology, Kakatiya University, Warangal- 506009.

ABSTRACT Article Received on 02 Dec 2015, The leaf gall infections caused by Trioza fletcheri minor are highly

Revised on 23 Dec 2015, virulent with considerable damage in leaf quality. Therefore an attempt Accepted on 12 Jan 2016 has been made to evaluate the efficient concentration of Dicrotophos in controlling the infestation through the studies on total soluble proteins, *Correspondence for free amino acids, total soluble sugars, total reducing sugars, secondary Author metabolites (Starch, Phenols, Phyticacid, Acid detergent fibre, total Dr. Lakshmi Marepally Post- Doctoral Fellow- chlorophyll, chlorophyll–a, chlorophyll- b and carotenoid in UGC, Department of Terminalia leaves. Results revealed that total soluble proteins and total Zoology, Kakatiya soluble sugars were increased in D1 (0.5% dicrotophos treated University, Warangal- Terminalia), D2(0.2% dicrotophos treated terminalia) and D3(0.1% 506009. dicrotophos treated Terminalia) batches over the healthy control but found decreased over infected control. In comparison with the healthy control, free amino acids and total reducing sugars were decreased in D1, D2 and D3 batches whereas a significant increase was noticed in these batches over the infected control. The starch content and ADF% of infected control found increased significantly followed by D3, D2 with least increase in D1batch. In comparison with the healthy control, the phenol, phytic acid content was reduced drastically in infected control with a least reduction in D1 batch. Photosynthetic pigments like total chlorophyll, chlorophyll–a, chlorophyll– b and carotenoids were decreased significantly in infected control whereas these got reduced at a lower level in D1 batch than D2 and D3 respectively. Based on the results obtained from the present study 0.5% Dicrotophos is found efficient in controlling the infestation. KEYWORDS: Secondary metabolites, photosynthetic pigments, Triozafletcheriminor, Terminalia arjuna, Dicrotophos.

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INTRODUCTION Terminalia arjuna is the primary host plant of the Tropical Tasar silkworm Anthereae mylitta drury in commercial sericulture. Silkworms feed on leaves during their entire larval period and utilize the leaf metabolites for the biosynthesis of silk. It is therefore clear that host plant plays a dominant role in cocoon production as a source of nutrition to the silkworms. However, foliage is prone to depredation by disease causing organism such as pathogens and pests. Among the several pests known to attack Tasar plants Trioza fletcheri minor (-) has attained a serious status. It has long been recognized that levels of proteins and carbohydrates in plants have significantly influenced by infection and metabolism alters accordingly by the rate of infection (Kirk and Allen,1965). Increase in protein concenteration was noted in the various tissues infected with root knot nematodes (Sailaja etal 2012).An elevated protein levels was also observed in giant cells of okra infected with M.incognita acrita (Narayanaswamy,2003). A significant increase in the total sugars and decrease in total reducing sugars of M5 mulberry leaves due to leaf roller infestation and mealy bug infection in hibiscus had been reported (Sailaja etal 2012). Decrease in the secondary metabolites like phenols and phytic acid and increase in starch and ADF (acid detergent fibre) has reported as a combined loss of total chlorophyll and carotenoids in the thrips - infested mulberry leaves(Das etal 1994). So the present experiment has taken up to identify the levels of primary, secondary metabolites and photosynthetic pigments in mealybug infected Terminalia arjuna a primary host plant of Tasar silk worm Anthereae mylitta drury. So, the present experiment has taken up to evaluate the efficient concentration of Dicrotophos in controlling the infestation through the studies on secondary metabolites (Starch, Phenols, Phyticacid, Acid detergent fibre (ADF)) and photosynthetic pigments (total chlorophyll, chlorophyll–a, chlorophyll- b and carotenoids) in Terminalia leaves.

MATERIALS AND METHODS The present experiment was conducted by collecting healthy and Trioza fletcheri minor infested Terminalia arjuna leaves. The leaves were cleaned and then oven dried at 80-90°C for several hours till drying completely. The dried leaves were ground into powder in an electric grinder (Bajaj Mixer grinder). The powdered leaf samples were used to analyze total free amino acid as (Moore and Stein,1948), total proteins (Lowry,1951), total soluble sugars and starch (Yemm and Willis,1954), total reducing Sugars (Miller,1972). Fresh Terminalia arjuna leaves were utilised to estimate the photosynthetic pigments (total chlorophyll, chlorophyll–a, chlorophyll – b) as Arnon(1949) and carotenoids (Litrell,1966),Phenols www.wjpr.net Vol 5, Issue 2, 2016. 1049

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(Bray,1954), Phytic acid (Wheeler,1971) and acid detergent fiber (ADF) (Vansoest,1991). To evaluate the efficienct concenteration of Dicrotophos in controlling Trioza fletcheri minor infestation in Terminalia arjuna, treatments were divided as Healthy control (healthy leaves), Infected control(Trioza fletcheri infested leaves) and Dicrotophos treated (Infected treated). Infected leaves were treated with three different concentrations of Dicrotophos like 0.5% (D1 Batch), 0.2% (D2-Batch) and 0.1% ( D3-Batch).

