Академический Документы
Профессиональный Документы
Культура Документы
Abstract
Methanolic extracts of forty plant species commonly growing across India were collected and have been
screened for antisporulant activity against Sclerospora graminicola (Sacc.) Schroet., the causative agent of
pearl millet downy mildew. The collection represented 38 genera of 30 families. The methanolic extracts
of nine species did not show any effect, whereas the activity of the extracts of Clematis gouriana, Evolvulus
alsinoides, Mimusops elengi, Allium sativum and Piper nigrum were commensurable to that of the marketed
botanical fungicides. The extracts of 11 species (Agave americana, Artemisia pallens, Citrus sinensis,
Dalbergia latifolia, Helianthus annus, Murraya koenigii, Ocimum basilicum, Parthenium hysterophorus, Tagetes
erecta, Thuja occidentalis and Zingiber offinale) exhibited remarkable antisporulant effect even after 10-fold
dilution of the crude extracts while in the case of remaining 15 plants the crude extracts loosed activity after
10-fold dilution. The antisporulant activity of commercialised Azadirachta preparation (Nutri-Neem) was
more pronounced than that of Reynutria based one (Milsana) and Sabadilla (veratrin), however, these
botanical preparations held off the extracts of C. gouriana and E. alsinoides and synthetic fungicides.
Keywords: Plant extracts, antifungal activity, Clematis, Evolvulus, downy mildew, pearl millet
Introduction
Present control technologies of downy mildews discouple the pathogen’s life cycle mainly in
two points of ontogeny. The applied chemicals either destroy spores, preventing the infection
or inhibit the biotrophic thallus, anticipating the formation of new infective propagules. The
preventive control of Sclerospora graminicola, the causative agent of pearl millet downy mildew
(PMDM) meets difficulties. The short vegetation reduces the remediative capacity of its host
plant, and as a consequence of it, pesticides are accumulated in the grain [1] challenging to
Correspondence: H.S. Shetty, Department of Studies in Applied Botany, University of Mysore, Manasagangotri, Mysore 570 006,
Karnataka, India. E-mail: hss_uom@hotmail.com
ISSN 0323-5408 print/ISSN 1477-2906 online # 2005 Taylor & Francis Group Ltd
DOI: 10.1080/03235400400007558
32 S. A. Deepak et al.
application of synthetic chemicals after formation of tillers. The resistant to PMDM cultivars
were selected and some of them have been cultivated widely. The pathogen shows high
natural variation in aggressivity [2]; even progenies of the same oospore could be classified
into distinct pathotype groups [3]. Although the downy mildew tolerance of pearl millet can
be enhanced by diverse methods [4, 5] the possibilities of biocontrol measures [6] as well as
enhancement of plant resistance with chemical treatment [7] were presented, none of these
approaches resulted the economically acceptable level of control. Thus, the vulnerability of
the resistance to the disease has been a major cause of concern as even 10% disease incidence
cause economic loss threatening the net return [8]. The only tool for creditable control of this
endobiotrophic peronospora is the use of systemically acting acylanilide derivatives. However,
the calculability of management of pearl millet downy mildew has been threatened by
emergence of acquired tolerance to this group of chemicals in India [9].
Losses caused by PMDM urges development of alternative control agents. One approach to
discover newer antimildew compounds is to search for their presence in natural sources [10].
The defence strategies of plants against their pathogens are multitudinous, including the use
of antimicrobial chemicals that either can be de novo formed during the pathogenesis
(phytoalexins) or are constitutive components (phytoanticipins) of cells [11]. On the other
hand, pathogens have evolved mechanisms to evade these barriers that demand application of
various pesticides in crop production. Microbial species or strains that do not invade the plant
are usually more sensitive to the components of performed barriers than a viable pathogen of
this plant [12]. Consequently, phytoanticipins and precursors of phytoalexins can represent a
prospective tool for PMDM management [13].
In the present investigation, we have studied the antisporulant activity of methanolic
extracts of plants growing at almost all locations throughout pearl millet producing areas of
India. The inhibitory effect of extracts against S. graminicola was compared to that of
marketed fungicides and phytochemicals.
Test organisms
The downy mildew pathogen S. graminicola (pathotype 1) was isolated from naturally infested
pearl millet (Pennisetum glaucum (L.) R. Br., hybrid HB3) in Bogadi village of Mysore district
Table I. Inhibitory effect of plant extracts on zoosporangium formation of Sclerospora graminicola.
