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J.Ecoroxicol.Environ.Monit.l3(4)249-254(2003),r t@ Palani Paramount Publications-Printed in India d

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ISSN: 097 1 -09 65-13-03-249

INDUCTION OF C C C ON GROWTH AND TUBERISATIOI{OF COLACASIA ESCULENTA

B ANNADURAII, S CHITRAI, K GOVINDARAJU 2 AND C RAJALAKSHMI 'l DEPARIMENT OF PLANT BIOLOGY AND BIOTECHNOLOGYC A H COLLEGE, MELVISHARAM, VELLORE 632 509 TAMIL NADU, INDIA

,DEPARTMENT OF ZOOLOGY THIRUVALLUVAR LINIVERSITY, VELLORE 632 OOl

TAMILNADU,INDIA

ABSTRACT

Effect of (2 chtoroethyl) trimethyl ammonium chloride(CCC) on the vegetative growth

and tuberisation of Colacasia esculenta (Linn.) and estimation of carbohydrate(RS, NRS, S) was

undertaken. C. esculenta was obtained from Vallambadugai near Chidambaram town and they

wereplantedinpots.TwodifferentconcentrationsofCCCviz.250and500ppmwereusedinthestudy. Areal spray was done at an interval of l5 days. Totally 5 sprays were given. Observations

were made on the vegetative growth and the corm yield. It was observed that there is a marked

reduction in the vegetative growth and increase in the contents of reducing sugar, non reducing

sugar and starch of treated plants. There is not much difference between 250 and 500 ppm

treatrnents.

Key words: CCC, Colacasia esculenta, Growth, Tuberisation, Starch, Induction.

INTRODUCTION

CCC is known:as Chlormequat. (2-chloroethyl) trimethyl ammonium chloride,

cyocoel and chlorocholine chloride. The structure of CCC is:

CHeI-

cH3-----T ----c2Haci

CH:

Its molecular weight is 158.1 Kd. It is soluble in water, decomposed in aqueous

solutions, stable but corosive to metals. It is anti gibberellin growth retardant. Analogue

of choline, used as a height shortened in wheat and poinsettias sugarcane ripened and

to reduce bulgiag in cereal grains (Dawson et al 1984). Application on rosette plants

reduces the endogenous levels of GA content. Plants of Samolus parttiflorus require

eight long days for flowering. Ifthese plants are treatedwith CCC fail to flowerbut on

the application of GA2 it is made to flower again. In the same way in almost all plants" '

!. .'' '

B ANNADURAIETAL

application of CCC flowering was suppressed due to the suppression of giberrellin

ii;ryr,rr".is (Riddell et al tiaz1. This-is also observed in curcurbits. When CCC is

applied before a weak of flowering resulted in the abortion of all developing fruits' It

was also observed CCC decre".., th. ethylene production (Leetham et al 1978)' This

may be due to reduction of auxin and dibenellin content in plants' Application of

ccc delays s€nescence ht Phaseolus wlgais.In the places of scarcity ofwater, wheat

plants die a week before than the ccc treated plants- In coleoptile of oats, ccc causes

irrt iuition of growth, reduces auxin content, inhibits GA synthesis and kaurenene

sinthetase enzyme, ccc are employed to restrict the vegetative growth of apple' In

o-u*"rrtul plants CCC application riduces vegetative growth and makes young plants

to bloom (Krishnamoorthy l98l; Annadurai lg75). In view of the importance of

colacasia esculenta is a perenial tuberose plant with large heart shaped leaf blades'

borne on long petioles 0.j-2 m, high arising from a group of underground farinaceous

.or.r, this ilnvestigation tr"s 6"i undertaken. The corms vary greatly in size and

shape, ihe central ones being big and conspicuous'

, tnt'AfERIALS AND METHODS

t. *" plant colacasia esculenta Linnaeus which was taken for study was purchased from

Vallambadugai where it thrives well and parts of the corn are purchased at Chidambaram vegetable

market. These corms were cultivated bothin pots and plots. In fots, lrst they were grown separately'

After it has reached the minimum amount of growth ii was separatef into three lines' First one is for

cbntrol line, secghd is for 500 and 250 pp* oi ccc was sprayed atlhe interval of 15 days once' The

selected plants were planted in well prepared pots in the -Botanic

Garden of Annamalai University'

Tamil Nadu, India on November Nn (Uonsoon period). Control pots consist 500 and 250 ppm pots'

each 6 pots and irrigation was done at proper intervals' )

CCC spray: 2 Chlora ethyl trimethy ammonium chloride 500 ppm was prepared in2Yotepal solution'

First spraying *as given after 45 days after planting. Control plants were sprayed with 2% tepal solution

only. No soil application of CCC was done. Aerial spray to the vegetative parts alone was done at an

interval of 15 days. v"f"tuti"" growttr and the estimation of reducing sugar, non-reducing sugar and

starch were measured at Proper intervals of 15 days'

