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INTRODUCTION
Spices constitute an important group of agricultural commodities, which is
considered as low volume and high value crops. Spices sector is one of the key areas
in which lndia has an inherent strength to dominate the global markets and plays a
significant role in our national economy ( P ~ t h i , 1998).
The fame of Indian spices is older than the recorded history (7000 BC), which
elucidates the extreme fascination of the rest of the world for the fabled wealth of'
lndia - the variety of spices. Indian spices chequered history, dis~overed or
destroyed lands, built or brought down kingdoms, won or lost wars, signed or
flouted treaties, sought or offered favours (Anonymous, 2002a). For this reason
lndia has earned various sobriquets like "The home of spices", "The spice bowl of
the world", and "The land of spices". The lure of exotic spices brought many
European navigators and explorers to the shores of lndia (Chezhiyan and Selvarajan,
2001). Many of the Indian spices are much valued in the world market because of
their intrinsic qualities; hence the world romance with Indian spices continues
unabated (Peter and Nybe, 2002).
An estimate of 500,000 tonnes of spices and herbs valued at US $1500 million
is now imported globally every year. An impressive quantity (46%) of this supply
comes from India. India stood first in the production and export of spices and its
related products. India with an expected population of 1.20 billion people by the
year of 2005 will also be the largest consumer of spices (Anonymous, 1999). About
8.5% of India's export earnings come from agricultural and allied products of which
spices constitute 1.24% (1999-2000). At present India produces 3.0 million tonnes
of spices annually from 2.5 million hectares. The lion share (90%) of the spices
produced is adsorbed in the domestic market and only a meagre share of 10% is
exported to more than 150 countries. India contributes of about 45-50% supply in
terms of volume or 25% in terms of value to international spice market, which earns
a sizeable foreign exchange of Rs.16.2535 lakhs (US $ 341.60 million)
(Anonymous, 2002a).
lndis commands a formidable position in the global spice market with broad
spectrum of spices. It is possible because India is endowed with wide range of agto-
climate regions specially suited for the cultivation of superior quality of spices, which
play an important role in the history of civilization (Rajesh el ol., 2002). Out of 109
spices listed by the IS0 (International Spice Organization) 63 are grown in India, of
which only 16 being very important viz., black pepper, cardamom, ginger, turmeric,
large cardamom, clove, chilli, garlic, saffron, celery, cumin, coriander, fennel,
fenugreek, ajwain and suwa. Based on their importance in foreign trade the black
pepper, cardamom, chilli, turmeric and ginger are grouped under major type and the
rest are considered as minor spices (Chezhiyan, 2000).
Turmeric is one of the major spices cultivated for its underground rhizome,
which is also called as "hidden Lilly" or "turmeric of commerce". In 1280, Mar Co
Polo described turmeric as a vegetable with the prope~lies of saffron and is not really
saffron (Suresh Muthukulam, 2001). This herbal plant is a native of southern Asia
(probably India) and is cultivated extensively throughout the warmer parts of the
world. It is g r o m on a large scale in India, China and East Indies. In India it is
cultivated in almost all the states particularly in Andhra Pradesh, Tamil Nadu,
Karnataka, Kerala, Orissa and to some extent in West Bengal, Maharashtra. Assam,
Bihar, Meghalaya. Goa, Tirupura, Uttar pradesh, Rajasthan, Arunachal pradesh and
Mizoram (Chadha, 2001).
lndia has been and continues to be undisputedly first in the production,
consumption and export of turmeric in the world (Narasimhudu and Balasubramanian,
2001a). Turmeric occupies about 6% of the total area under spice and condiments
cultivation in India. 'The area and production of turmeric in the country showed an
increasing trend during the last five decades. Turmeric is traded as whole dried type,
turmeric powder besides value added products like curcumin, turmeric oleoresin and
turmeric oil (Chadha, 2001).
Turmeric has versatile uses in flavoring, dye making, drug preparation,
cosmetics and medicine (Kallupurackal and Ravindran, 2002; Deeksha Dixit et al.,
2002). Turmeric is officially entered in the Ayurvedic Pharmacopoeia of lndia,
Pharmacopoeia of the People's Republic of China and in Japanese standards o f
herbal medicines (Usha, 2001).
