SCIENTIFIC CORRESPONDENCE

Table 2. Results from F-1 inbreed and backcrosses of F-1 with B. sphaericus resistant and susceptible strains of An. stephensi

No. of larvae/families No. of larvae alive No. of larvae dead

Cross no. exposed to B. sphaericus after 48 h (expected*) after 48 h (expected*) χ2

1 R/S R/S 1016/7 243 (254) 773 (762) 0.2034**

2 S/R S/R 1139/8 287 (285) 852 (855) 0.001**
3 R/S R/R 1338/9 658 (669) 680 (669) 0.027**
4 R/R R/S 1420/11 704 (710) 716 (710) 0.0071**
5 R/S S/S 1551/12 0 (0) 1551 (1551) 0
6 S/S R/S 1297/11 0 (0) 1297 (1297) 0

R, B. sphaericus resistant; S, B. sphaericus susceptible.

*, Based on the hypothesis that resistance is due to a single major factor, expected number of larvae that would die at a concentra-
tion of 40 mg/l within 48 h.
**, Not significant (P > 0.05).

receptors of larval midgut brush border
membrane in the resistant strain of Cx.
quinquefasciatus10. Both these toxins re-
quire only single class of receptors, one
helping in binding and the other perform-
ing toxic actions11.
Bioassay tests with Bt H-14 against 3rd
instar larvae of B. sphaericus resistant and
susceptible strains of An. stephensi gave
LC50 values of 0.117 and 0.136 mg/l.
These results show that resistance to B.
sphaericus in An. stephensi does not show
any cross-resistance to Bt, probably be-
cause of the different modes of action of
two the bacterial toxins12. Thus in case of
B. sphaericus resistance, Bt can be used
to control the resistant mosquito larvae.
1. Indranil, K., Eapen, A., Ravindran, K. J.,
Chandrahas, P. K., Appavoo, N. C., Sada-
nand, A. V. and Dhanraj, B., Indian J.
Malariol., 1997, 34, 25–36.
2. Kumar, A., Sharma, V. P., Thavaselvam,
D. and Sumodan, P. K., J. Am. Mosq.
Contr. Assoc., 1995, 11, 86–89.
3. Kumar, A., Sharma, V. P., Sumodan, P. ACKNOWLEDGEMENT. Assistance provi-
K., Thavaselvam, D. and Kamat, R. H., ded by Sh. Brij Mohan and Sh. Rajender Singh
J. Am. Mosq. Contr. Assoc., 1994, 10, is acknowledged.
535–539.
4. Mittal, P. K., J. Vector Borne Dis., 2003,
40, 20–32. Received 4 April 2005; accepted 28 April 2005
5. Mittal, P. K., Adak, T. and Sharma, V.
P., Indian J. Malariol., 1993, 30, 37–41.
6. Mittal, P. K., Adak, T., Batra, C. P. and
Sharma, V. P., Indian J. Malariol., 1993, P. K. MITTAL1,*
30, 81–90. T. ADAK1
7. Adak, T., Mittal, P. K., Raghavendra, K., S. K. SUBBARAO2,3
Subbarao, S. K., Ansari, M. A. and
Sharma, V. P., Curr. Sci., 1995, 69, 695–
1
698. Malaria Research Centre,
8. Mittal, P. K., Adak, T. and Sharma, V. 2, Nanak Enclave,
P., Indian J. Malariol., 1998, 35, 178–183. Radio Colony,
9. Aslamkhan, M., Pak. J. Zool., 1973, 5, Delhi 110 009, India
127–130. 2
Malaria Research Centre,
10. Nielsen-Leroux, C., Charles, J. F., 22, Sham Nath Marg,
Thiery, I. and Georghiou, G. P., Eur. J.
Delhi 110 054, India
Biochem., 1995, 228, 206–210. 3
11. Baumann, P., Clark, M. A., Baumann, L. Present address:
and Broadwell, A. H., Microbiol. Rev., Indian Council of Medical Research,
1991, 55, 425–436. New Delhi 110 029, India
12. Porter, A. G., Parasitol. Today, 1996, 12, *For correspondence.
175–179. e-mail: pk_mittal52@yahoo.co.in