STATISTICAL ANALYSIS: Each assay was replicated 3 times. Values were expressed as mean ± SE of replication and Student’s t-test was applied to locate significant (P ≤ 0.05) differences between treated and untreated leaves. Critical differences (CD5%) were analyzed by Tukeys post hoc procedure.

RESULTS AND DISCUSSION Table 1 indicate the performance of Dicrotophos in controlling Trioza fletcheri minor infestation in Terminalia arjuna and its influence on total soluble proteins,free amino acids, total soluble sugars and total reducing sugars. In comparison with the healthy control and infected control, total soluble proteins in D1, D2 and D3 batches found increase by 4.5,9 and 10.3% respectively and a decrease by 14.5, 11.8 and 9.8% respectively. It is observed that total soluble proteins were recorded low in D3 batch compared to D1 and D2 which may be due to the resistance attained against infection. High protein content correlates with activation of several lines of defense mechanism and release of disease resistant proteins in fungal infected plants(Ashraf etal 2011). Increased rate of synthesis of proteins can be correlated with the increase in metabolic activities in infected tissues and addition of viral proteins (Charitha devi and Radha,2012). Mesta plants infected with yellow vein mosaic disease have shown an increase of disease related proteins because plant pathogens elicit the synthesis of host proteins which prevents the multiplication of pathogens in the host (Arpita Chatterjee and Subrata Ghosh 2008). It is evident from the results that the free amino acids of D3, D2 and D1 batches were decreased by 7.5, 9.9 and 30.46% in comparison with the healthy control whereas increased by 55.2,36 and 16.5% over infected control. Decrease in the amino acids can be correlated with the increase in protein synthesis. The amount of free amino acids also decreased with time with grown gall infection in sunflower (Kerry etal 2014). In comparison with the healthy control and infected control, total soluble sugars in D1, D2 and D3 batches found increase by 6, 23.3 and 38.6% respectively and decrease by 30.5, 19.2 and 9.2% respectively. Alteration in the sugars may be due to reduction in leaf lamina and

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malformation of leaves in pest affected plants resulting in less productivity (Raithak and Gachande,2012). It is found that the D1, D2 and D3 batches have shown a decrease in total reducing sugars by 15, 27.3 and 36.8% respectively than healthy control batch whereas an increase of 76.6%, 50.7% and 31% have shown over infected control. A significant reduction in the total reducing sugars in the leaves of K2, S30, S36, S40 and S54 mulberry varieties infested by M. hirsutus was also observed. Decrease of total reducing sugars in a highly infected host tissue is due to the excess utilization of sugars by the organisms for their growth and sustainability (Shree andUmesh,1989). Decrease in carbohydrate content in the highly infected plant can be correlated with the increase in glucose6phosphate dehydrogenase activity and 6-phosphogluconate dehydrogenase in infected tomato (Raithak and Gachande, 2012).

Table 2 explains that in comparison with the healthy control, infected terminalia had shown a significant increase (56%) in the starch content. A significant increase of 8, 16.8 and 32% starch content was reported in D1, D2 and D3 batch terminalia over the healthy batch. The results also show that in comparison with the infected control, starch content in D1, D2 and D3 batch terminalia had reduced by 30.8, 25.13 and 15.4% respectively. Results show that a significant reduction (43%) of phenol content was observed in infected control than healthy control. When compared with the healthy control and infected control, phenol content of D1, D2 and D3 batches was low by 6.2, 12.3 and 30.8% and was high by 65, 54 and 21.6% respectively. Significant decrease in total phenolic compounds was also reported in mulberry species attacked by leaf roller. High phenol content in the healthy terminalia leaves is due to high level of resistance as phenols supports the defence mechanism (Narayanaswamy,2003). Phytic acid content found decreased drastically by 73.8% in infested Terminalia. A significant decrease of 7.3, 18.7 and 22.7% phytic acid content was reported in D1, D2 and D3 batch terminalia over the healthy batch. The results also show that in comparison with the infected control, phytic acid content in D1, D2 and D3 batch terminalia had increased by 25.8, 10.2 and 4.8% respectively. The reduction in phytic acid content can be correlated with the decrease in proteolytic activity thus an increase in protein content and decrease in amino acids (Megat Rusydi and Azrina 2012). Results also show that in comparison with the healthy control, infected terminalia had shown a significant increase (65.5%) in the ADF%. The results also show that in comparison with the healthy control, ADF% in D1, D2 and D3 batch terminalia had increased by 6.1,15 and 31% respectively. A significant decrease of 70, 75, 85.8% ADF was reported in D3, D2 and D1 batch terminalia over the infected batch. www.wjpr.net Vol 5, Issue 2, 2016. 1051

