1. 625
IMPROVED GERMINATION OF GYMNACRANTHERA CANARICA
F O R M A T T E D P R O O F
Tambat, B., Vishwanath, K., Chaithra, G.N. and Hareesh, T.S. (2006), Seed Sci. & Technol., 34, 625-630
Improved germination of Gymnacranthera canarica Warb. an
endangered, endemic tree species of Myristica swamps, Western
Ghats, India
B. TAMBAT1
, K. VISHWANATH2
, G.N. CHAITHRA3
AND T.S. HAREESH4
1
Department of Crop Physiology and University of Agricultural Sciences, Bangalore, India
1
(E-mail: btambat@yahoo.com)
2
Department of Seed Science and Technology, University of Agricultural Sciences, Bangalore, India
3
College of Forestry, Sirsi, Uttara Kannada, Karnaraka, India
4
Bio-Science Department, University of Mysore, Hemagangothri, Hassan, Karnataka, India
(Accepted April 2006)
Summary
Gymnacranthera canarica Warb, an endangered, endemic tree species of Western Ghats, India. The species is
exclusively associated with swampy conditions and habitat destruction seems to be the major threat. Although
seed viability was 98 per cent confirmed by TZ test, initial germination was only 40% suggesting dormancy
problem in this species. Among the 14 treatments, seed coat removal itself enhanced germination by 20 per cent
(60%) over control (40%) showing partial permeability of seed coat to water. GA3 @ 50,100 and 250 ppm with
partial removal of seed coat showed significantly higher germination by 39, 33 and 30 per cent respectively
over control. However, higher concentration of GA3 @ 500 ppm showed inhibitory effect (10%). With respect
to IAA we fail to observe any pattern in this species. Further, nil germination in boiling water, cold, boiling
water+ hormonal treatment confirming the dormancy of seeds. Our results provide means to enhance the seed
germination and helps for restoration and conservation of an endangered species in Western Ghats.
Introduction
Gymnacranthera canarica Warb. is one of the threatened tree species (IUCN, 2002) that
is associated with swampy habitat in the Western Ghats, a global mega diversity hot spot
in India (Myers et al., 2000). It belongs to family Myristicaceae (or nutmeg), a family one
among the primitive flowering plants and mostly known for its mace and seeds, which
are of high medicinal importance (Purseglove et al., 1981; Yoganarasimhan, 2000). The
fruits are globose with thick and fleshy pericarp. The seed conforms to the fruits, testa
woody with bright red colored aril (Gamble, 1935). The seeds contain much fat, probably
as an adaptive mechanism for dispersal and survival under swampy conditions. In past
few decades due to conversion of swamps into areca gardens and diversion of water
for agricultural purpose, regeneration and survival of G. canarica is drastically affected
(Chandran et al., 1999; Chandran and Mesta, 2001; Tambat et al., 2004). The field study
by Tambat et al., (2004) has demonstrated that species is exclusively associated with
swampy conditions with few meters away from the swamps the species disappear. Thus,

2. 626
B. TAMBAT, K. VISHWANATH, G.N. CHAITHRA AND T.S. HAREESH
F O R M A T T E D P R O O F
habitat destruction, which influences the local microclimate, could be one of the major
cause for reduced germination and survival of G. canarica. As for as seed germination is
concerned the species possess hard seed coat and shown lower germination under nursery
conditions (unpublished). It is likely that the species may have seed coat induced dormancy,
as observed in few forest species (Naidu et al., 2000; Tigabu and Oden, 2001).
The present study aims to assess the effective seed treatment that would enhance
seed germination under laboratory conditions and thereby facilitate conservation of G.
canarica an endangered tree species through re-introduction or enrichment planting.
The reintroduction of species in to its natural habitat is one of the best strategies for the
conservation of critically endangered endemic species, as it would act as a functional
bridge between the ex situ and in situ conservation methods (Frankel et al., 1995).
Materials and methods
Mature seeds of Gymnacranthera canarica Warb. were collected from Kathlekan
forest, Siddapura range (14° 16' 250, 74° 44' 880) of Uttara Kannada in Karnataka
during October and November 2004. The seeds were washed in water and bulked before
imposing any treatment. The seed viability was confirmed by following 2,3,5-Triphenyl
Tetrazolium Chloride (TTC) test using randomly selected 300 seeds of 100 seeds in each
replications from the seed lot (Agrawal and Dadlani, 1995). The seeds were allowedto
germinate in pots containing sand under ambient conditions, as Bhat and Kaveriappa,
(2002) have shown better germination by sand method than blotter in laboratory condition
(25-28°C) in Gymnocranthera farquhariana Hook. (Synonymous with Gymnacranthera
canarica Warb., Gamble 1935). Totally 14 treatments were imposed including control
with three replications of 50 seeds and details as follows; (1) Control (without any
treatment), (2) Entire seed coat removal, (3) Diaphragm aperture + GA3 @50 ppm (16hrs),
(4) Diaphragm aperture + GA3 @100 ppm (16hrs), (5) Diaphragm aperture + GA3 @250
ppm (16hrs), (6) Diaphragm aperture + GA3 @500 ppm (16hrs), (7) Diaphragm aperture +
IAA @50 ppm (16hrs), (8) Diaphragm aperture + IAA @100 ppm (16hrs), (9) Diaphragm
aperture + IAA @250 ppm (16hrs), (10) Diaphragm aperture + IAA @500 ppm (16hrs),
(11) Boiling water (100°C for 15 sec), (12) Boiling water (100°C for 15 sec)+ IAA
@100 ppm (16 hrs), (13) Boiling water (100°C for 15 sec) + GA3 @100 ppm (16 hrs) and
(14) Cold (-12°C for 12 hrs).
The number of seeds germinated was recorded by taking out the seedling from the
sand bed. Per cent germination was computed based on normal seedling basis. The data
of germination was subjected to arcsin transformation for statistical analysis. Following
Completely Randomized Design (CRD) and adopting “Fischer’s Analysis of Variance
Technique” critical difference values were calculated at 5 per cent probability level (where
“F” test was significant). Dunnett’s “t” test was then performed to compare control with
all other treatments (Panse and Sukhatme, 1967).

