Journal of Tropical Forest Science 12(2):221-226 (2000)

ROL FOLIAF O E R PROTEI POLYPHENOD NAN D LAN THEIR RELATIONSHIP TO CLONAL RESISTANCE IN TEAK AGAINS E LEATTH F SKELETONISER, PALIGA MACHOERAL1S WALKER (: PYRALIDAE)

Avinash Jain . RoychoudhuryN , Alk*& a Bhargava

Chemistry of Forest Produce Division, *Forest Entomology Division, Tropical Forest Research Institute, Jabalpur 482021 (M.P.), India

Received June 1997______

JAIN, A., ROYCHOUDHURY, N. & BHARGAVA, A. 2000. Role of foliar protein and polyphenol and their relationship to clonal resistance in teak against the leaf skeletoniser, Paliga machoeralis Walker (Lepidoptera: Pyralidae). Leaf protein and polyphenol contents were estimate r selectefo d d resistant (ORANR-2, ORANR-3, ORANR-4, ORANP-7 and MHSC-A3) and susceptible (APT-8, APT-14, TNT-2 and ORPB-15) clones of teak to the leaf skeletoniser, Paliga machoeralis. The results were correlated wit degree hth f resistance/susceptibilityo e e protei Th .polypheno d nan l contents of clonal leaves were directly and inversely proportional respectively, to amoune th leaf o t f damage e rati Th f .proteio o o polyphenot n l playe decisiva d e role in determining the degree of resistance. Alow ratio indicates higher resistance vicd an e versa.

Key words: Teak - clonal resistance - skeletoniser - leaf - protein - polyphenol

JAIN, A., ROYCHOUDHURY, N. & BHARGAVA, A. 2000. Peranan protein dan polif enol daun serta kaitannya dengan kerintangan klon dalam pokok jati terhadap ulat perangka daun, Paliga machoeralis Walker (Lepidoptera: Pyralidae). Kandungan protein dan polifenol dianggarkan bagi klon rintangyang terpilih (ORANR-2, ORANR- 3, ORANR-4, ORANP-7 dan MHSC-A3) dan klon rentan (APT-8, APT-14, TNT-2 dan ORPB-15) bagi jati terhadap perangka daun, Paliga machoeralis. Keputusan adalah seimbang masing-masing secara langsun secarn da g a songsang dengan jumlah kerosakan daun. Nisbah protein kepada polifenol memainkan peranan yang penting dalam menentukan darjah kerintangan. Nisbah yang rendah menunjukkan kerintangan yang tinggi, dan nisbah yang tinggi menunjukkan kerintangan yang rendah.

Introduction

Host resistance has provided a highly practical approach to solving problems (Ortman &: Peters 1980, Smith 1989, Roychoudhury et al 1995a). Clonal resistanc n teaei k (Tectona grandis L.f.) (family Verbenaceae) agains s primit t e defoliator bees ha sn well established (Tewari 1992 t informatiobu ) n still remains limited regarding the phytochemicals influencing the dynamics of insect-plant interactions (Hanover 1975, 1980, Norri Kogas& n 1980, Matso Scriben& r 1987) whicd an decisivs hi impartinn ei g resistance against (Saxena 1986). Foliar

221 2 22 Journal of Tropical Forest Science 12(2) :221-226 (2000)

constituents have a significant role in population dynamics, insect behaviour and the physiology of defoliating insects in a number of ways (Jain et al. 1995, Roychoudhury et al. 1995b).The present study is an attempt to examine the role of leaf protein and polyphenol in clonal resistance of teak against the leaf skeletoniser, Paliga machoeralis Walker (Lepidoptera:Pyralidae), a devastating defoliator of teak. Preliminary screenings, were conducted on 167 clones of teak from ten Indian states, to identify teak clones resistant to P. machoeralis, at the National Teak Germ Plasm Bank, Chandrapur (Maharashtra) Roychoudhury b , Joshd yan i (1996). Further studies were carrie selecten o t dou d teak clones through field observations and feeding bioassays in the laboratory (Roychoudhury et al 1995c). All these studies suggest that teak clones exhibit resistance (antixenosis/non-preference) to the leaf skeletoniser.