Oryza rufipogon: A possible source of novel resistance specificities

against rice blast (Magnaporthe grisea)
Blast disease caused by Magnaporthe
grisea is a serious production constraint
for rice world wide. In the past, rice breeders
have successfully tapped sources of resis-
tance available in cultivated rice germ-
plasm for developing disease-resistant
varieties. To date more than 40 genes
conferring resistance to blast have been
identified from cultivated rice lines1. Be-
sides the cultivated types, the wild and
weedy species within the primary gene
pool of Oryza represent a potential source
of easily transferable novel pest and disease
resistance2. Among the most successful
examples of utilization of wild germ-
plasm in rice resistance breeding has
been the use of gene(s) of an accession of
O. nivara from northwestern Himalayas,
to provide long-lasting resistance to grassy
stunt virus3. The hilly state of Himachal
Pradesh (HP), representing the north-
western Himalayan region of India, is
one of the centres of diversity of cultivated
rice4. This region abounds in diversity of
wild rice varieties identified as types
within wild rice O. rufipogon, which is
believed to be the progenitor of Asian
cultivated rice (O. sativa)5.
Since, O. rufipogon – M. grisea patho-
system has a long co-evolutionary history
than the O. sativa – M. grisea system, we
believe considerable diversity for R-genes
must have evolved in the wild rice com-

CURRENT SCIENCE, VOL. 89, NO. 3, 10 AUGUST 2005 443

SCIENTIFIC CORRESPONDENCE
Table 1. Geographic origin of 53 accessions of wild rice (Oryza rufipogon) from Himachal Pradesh

Place of collection Sample size Accession no.

Bodal 11 WRA-1, WRA-2, WRA-3, WRA-4, WRA-5, WRA-6, WRA-7, WRA-8, WRA-9, WRA-10, WRA-11
Chauntra 3 WRA-12, WRA-13, WRA-14
Chimbalhar 2 WRA-15, WRA-16
Gorta 1 WRA-17
Kukaina 1 WRA-18,
Lahad 2 WRA-19, WRA-20
Lunapani 1 WRA-21
Mahakal 3 WRA-22, WRA-23, WRA-24
Mehnja 2 WRA-25, WRA-26
Nagrota 3 WRA-27, WRA-28, WRA-29
Panchrukhi 3 WRA-30, WRA-31, WRA-32
Paprola 3 WRA-33, WRA-34, WRA-35
Rani Sidhpur 3 WRA-36, WRA-37, WRA-38
Salyana 3 WRA-39, WRA-40, WRA-41
Sulah 5 WRA-42, WRA-43, WRA-44, WRA-45, WRA-46
Unknown 2 WRA-47, WRA-48
Kunsal 1 WRA-49
Usterh 4 WRA-50, WRA-51, WRA-52, WRA-53

Incompatabiltiy of all the lines of cluster with blast isolates

Cluster No of isolates to
WRA-1 which the lines of
WRA-2 the cluster are Pg-1 Pg-2 Pg-3 Pg-4 Pg-5 Pg-6 Pg-7 Pg-8 Pg-9 Pg-10
WRA-6
WRA-7 resistant
WRA-41
WRA-16
R-575
WRA-26
WRA-39
WRA-40
I
WRA-22
WRA-49 0-2 + + + + + + + + + +
WRA-17
WRA-25
WRA -3
WRA-11
WRA-33
WRA-34
WRA-35
WRA-23
II 1-4 + + + + + + + + + +
WRA-24
WRA-30
WRA-9
WRA-10
WRA-15
WRA-19
WRA-29
WRA-37
III 2-5 + + - + + + + + - +
WRA-38
WRA-20
WRA-36
WRA-42
WRA-43
WRA-45
WRA-46 IV 3-6 + + - + + + + + + +
WRA-51
WRA-4
WRA-31
WRA-32
WRA-50
WRA-12
WRA-44 V
WRA-13
WRA-14
WRA-53
5-7 + + + + + + + + + +
China-988
WRA-52
WRA-5
WRA-18
WRA-21
Palmadhan-957 VI 6-10 + - - + + - + - + -
WRA-47
Himdhan
WRA-48
WRA-8
WRA-28
HPU-741
+ - + - + + + + - +
Himalaya-2
WRA-27 VII 3-6
0.25
0.25 0.50
0.50 0.75
0.75 1.0
1.00
Similarity(%)

Figure 1. Cluster analysis of wild rice accessions and rice cultivars based on their reaction to M. grisea isolates. Cophenetic correlation coeffi-
cient (r) of the dendrogram is 0.95. ‘–’, All the lines of the cluster show incompatible reaction with given M. grisea isolate; ‘+’, All or few lines of
the cluster show compatible reaction with given M. grisea isolate.