Common Names
Asabi-e-safr. Avea, kurkum, Aurukesafur, Urukesabaghin. Arabic countries Urukessubr, Urukesufr, Zarsud.
I
Country
Acafrao, Ango. Ango hina, Turmeric
1 B e s q cago Curcuma. Haledo, Kalo haledo. turmeric / Nepal I
Brazil
1 Curcuma, Turmeric, Zardchoobeh i
1
Dilau, Nisha, Pasupu, Rajani, Turmeric, Ukon, Zholti, Manjal, Pitras. Mannal, Marinalu, Pampi. Aneshta, Bahula, Dirgharaga, Gandha palashika, Gauri, Gharshani. Mangalaprada, Mehaghni, Kshapa Lakshmi, Pinga. Pitavaluka, Ratrinamika, Shobhana, Shyma, Varangi, Varandatri, Yuvati, Arishina, Haldil. Haldar, Haldhar, Haldi, Halodhi, Halud, Halada, Halede, Halja, Haladi, Haridara, Harita, Hemaragi, Arishina, Lidar, Varavat Ela.
Dilaw
India
Philippines
Goeratji, Javanese turmeric, Kakoenji, Oendre, Koening, Koenir. Koenjet, Kondin, Rame, Temu-lawak / Indonesia !
1 Hardi, Haldi 1 Fiji I 1 I Harindra, Huang ch~ang, Temu kunyit, Turmeric, Wong keong, 1 Wong keung
Malaysia
1 Kerqum ~ o r o c w i Khamin chan, turmeric Kiko eka, Turmeric
Kurcum, Warse
Nighe
Thailand Marquesas islands
Oman
Vietnam
Mena
Rerega, Renga
S&an vert, Safran, Turmeric
Safran, Tale's, Temow lawak
(Dana and Mukeji, 1950; Chopra el al , 1959; Ross, 1999; Kaliupurackal and Ravindran, 2002).
Rotuma
Cook islands
Mauritius
Rodrigues islands
Turmeric, Ukon
Turmeric
Turmeric, Ukon
Turmeric
UI Gum
Yushin
Curcuma
Geelwortel
Curcuma
Gurkmeja
Kurkuma Gelbivaml
Curcuma
Acfrao-da
lmbir
Around 100 active constituents have been recorded from turmeric. They are:
2-hydroxy-methyly anthraqulnone, 4-hydroxy bisabola-2-10-dien-9-one, 4-hydroxy
cinnamoyl-(femloy1)-methane, 4-hydroxy-cinnamoyl methane, 4-hydroxy-fe~loxyl
methane, 4-methoxy bisabola-3-IO-dien-2-one, 5-hydroxy bisabola-2-10-dien-9-one
5-hydroxy procurcumenol, 5'-Methoxy curcumin, Alpha atlantone, Alpha curcumene,
Alpha phellandrene, alpha pinene, Alpha turmerine, Alpha turmerone, Beta
bisabolene, Beta pinene, Beta sesquiphellandrene, Beta sitosterol, Beta tutmerone,
Sri lanka
Taiwan
U.S.A.
