Das & Bajpai - Growth Patterns and Light Induced Kinetics of the Fungi C. sacchari on Agar...

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I

Indian Joual of CemstryVo139A, July 2 pp 697-702

Growth pattes and light induced kinetics of the fungiCphalosporium sacchari on agar pates

Iswar Das* & Alpana 8ajpaiCemistry Department, DD U Goakpur University

Gorakpur 273 9, India

R 15 Dmb 1999; s 1 Ma

Grow pattes and gowt kinetics of te fungi haosom saha on agar surface ave been investigated. emycelium offungi grows simi ar to noniving systems e grow pates ave been deveoped on diffeen cuture media ekinetic data obey te simple equation mt + were m and are sope and intecep respecivey and t is te time. Te growwas found to be lig induced Dependence of of media on morpology and grow of te microorganism as been studiedo simlar and dissimilar myceia discs ave been inoculaed togeter at a fxed distance and te infuence on pates ave

been studied Te contact zone as been caacterised by a lack of spores abbeviated as inibiion pate. Te suppessedspouaion at te contac zone is due to competition over nurients

Chmia aion a om quiibium may poduining mpoa and paia u uh a hmi-a wa and piodi pipiaion po 1 1 1 , oiaoy hmia aion and mu ip aionay a 12 .Piodiiy ha bn obd in a ga aiy uh ahyhmi band in aga, onni ay in gaon,piodi dpoi ion in poymi ym, andhyhmiiy i n baia gowh 1 Th po o hyhmi gowh and poduion i ao impoan oogiay. Maoopi onaion pan in uu oiamnou ngi a known and pod in h ia-tu 1 Fungi a o onidab in du o hiu in mdiin inuding h ynhi o iaminanibioi, poduion o pai maia diono ogani wa and u iiaion o ungi a ood 1 andpoing o ood podu Fiamnou ngi d ohi abiiy o ag amoun o poin a dxniy o h poduion o induia nym .In paia onni ing and pia ha bn pod o tain ungi .g. Neceria ciabaria adPeicilliu dieruI7IK imia o ho obd innoniing ym. In iw o hi wid aiy o appiaion, in in nonquiibium gowh pa and imiaii wih baia gowh, w pot inh pn pap h mophoogi and owh o hngi Cephalopori acchari whih ha anom h iw poin o tain dia o ugaan adin xpimna ondiion inding H o hmdium, igh and inaion o oh myim pnin i nighbouhood.

Materials and Methods

Prepario of culure edia : Fi din mdia wppad wih h oowing ompoiion (i ) poaodxoaga (PDA) mdium wa ppad by boiing poao (250 g) in wa onaining dxo (20 g) andaga aga ( 5 g) ; ( ii) omaodxoaga (TDA) m-dium onaind omao (250 g) in pa o poao, ohingdin w h am; (ii i) mai mdium onaind

mai(60

g) and aga aga(20

g) in wa; ( i) apkmdium wa ppad by dioing h oowing inwa NaN03 (30 g), HP0 ( 0 g), MgS07H(05 g) KC (05 g), FS0.7H (00 g), uo(30 g) and aga aga ( 5 g) H wa adjd o 7.and () oa ma aga onaind oa ma (60 g) andaga aga (20 g).

A ingdin o abo mdia w mixd hooughyand iid in an auoa o 30 minu A i-iaion, mdia w aowd o oo, anibioi pnii -in (ampiiin ) wa addd o ah mdium and podino pi dih.

Deelope of growh pae i dere ediaDi o 40 mm diam w u om h my-

ium. Eah di wa and o a ga pidih (90mm diam) onaining 20 m!. o mdium and pu inan inbao mainaind a 300 "c. Expimn wpomd in ipia. Gowh pa obd whown in Fig .


