1. 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 PALAE
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.Theincreascof
concentration 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 METHODS
Mycelial 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

3. 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 of
enzylne lvere incubated at 32 + lo C lbr lhour. The reaction was stopped by addirrg 2.0 ml of
alkaline copper reasent' The tubes were kept in a boiling water bath for'30 rninLrtes. ihe control
treatluetrts were carri*d otrt in the same manrler except was added after aclcling the alkaline
copper reagenl. tr-he tubes were tlten cooleil to rooni temperature (32 +loC) ancl l.0rnl of
ArsenonrolybCate reagent was added and then it rvas read at -(30nrr in uv 260 ShirnadazLr
spectrophotoirreler.
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 the
nrethod of Sreekantiah (197 6).
20rnl of the pectin rrediurn was supplernented rvith lo/oto 5ok (w/v) of groundnut meal
eacii 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 the
nledium in a sterile chamber. The rnediunr containing the fungus *u. !ro*n for l6 clays. After l6
days" 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. All
collcentration of groundnut meal from 0.05% to 0.5% increase the mycelial growth where as
endoPG activity is not significant. Protein content significantly increased Ay O.ZN to 0..5% at
0.1% level.
Data presented in Table I reveals the
activity. It indicates that there is a decrease in
groundnut meal increases.
effect of groundnut meal on endopG
EPG 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, for
researclr grant.
REFERENCES
l' Ayers,W.A and Papavisas,G. C. 1955. An exocellular pectolytienzymes of Aphcntontvces
c ute i c hes " Plrytopathologt, 5 5 : 249 -253.
Bulletin of Pure and Apptied Sciences.Vol.2jB(No.t)2004
59

4. r15
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 Pvt
Ltd.. New Delhi : 27 4-297 .
3. Cooper,R. M. and Wood, R. K. S.1975. Regulation of synthesis of cell rvall degrading
enzymes by Verticilliunt alboatrum and Fusarium oxysporum fsp.lycopersici. Physiol Pl
Puthol.5: 135-156.
4. Grant 1985.The effect of Pectin and Related Compounds <ln Encystment and Germination of
P 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 Protein
nreasurement with the Folin phenol reagerft.J Biol Chem, 193:265-295.
6. Nelson,N. l944.APhotometricadaptationofthesornogyi methodforthedeterminationof
Glucose. 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 of
strains 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, University
of Mysore
Bulletin of Pure and Applied Sciences.Vol.23B(No.1)2004
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