South Korea
China
Spain
Dutch
France
Sweden
German
Italy
Protugese
Russia
Bis-(4-hydroxy-cinnamoyl), Bis-@ara-hydmxy+imamoyl) methane, Bis-demethoxy
curcumin, Bisabola-3-10-diene.2-S-dihydmxy, Bisabolene. Bisacumol, Bomeol,
Caffeic acid, Campesterol, Camphene, Camphor, Caryophylene, Cholesterol, Cineol,
Curcumene, Curcumenol, Curcumenone, Curcumin, Curdione, Curlone,
Curzerenone-C, Curzerenone, cyclocurcumin, Dehydro cr.rdione, Demethoxy
curcumin, Di-para-coumaroyl methane, Di-femloyl methane, Epi-procurcumenol,
Di-para-coumaroyl methane. Epi-pmcurcumenol, Eugenol, Feruloyl-para-coumaroyl
methane, Gamma atlantone, Germacmn (4S',SS)-epoxide, Germacron-13-al,
Germacrone, Germacrone. 4(S)-S(S)epoxy, Guaiacol, Hepta-l-4-6-niene-3-one,l-7-bis-(4-
benzenoid-hydroxy-phenyl), hepta-l-6diene-3-5dione.l-(4-hydmxy-3-methoxy-phayl)-
7-(3-4dihydroxy-phenyl), Iso-bomeol, Iso-pmmumenol, Limonene. Linalool, Mono-
demethoxy curcumin, Ortho coumaric acid, Para coumaric acid, Para cymene, Para-
hydroxy-cinnamoyl feruloyl methane, Para-tolyl-methyl-carbinol, Penta-trans-l -trans-4-
dien-3ane,l-5-bis-(4-hydmxy-3-methoxq-pheny), Penta-trans-4dien-3-one,l(4-hydroxy-3-
methoxy-pheny1)-5-(4-hydroxy-phenyl), Pmurcumenol, Protocatechuic acid, Sabinene,
Saturated fatty acids, Stigmasterol, Syringic acid, Terpinene, Terpineol, Tolyl-methyl-
carhinol, Turnerin, Turmerone AR, Turmerone,RT, Turmeronol A. Turmeronol B,
Ukonan A, Ukonan B, Ukonan C, Ukonan D. Unstaurated fatty acids. Vanillic acid,
Zedoarondiol, zingiberene etc. (Ross, 1999; Geoffrey et al., 1998).
Turmeric finds its application in Abortifacient effect, Adrenal hypertrophy effect,
Alkaline phosphatase inhibition, Alkaline phosphatase stimulation, Allergenic activity,
Antiamoebic activity, Antiasthamatic activity, Antibacterial activity. Anticoagulant
activity, Anticomplement activity, Anticonwlsant activity, Anticmstacean activity,
Antiedema activity, Antifungal activity, Antihepatotoxic activity, Antihyper-
cholesterolemic activity, Antihyperglyceridemia effect, Antihypzrlipemic activity, anti-
implantation effect, Anti-inflammatory activity, Anti-ischemic effect. Antimutagenic
activity, Antimycobacterial activity, Antinematodal activity. Antioxidant activity.
Antispasmodic activity, Antispermatogenic effect, Antitumor activity, Antiulcer activity,
Antiviral activity, Antiyeast activity, Apoptosis induction. Arachidonate metabolism
inhibition, Ascaricidal activity. Carcinogenesis inhibition, Cardiotonic activity, Catdase
stimulation, Choleretic activity, Chromosome abenations induced, Clastopnic activity.
CNS depressant activity, Cytochrome B-5 increase, Cytochrome B-5 inhibition.
Cytochrome P-450 induction, Cytochrome P-450 inhibition, Cytotoxic activity,
Desaturase-Delta-5 inhibition Diuretic activity, Embryotoxic effect. Food consumption
reduction, Gastric secretory inhibition, Gastrointestinal diwrders, Genotoxicity activity.
Glutamate oxdoacetate transaminase inhibition, Glutamate oxaloacetate transaminase
stimulation, Glutamate pyruvate Uansaminase inhibition, Glutamate pyruvate
transaminase stimulation, Glutathione- formation induction, Glutathione peroxidase
stimulation, Glutathione-S-transferase induction, GRAS status, Hyaluronidase
inhibition, Hypglycemic activity, Hypothermic activity, lmmunostimulant activity,
Immunosuppressant activity, insect repellent activity, Insecticide activity, Interferon
induction stimulation, Intestinal absorption inhibition, Lactate dehydrogenase
stimulation, Leukopenic activity, Lipid peroxide formation inhibition, Liver regeneratio'n
stimulation, Mutagenic activity, Myocardial uptake of B6-RB enhanced, Necrotic effect,
Nematocidal activity, Nitrosation inhibition, Ovulation inhibition effect. Phagocytosis
capacity increased, Plasma bilirubin decrease, Platelet aggregation inhibition, protease
(HN) inhibition, Radical scavenging effect, RBC synthesis antagonist, Spasmogenic
activity, Spasmolytic activity, Sulthydryl-containing compounds increased, Superoxide
dismutase stimulation, Taenicide activity, Teratogenic activity, Thromboxane B-2
synthesis inhibition, Tyrosinase inhibition, Uterine stimulant effect, WBC-macrophage
stimulant, and weight gain inhibition (Ross.1999; Khanna, 1999; Geoffrey er a/., 1998;
Torres et al., 1998; Palaniswamy, 2000; Kelloff el al., 2000; Graf, 2200; Dorai et a/ . ,
2000; Polassa el al., 1992; Selvam et al., 1995; Van Dau et a!., 1998).