pared to cultivated rice. Resistance speci- novel. We report here, results on explor- genetic base for blast resistance in culti-
ficities in wild rice must be evolving in- ing the possibility of identifying novel vated rice. O. rufipogon accessions col-
dependently even after domestication of resistance specificities in O. rufipogon lected from different regions of HP,
cultivated rice, and therefore these could be types for eventual diversification of the together with five elite rice cultivars as

444 CURRENT SCIENCE, VOL. 89, NO. 3, 10 AUGUST 2005


control, were screened against M. grisea
isolates endemic to the region.
A total of 53 O. rufipogon accessions
numbered WRA-1 to WRA-53, were col-
lected from 18 rice-growing localities of
HP (Table 1). For each accession, seeds
were collected from a single panicle and
stored at 50°C for 5 days to break the
dormancy. Seeds of all the accessions
and six elite rice cultivars, China-988,
Himalaya-2, Himdhan, HPU-741, Palm-
dhan-957, and R-575 were sown in plastic
trays (20 × 15 × 4 cm) filled with potting
mixture (soil : sand :: 3 : 1). Two rows of
seeds of each line were planted in each
tray using 5–6 seeds per hill and trays
were kept in a greenhouse at 25 ± 1°C
for 15 days. All the lines were tested
twice against each isolate and for a few
lines showing ambiguous reactions, the
experiments were repeated until consis-
tent reactions were obtained.
Ten M. grisea isolates of varied viru-
lence spectrum were collected from rice
blast screening nursery, sown with lines
of different resistance specificities (Table 2).
Single spore cultures of the isolates were
established on oatmeal agar. Mycelia
from ten-day-old slants were macerated
Table 2. Origin of M. grisea isolates used in the study
Isolate no. Rice genotype of origin
Pg-1 Unknown
Pg-2 Dular
Pg-3 Himalaya-799
Pg-4 C102 PKT
Pg-5 HPU-741
Pg-6 Sha-tiao-tsao
Pg-7 Himalaya-2
Pg-8 C101A51
Pg-9 Usen
Pg-10 Zenith
? Unknown resistance gene(s).
Figure 2. Reaction of cultivated rice (HPU-741) and O. rufipogon accessions WRA-18 and
WRA-21 to M. grisea.
CURRENT SCIENCE, VOL. 89, NO. 3, 10 AUGUST 2005
SCIENTIFIC CORRESPONDENCE
in 5 ml distilled water and plated onto
Mathur’s medium6 for sporulation. After
8 to 10 days of incubation at 25 ± 1°C,
plates were washed with 10 ml distilled
water to make a spore suspension. Each
spore suspension was filtered through
two layers of muslin cloth and the spore
concentration was adjusted to 40–50 co-
nidia per microscopic field (40x). About
30–40 ml of the spore suspension con-
taining Tween-20 (0.2%) was sprayed
onto 15-day-old seedlings using a glass
atomizer. Inoculated seedlings were kept
in a humidity chamber maintained at 25 ±
1°C for 24 h and thereafter sprayed three
to four times a day with distilled water to
maintain high humidity. Disease reaction
of each line was recorded 6 days after
inoculation using a 0–5 disease scoring
scale7, i.e. 0, No evidence of infection; 1,
Brown specks smaller than 0.5 mm in dia-
meter, no sporualtion; 2, Brown specks
about 0.5–1.0 mm in diameter, no sporu-
lation; 3, Roundish to elliptical lesions
1–3 mm in diameter, grey centre sur-
rounded by brown margins; 4, Typical
spindle-shaped blast lesions capable of
sporulation, 3 mm or longer in diameter;
5, Lesions as in 4, but about half of 1–2
Resistance gene(s) reported in the host genotype
? ness of the dendrogram. On the basis of
Pi-ka+?
? were grouped into seven clusters (I–VII)
Pi-4a
? ferent clusters were effective against 0 to
Pi-ks
?
Pi-2
Pi-a+?
Pi-z, Pi-a, Pi-i
leaf blades killed by coalescence of lesions.
Genotypes exhibiting reaction types 0, 1,
2 and 3 were rated as resistant, while those
showing reaction types 4 and 5 were con-
sidered susceptible.
Cluster analysis of the lines based on
their reaction to ten M. grisea isolates
was done using the NTSYS-pc package8.
A binary matrix of resistance spectrum
of lines based upon their reaction to 10
M. grisea isolates was constructed by
coding resistance as ‘1’ and susceptibility
as ‘0’. The binary matrix was then used
to derive a similarity matrix based on the
simple matching coefficient (SM = m/n,
where m is the total number of isolates to
which two lines in a pair are resistant or
susceptible, and n is the total number of
isolates) using the SIMQUAL program
of the NTSYS-pc package. To test the
phenetic robustness of the dendrogram,
cophenetic correlation coefficient (r) was
computed by comparing the similarity
matrix with cophenetic matrix using ma-
trix comparison (MXCOMP) program of
NTSYS-pc package.
The virulence frequency of M. grisea
on different wild rice accessions ranged
from 0 to 100%, while on cultivated rice
genotypes 33.33–100% of the isolates
were virulent (Table 3). Cluster analysis
of virulence data generated a dendrogram
with high cophenetic correlation coefficient
(r = 0.95), thus substantiating the robust-
their resistance spectrum, all the lines
(Figure 1). The constituent lines of dif-
10 pathogen isolates. Genotypes falling
in cluster I were least effective against the
pathogen population as the constituent
lines were resistant to only 0 to 2 isolates.
Local rice cultivar R-575 and two wild
rice lines, WRA-16 and WRA-26 falling
in this cluster were susceptible to all the
isolates tested. Genotypes in clusters V
and VI exhibited the broadest resistance
spectrum, as individual lines of these
clusters were resistant to 5 to 7 and 6 to
10 isolates respectively. Two accessions
within the cluster VI, namely WRA-18
and WRA-21 collected from Kukaina and
Luna Pani, were resistant to all the iso-
lates tested. Interestingly, these acces-
sions consistently exhibited ‘0’ and ‘2’
type reactions to all the pathogen isolates
(Figure 2). The wild rice accessions from
one location (Bodal) exhibited greater
diversity for blast resistance by exhibiting
incompatibility with 1 to 8 isolates. These
445
SCIENTIFIC CORRESPONDENCE
Table 3. Virulence spectrum of rice limited isolates of M. grisea on wild rice (O. rufipogon) accessions and cultivated rice genotypes

