Effect of organic manures on biochemical components of pigeon pea, Cajanus cajan (L.) Millsp. and their impact on the incidence of insect pests

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1 Effect of organic manures on biochemical components of pigeon pea, Cajanus cajan (L.) Millsp. and their impact on the incidence of insect pests B. Bommesha 1, M.I. Naik 2, G. P. Mutthuraju 2* Arati Pannure 2 Syed Imran 2 and C. Prashantha 2 1 Department of Agricultural Entomology, College of Agriculture, Shimoga, Karnataka, India 2 Department of Agril. Entomology, University of Agricultural Sciences, GKVK, Bangalore, India * mutthuwithu@yahoo.co.in ABSTRACT Effect of biochemical components in leaves and green pods of pigeon pea on incidence of insect pests as influenced by application of organic manures was studied during Kharif At 45 and 75 days after sowing, the green leaves of pigeon pea under neem cake treated plots had higher phenol content (4.29 and 5.29 mg/g) and lower total chlorophyll (1.829 and mg/g), protein (10.83 and 11.45%), reducing sugars (1.20 and 1.71%) and total sugars (1.77 and 2.35%), respectively. Green pods had higher phenol content (7.97 mg /g), lower protein (21.60 %) and lower reducing sugar (1.42 %) in neem cake (NC) treated plots and lower total sugar (3.42 %) in poultry manure (PM) treated plots. The early incidence of pests viz., leaf hopper and leaf roller showed a significant positive relationship with total chlorophyll (r=0.73** and 0.87**), protein (0.82** and 0.94**), reducing sugars (0.68* and 0.75**) and total sugars (0.65* and 0.84**) whereas exhibited a significant negative relationship with phenols (r= -0.85** and -0.86**). The infestation of gram pod borer was significantly positive with protein (0.64*) and negatively correlated with the phenol content (-0.74**). There was a significant positive correlation between the incidence of spotted pod borer (0.60*), blue butterfly (0.65*), gram pod borer (0.63*) and pod bugs (0.64*) with the protein and also the spotted pod borer (0.63*) and gram pod borer (0.59*) with the reducing sugars. While the spotted pod borer (-0.73**), blue butter fly (-0.67*), gram pod borer (-0.77**), plume moth (-0.68*), tur pod fly (-0.68*) and pod bugs (- 0.76*) were negatively correlated with the phenol content. The low content of protein, reducing sugar, total sugars and high phenol content offers resistance against insect pest incidence in pigeon pea which were induced by the application of organic manures. KEY WORDS: Biochemical components, insect pests, organic manures, pigeon pea INTRODUCTION Pigeon pea [Cajanus cajan (L.) Millsp.] commonly known as arhar or red gram or tur is an important legume crop of rainfed agriculture in semi-arid tropics. In India it is mainly a kharif crop and arrivals start from October and extend till December. It is the second most important pulse crop after chickpea accounting about 11.8 per cent of total pulse area and 17 per cent of total pulse production of the country (Shah and Agarwal, 2009). Among the various constraints for low productivity in pigeon pea the infestation of insect pests is the 171

2 major one. Pigeon pea is damaged by about 300 species of insect pests infesting at various growth stages (Lal, 1998). Several biochemical factors are known to be associated with insect resistance in pigeon pea. The biochemical constituents present in quantities and proportions to each other in host plants have been reported to exert profound influence on growth, development, survival and reproduction of insects in various ways (Painter, 1958). Inducing resistance through organic means has become more viable. Induced resistance is the qualitative and quantitative enhancement of plant defence mechanisms and is the non heritable resistance where host plants are induced to impart resistance to tide over pest infestation. Information on effect of biochemical constituents of plants on the incidence of pests of pigeon pea in organic treated field is very much limited. Therefore in the present investigations an effort was made to establish a possible correlation between biochemical parameters with pest incidence. MATERIALS AND METHODS A field experiment was laid out in randomized complete block design with 12 treatments and three replications during kharif, 2010 at College of Agriculture, Shimoga, Karnataka (latitude of 14 0' N to 14 0' N, longitude of 75 40' to 75 42'E and 650 m AMSL). Pigeon pea variety ICPL- 87 was grown following all package of practices except fertilizers and plant protection measures. The treatments included application of vermicompost (VC), neemcake (NC), poultry manure (PM) and their combinations with farm yard manure (FYM) and recommended dose of fertilizer (RDF) on the basis of nitrogen requirement of soil (25:50:25 kg/ ha). The organic manures were applied in respective plots as basal applications. The changes in the biochemical components viz., total chlorophyll, phenol, protein, reducing sugars and total sugars were assessed by using the collected fresh leaf samples of five randomly selected plants at 45 and 75 days after sowing (DAS) and also in immature pod (green pods) at 90 DAS. Estimation of total chlorophyll, phenols, proteins, reducing sugars and total soluble sugars in leaves were estimated as per the method suggested by Mahadevan and Sridhar (1986), Mallik and Singh (1980), Lowry et al. (1951), Nelson (1944) and Dubois et al. (1956), respectively. RESULTS AND DISCUSSION Total chlorophyll content in leaves of pigeon pea showed significant variation among treatments which ranged from to and to mg per g of samples collected at 45 and 75 DAS, respectively (Table 1). The maximum chlorophyll content of 2.286, mg/g at 45 and 75 DAS, respectively was recorded in RDF received plots which was slightly higher than the standard check where inorganic fertilizers were used (2.237 and at 45 and 75 DAS, respectively) as compared to minimum of and mg/g at 45 and 75 DAS, respectively in untreated control. At 45 DAS, among the 172