Turmeric prefers a warm humid climate with a rainfall of 1,500 mm and
temperature of 20-30°C. It thrives well up to 1,200 m above mean sea Icvel. Well-
drained sandy or clayey loam or red loamy soils having acidic to slightly alkaline pH
are ideal for its cultivation. Turmeric is either planted on raised bed on ridges and
furrows or in flat system. Turmeric can also be grown as intercrop in coconut gardens,
ginger field or as mixed crop with red gram, chili, colocasia, vegetables, maize, ragi
and onion. Turmeric also comes up well under sparse shade (Chadha, 2001).
This herbal plant is highly prow to several fungal diseases (Naidu, 1988;
Rangaswami and Mahadevan, 2001: Pruthi, 1998). The major diseases and causal
organisms of turmeric plant are:
Leaf blotch : Taphrrnu maculans
Leaf spot : CaNelotrichum capsici
Leaf blight : Hhizocronia solani
Rhizome and Root rot : Pyhium graminicolum
Basal rot : Sclerotium rolfsi
Leaf spot : Curvularia lunala
Storage rot : Macrophominaphoseolina
Cladosporium cladosporioides
The devastating leaf spot disease of turmeric is caused by Colletorrichum
capsici (Syd.) (Butler & Bisby). This disease was first reported in Coimbatore district,
India by Mc Rae in 1917 (Ramakrishnan, 1954). It results in the yield loss between
1540% (Joshi and S h m a , 1980; Pruthi, 1998; Narasimhudu and Balasubramanian,
2001b). The dry weight of the rhizome is also reduced to 62% approximately.
There are nearly 1000 form species of Colletotrichum spp. that have been
described on the basis of the disease caused on various hosts. C. capsici were also
reported as unspecialized parasite from more than 100 plants in a wide range of
angiosperm families. Although the disease is found throughout India, it is more severe
in the southern states. (Gomathi, 2001).
The disease was described by Ramakrishnan (1954) as follows: The disease
manifests itself on the leaves in the form of elliptic or oblong spots, variable in size.
In the initial stages, they are small but very soon many of them increase in size and
may measure one and a half to two inches in length and about an inch to an inch and a
half across. Two or more spots may coalesce developing into irregular patches often
involving a major portion of the leaf, which eventually dries up. Each individual spot
has a characteristic appearance. The centre is greyish white and thin with numerous
black, dot-like acewuli on both surfaces. These are arranged in concentric rings.
Beyond the greyish white portion is a brown margin all round the spot. On this also
the a c e ~ i l i may be seen. Outside this is an indefinite yellowish region forming a halo
round the spot. The spots, though visible on both surfaces, are more marked on the
upper surface in the fresh leaves. But on drying they are equally conspicuous on both
surfaces. Invariably, the lesions are on the leaf blade. But sometimes stromata or
acervuli may be seen on the leaf sheaths also. When the incidence o f the disease is
heavy. most of the leaves dry up and the field presents a parched-up appearance. The
central region of the spot may become papery and easily tom. Owing to the
destruction of the chlorophyll-bearing leaf area, the production of rhizomes is reduced
sometimes to less than 50%. Stromatoid bodies are formed even on the scales of the
rhizon~es.
The acewuli are located intra-epidermally. The hyphae are septate and hyaline in
the initial stages passing between and inside the cells of the mesophyll. Later, pale
brownish h p h a e accumulate inside the epidermal cells and form the basis of stroma
development. The stromata are made up of light to dark brown pseudoparenchymatous
cells. l'he outer wall of the epidermis is ruptured and the setae and conidiophores are
exposed. Setae are formed from all over the acervulus and are not confined to the
margin. Crescent-shaped conidia are bome on the conidiophores. The stromata are
iariable in size. ranging from 6 0 - 1 2 0 ~ in diameter. The setae are b r o w , septate and
tnpcrlng l'he length of the setae is variable and ranges from 50-145~.
