B ulletin of Pure ancl Appliett Sciences.Vol.23 B(No' I ) 2004 : P' 5 7' 60'

EFFECT OF GROUNDNUT MEAL ON PR'ODUCTION OF

f,NDOPOLYGALACTI.IRONASE AF A LTE RNARIA CE PA LAE

B. Annadurai , N. Nandakumar " and D.B"Motlag

Department of Plant Biology and Biotechnology, C'Abdul Hakeem College,

Melvishararn -632 509, Vellore District, 'Iamilnadt', India

ABSTRACT

Effect of various concentration ol'Groundnut in Pectin mediunr on the grorvlh of

AlternariacepttlaeandproductionofenitoPGwasgstinrated.Theincreascofconcentration of groundnirt nreal increases the growth and decreases the endol'G

Production'

Key words: A.<'epulae' Groundnut mcal. IintloP[i

INTRODUCTION

Endopolygalacturonase (Poly, o, I.4 galacturonide glycanohydrolase, ( EC 3.2.1 ' l5) plays

a significant - -role ilt path,rgenesis of nany plant diseases (cooper and

WooajS7S).gndopolygalacturLrnase is one of the prime lnacerating erzyme produced by

Altc:rti,ariu cepu!,*...iu.iiig lccfbligirt disease of onion ( Ponnappa et al' 1977)'

Endopolygalacturorlase is proiluced both constitutively and adaptively by different

nticroorganisms. Ii most instanees. tie enzymes are produced adaptively rather than costitytively'

Fuslrriu"nr moniliforme ant) Scleroliuu, ,rilltii produce it adaptively in the presence of pectic

strbitrates ( Bilgrarni,lgS2 ). A snrall number of pathogens like Piriculariafi-lometosct ( Ayers and

papavizas,'l96i) and Collitotrichutnful,cgtnm (Singh and Hussain,l964). Verticillium alboqtrunt

at'td Aphanornycqs ctttcices ( Ayers arrd Papavisas,l965 ) are ktlowtl to produce polygalacttrronase

in a coustitutive manner.

,

Pectin induces greater enzyme production in adaptive enzymes (crant, 1985).Tlte

pathogen i,i i;trr" confrints witS pectin iubstances npt in isolation, ,but

in cornbination with

other*carbohydrates. These carboliydrates are report{d ,to .control the production of pectic

enzyrxes (Ivlaldona,l986). Catabolic repression of the synthesisbf pectic enzymes by sugars is a

.o,lrn1on ph.no,r,.no,, in many phytopatliogenic organisrns(Cooper and Wood'1975)'

The endopG levels in infected opion leaves and in pectin rnedium were ascertaitled.

MATERIALSAND METHODSMycelial dry weight determination

. After 16 days of inoculation of Alternaria cepulae in different physicoci -.,rical

environrnent the contents of the erlenmeyer flask was filtered through a glass funnel fitted with a

coarse grade sintered glass filter and waihed thoroughly with water' The mat was pressed in filter

paper tJ remove the eicess of moisture. This was transferred to a previously weigheci filter paper'

It was dried in an oven at70"c overnight. It was cooled to room temperature ( 32 t l0 oc) ilt a

desicator and weighed.

*Depaftment ofBiotechnology, Central Leather Research lnstitute, Adyar, Chennai-600 020' India

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B. Annadurai , N. Nandakumar and D.B.Motlag

Bulleth of Pwre and Applied Sciences.Vol.23B(Na.l)2004

58

Effect of groundnut mear on production of endopolygaracturona se of Arternaria ...

Estimation of endopolygalacturonase

EndoPG activity was estimated by reducing sugar rrethod according to Nelsorr (1944)and Sornogyi ( 1952)"

The incubation mixture containing 1.0 ml of Sodiurr polypectate at pH 5.0. 0.5 nrl ofenzylne lvere incubated at 32 + lo C lbr lhour. The reaction was stopped by addirrg 2.0 ml ofalkaline copper reasent' The tubes were kept in a boiling water bath for'30 rninLrtes. ihe controltreatluetrts were carri*d otrt in the same manrler except was added after aclcling the alkalinecopper reagenl. tr-he tubes were tlten cooleil to rooni temperature (32 +loC) ancl l.0rnl ofArsenonrolybCate reagent was added and then it rvas read at -(30nrr in uv 260 ShirnadazLrspectrophotoirreler.