3 organic treatments higher chlorophyll content of to was recorded in VC, PM, VC+PM, NC+VC and FYM+VC which were statistically on par with each other whereas lower chlorophyll content of to was recorded in NC, PM+NC, FYM+NC and FYM+VC which were on par, At 75 DAS, among the organic treatments higher chlorophyll content of , and was recorded in FYM+VC, VC and VC+PM, respectively followed by PM, PM+NC, NC+VC and FYM+PM which were statistically on par ( to ) whereas lower chlorophyll content of and recorded in NC and FYM+NC, respectively. The present findings are in line with the findings of Suresh et al. (2007), who reported that the total chlorophyll content was maximum in NPK applied plants compared with untreated control irrespective of crop duration. The plants treated with organics recorded minimum level of total chlorophyll content. Similar results are reported by Prasanna (2009) who reported significantly highest chlorophyll in chemical fertilizer plots compared organic amendments plots. There was a significant positive relationship between total chlorophyll with leaf hopper (r=0.73**) and leaf roller (r=0.87*) incidence, whereas gram pod borer incidence had non significant positive relationship (r =0.39) (Table 3). The present findings are in agreement with Prasanna (2009) who observed that sucking pest incidence is positively correlated with chlorophyll content in leaves of cotton. The protein content in leaves of pigeon pea did not exhibit much difference among organics during all periods of observation on 45 or 75 DAS. However, lowest protein content of 10.61% in PM+NC and highest of in RDF treatments, followed by the standard check (12.62%) was observed at 45 DAS. The similar trend was observed during 75 DAS. The protein content in green pods was and per cent in NC and RDF treatments, respectively (Table 2). Among organic treatments the lowest protein content was recorded in PM, FYM+NC, FYM+PM and PM+NC which ranged from to per cent. Though adequate information on the relationship between protein content and pod borer incidence in pigeon pea is not available, Murkute et al. (1993) observed higher protein in pigeon pea cultivars being susceptible to pod borer damage compared to the resistant cultivars. Similarly, Hardwick (1956), observed the feeding preference of Helicoverpa armigera larvae towards the flowers and pods of chickpea having high protein content. Aphids showed significant positive correlation with protein content (r= 0.58*) whereas leaf hopper, thrips, whiteflies and mites showed non significant positive correlation at 30 DAS (Table 3). The significant positive relationship was observed between protein and spotted pod borer (r=0.60*), blue butter fly (r=0.63*) gram pod borer (r=0.63*) and pod bugs (r=0.64*) (Table 4). The present findings are in line with the findings of Suresh et al. (2007) who reported that protein content was maximum in NPK applied plants as compared to untreated control irrespective of crop duration