Methanolic extracts4
No. Species (Family)1 Origin2 Part used3 crude 1:9 1 : 99 Reported activity against fungi5
Strong
1 Clematis gouriana Roxb. (Ranunculaceae) MY T + + ++ + F36, F42, F43
2 Evolvulus alsinoides L. (Convolvulaceae)a,b HA T + + + +
3 Allium sativum L. (Liliaceae)a,b MY B + + ++ 7 F16, F23
4 Piper nigrum L. (Piperaceae)a,b MY S + + ++ 7 F16, F6, F9, F10, F20, F27, F30, F36, F38,
F42, F43
5 Mimusops elengi L. (Sapotaceae)b MY F ++ ++ 7
Remarkable
6 Agave americana L. (Agavaceae)a MY L + + + 7
7 Thuja occidentalis L. (Cupressaceae) MY L + + + 7
8 Artemisia pallens Wall. (Asteraceae) CH L + + + 7
9 Helianthus annuus L. (Asteraceae) MY S + + + 7
10 Parthenium hysterophorus L. (Asteraceae) MY L + + + 7
11 Tagetes erecta L. (Asteraceae) MY L + + + 7
12 Ocimum basilicum L. (Lamiaceae)a,b MY L + + + 7 F5, F12, F21, F25, F27, F41
(continued).
33
34
Table I. (continued).
S. A. Deepak et al.
Methanolic extracts4
No. Species (Family)1 Origin2 Part used3 crude 1:9 1 : 99 Reported activity against fungi5
(Karnataka state, India) during 1970 by Shetty. The strain was maintained on greenhouse
grown pearl millet plants before being used as inoculum for the experiments. The method was
described in detail by Safeulla [14].
Dimethomorph A – + + ++ ++
Metalaxyl A – + + ++ ++
Mikal B – – + + ++
Digitonin A – – + + ++
Podophyllotoxin A – – – + ++
Veratrin A – – – + +
Nutri-Neem B – – – + +
Milsana B – – – + ++
The antisporulant activity was evaluated by following scale; full inhibition (+ +), partial inhibition (+) and no
inhibition (7). 1A = 25% methanolous stock solution of active ingredients containing 1% of Tween 20 was used for
preparing dilution series. The methanol and Tween 20 did not exhibited any inhibitory effect alone when applied at
maximum doses (5 and 0.2%, respectively). B = Commercial preparations were used.
accordance with our data. Leaf extracts of M. koenigii could control Pythium damping off at
67% when applied via soil in tomato [47] as well as the efficacy of bark-debris of Eucalyptus
against Phytophtora sp. was demonstrated [49]. Broad spectrum of antifungal activity was
reported in the case of several test plants (9, 16, 21, 22, 26), however, in our tests only Z.
officinale and O. basilicum exhibited remarkable antisporulant effect while E. globosus, A. indica
and O. sanctum acted weakly. Analysing the activity scores of different plant extracts against S.
graminicola (Table I) in relation to their effects reported against various phytopathogenic
fungi, there was clear that the sensitivity spectrum of S. graminicola is entirely different.
Moreover, no relationship was revealed between taxonomic position of plants and
antiperonospora activity of their extracts.
The role of phytoalexins in defence mechanisms was intensively studied meanwhile to
constitutive compounds has been paid less attention [13]. The term ‘phytoanticipin’ was
proposed for description of the latter group in 1994 and includes all types of low molecular
weight antimicrobial metabolites other than phytoalexins that are supposed to play a role in
disease resistance of plants [11]. In our case tissues of healthy plants were collected. It can be
presumed that the extracted constitutive compounds of plants were responsible for
antisporulant effect, which was manifested in our experiments. Nevertheless, it is often
difficult to determine whether a molecule is constitutive or induced, as some of them may
normally be present in hardly detectable quantities, but dramatically increase in concentration
after infection. Moreover, the same compound may be preformed antifungal substance in one
species and a phytoalexin in another [13].