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698 NDAN CHM SC. A, UY 2

a b c

g - Gro pattes ofCehalosoum saha developed on different media at 37 "Ca) Capek medum (b) PDA medum; and )TDA medium

Table - Values of sope m), interep (c and oreation oeffiient R) used in te equaion = m t obtaned at dfferen expemental ondtons a 37C

Medium H Slopem)m/day

Poato dextrose agar agar 39

Tomao dexrose-agar agar .357

Capekagar agar .334

Maize-agar agar .332

Oa mealagar agar 68 .3396* .5586.8* 69 * .576

*Observations aken at 3 C

Gwth kieticAe inocuion o ungi Cephaloporium acchari

ino dieen medi, gowh kineic wee udied wihhe hep o veing micocope ccue o 0.00cm. Reu e ecoded in Fig.

uece of p o the morphology ad growth kieticInuence o pH o he medium on gowh pe

nd kineic h een udied. OMA medium w e-leced o hi pupoe Expeimen wee cied ou on

Inerep Correlaon(c oefient R)m

-8 996

. 8 998

-9 998

53 998

993384 99-.4 996-484 .995

OMA medium o vioupH vi .96, 6.8, 80 nd 1 0pH o culue medium w meued wih digil pHmee. Rdii o he cicul gowh envelop w me-ued dieen ime inev wih he hep o eling micocope nd gowh pe e ecoded in Fig nd 4.

uece of LightLigh i oluey eeni o he omion o vi

ou ype o epoducive ogn To udy he inluence


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DAS et al. GROWT KINETCS OF Cepalosporim saari ON AGAR PLATE 99

[

E

'0

2.0.(

.0

0.02

3 5

6 7

8 9

Ii a

ig 2 - Gw kinics daa fr Cepaosporm saar n diffrn mdia.[TDA [ PDA [OMA [ Maiz aga I Czapk mdia

of light o the gowth ehaviou ad patte of th fugi,TDA medium wa take ito peti di he iouated adkept i omplete dake, while othe peti dih wa

kept i the peee of vi ie ight at the ame tempeatue. Gowth wa meaued employig the uual teh-ique a deied ealie. Reut ae eoded i Fig. 5.

hibitio patteTo tudy the depedee of hypha gowth ad dif

feetiatio o adjaet hyphae, iteatig hyphae aeexpeimetally geeated y hooig two ioulatigete at a fixed ditae Simia ad diiia mi-oogaim di wee ioulated o a plate at a 30m. apat. Gowth patte wee oeved ate 7 dayad photogaphed. Reult ae eoded i Fig.6

Results and Discussion

We have examied the oe of expeimetal oditio with epet to maoopi patte fomatio ofthe gi Cephaloporium acchari o aga pate otaiig ( i potato ad dextoe (ii tomato ad dextoe(iii NaN03, K2HP0, MgS0.7H FeS0 .7H, KCad uoe (Capek' medium(iv maie ad (v oat

meal. The media otaied eetia ly ao oue admal amout of iogai utiet. Cao ompoudeve two eetia tio i the metaolim ofgi:(i t upplie the ao eeded fo the ythei of po-tei, ue i aid, el wal memae, eeve food et ad (i i the ole oue of ppeiae amout of eegyi the ox idatio of ao ompoud additio to a-o oue, etai iogai mioutiet ae alo equied depedig o the utiet upplied. Fugi apodue a geat vaiety of ompex oai ompoudTypial gowth patte deveoped o PDA, TDA, adCapek aga media ae how i Fig . 1 . Duig the myelium developmet, utiet ae take up ad fuguhage the io i ad moleula pofile i the uoud-ig medium Peiodiity i oeved i amot al ae.The gowth ou ieay a evidet y the gowth kieti eult how i Fig. . The data oey imple em-piial equatio d = m t +c whee d i the di tae othe hyphal tip fom the ete (adiu of the iulaevelop whih vaie with tie t ad c i the iteeptof the taight ie o the y axi Vaue of ope (mieept ( ad oelatio oeffiiet (R fo diffeetmedia ae eoded i Tale 1 Figue how the e