Isolate no.
Virulence
Genotype/isolate no. Pg-1 Pg-2 Pg-3 Pg-4 Pg-5 Pg-6 Pg-7 Pg-8 Pg-9 Pg-10 frequency (%)
Oryza rufipogon
WRA-1 4 4 4 0 4 0 5 4 5 4 80
WRA-2 5 4 4 0 4 4 4 4 4 4 90
WRA-3 4 4 1 4 4 4 0 4 4 4 80
WRA-4 0 4 0 0 0 4 0 0 4 4 40
WRA-5 4 0 2 0 0 2 4 0 1 0 20
WRA-6 4 4 5 0 4 4 5 4 4 4 90
WRA-7 4 4 4 0 4 4 4 4 4 4 90
WRA-8 4 1 4 3 0 4 4 4 1 0 50
WRA-9 4 4 0 4 5 4 4 4 0 2 70
WRA-10 4 4 0 4 4 4 4 4 0 0 70
WRA-11 4 4 0 4 4 4 4 5 4 4 90
WRA-12 0 – 2 0 0 4 0 3 4 0 22
WRA-13 0 4 0 4 0 4 0 0 0 0 30
WRA-14 0 5 1 4 4 4 2 0 4 0 50
WRA-15 4 4 0 4 4 4 4 4 3 0 70
WRA-16 4 – 4 5 4 4 4 4 4 4 100
WRA-17 4 4 4 4 4 4 4 0 4 0 80
WRA-18 0 0 0 0 0 0 0 0 0 0 0
WRA-19 4 4 0 0 4 5 4 4 0 0 60
WRA-20 4 4 2 4 0 3 4 4 0 0 50
WRA-21 2 2 2 2 2 2 2 2 2 2 0
WRA-22 4 4 4 5 5 0 4 0 4 4 80
WRA-23 4 4 3 4 4 1 0 0 4 4 60
WRA-24 4 4 4 4 4 4 2 0 0 0 60
WRA-25 0 4 4 4 0 4 4 4 4 4 80
WRA-26 4 4 4 5 4 4 4 4 4 4 100
WRA-27 0 3 4 0 4 1 4 0 3 4 40
WRA-28 4 0 4 0 4 4 4 4 0 4 70
WRA-29 0 4 0 – 4 5 4 4 0 0 56
WRA-30 4 4 4 4 4 4 0 4 0 4 80
WRA-31 1 4 0 2 0 4 3 0 0 4 30
WRA-32 0 4 0 0 0 4 0 3 0 4 30
WRA-33 4 4 0 4 4 4 3 4 4 0 70
WRA-34 4 4 0 4 4 4 0 4 4 0 70
WRA-35 4 4 2 4 4 4 0 4 4 0 70
WRA-36 0 4 0 4 4 2 4 4 4 4 70
WRA-37 4 – 0 4 0 4 4 4 0 5 67
WRA-38 4 – 0 4 5 4 4 4 2 4 78
WRA-39 4 4 4 4 4 3 4 4 4 4 90
WRA-40 4 4 4 4 4 0 4 5 4 4 90
WRA-41 4 4 4 0 4 4 4 4 4 4 90
WRA-42 0 5 0 0 4 3 4 4 4 4 60
WRA-43 0 4 0 0 4 4 4 4 4 4 70
WRA-44 0 4 0 2 4 4 4 0 0 4 50
WRA-45 0 4 2 0 4 0 0 4 4 0 40
WRA-46 0 4 0 0 4 1 0 4 4 0 40
WRA-47 4 0 0 4 4 0 4 1 0 0 40
WRA-48 0 0 2 4 4 0 4 0 0 0 30
WRA-49 4 5 4 0 4 4 4 0 4 4 80
WRA-50 0 4 0 2 4 4 0 2 5 4 50
WRA-51 4 4 0 0 4 0 4 4 4 0 60
WRA-52 0 0 4 0 0 4 4 0 4 0 40
WRA-53 4 0 0 0 1 5 0 2 0 4 30
Oryza sativa
China-988 4 – 1 0 0 4 1 4 0 2 33.33
Himdhan 4 0 0 4 4 0 4 0 0 0 40
HPU-741 5 1 4 2 4 0 4 4 0 0 50
Palamdhan-957 4 0 0 0 4 0 2 0 4 2 30
R575 4 4 4 4 4 4 4 4 4 4 100
Himalaya-2 4 0 0 2 4 0 4 4 3 0 40
a
Computed as percentage of total isolates that produce compatible reaction (reaction type 4–5) on a given genotype; ‘–’, reaction not determined.