The results of the present work indicate that some of the tested plants are promising
candidates for PMDM management. The transmission of S. graminicola to new areas by wind
can be successfully stopped by inhibition of zoosporangium formation [52]. The species
providing active extracts (A. sativum, P. nigrum, C. gouriana, E. alsinoides and M. elengi) are
found in pearl millet growing areas and are well known plants as most of them are used for
various medical purposes [11]. Their utilization makes possible the efficient pest management
exploiting the local natural resource base. The use of extracts possessing with broad spectrum
of activity (species 1, 3 and 4) can inhibit whole pathogen complex [53] associated to pearl
millet as well. Moreover, the costs of treatment are low and the contamination with residual
amounts of pesticides can be avoided [54].
Antimildew activity of plant extracts 37
The use of various extracts of plants in agricultural practices has advantages. The sources
can be find in millet producing areas and the extracts do not need further bioremediation.
Although utilisation of toxic solvent in preparations made on-farm is not advisable, the
technology of manufacturing is simple and the resulted detoxified preparation easy to transfer
to the farmers, and thus to promote sustained millet production [53]. The analysis of
chemical composition of methanolic extracts can direct to discovery of new antimildew
substances which might be promising lead compounds for development of new synthetic
molecules with antiperonospora activity.
Acknowledgement
This research was supported by the co-operation project ‘‘ Theoretical base of integrated pest
control methods’’ between Indian National Science Academy and Hungarian Academy of
Sciences (Grant No. 11). The authors thank the Project Coordinator (ICAR-AICPMIP
Project, Jodhpur, India) for laboratory and field facilities provided.
References
[1] Reddy MVB, Shetty HS, Reddy MS. Residue free treatments of metalaxyl for effective control of pearl millet
downy mildew. Disc Innovat 1996;8:53 – 57.
[2] Ball SL, Pike DJ. Intercontinental variation of Sclerospora graminicola. Ann Appl Biol 1984;104:41 – 51.
[3] Thakur RP, Pushpavathi B, Rao VP. Virulence characterisation of single-zoospore isolates of Sclerospora
graminicola from pearl millet. Plant Dis 1998;82:747 – 751.
[4] Desmukuh SS, Mayee CD, Kulkarni BS. Reduction of downy mildew of pearl millet with fertilizer management.
Phytopathology 1978;68:1350 – 1353.
[5] Wilson JP, Gates RN, Panwar MS. Dynamic multiline population approach to resistance gene management.
Phytopathology 2001;91:255 – 260.
[6] Umesha S, Dharmesh SM, Shetty SA, Krishnappam M, Shetty HS. Biocontrol of downy mildew disease of pearl
millet using Pseudomonas fluorescens. Crop Prot 1998;17:387 – 392.
[7] Shailasree S, Sarosh BR, Vasanthi NS, Shetty HS. Seed treatment with beta-aminobutyric acid protects
Pennisetum glaucum systemically from Sclerospora graminicola. Pest Manag Sci 2001;57:721 – 728.
[8] Thakur RP, Rai KN, Rao VP, Rao AS Genetic resistance of pearl millet male-sterile lines to diverse Indian
pathotypes of Sclerospora graminicola. Plant Dis 2001;85:621 – 626.
[9] Anonym. Report of the All India Co-ordinated Pearl Millet Improvement Project. Indian Council of Agricultural
Research, Project Co-ordinating Unit, Agricultural Research Station, Mandor, Jodhpur, India, 2001.
[10] Dixon RA. Natural products and plant disease resistance. Nature 2001;411:843 – 847.
[11] VanEtten HD, Mansfield JW, Bailey A, Farmer EE. Two classes of plant antibiotics: Phytoalexins versus
‘‘phytoanticipins’’. Plant Cell 1994;6:1191 – 1192.
[12] Prusky D. Pathogen quiescence in postharvest diseases. Annu Rev Phytopathol 1966;34:413 – 434.
[13] Grayer RJ, Kokobun T. Plant-fungal interactions: the search for phytoalexins and other antifungal compounds
from higher plants. Rev Phytochem 2001;56:253 – 263.
[14] Safeulla KM. Genetic vulnerability: the basis of recent epidemics in India. In: Day PR, editor. Genetic basis of
epidemics in agriculture. Annals of New York: Academy of Science; 1976. pp 72 – 85.
[15] Chopra RN, Chopra IC, Handa KL, Kapur LD. Indigenous drugs of India. 2nd rev ed. Calcutta, India: UN
Dhur and Sons Private Ltd.; 1958.