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7 NDAN CHEM SEC A JULY 2

c

Fg 3 Gowth atte of ehalospom saha develoed on OMA late contanng meda of dfferentp a 3C p [] 39 , [ 8 and [c)

T la oys)

F' 4 - Growh knetc data for ephalosom saha onOMA late contanng meda of dfferent [0 ] 39[] 8 ] I

suts obtained on different culture edia Results recorded in Tabe shows that the growth is axiu inPDA ediu

Growth patts at different H of the OMA ediuare shown in Fig 3 Plates i-iii show the orphology at a

lower H vaue 39 nral value 8 and t rlatively higher vue 0 respectively Dense radal lecolony is observed at ow and noral H (plates a,b)while sharp trnsition to rhythic patte is noticed atH (plate c). It grows best in th

e ediu

having Hin the noral range and declines the aCdC 3.9 orin alkaline ediu (H .0). Growth kinetic data atdifferent H are recorded in Fig 4. The data obey thesiple relation +c as evident by corelation co-

4

3,

' 2

iu

,

ie (td

Fg 5 Growth knetc data forephalospoim sahain reence of lght [0] and n the comlee darne [ ]at 3C

efficient values The values of m and c depend on H ofediu Results are recorded in Table 1 .

Growth kinetics of the ngi Cehaooriu accharion DA ediu in the presence and absence of light

have been studied and results are shown in Fig5 Results show that the growth is linear and light induced.Further the zonation is observed in the presence of lightat the given experientl condition. It suggests that thefungus sporulats relatively faster in the light

When petridishes were inoculated with two similaror dissiilar ycelia at a certain distance on a petridishthey grow and eventually eet each other as shown inFig.6 Plates show the growth pattes when the two in


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DAS et al.: GROWT KNETCS O Cephalospoim sahai ON AGAR PLATE 701

c

ig nibiin parns Cephalospoim sahai n TDA mdium n inculad i a Cephalospoimsahai b Fsaim ospom and c) Cvaia pallesens a a xd disanc 3.0 cm

olating diss wee of simila pathogen, phalosporium sacchari (Plate i) o the two diss of dissimilapathogens phalosporium sacchari-Fusariumoysporum and phalosporium sacchari - urvulariapallscns espetivey (plates ii and i ii, Fig 6 The ontat zone is haateised y a ak of spoe de to ompetition ove ntients Plate (ii i) does not show sh aak of sporlation at a ontat zone, poaly thee isno sh ompetition fo ntients.

Inflene of additiona hypha on the deveopment ofa myelim was stdied. Inolating a petidish with twoo moe diss is giving ise to two o moe myelia gow

ing towads and eventally meeting eah othe The ontat zone is haateised y a lak of spoe (inhiitionpatte) . The sppessed spoation is de to ompeti-tion ove ntients.

Physical xplanation of ringtyp structurRings of this geneal type have een oseved in geat

variety whih inde periodi peipitation of spaingysolle salts ?, gowth of the ateria Pr vulgaris1 3 andseveal othes. Explanation fo the peiodi peipitation phenomena was gi ven y seveal wokes ased ona) the spesatation theory 9, ) ompetitive patilegowth mode and ) so oaglation mode Aoding to the spe satation theoy whih is most om-mon amongst the thee, the two eatants A and B dif-se fom opposite dietions and oexist in the gel nti the solility pod eahes a itia vae aove whihthe neation os aoding to the eation

A +B AB (solid)

One the nleation hs sted, a depletion of onentation of A and B os in the soding Heneat this stage, the formation of peipitate stops neation in the neighohood, peipitation i s also stoppedas diffsion ontines fthe. This is folowed y thefomation of lea spaes. Difsion of ions pesists andafte a ertain stage, the itia spesatation exeedsand anothe nd is fomed in a simila manne Thepoess is epeated nti the spply of one of the eetolytes is exhasted

n a moe eent vesion of the theory22 the pite ismodified The two speies A and B ae ssmed to eat

to pode a new speies C whi h aso diffses in to thegel. Sseqenty nlei ae fomed folowed y ys-tallisation aoding to the following sheme,

A + B CC nei (n)nlei rystal (D)

when the lo onentation of C ehes some thesh-old vale, nleation os and sseqenty D s-tes ae formed. Modifiation of the aove model wasmade y Poezhaev and Mle .