446 CURRENT SCIENCE, VOL. 89, NO. 3, 10 AUGUST 2005


results indicated that there is a great di-
versity for blast resistance in the native
O. rufipogon populations.
Since the northwestern Himalayan re-
gion is one of the centres of diversity for
Oryza spp., a prolonged co-evolution of
host–pathogen coupled with mixed mating
system of wild rice9, has probably enhan-
ced the diversity for resistance in the na-
tive wild rice populations. The present
data also indicate that some wild rice varie-
ties share common resistance specificities
with cultivated rice varieties. For example,
all the accessions in cluster VI, including
cultivated rice genotypes, Palmdhan-957
and Himdhan, shared common resistance
specificities to pathogen isolates Pg2,
Pg3, Pg6, Pg8 and Pg10. Likewise, all
the accessions in cluster VII, that also
includes rice cultivars HPU-741 and Hima-
laya-2, shared resistance specificities to
isolates Pg2, Pg4 and Pg9. These com-
mon specificities could either be the re-
sult of recent transfer from cultivated to
wild types or might be the relics of ancient
diversity that existed in cultivated rice since
its domestication from wild rice. Never-
theless, wild rice germplasm represents a
potential source of new blast-resistance
specificities for broadening the genetic
Rapidly in vitro multiplied Drosera as reliable source for plumbagin
bioprospection
Drosera (Droseraceae) is the largest group
of insectivorous plants. It comprises of
170 species and is cosmopolitan. All spe-
cies of Drosera contain plumbagin (5-
hydroxy-2-methyl-1,4-napthoquinones) or
ramentaceone (7- methyljuglone) or both1,2.
Plumbagin exhibits a variety of therapeu-
tic functions such as anti-microbial, anti-
tuberculosis, bronchial infection, whooping
cough, anti-asthma, antispasmodic, anti-
cancer, enhances in vitro phagocytosis of
human granulocytes, antileprosy, antiferti-
lity, abortifacient, chitin synthetase inhibitor,
immunomodulator, cosmetic, aphrodisiac,
used against old age, arteriosclerosis, ex-
tract used in certain sweets, antifeedant,
anti-malarial, hyperglycaemic and other
properties3–6. In European countries,
Droseras is included in pharmacopoeias
and dried plants are marketed as ‘Herba
Droserae, Herba Drosera rotundifoliae’
CURRENT SCIENCE, VOL. 89, NO. 3, 10 AUGUST 2005
SCIENTIFIC CORRESPONDENCE
base for blast-resistance in rice. For ex-
ample, wild rice accessions WRA-18 and
WRA-21 exhibited broad resistance spec-
trum to all the isolates tested, including
Pg-1 that infects all the cultivated rice
genotypes tested. Since O. rufipogon with
AA genome is sexually compatible with
cultivated rice, these two accessions can
be used as potential donors of easily trans-
ferable blast resistance in rice breeding
programmes. Further investigations on
type of gene action and allelic relationship
of these resistance sources with already
known blast resistance genes that are
highly effective against rice blast popula-