The conidia are crescent-shaped, hyaline and one-celled measuring 25 x 3 p
( 1 7-3 1 x 3-4). A conspicuous vacuole is present in the center of the conidium. But as
it grows old, the contents become more granular or highly vacuolated and the solitary
iacuole is not evident. The conidial masses are of pink, though individual spores are
hyaline The spots are embedded in a gelatinous substance and on drying they are all
held together. But when the mass is floated in drops of water, the spores readily
separate. The conidia germinate readily producing germ tubes from either or both
ends. A septum is also developed dividing the cell into two compartments. Olive-
brown, round or irregular thick-walled appressoria are formed at the ends of the germ
tubes or from the tip or the spore irself (Sundararaman. 1925: Kamakrishnan, 1941).
The fungus was brought into pure culture and its growth on different media was
studied. On oat agar, the isolate exhibited good grouth producing pale. olive grey
aerial mycelium and numerous buff-pink acervuli in the course of a ueek. The
acervulus has a black stromatic base on which masses of conidia are produced to the
aerial growth to give buff-pink colour to the fructification. On French bean agar, the
aerial growth is a mixture of white and grey. Black stromata and pink acervuli are
developed in large numbers in six to seven days. When the isolates are maintained in
agar media for several generations, there is a reduction in the quantity of aerial
mycelium produced. The surface of the slant becomes studded with numerous
stromata and acewuli. Chlamydospores are formed in large numbers, especially at the
junction of the surface of the slant and the sides of the test rubes (Rsmakrishnan,
1954).
Several works have been conducted to control (' ci~psfci by using fungicides of
chemical and biological origin. The current methods of agr~culture and horticulture
rely heavily upon the use of chemlcal fungicideslpestic~des. which are very needy and
its usage has become unavoidable for higher effectwe yield and crop protection. The
first recorded use of chemicals to control pests dates back to 2500 BC (Hock c v a / .
1991). The chemical pesticides have been w~dely used for the past three decades
(Chandrasekhara Rao and Murthy. 1999). Around 100,000 chemical pest~cides are
now in commercial use worldwide with more than 2.5 million tonnes were applied to
the field annually (Anonymous, 2000~).
Although the usage of chemical pesticides became a paramount need for the
crop protection and prodigious yield, there are many disadvantages such as causing
resurgence, resistance in pathogens and posing residue problems to the consumers o f
treated products, besides causing environmental contamination (Chandrasekhara Rao
and Munhy. 1999).
The constant and continuous use of the fungicides may develop resistance in
the pathogen against fungicidal action of the fungicides (Kore and Apes, 1989;
Sariah, 1989; Griffiths, 1981; Bollen, 1971). They were reported to inhibit the
photosynthetic process (Querns el al.. 1998) and reduce plant growth and biomass
(Shilling el al.. 1994). Use of chemical fungicide was proved to suppress the non-
target beneficial organisms and thus they might facilitate the pathogen to spread
the disease quickly in their respective hosts and the reduction of these beneficial
organisms can also result in changes in the natural biological balances (Knauss,
1972, 1977). "Iatrogenic diseases" were also reported due to the application of
chemical fungicides. The use of Fytolan causing iatrogenic effect in turmeric was
reported by Verma (1986). Various ill effects of synthetic fungicides on crop and
ecosystems were described by Byrdc (1991). They were reported to affect the soil
microfauna, animals and human population. 672 million birds were exposed to
pesticides every year and 10% (67 million) of them die annually (Anonymous,
2 0 0 0 ~ ) .
The hazards of pesticides are now widely recognized worldwide, although
statistics are hard to gather The world health organization estimates that at least
three million people are poisoned by pesticides every year and more than 200,000
died are attributed to pesticides. It is estimated that up to 25 million agricultural
workers are poisoned every year (Anonymous, 2000a,b). The pesticides poisoning
the food in the form of residues is reported in most of the vegetable and fruits that
are used in our day today life. The residue problems of the chemical usages also
affect the livestock, waterways and the general environment. Every person on the
earth has absorbed at least 250 synthetic chemicals into their body due to pesticide
residue (Anonymous, 2000~) . The spiraling and fluctuating cost of chemical
pesticides demands for alternative. Chemical pesticides are akin to environmental
pollution in producing hazardous consequences In terms of soil pollution thus
reducing the quality of arable land (Bezbamah, 2000). In Indian scenario the thrust
for more use of pesticides in intensively cultivated areas led to the developnlent of
resistance by the pathogen against the particular pesticide used. Ground water
contamination by leached chemicals can occur in high use areas if persistent
products are used.