[16] Afolayan AJ, Grierson DS, Kambizi L, Madamombe I, Masika PJ. In vitro antifungal activity of some South
African medicinal plants. S Afr J Bot 2002;68:72 – 76.
[17] Mohamed S, Saka S, ElSharkawy SH, Ali AM, Muid S. Antimycotic screening of 58 Malaysian plants against
plant pathogens. Pest Sci 1996;47:259 – 264.
[18] Ramezani H, Singh HP, Batish DR, Kohli RK. Antifungal activity of the volatile oil of Eucalyptus citriodora.
Fitoterapia 2002;73:261 – 262.
[19] Govindachari TR, Suresh G, Gopalakrishnan G, Banumathy B, Masilamani S. Identification of antifungal
compounds from the seed oil of Azadirachta indica. Phytoparasitica 1998;26:109 – 116.
[20] Fabry W, Okemo P, Ansorg R. Fungistatic and fungicidal activity of East African medicinal plants. Mycoses
1996;39:67 – 70.
38 S. A. Deepak et al.
[21] bin Jantan I, Yassin MSM, Chin CB, Chen LL, Sim NL. Antifungal activity of the essential oils of nine
Zingiberaceae species. Pharm Biol 2003;41:392 – 397.
[22] Zollo PHA, Biyiti L, Tchoumbougnang F, Menut C, Lamaty G, Bouchet P. Aromatic plants of tropical central
Africa. Part XXXII. Chemical composition and antifungal activity of thirteen essential oils from aromatic plants
of Cameroon. Flavour Fragr J 1998;13:107 – 114.
[23] Rajesh SGL. Studies on antimycotic properties of Datura metel. J Ethnopharmacol 2002;80:193 – 197.
[24] Dixit SN, Tripathi SC, Upadhyay RR. Antifungal substances of rose flowers (Rosa-indica). Econ Bot
1976;30:371 – 374.
[25] Meir S, Droby S, Davidson H, Alsevia S, Cohen L, Horev B, Philosoph-Hadas S. Suppression of Botrytis rot in
cut rose flowers by postharvest application of methyl jasmonate. Postharvest Biol Tec 1998;13:235 – 243.
[26] Moline HE, Locke JC. Comparing neem seed oil with calcium chloride and fungicides for controlling
postharvest apple decay. Hortsci 1993;28:719 – 720.
[27] Damayanti M, Susheela K, Sharma GJ. Effect of plant extracts and systemic fungicide on the pineapple fruit-
rotting fungus, Ceratocystis paradoxa. Cytobios 1996;86:155 – 165.
[28] Tewari SN, Nayak M. Activity of 4 plant leaf extracts against 3 fungal pathogens of rice. Trop Agr 1991;68:
373 – 375.
[29] Anthony S, Abeywickrama K, Dayananda R, Wijeratnam SW, Arambewela L. Fungal pathogens associated with
banana fruit in Sri Lanka, and their treatment with essential oils. Mycopathologia 2004;157:91 – 97.
[30] Singh HNP, Prasad MM, Sinha KK. Efficacy of Leaf extracts of some medicinal-plants against disease
development in banana. Lett Appl Microbiol 1993;17:269 – 271.
[31] Taechowisan T, Lumyong S. Activity of endophytic actinomycetes from roots of Zingiber officinale and Alpinia
galanga against phytopathogenic fungi. Ann Microbiol 2003;53:291 – 298.
[32] Fiori ACG, Schwan-Estrada KRF, Stangarlin JR, Vida JB, Scapim CA, Cruz MES, Pascholati SF. Antifungal
activity of leaf extracts and essential oils of some medicinal plants against Didymella bryoniae. J Phytopathol
2000;148:483 – 448.
[33] Gupta VP, Govindaiah AKB, Datta RK. Plant extracts: A non-chemical approach to control Fusarium diseases
of mulberry. Curr Sci 1996;71:406 – 409.
[34] Harris JC, Cottrell SL, Plummer S, Lloyd D. Antimicrobial properties of Allium sativum (garlic). Appl Microbiol
Biot 2001;57:282 – 286.
[35] Rai MK, Qureshi S, Pandey AK. In vitro susceptibility of opportunistic Fusarium spp. to essential oils. Mycoses
1999;42:97 – 101.