4 Anantharaju and Muthiah, (2008) reported the positive association of crude protein (r=0.232 and 0.273), total free amino acids (r= and 0.609) with incidence of blister beetle and spotted pod borer, respectively, suggesting that the lines with better protein /amino acid content do suffer most. There was significant difference observed in phenol content in leaves of pigeon pea crop due to imposition of various organic manures, which ranged from 3.54 to 4.49 and 4.23 to 5.21 mg /g of 45 and 75 DAS, respectively. Among organics, neem cake recorded significantly high phenol (4.49 mg/g) which was on par with rest of the organic treatments. The next best treatments like PM+NC (4.27 mg/g), FYM+NC (4.23 mg/g), PM (4.15 mg/g), NC+VC (4.13 mg/g), FYM+PM and VC (4.12 mg/g) which were statistically on par, but these were superior over RDF (3.90 mg/g), standard check (3.89 mg/g) and untreated control (3.54 mg/g). Similar trend was observed at 75 DAS. With respect to the phenol content in green pods there was no significant difference recorded among the organics. However treatments NC recorded highest phenol content (7.97 mg/g) followed by PM+NC (7.89 mg/g) which was on par with all the organics and significantly superior over RDF (6.94 mg/g) and standard check (7.04 mg/g). The present findings are in close agreement with Lyashenko et al. (1982), who reported that increased level of leucoanthocinins, catachins, flavanolglycocides and phenol-carboxilic acids in plants that received organics and Prasanna (2009) who reported that positive impact on phenol content in leaves of cotton compared to inorganic plots. The results indicate that significant negative correlation was observed between leaf hopper (r=-0.85**), leaf roller (r=- 0.86**), gram pod borers (r=-0.74*8) with the phenol content in leaves (Table 3) and spotted pod borer (r=-0.73**), gram pod borer (r=-0.77**) and pod bugs (r=0.76**) phenol content in green pods of pigeon pea (Table 4). The present findings are in confirmity with findings of Rao (2003), who reported the significant negative correlation of population of Spodoptera litura (Fab.) (r= -0.70*) and Helicoverpa armigera (Hub.) (r= -0.76*) with phenol content of leaves of ground nut. Sahoo and Patnaik (2003) also observed a significant inverse relationship between the phenol and incidence of most pod borer species in pigeon pea. Total phenolic content is non significant and negatively correlated (r=-0.378) with spotted pod borer incidence and highly significant negative correlation (r=-0.594) with the incidence of blister beetle (Anantharaju and Muthiah, 2008). Chemical analysis further revealed that the reducing sugar was high in leaves collected from RDF treatment, followed by the standard check with reducing sugars of 1.87, 2.48 and 1.98, 2.41 per cent on 45 and 75 DAS respectively, while the lowest among organics was recorded in NC, PM and VC+PM and other organic treatments were superior to RDF and standard check. Whereas, in green pods of all organics treatments (T 2 to T 10 ) low reducing sugars were recorded compared to NPK applied (T 1 and T 11 ) treatments. A significant positive combination of FYM, PM and NC had 174

5 correlation observed between pest incidence and reducing sugar content indicating that higher levels of reducing sugars responsible for high pest incidence in NPK applied plots compared with organic treatments. The results collaborated with earlier findings of Rani (2005), who reported that total proteins, total reducing sugars were comparatively lower in flower buds, pods, and seeds in the tolerant varieties with lower pod borer damage than in the susceptible varieties. However, Gowda et al. (1996), opined that per cent reducing sugar, total sugar, silica and potassium content did not had any influence on incidence of shoot fly in sorghum. It is clearly evident from Table 1that among the treatments all organics (T 2 to T 10 ) had significantly lower total sugar content compared to NPK applied plots (T 1 and T 11 ) at 45 DAS. However, at 75 DAS highest total sugar was recorded in standard check (3.22%) followed by RDF (3.03%), while lowest in NC, VC+PM, FYM+PM and FYM+NC with 2.35, 2.49, 2.49 and 2.40 per cent sugars, respectively and were on par with rest of the organics. Lowest total sugar in green pods of pigeon pea was found in PM, NC and VC+PM with 3.42, 3.44 and 3.50 per cent, respectively and these were superior over RDF (4.03%), standard check (4.11%) and on par with the untreated control (3.75%). The results indicate that lower content of total sugar recorded in organics compared to application of inorganic fertilizers plots. Thus a preference by the pod borers to damage the pods having more sugar content in the pods was indicated in Table 4, it revealed that the incidence leaf hopper, leaf roller and non significant correlation with gram pod borer incidence with total sugars in leaves of pigeon pea. However, non significant positive correlation was found between spotted pod borers, blue butterfly, gram pod borer, plume moth and pod fly populations with total sugars in green pods. The present findings are in conformity with the findings of Murkute et al. (1993), who reported that significant positive correlation between pod borer damage and total sugar content of pigeon pea varieties, while Thimmaiah (1992), reported that the total sugar content in the leaves, stem tip, squares and boll rind of cotton genotypes reveals that vegetative parts of susceptible genotypes to insect pests invariably have higher levels of sugars, whereas the resistant genotypes have comparatively lower levels of sugars, besides cotton genotypes susceptible to insect pests have shown higher level of nitrogen and crude protein content at all the stages of crop growth. ACKNOWLEDGEMENT The authors are thankful to Dr. A. G. Shankar, Professor, Department of Crop Physiology, UAS, GKVK, Bangalore for providing lab facility for biochemical analysis. significant positive relationship between the 175