Material methodd san s role proteif th e fino t o T polyphenod ou nan clonan li l resistanc teaf eo k against P. machoeralis, nine clone teakf so , viz. ORANR-3, MHSC-A3, ORANR-2, ORANP-7, ORANR-4, APT-8, TNT-2, ORPB-15 and APT-14, were ranked as most resistant, highly resistant, resistant, moderately resistant, susceptible, highly susceptible and most susceptible respectively based on our earlier work (Roychoudhury & Joshi 1996, Roychoudhury el 1995c)la . Leave f intermediatso e maturit month3 y( s old) (Roychoudhury et al 1995b) were collected from one-year-old plants during late August (coinciding with the outbreak of the teak leaf skeletoniser) and , pulverise°C 0 drie 4 subjected t dan a analyseso dt e proteiTh . n contens wa t estimated at 660 nm on a PC-based UV/VIS spectrophotometer by the extraction technique described by Lowry (1951). The polyphenol content was determined using the method given by Folin and Ciocalteu (1927) with some modifications (Singleton 1974) e readingTh . s wer agaim n e n0 take wit 70 PC-base e t nha th d UV/VIS spectrophotometer. The data were based on three trials, each containing three replication eacr sfo h clone. These were subjecte statisticao dt l analysey b s ANOVA using CRD design by SPSS statistical package and the results correlated with degree of resistance. Data were angular transformed to conform to normal distribution.

Results and discussion

Leaf protein

e proteiTh n conten f contributoro t y leave f selecteo s d teak clones exhibited significant (F=7.632, p <0.01; d.f. 8,18) variation in their mean values (Table 1). Clonal leaves revealed gradual decreas protein ei n content with increased degree of resistance mose Th . t resistan susceptibld an t e clones, ORANR- APT-14d 3an , showed the lowest and highest amounts of protein (p < 0.01) respectively compared to other clones in this work. The protein content of clones MHSC-A3, ORANR-2, Tabl Dat. e1 protein ao polyphenod nan l content f contributorso y leave selectef so d teak clone theid san r ratio s 2 6

K> NO

XIo o o

Data based on pooled mean of three trials. Range values are inside parentheses. MRl=most resistant, HR=highly resistant, R=resistant, MR2=moderately resistant, MSl=moderately susceptible, S=susceptible, HS=highly susceptible, MS2= most susceptible. SEm=standard error of mean. CD=critical difference. NO IxO 05 224 Journal of Tropical Forest Science 12(2) :221-226 (2000)

ORANP-7, ORANR-4, APT-8 and TNT-2 failed to exhibit conclusive results (p>0.05). Further, clones TNT- ORPB-1d 2an 5 showe significano dn t (p>0.05) difference among themselves. However, clone APT-14 revealed significant (p<0.05) difference when compared with others except clone ORPB-15, where conclusivo n e difference existed.

Leaf polyphenol

polyphenoe Th l conten f selecteo t d clonal leave teaf so k revealed significant ( 23.091F= 0.01< p , ; d.f. 8,18) difference thein i s r mean values (Tabl Leave. e1) s of teak clones showed gradual increase in polyphenol content in relation to increase in degree of resistance. The most resistant clone, ORANR-2, showed a significantly higher concentration (p<0.01) compared to others, except clone MHSC-A3 wher differenco en 0.05 recorded> s p ( e)wa polyphenoe .Th l content of clones ORANR-2, ORANP- ORANR-d 7an 4 showe significano dn t difference among themselves (p>0.05). Further, clones APT- d TNT-8an 3 exhibiteo dn conclusive difference (p>0.05) e mosTh . t susceptible clone, APT-14, revealed significandy (p<0.05) the lowest content of polyphenol compared to the others except clone ORPB-15, wher significano en t differenc observes ewa 0.05)> p d( .