EstimationofreducingsugarsandcarbohydrateswasestimatedaccordingtoPhenolsulphuricacid calorimetric method Dubois er al (1956), Non-reducing sugars was done according to the procedure

of Nelson (1941) and somogyi (1952) and starch is by the method of Macrae (1971)'

Procedure for extract: One hundred gms of oven dried material was taken' It was extracted with 80%

alcohol. The alcohol extract was evapolrated to 10 ml on a water bath and cooled it to room temperature

(32+l"c). Then 5 ml of satiratedneutral lead acetate is added to precipitate the proteins' Above 100 ml

of saturated aqueous disodium phosphate was added to precipitate the excess of lead' About 300 gms

of activated carbon was added. it wai shaken well at an interval of 30 minutes and filtered' The filtrate

.JournalofEcotoxicology&EnvironmentalMonitoring.Vol.13(2003)

INDUCTION OF C C C ON CnOWTU AND TUBERIZATION OF COLACASIA ESCULENTA

diluted to 100 ml in a volumetric flask. This solution is used for estimation of rcducing sugars and

Non-reducing sugars. The residue left after alcoholic extraction was taken for starch estirnation.

Estimation of reducing 6ugar: lstimatiop of reducing sugars and carbohydrates was estimatedaccording to Phenol sulfuric acid calorimetric method of Dubois et al (1956).

Non-reducing sugar: Non-reducing sugars was measured according to the procedure of Nelson(1941) and Somogyi (1952).

\Estimation of starch: Starch was estimated based on the method of Macrae '(197 l).

Statistical procedure: The mean (i)and the standard deviations (o) were calculated from the

determined values by using the standard procedure. In order to examine whether the difference inresults obtained was significant or not the student's test was employed. The level of significance (P-

value) between x, and x2 was determined by using the students 't' distribution table of ftactiles and

critical values.

RESULTS AND DISCUSSION

Changes occurred on the number of leaves, length of leaves and petiolar lengthand breadth of leaves after the aerial application of CCC from I month of applicationto 5 months of application. Hormonal role in signal transduction and variousmetabolism is of greator interest (Crapo 1985). They are produced in very minutequantities in the growing regions and translocated to the region of requirement. Inplants there is no specialised glands as seen in animals. They are now regarded as

phytohormones or growth regulators. IAA, GA and Cytokinins are courmonly presentin wide range of plants. Following th€ir diseovery and wide range of their commercialapplication in tissue culture and bioteehnology (Norman & Litwack 1987) so manyderivatives like NAA, IBA, IPA, GAI to€A23 and retardants like morphactins, AB4.Ethylene, MH, CCC, Alar 85, AMO 1618, Phosphan D, Coumarin, Scopoletin and pq-

Ascorbic acid have been added to this branch of hormonal chemistry. Tolbert (1960)has recognized a group ofquaternaryammonium compounds, which can act as growthretardant. Cathey (1960) has reviewed various growth retardant. 2-chloroethyltrimethyl ammonium chloride was the most active and extensively used growthchlorocholine chloride.

Table 2 reveals the effect of CCC on the reducing sugar, Non-reducingsugar and starch contents of C. esculenta.

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Journal of Ecotoxicology & Environmental Monitoring. Vol. 13 (2003)

I Month II Month III Month IV Month V Month

c 250 500 c 250 s00 cppm ppm ppm ppm

2s0 500 c 250 500

ppm ppm ppm ppmc 2s0 s00

ppm

444

333

966

32.6 29.7 28.

17.2 74,6 13.

t4.4 13.2 ll.

46.8 45.2 44.

9.7 8.2 8.0

Nurnberbf 4pots

Number of 3'plantsNumber of 8

leavesLength of 23.7petioleLength of 14.9leafBreadth'of 10.8leafWeight ofthe cormLeafweight

44444444444

33333333333

66866966966

2t.8 20.7 24.5 '.r3.4 22.8 24.9 23.8 229 28.5 26.8 23'4

10.2 9.8 15.8 11.7 10.8 15.9 12.6 11.8 16.6 l3.4.lZ'9

9.g 8.1 12.8 11.3 8.7 13.5 11.9 9.5 t3.8 12.7 10.2

252 B ANNADURAIETAL

Table 1 Effect of ccc on growth and tuberization of c. esculenta

c = contol; values given are the mean value (X) ! 6 values; d.f. : degrees of freedom : n-11

Significance 11- : p<0.001; + : p<0.05; NS : Not significant

Table 2 Effect of CCC on the RS, NRS and S content af C. esculentaLrna'.