6 Table 1: Effect of different organic manures on biochemical parameters of pigeon pea leaves at different plant growth stages Treatments Total chlorophyll (mg/g) Protein (%) Phenol (mg/g) Reducing sugars (%) Total sugars (%) 45DAS 75DAS* 45DAS 75DAS 45DAS 75DAS 45DAS 75DAS 45DAS 75DAS T 1 RDF 2.28 a 2.37 a a ab 3.90 bc 4.34 cd 1.98 a 2.41 a 2.43 a 3.03 ab T 2 VC 1.93 bc 2.09 abc abc bc 4.12 ab 4.78 bcd 1.67 abc 2.16 ab 1.99 b 2.56 bc T 3 PM 1.93 bc 2.01 bc c c 4.15 ab 4.81 abc 1.27 bc 1.83 bc 1.93 b 2.68 bc T 4 NC 1.82 c 1.90 c c c 4.49 a 5.21 a 1.20 c 1.71 c 1.77 b 2.35 c T 5 VC+PM 1.96 bc 2.08 abc bc bc 4.08 ab 4.79 abc 1.51 abc 2.11 abc 1.97 b 2.49 c T 6 PM+NC 1.85 c 1.97 bc c c 4.27 ab 4.81 abc 1.59 abc 2.24 ab 1.90 b 2.57 bc T 7 NC+VC 1.91 bc 1.98 bc c bc 4.13 ab 4.83 abc 1.69 ab 2.23 ab 1.95 b 2.62 bc T 8 FYM+NC 1.84 c 1.91 c c c 4.23 ab 4.93 ab 1.61 abc 2.17 ab 1.89 b 2.49 c T 9 FYM+PM 1.87 c 1.97 bc c c 4.12 ab 4.83 abc 1.65 abc 2.24 ab 1.85 b 2.40 c T 10 FYM+VC 1.96 bc 2.09 abc abc bc 4.06 ab 4.76 abc 1.77 a 2.32 a 1.99 b 2.65 bc T 11 Standard check 2.23 ab 2.27 ab ab a 3.89 bc 4.44 bcd 1.87 a 2.48 a 2.51 a 3.22 a T 12 Untreated check 1.73 c 1.86 c abc ab 3.54 c 4.23 cd 1.71 ab 2.22 ab 2.01 b 2.55 bc S.Em.± CD at 5% CV (%) * At flowering DAS- Days after sowing; RDF- Recommended dose of fertilizer; VC-Vermicompost ; PM-Poultry manure; NC - Neem cake; FYM- Farm yard manure; In a column means followed by same letter(s) are not significantly different at p=0.05 as per DMRT 176

7 Table 2: Influence of different organic manures on biochemical parameters of immature pods (green pods) of pigeon pea Treatments Protein (%) Phenol (mg/g) Reducing sugars (%) Total sugars (%) T 1 RDF a 6.94 cd 2.69 a 4.03 ab T 2 VC bc 7.43 abc 2.34 bcd 3.61 bc T 3 PM c 7.77 ab 1.94 d 3.42 c T 4 NC bc 7.97 a 1.92 d 3.44 c T 5 VC+PM c 7.46 abc 2.27 bcd 3.50 c T 6 PM+NC c 7.89 a 2.18 cd 3.56 bc T 7 NC+VC c 7.43 abc 2.40 abc 3.62 bc T 8 FYM+NC c 7.64 abc 2.25 cd 3.56 bc T 9 FYM+PM c 7.48 abc 2.32 bcd 3.52 bc T 10 FYM+VC bc 7.42 abc 2.41 abc 3.60 bc T 11 Standard check ab 7.04 bcd 2.79 a 4.11 a T 12 Untreated check ab 6.57 d 2.56 abc 3.75 abc S.Em.± CD at 5% CV (%) RDF- Recommended dose of fertilizer; VC-Vermicompost ; PM-Poultry manure; NC - Neem cake; FYM- Farm yard manure; In a column means followed by same letter(s) are not significantly different at p=0.05 as per DMRT 177