Considerable attention was bestowed to critically evaluate the disadvantages
of pesticide, as it now became a central issue, which must be addressed properly.
The crucial problems confronted about chemical pesticide usage needs a
recommitment to science and research that the biod~versity and environment safety
are not excluded and undermined. Strategies for raising safety measures against
the cause or impediments of pesticides hazards must be developed on a war
footing.
The sensitive issue regarding the demerits of the chemical pesticides has
demanded greater attention towards the development of safer methods of crop and
environment protection. To sustain and also to augment the yield, it is necessary to
adopt strong mop protection strategies without altering the ecological balance of the
cultivated land and environment. These unavoidable reasons compelled the researcher
to opt for biological control methods, which are now considered as the apt substitute
for the chemical pesticides.
Biological control of plant pests and in particular diseases caused by soil
borne plant pathogens, by introduced microorganisms has been the focus of
study for over 75 years (Baker, 1987). Biological control is a natural
phenomenon, which is defined as any condition under which or practice whereby
the survival activity of a pathogen is reduced through the agency of any other
living organism with the result that there is a reduction in the incidence of the
disease caused by the pathogen or the reduction of inoculum density or disease
producing activities of a pathogen or parasite in its active or dormant state, by
one or more organisms, accomplished naturally or through manipulation of the
environment, host or antagonists, or by mass introduction of one or more
antagonists (Suseela Bhai, 2000). At the end of 2001, there were approximately
195 registered biopesticide active ingredients and 780 products (Anonymous,
2002b).
The biopesticides have several distinct advantages over chemical
fungicides (Suseela Bhai, 2000; Singh, 1993: Singh, 1999). Biopesticides are
usually inherently less toxic than conventional fungicides. They generally affect
only the target pathogenic organisms, in contrast to broad spectrum,
conventional fungicides that may affect organisms as different as birds, insects
and mammals. Also they are effective in very small quantities and often
decompose quickly, thereby resulting in lower exposures and largely avoiding
the pollution problems caused by conventional fungicides. When used as a
component of Integrated Pest Management (IPM) programs, biopesticides can
greatly decrease the use of conventional fungicides, while crop yields remain
high. The biopesticides are safe to manufacturers, users and consumers of treated
crops and all aspects of the environment. They have a very specific mode o f
action and relatively critical application times. They have limited field
persistence and a short shelf life safer to humans and the environment and
present no residue problems. In fact. new biopesticides are often registered in
less than a year. compared with an average of more than 3 years for conventional
pesticides. They are easy to formulate and quicker to use in the field application
(Anonymous, 2002b).
Hence in reccnt years, the biopestic~des of plant and microbial origin are being
used as an alternative for chemical fungicides (Asha. 1999; Gomathi, 2001).
Biological control of Colleioir~chum spp. was reported by many workers
(Rajathilagam, 1999; Sourechc @ Venguidaragavane ei a / . , 2001; Rajavel, 2000;
Jeyalakshmi and Seetharam, 1998: Muthuraj. 1998).
Objectives
The present study was undertaken to find out the efficacy of four antagonistic
agents of microbial origin viz. Prchodcrma vrride, T horzranum, G!iocladrum
virens and Pseudomonas ,Jluore.scens in controlling C capsici causing leaf spot of
turmeric.
O To evaluate the efficacy of the above mentioned biocontrol agents on the
growth and physiology of C capsici infecting turmeric.
-3 To investigate the effects of the biocontrol agents in reducing the disease
intensity in the host plant caused by C. capsic;.
*3 To study the host-pathogen interaction by analysing the biochemical
changes in the host plant under healthy, infected and treated condit~ons.
To study the effect of biocontrol agents on the antioxidant enzyme
metabolism of the host.
4 To evaluate the yield of the host plant under healthy, infected and treated
conditions.
+:. To find out the best candidate out of four biocontrol agents for controlling
C capsici.