tion of HP are urgently needed10.
1. Nagato, Y. and Yashimura, A., Rice
Genet. Newsl., 1999, 16, 8–14.
2. Li, R.B., Int. Rice Res. Notes, 1994, 19,
8–9.
3. Plucknett, D. L., Smith, N. J. H., Williams,
J. T. and Anishetty, N. M., In Gene
Banks and the World’s Food (eds Smith,
N. J. H., Williams, J. T. and Anishetty,
N. M.), Princeton University Press,
Princeton, NJ, 1967, pp. 171–185.
4. Sharma, R. D., Sci. Rep., 1998, 9–15.
5. Khush, G. S., Plant Mol. Biol., 1997, 35,
25–34.
which is difficult to obtain from natural
habitat. Hence, Drosera from the southern
hemisphere, such as D. indica, D. bur-
manii, D. peltata, D. ramentacea and D.
madagascariensis is being exported to
European countries7. In India D. indica L.,
D. burmanii Vahl. and D. peltata J.E.Sm.
ex Willd have been reported from differ-
ent parts. They are used as vital compo-
nents in an Ayurvedic preparation called
‘Swarnabhasma’ (Golden ash). Macera-
ted D. indica is used to remove corns and
has been categorized under vulnerable
medicinal plants8,9.
We have collected D. indica and D.
burmanii from the following locations in
Andhra Pradesh (district name mentioned
in parenthesis) Gachibowli (Ranga Reddy),
Narsapur (Medak), Anjodigadda (Visak-
hapatnam), Talakona (Chitoor), Tada
(Nellore), Mulakanuru (Kharmnagar),
6. Mathur, R. S., Barnett, H. L. and Lily, V.
G., Phytopathology, 1950, 40, 104–114.
7. Mackill, D. J. and Bonman, J. M., Phyto-
pathology, 1992, 82, 746–749.
8. Rohlf, F. J., NTSYS-pc: Numerical Taxo-
nomy and Multivariate Analysis System
Version 1.80. Exeter Software, Setauket,
New York, 1993.
9. Gao, L. Z. and Hong, S. G. D., Theor.
Appl. Genet., 2000, 101, 494–502.
10. Rathour, R., Singh, B. M. and Sharma, T.
R., J. Phytopathol., 2004, 152, 304–312.
Received 23 November 2004; revised accep-
ted 20 May 2005
R. RATHOUR1,*
V. S. GAUR1
R. P. KAUSHIK2
R. S. CHAUHAN1
1
Advanced Centre of Hill Bioresources
and Biotechnology,
CSK H.P. Agricultural University,
Palampur 176 062, India
2
Regional Rice Research Station,
CSK H.P. Agricultural University,
Malan 176 047, India
*For correspondence.
e-mail: rrathore@hillagric.org
Kondaparthi (Warangal); D. indica: Chel-
vaye (Warangal); Panchamatalu (Kurnool);
Arakuvally (Visakhapatnam), Pakhal
lake (Warangal); Srisailam (Kurnool),
and D. burmanii: Motugudem (Khammam);
Tirumala (Chitoor). Eriocoluan, Utricu-
laria and Sphagnum were the associated
species in most of the investigated locations.
The microhabitat in all these locations
comprises wet slopes and poorly drained
rocky outcrops with sand and clay soils,
often with moss and peat. Urbanization,
drought, agricultural pollutants, soil ero-
sion, habitat conversion and fragmenta-
tion are the primary causes of threat.
We initiated research on Drosera as
part of ex situ conservation and established
in vitro protocols for rapid multiplication.
Shoots of D. indica and D. burmanii
were cultured on MS basal medium with
3% sucrose, 0.3% agar at pH 5.7. The
447