Reducing sugar Non-reducing sugar Starch

c 250 500 c 250 500 c 250 500

% Month s.26 6.15 7.33 26.87 30.84 36.70 5642 6245 7560

C : Control; Values given are the mean value (I) * 5 values; d.f. : degrees of freedom : n-1;

Significance ++ : p<0.001; + : p<0.05; NS : Not significaut , '

The present work on corms of C. esculenta sp. were obtained from

Vallambadu gai; a nearby village of Chidanibaram town in the fields of Botany

departrnent. ihese materials were planted in well-prepared pots in the Botanical Garden

of the Botany department. First set ofplant has kept as control while the other two sets

were subjectedto aerial spray of 250 and 500 ppm. Results clearly indicates that there

is a reduition in the vegelative growth of the treated plants. It is also noted from the

Journal of Ecotoxicology & Environrnental Monitoring' Vol' 13 (2003)

dr rnnn rz ^ rr^\i AE r1/-r ' ill

INDUCTIoNoFCCCoNGRowTHANDTUBERIZATIoNoFC2LACASIAESCULENTA')

tabte 1 that there is a difference'between on the vegetative growth on the application

of 250 and 500 ppm CCC treated plants' /

Finally, the ccc treated plants removed from the pots were weighed indicfes

that there is a marked retardation in the vegetative growth, which might have influ#ced

the corm formation. rnis retarding effect is similar to the results obtained by Srinivasan

(1g75)on sweet potato and by .qoar", Peter (1975) on curcuma longa and Riddell er

al (1962).neducinf sug*, non-r"ducing sugar and starch content (Table 2) estimated

was found to be encouraging ana givesldea on the mechanism of tuberization'

In underground tubers, bulbs and corms it is necessary to store and transport

throughouttheyearforcommercialpurpose.slgrins.withrefrigeratorandlAAand NAA become a cbstly irruo.Hence c-cc at 250 and 500 ppm concentration is

tried. In ornamental and Lofticulture plants vegetative growth has to be suppressed

and control by pruning is usually has been dott*. Hence, application of ccc in

controtling vegetative growth has been suggested here'

, OCKNOWLEDGEMENT

TheauthorrecordadeepsenseofgratitudetolateProfessorDr.K.RangaswamiAyyangar,Head of &e Department of Botany, err,u*utuiuniversity for his kind suggestions of this project work

and indebted to lute prof. C. Srinivasan, Retd. Head, Departnent of Botany' Annamalai University for

their guidance'

REF'ERENCES

Andren Peter M and Pannerselvam S 1975 Effect of ccc on growth and tuberisation of curcuma

longa,M.Scdissertation, Annamalai Universibz' ^ -^t-^^^:- ^-^.,to-tn t int

Annadurai B 19-75 Effect of ccc on growth and tuterization of colacssia esculenta Linn' M'Sc'

--- dissertation, Annamalai University'

cathey H J 1964 Physiology of growth retarding chemicals' Ann' Rev' Plant Physiol'1 : 271-302'

Crapol 1985 Hormones' Freeman, London'- .,1 n r^<( n,Dubois M, Gilles K A, Hamilton J K, Rebers P A and smith F 1956 Phenol Sulphuric acid calorimetric

rnethodforestimationof'reducingsugarsandcarbohydrates.Anal.Chem28:350.Dawson R M C, Elliot W H and Jones f M iSS+ Data for biochemical research, 3rd ed' Clarendon

Press, OxfordKrishnarrioorthy te8i Plant growth and substances including application in agriculture' Tata'

McGraw-Hill Publishiig Company Limrted' New Delhi'

Leetham D.S, Goodwin P B and Higgins T J (eds) 1978 Phytohornones and related compounds: A

comprehensive treatise. vot. tt. Phytohormones und the development of higher plants' Elsevier

i North Holland, Amsterdam, Oxford' New York' p'648'

Jour-nal of Ecotoxicology & Environmental Monitoring' Vol' 13 (2003) '

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254 B ANNADURAIETAL

Macrae J C 1971 euantitative measurement of starch in very small amounts of leaf tissue. Planta 96 :

101.

NelsonN tg44 Aphotometric adaptation of the Somogyi method for the determination of Glucose' J'

Biol. Chem. 153 : 375-380.

Riddelt J A, Hageman H A, Anthony c M J and Hubbard w I 1962 Retardation of plant growth by a

new group of chemicals. Science 136 : 391'

somogyi Ia tqsz Notes on sugar determination. J. Btol. chem. 195 : 19-23.

Srinivasan p.and panneersetvam S 1975 Effect of CCC on growth and tuberisation of sweet potato'

M.Sc dissertation, Annamalai University'

Tolbert N E 1960 (2-Chloroethyl) tirnethyl-ammonium chloride and related compounds as plant growth

substances, I, Chemical structure and bioassay. J. Biol. chem. 235 475-79.

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