[36] Owolade OF, Amusa AN, Osikanlu YOK. Efficacy of certain indigenous plant extracts against seed-borne
infection of Fusarium moniliforme on maize (Zea mays L.) in south western Nigeria. Cer Res Comm
2000;28:323 – 327.
[37] Coventry E, Allan EJ. Microbiological and chemical analysis of neem (Azadirachta indica) extracts: New data on
antimicrobial activity. Phytoparasitica 2001;29:441 – 450.
[38] Amadioha AC, Uchendu PN. Post harvest control of tomato fruit rot caused by Fusarium solani with extracts of
Azadirachta indica. Discov Innovat 2003;15:83 – 86.
[39] Singh R, Rai B. Antifungal potential of some higher plants against Fusarium udum causing wilt disease of Cajanus
cajan. Microbios 2000;102:165 – 173.
[40] Bouzouita N, Kachouri F, Hamdi M, Chaabouni MM. Antimicrobial activity of essential oils from Tunisian
aromatic plants. Flavour Fragr J 2003;18:380 – 383.
[41] Hakulinen J, Julkunen-Tiitto R. Variation in leaf phenolics of field-cultivated willow (Salix myrsinifolia) clones in
relation to occurrence of Melampsora rust. Forest Pathol 2000;30:29 – 41.
[42] Caccioni DRL, Guizzardi M, Biondi DM, Renda A, Ruberto C. Relationship between volatile components of
citrus fruit essential oils and antimicrobial action on Penicillium digitatum and Penicillium italicum. Int J Food
Microbiol 1998;43:73 – 79.
[43] Singh G, Upadhya RK, Narayanan CS, Padmkumari KP, Rao GP. Chemical and fungitoxic investigations on
the essential oil of Citrus sinensis (L) Pers. J Plant Dis Prot 1993;100:69 – 74.
[44] Suresh G, Narasimhan N, Masilamani S, Partho PD, Gopalakrishnan G. Antifungal fractions and compounds
from uncrushed green leaves of Azadirachta indica. Phytoparasitica 1997;25:33 – 39.
[45] Du ZZ, Zhu N, Shen YM. Two novel antifungal saponins from Tibetan herbal medicine Clematis tangutica.
Chinese Chem Lett 2003;14:707 – 710.
[46] Kamalakannan A, Shanmugam V, Surendran M. Effect of plant extracts on susceptibility of rice seedlings to
blast disease and consequent biochemical changes in rice plants. J Plant Dis Prot 2001;108:536 – 543.
[47] Pandey VN, Dubey NK. Antifungal potential of leaves and essential oils from higher plants against soil
phytopathogens. Soil Biol Biochem 1994;26:1417 – 1421.
Antimildew activity of plant extracts 39
[48] Agarwal M, Walia S, Dhingra S, Khambay BPS. Insect growth inhibition, antifeedant and antifungal activity of
compounds isolated/derived from Zingiber officinale Roscoe (ginger) rhizomes. Pest Manag Sci 2001;57:289 –
300.
[49] Hardy GES, Sivasithamparam K. Effects of sterile and non-sterile leachates extracted from composted
eucalyptus bark and pine-bark container media on Phytophthora spp. Soil Biol Biochem 1991;23:25 – 30.
[50] Annapurna J, Amarnath PVS, Kumar DA, Ramakrishna SV, Raghavan KV. Antimicrobial activity of Ixora
coccinea leaves. Fitoterapia 2003;74:291 – 293.
[51] Singh KV, Pathak RK. Effect of leaf extracts of some higher plants on spore germination of Ustilago maydis and
Ustilago nuda. Fitoterapia 1984;55:318 – 320.
[52] Weston WH. Nocturnal production of conidia by Scleospora graminicola. J Agr Res 1924;27:771 – 783.
[53] Wilson JP. Pearl millet diseases: A compilation of information on the known pathogens of pearl millet,
Pennisetum glaucum (L.) R.Br. Agriculture Handbook No. 716. USDA, Agricultural Research Service; 2000.
[54] Andersen HR, Vinggaard AM, Rasmussen TH, Gjermandsen IM, Bonefeld-Jorgensen EC. Effects of currently
used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro. Toxicol Appl Pharm
2002;179:1 – 12.
[55] Nyemba JA. Sustainable food-crop production in the semi-arid tropics: Strategy for technology transfer in millet
research. J Sustain Agr 1997;9:25 – 47.