Ratio of leaf protein to polyphenol

ratie foliaf Th oo r protei polyphenono t testef lo d teak clones showed significant (F=13.860,p< 0.01; d.f. 8,18) variatio thein i r mean values (Tablexhibited an ) e1 d correlation with regard to the degree of resistance. It was found to be inversely proportiona resistanceo lt lowe.A r rati recordes mosoe wa th r t dresistanfo t clone, ORANR-3, wherea mose sth t susceptible clone, APT-14, exhibite dhighea r ratio. presene Th t work demonstrates tha foliae tth r protein conten f selecteto d teak clone inversels si y proportiona degree th o resistancelt f eo . These findingn i e sar agreement wit observatione hth s ofjayaraj (1967 Uthamasamd an ) l (1971)a t ye , who have also recorded increased protein content in varieties of castor and bhendi susceptibl o leat e f hoppers Empoasca flavescens d Amrascaan devastans (Jassidae) respectively e polyphenoTh . l conten f clonao t l leaves exhibitn a s increasing trend in relation to resistance. Although, contradictory opinions exist regardin role gf th polyphenole o dynamice th n si f insect-plano s t interactions (Porte Hemingwar& y 1989, Roychoudhur l 1995d)a t ye presene ,th t study clearly suggest e allomonith s c effect f foliao s r polypheno tean li k clones againse th t leaf skeletoniser similaA . r notio alss noha been envisage somr dfo e other insect- host interactions (Reese 1979). Apart from presenthesee th n i , t work gooa , d correlation, (r = - 0.779, p < 0.01), was observed between proteins and polyphenols, the ratio of which strongly influences the degree of resistance in teak clones. The ratio between these two is more important than their absolute concentrations. A lower ratio indicates higher resistance whereas a higher ratio is an indicator of susceptibility. The ratio of protein to polyphenol is considered to be one parameter Journal of5 Tropical22 Forest Science 12(2):221-226 (2000) nutritionae th f o l qualit leaa f yfo (McManus el 1983)ta lowe;a r rati hosa on i t plant bees ha n foun affeco dt l aspectal t performancf so e including feeding guildn so Diacrisia casignetum (Banerje Haqu& e e 1985) wholea s e A presen. th , t study suggests thae contentth t f leao s f protein, polypheno d theian l e r b rati n oca considere determinants da clonaf so l resistanc tean ei k agains . machoeralis.tP

Acknowledgements

e authorTh . Gupta gratefue N . sar B ,o t Directol r General, Indian Councif o l Forestry Researc Educationd han , Dehradun . LalB , . R Director d an , , Tropical Forest Research Institute, Jabalpur (MP), for their encouragement and providing research facilities.