8 Table 3: Correlation coefficient of the sucking/early pests with biochemical constituents in leaves of pigeon pea (pooled data) Correlation coefficient Early insect pests Total Reducing Total Protein Phenol chlorophyll sugars sugars Leaf hopper, Empoasca kerri 0.73** 0.82** -0.85** 0.68* 0.65* Leaf roller, Caloptilia soyella 0.87** 0.94** -0.86** 0.75** 0.84** Gram pod borer,helicoverpa armigera * -0.74** * Significant at 5%; ** Significant at 1%; r values =0.576 and = Table 4: Correlation coefficient of the pod borers incidence with biochemical constituents of the immature pods (green pods) in pigeon pea Pod borer species Protein Correlation coefficient Phenol Reducing content sugars Total sugars Spotted pod borer, Maruca testulalis 0.60* -0.73** 0.63* 0.47 Blue butter fly, Lampides boeticus 0.63* -0.67* Gram pod borer, Helicoverpa armigera 0.63* -0.77** 0.59* 0.52 Plume moth, Exelastis atomosa * Tur pod fly, Melanagromyza obtusa * Pod bugs, Riptortus pedestris, Riptortus linearis and Clavigralla gibbosa 0.64* -0.76** * Significant at 5%; ** Significant at 1%; Table r values =0.576 and =

9 REFERENCES Anantharaju, P. and Muthiah, A. R., 2008, Biochemical components in relation to pests incidence of pigeon pea spotted pod borer (Maruca testulalis) and blister beetle (Mylabris spp.). Legume Res., 31: Dubois, M., Gilles, K. A., Hamilton, J. K., Robers, P. A. and Smith, F., 1956, Estimation of total soluble sugars. Anal. Chem., 26: 350. Gowda, K., Jagadish, P. S., Muniswamygowda, K. N. and Ramesh, S., 1996, Biochemical basis of resistance to shoot fly (Atherigona pulla Wiede) in little millet. Karnataka J. Agric. Sci., 9: Hardwick, D. F., 1956, The corn ear worm complex, Memorial Ent. Society, Canada, 40:247. Lal, S. S., 1998, Host plant resistance in pigeon pea against the podf1y, Melanagromyza obtusa Malloch. Indian J. Pulses Res., 11: Lowry, O. H., Brough, R. N. J., Farr, A. L. and Randal, R. J., 1951, Plant biochemical analysis. J. Biol. Chem., 193: 265. Lyashenko, N. I., Solody, U. K. G. D., Godovany, A. A., Verzhitskii, V. I. and Chuk, M. N. I., 1982, The effect of fertilizer on phenol metabolism in hopes on plants resistant to aphids. F. R. astenil, 14: Mahadevan, A. and Sridhar, R., 1986, Methods of physiological Plant Pathology, Sivakami Publications, Madras, p Mallik, C. P. and Singh, M. B., 1980, Plant Enzymology and Histo Enzymology, Kalyani Publishers, New Delhi, p Murkute, G. R., Dhage, B. B., Desai and Kale, A. A., 1993, Biochemical parameters associated with pod borer damage as influenced by maturity group and growth stages of pigeon pea, Cajanus Cajan (L.) Millsp. Legume Res., 16: Nelson, N., 1944, A Photometric adoption of the Somogyi method for the detection of glucose. J. Biol. Chem., 153: Painter, R. H., 1958, Resistance of plants to insects. Ann. Rev. Ent., 3: Prasanna, S. O., 2009, Influence of soil amendments on the incidence of sucking pests infesting Bt cotton. M.Sc. (Agri) Thesis, Univ. Agric. Sci., Bengaluru, India. Rani, S. S. N., 2005, Biochemical basis of tolerance in pigeon pea to the pod borer (Helicoverpa armigera) infestation. M. Sc. (Agri.) Thesis, University of Agricultural Sciences, Bengaluru, India. Rao, R. K., 2003, Influence of host plant nutrition on the incidence of Spodoptera litura and Helicoverpa armigera on groundnut. Indian J. Ent., 65:

10 Sahoo, B. K. and Patnaik, H. P., 2003, Effect of biochemicals on the incidence of pigeon pea pod borers. Indian J. Pl. Prot., 31: Shah, K. and Agarwal, A., 2009, India agriculture review: Ghana outlook, http//.wwwstockmarketsreviewcom. Suresh, K., Rajendran, R. and Rani, U. B., 2007, Eco-friendly approaches for managing major sucking pests of brinjal. J. Ent. Res., 31: Thimmaiah, K. K., 1992, Physiological and histo-chemical basis of insect pest resistance in cotton (Gossypium spp.) genotypes. Ph.D. Thesis, Univ. Agric. Sci., Dharwad, India [MS received 03 July 2012; MS accepted 28 August 2012] Disclaimer: Statements, information, scientific names, spellings, inferences, products, style, etc. mentioned in Current Biotica are attributed to the authors and do in no way imply endorsement/concurrence by Current Biotica. Queries related to articles should be directed to authors and not to editorial board

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