Estimation of protein

Protein in the culture filtrate was estir:rated by errploying the rnethod of Lowry et ctl( 195 l) using crystalline Bovine Serum Albumin as standard,

Inlluence of groundnut meat on Endo pG activity

The influence of groundnut meal on Endo PC activity was carried out accorcling to thenrethod of Sreekantiah (197 6).

20rnl of the pectin rrediurn was supplernented rvith lo/oto 5ok (w/v) of groundnut mealeacii irr loOrnl Erlenmayer flasks. The flasks were sterilized at l5 lb pr.rr,,r. for 20 mirrutes.T'hen they were cooled to room temperature at 32 i luc. The fungus was inoculate into thenledium in a sterile chamber. The rnediunr containing the fungus *u. !ro*n for l6 clays. After l6days" the enzyme preparation was done using the same procedure desciibecl above.

RESULTS AND DISCUSSION

The effect of groundnut meal on Endo PG activity is indicated in Table-1. Allcollcentration of groundnut meal from 0.05% to 0.5% increase the mycelial growth where asendoPG activity is not significant. Protein content significantly increased Ay O.ZN to 0..5% at0.1% level.

Data presented in Table I reveals theactivity. It indicates that there is a decrease ingroundnut meal increases.

effect of groundnut meal on endopGEPG activity when the concentration of

ACKNOWLEDGEMENT

^- TIre author B.A is grateful to Late Dr.S.C.Dhar Scientist at the Departnient of Biorechnology,

CLRI, Chennai , for Laboratory facilities and useful suggestion and UCIC, Nerv Delhi, forresearclr grant.

REFERENCES

l' Ayers,W.A and Papavisas,G. C. 1955. An exocellular pectolytienzymes of Aphcntontvcesc ute i c hes " Plrytopathologt, 5 5 : 249 -253.

Bulletin of Pure and Apptied Sciences.Vol.2jB(No.t)2004

59

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t.*,

B. Annadurai , N. Nandakumar and D.B.Motlag o , t

2. Bilgrami ,K.S and Verma, R. N. 1982. Physiology of Fungi, Vikas Publishing House PvtLtd.. New Delhi : 27 4-297 .

3. Cooper,R. M. and Wood, R. K. S.1975. Regulation of synthesis of cell rvall degradingenzymes by Verticilliunt alboatrum and Fusarium oxysporum fsp.lycopersici. Physiol PlPuthol.5: 135-156.

4. Grant 1985.The effect of Pectin and Related Compounds <ln Encystment and Germination ofP hy t op t hora pal miy ora zoospores. J G en Micro b iol,l 3 | :669 -67 6.

5. Lowry, O. H., Rosebrough, N. J., Farr, A. L. O. and Randall, R. J. 195 l. J Proteinnreasurement with the Folin phenol reagerft.J Biol Chem, 193:265-295.

6. Nelson,N. l944.APhotometricadaptationofthesornogyi methodforthedeterminationofGlucose. J Biol Chem, 155 : 375-380

7. Ponnappa, K.M., Anil kumar, T.B, Sulladmath ,V.V. and Hiremath, P. C. 1977.Polygalacturonase production by Alternaria cepulae,the causal agent of leaf blight of onion.IndJ oJ l{ycol and Pl Pathol, l:6J.

8. Singh,G.P and Hussain, A. l964.Relation of hydrolytic enzyme activity with virulence ofstrains of Col/e totrichum falcatum.P hytop atholo gt, 54 : I I 00- I I 0 I .

9. Somogyi, M. 1952. Notes on Sugar determi nation, J. Biol.Chem,l9l :19.

10. Sreekantish, K. R. 1976. Studieson Pectin degradingstrain of Fungi, Ph.D thesis, Universityof Mysore

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