References

BANERJEE, T. C. & HAQUE, N. 1985. Influence of host plants on development, fecundity and egg hatchability of the arctiid , Diacrisia casignetum. Entomologia Experimmtalis Applicata 37:193-198. FOLIN CIOCALTEU& . O , . 1927n tyrosinV ,O . d tryptophanean e determination proteinsn si . Journal of Biological Chemistry 73:627. HANOVER, J. W. 1975. Physiology of tree resistance to insects. Annual Review of Entomology 20:75-95. HANOVER, J. W. 1980. Breeding forest trees resistant to insects. Pp. 487-511 in Maxwell, F. G. & Jennings, P. R. (Eds.) Breeding Plants Resistant to Insects. John Wiley & Sons, New York. JAIN , ROYCHOUDHURYA. , JOSHI. & 1995C . , . N ,K , Population outbrea f defoliatorko , Hyblaea puera Crame skeletonizerd an r , Eutectona machaeralis Walke teakn ro relation i , o leant f chemical status. Myforest 31:49-54. JAYARAJ, S. 1967. Effect of leafhopper infestation of carbohydrate and nitrogen in castor varieties in relation to their resistance to Empoasca flavescens (F.) (Homoptera:Jassidae). Indian Journal of Experimental Biology 5:156-162. LOWRY, ROSEBROUGHH. . O , RANDALL& , FARR. J. L . . . 1951,J N . A , ,R . Protein measurement wite th h folin phenol reagent. Journal of Biological Chemistry 193:265-275. MATSON, W. J. & SCRIBER, J. M. 1987. Nutritional ecology of insect folivores of woody plants: water, nitrogen, fibrminerad an e l considerations . 105-14.Pp Slansky6n i Rodriguez,J.(Eds.& . ,F ) Nutritional Ecology of Insects, Mites Spiders.& John Wile SonsYorky& w Ne ,. McMANU , HASLAMLILLEY& J. . S H . . 1983,T E , . Plant polyphenol theid san r associations with proteins. Pp. 123-137 in Hedin, P. A. (Ed.) Plant Resistance to Insects. American Chemical Society, Washington. NORRIS, D. M. & KOGAN, M. 1980. Biochemical and morphological bases of resistance. Pp. 23-62 in Maxwell, F. G. &Jennings, P. R. (Eds.) Breeding Plants Resistant to Insects. John Wiley & Sons, New York. ORTMAN, E. F. & PETERS, D.C. 1980. Introduction. Pp. 3-13 in Maxwel, F. G. & Jennings, P. R. (Eds.) Breeding Plants Resistant to Insects. John Wiley & Sons, New York. PORTER, HEMINGWAYL.J& . , R.W. 1989. Significanc condensee th f eo d tannins . 988-102.Pp Rowe7n i , J. W. (Ed.) Natural Products of Woody Plants II. Springer-Verlag, Germany. 1243 pp. REESE . 1979C . . J , Interaction f allelochemicalso s with nutrient herbivorn si e food . 309-33Pp . n 0i Rosenthal Janzen & . (Eds.. H A . . D ,) G , Herbivores : Their Interactions with Secondary Plant Metabolites. Academic Press Yorkw Ne , . ROYCHOUDHURY, N..JAIN, A., JOSHI, K. C. & GUPTA, B. N. 1995a. Concept of natural resistance of trees insectso t . Advances Forestryn i Research Indian i 13:210-220. 226 Journal of Tropical Forest Science 12(2):221-226 (2000)

ROYCHOUDHURY, N., JAIN, A. & JOSHI, K. C. 1995b. Altenltion of growth and development in leaf skeletonizer, Eutectona machaeralis Walker, due to variations in teak leaves of different maturity. Indian Journal of Experimental Biology 33:227-229. ROYCHOUDHURY, N., JAIN, A. &JOSHI, K. C. 1995c. Resistance in teak clones against leaf skeletonizer, Eutectona machaeralis Walker (Lepidoptera:Pyralidae). Advances in Forestry Research in India 13:140-157. ROYCHOUDHURY BHARGAVA & , JAIN , N. TOSHI. , C A. . . . K 1995d, A , . Insect-host interactions: rolf eo * * J * » J * ' polyphenols. Tropical Sylvics 3:1-2. ROYCHOUDHURY JOSHI . JAIN, 1996& C . N. , , A ,K ,. Searc r naturahfo l resistanc tean ei k clones against Eutectona machaeralis Walker (Lepidopter aPyralidae): . Indian Journal of Forestry 19:205-213. SAXENA . 1986C . R ,. Biochemical base f inseco s t resistanc n rici e e varieties . 142-15Pp . Green9n i , M. B. & Hedin, P. A. (Eds.) Natural Resistance of Plants to Pests: Role of Allelochemicals. American Chemical Society, Washington. SINGLETON, V. L. 1974. Analytical Fractionation of the Phenolic Substances of Crapes and Wine and Some Practical Uses of Such Analysis. Advances in Chemistry Series 137. American Chemical Society: 184. SMITH, C. M. 1989. Plant Resistance to Insects : A Fundamental Approach. John Wiley & Sons, New York. 28. 6pp TEWARI . 1992N . MonographA D ., Teakn o (Tectona grandis Linn.f). International Book Distributors, Dehradun. 479 pp. UTHAMASAMY, S..JAYARAJ SUBRAMANIAN& . S , . 1971R . T ,. Studie varietae th n so l resistanc f bhendieo , Abelmoschus esculentus (L.), to the leafhopper, Amrasca devastans (Dist.) (Homoptera: Jassidae). South Indian Horticulture 19:53-59.