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Table 3 explains that total chlorophyll content reduced by 55.6% in the infected leaves when compared with the healthy control. It is evident from the results that total chlorophyll content of D1,D2 and D3 batches have decreased by 9,13 and 32.2% respectively when compared to that of healthy control. A significant increase of 64, 56.9 and 24.4%respectively total chlorophyll was reported in D1, D2 and D3 batch terminalia over the infected batch. Present results show that a drastic reduction of 41% chlorophyll-a was found in the infected Terminalia over the healthy control. In comparison with the healthy control chlorophyll-a found decreased by 8.2, 24.5and 33.3%respectively in D1, D2 and D3 batches. A significant increase of 55,27.6 and 13.8% chlorophyll-a was reported in D1, D2 and D3 batch terminalia over the infected batch. The chlorophyll-b of infected Terminalia was found decrease significantly by 45.5% over the healthy control. When compared with the healthy control the chlorophyll-b content of D1,D2 and D3 batches was low by 19, 25.4 and 31%respectively whereas it got increased by 80,64 and 52% respectively over the infested control. Results reveal that infected terminalia has reduced by 53.3% in its carotenoids content over the healthy control. A reduced amount of carotenoids in D1, D2 and D3 batches by 90.5, 74.3 and 62% respectively was observed in comparison with healthy control. It is found that the D1, D2 and D3 batches have increased in carotenoids by 70, 39.3 and 16.1% respectively than infected control. The significant decrease of carotenoids in leaf roller infested mulberry variety was due to malformation of leaves( Mahadeva and Nagaveni,2011). Decrease in chlorophyll and carotenoid contents in infested leaves may be due to loss in chlorophyll synthetase activity (Neog,2011).

Thus in conclusion 0.5% Dicrotophos is found efficient in controlling Trioza fletcheri minor infestation and the treatment improves primary and secondary metabolites.

Table 1: Effect of Trioza fletcheri minor infestation on biochemical components of Terminalia arjuna leaves Total soluble Free amino Total soluble Total reducing Treatment proteins (mg/mg) acids(µg/g) sugars (mg/g) sugars(mg/g) Healthy control 113.5±3.45 12.15±1.15 2.15±0.05 135.25±2.68 Infected control 138.65±4.55 7.25±1.05 3.28±0.08 65.26±4.26 D1Batch(0.5%) 118.6±2.25 11.25±1.04 2.28±0.04 115.24±2.65 D2Batch(0.2%) 123.8±3.28 9.86±1.24 2.65±0.05 98.35±4.65 D3Batch(0.1%) 125.2±2.45 8.45±0.8 2.98±0.06 85.45±3.68 CD5% 0.54 0.28 1.65 1.54 CD: Critical difference. All the values are the mean values of five replications

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Table 2: Effect of Trioza fletcheri minor infestation on secondary metabolites of Terminalia arjuna leaves Treatment Starch (mg/g) Phenols (mg/g) Phytic acid (mg/100g) ADF (%) Healthy control 1.25±0.08 3.25±0.08 2525.15±0.05 21.25 Infected control 1.95±0.06 1.85±0.06 1863.28±0.08 32.45 D1Batch(0.5%) 1.35±0.04 3.05±0.05 2342.18±0.04 22.55 D2Batch(0.2%) 1.46±0.05 2.85±0.04 2052.65±0.05 24.45 D3Batch(0.1%) 1.65±0.08 2.25±0.08 1952.28±0.06 27.85 CD5% 0.44 0.26 2.65 0.54 CD: Critical difference. All the values are the mean values of five replications

Table 3: Effect of Trioza fletcheri minor infestation on photosynthetic pigments of Terminalia arjuna leaves Total chlorophyll Chlorophyll-a Chlorophyll-b Carotenoids Treatment (mg/ml) (mg/ml) (mg/ml) (µg/ml) Healthy control 2.25±0.08 2.45±0.06 0.55±0.05 1.05±0.02 Infected control 1.25±0.06 1.45±0.04 0.25±0.06 0.56±0.04 D1Batch(0.5%) 2.05±0.04 2.25±0.05 0.45±0.04 0.95±0.06 D2Batch(0.2%) 1.96±0.05 1.85±0.04 0.41±0.05 0.78±0.08 D3Batch(0.1%) 1.55±0.08 1.65±0.08 0.38±0.08 0.65±0.05 CD5% 0.45 0.28 0.04 0.05 CD: Critical difference. All the values are the mean values of five replications

ACKNOWLEDGEMENTS The author Dr.Lakshmi Marepslly is thankful to University Grants Commission, New Delhi for providing financial assistance under Post- Doctoral Fellow for Women (PDFW).

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