Coonies of Pr vulgaris wee also oseved to pos-sess onenti ings. Howeve, a diffeent mehanismwas sggested. The ateia whih i s normally spaselyflagellated ates i tself to aqie arge nme of flagean addition thee is essation of e divi sion de to inhi-ition of septm formation eading to formation of mti-nleate, densely flagelate, nonseptate ells . The gowthold also e thoght of pimaily as a onseqene ofntient diffsion and loa exhastion.


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72 INDIAN J CHEM SEC A ULY 2

Bacteria frst spread n search of nutrient contained

in the medium and would consume that nutrients not only

wthin the ci rcle of resdence but by creating a dfson

gradient, within a ring of certain thickness Witin thatring bacteria could not subsequently floursh. The proc

ess would then be repeated.

In te case offungi the growth occurs only at hyphal

tips A hyphal element arises from te out growth of a

single spore Upon germination the growt of te

Spe Germube

Densey banchedhyphal eemen

Aggmaae hyphal eemen

spore rapdly results n the formation of a tube wch

extends ts lengt by growth at the tip The germ tube

develops into a hypa, exceeds a certan length branches

are formed resultng in a hyphal element consisting of a

branced network Macroscopic morphology is deter

mned by the growth kinetics of the ndividual hypal

elment witin he pellets.

We have observed similar zonaton pattes duing

te fungal growth. Various ypoteses were tested witthe help of a cellular automation model wich mimicksgrowth brancing and differentiaton of a fungal myc

elium. A cellular automation is a teoretical concept tat

can be applied to certain spatially dstributed systems n

wc macroscopic behaviour results Differentiation

may be incorporated by means of various activator-inibitor mechanisms .

Acknowdgmnt

Tanks are due to the U.G.C, New Delhi for sup

portng the investigation Authors also wish to thank Prof.

N. B Sng Head Cheistry Department for providingnecessary faclities and Dr. G Rao Sugar Cane Institute Kuraghat, Gorakhpur for providing te patogens

and helpful d scussions

Rfrncs

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729.5. Das I Lal R S & Pushkaa A J hys he, 9 987 747. Das I Pushkarna A & Chand S J olloid iteae Si 50

992 78.7. Das I Das S S Pushkarn A & Chand S J olloid itea

Si, 3 989 7.8. Taanque Y J hem Edu, 71 ( I ) 994 589. Henisch H K ystals i gels ad Liesegag igs Cam

bridge Univ. Pess Ne Yrk) 988.1 . Muer S C Osillatios admohologies edited by L Rensing

Marcel Dekke NeYk) 993p 43.I Pezhaev A A & Mule r S C HAOS 4 994 3 .

1 2. Das I Lal R S & Pushkarna A J st Gowth 84 987 23 . 3 Das Sharma A & Sing U K Idia J ex Bio, 29 991

1 09 4 Nielsen J TIBTEH, 4 99 43815 . Gray W D The elatio offugi to hua aais Heny Hd

Ne Yk) 1 959.

Physiology of fugi edied by Y W Cchrane hn WieyLndn) 1958.

7 Deusch A Dress A ensing L Mehaism of develomt44 1993 17

1 8. Bure J A Lincn R G Carpenter B H Siee 9973.

9. Pager S J hem Phys, 25 1 95 2792 licke M&Rss J J hem Phys, 1 974 34582 . Dha N R & Chaeee A C Kolloid Z 39 925 222 Chpard B Luhi P & Dr M J Statist Phys, 7 994

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