3. 627
IMPROVED GERMINATION OF GYMNACRANTHERA CANARICA
F O R M A T T E D P R O O F
Results and discussion
From Tetrazolium test it was confirmed that viability of seeds selected for imposing
dormancy treatments was 98 per cent and were showed deep red embryonic axis and
cotyledons. Only two per cent of seeds were showed unstained embryonic axis and pale
red staining with unstained areas in cotyledons. However, we could observe only 40 per
cent germination under ambient condition, emphasizing the presence of dormancy in these
seeds.
Pre sowing treatments have been shown to enhancing seed germination in many
species (Kattimani et al., 1999; Pandey et al., 2000; Joshi and Dhar, 2003). The effect
of seed treatment on germination of Gymnocranthera canarica after four months has
been shown in figure 1. Per cent germination of seeds with 14 treatments varied from 0
to 79 per cent. As for as improved germination, seed coat removal alone increased the
germination by 20 per cent indicating the partial permeability of seed coat to the water.
Such results were also noticed by Naidu et al., (2000) in Sapindus trifoilatus and Tigabu
and Oden, (2001) in Albizia sp.
The application of GA3 along with partial seed coat removal further enhanced the
germination (40% over control). However, the per cent germination varied with the
concentration of GA3; we observed maximum germination (79%) in seeds treated with
100 ppm GA3 then that of other treatments (i. e. 73 % @ 50 ppm and 70% @ 250 ppm).
Forest species are known to contain certain inhibitory compounds in seed coat and (or)
Figure 1. Mean seed germination (%) as influenced by the treatment in Gymnacranthera canarica. (Note:
Hormonal treatments was imposed by soaking the seeds in different concentrations of hormone solution after
diaphragm aperture / partial removal of seed coat for 16 hours).
* 1
- Significantly superior over control
* 2
- Significantly inferior over control
NS - Non significant over control
0
10
20
30
40
50
60
70
80
90
Control
Entireseedcoatremovel
HormoneGA350(16hr)
HormoneGA3100(16hr)
HormoneGA3250(16hr)
HormoneGA3500(16hr)
HormoneIAA50(16hr)
HormoneIAA100(16hr)
HormoneIAA250(16hr)
HormoneIAA500(16hr)
Boilingwater(100oC-15sec)
Bolingwater+IAA100
Bolingwater+GA3100
Cold(-12oC-12hrs)
MeanGermination(%)
LSD(P<0.05): 6.83
*
1
*
1
*
1
*
1
*
2
*
2
NS
*
2
NS
*
2
*
2
*
2
*
2

4. 628
B. TAMBAT, K. VISHWANATH, G.N. CHAITHRA AND T.S. HAREESH
F O R M A T T E D P R O O F
endosperm, which affects the germination process; external supply of GA3 neutralizes
such effect and facilitates germination (Evenari, 1949; Mary, 1972). Earlier studies on
forest species have showed GA3 enhanced seed germination in Cassia (Padma et al.,
1996), Annona squomosa (Pawshe et al., 1997), Pittosporum floribundum (Subodh et
al., 1998), Echinacea purpurea (Kochankov et al., 1998), Albizia sp. (Tigabu and Oden,
2001), Arbatus andrachne (Karam and Al-Salem, 2001).
However, germination was reduced drastically to 10 per cent as GA3 concentration
increased to 500 ppm, which may possibly due to imbalanced cell activity or toxic effect
of GA3 at higher concentration (Takashashi et al., 1991; Deno, 1993). Such results were
also noticed by Naidu et al., (2000) in Sapindus trifoliate, Tigabu and Oden (2001) in
Albizia sp. and Karam and Al-Salem (2001) in Arbutas andranchne.
Indole acetic acid (IAA) a phyto-hormone which is also known to enhance the
germination in certain species such as Aleurites fordii (Chatterjee, 1960), Hibiscus
esculent (Omran et al., 1980) and Sapindus trifoliates (Naidu et al., (2000). However, in
the present study irrespective of concentrations gradient we fail to observe such enhanced
germination. IAA @ 100 and 500 ppm the germination was found to be on par with the
control, where as IAA @ 50 and 250 ppm there was significantly lower germination than
that of control. Since, there is no clear pattern we feel there is a need to understand the
role of IAA in Gymnocranthera canarica during germination with further studies.
Other treatments such as boiling water, cold, boiling water along with hormones
showed nil germination and were found to be significantly inferior over control. No
germination also indicates dormancy problems in this species. These results might also
indicate temperature sensitive of the species which is reported in Myristica malabarica
Lam. a close relative of G. canarica (Anil Kumar et al., 2002). Thus, in this regard
further studies are needed.
In conclusion, Gymnacranthera canarica is an obligatory swampy species thus
may contain certain inhibitory compounds in the seeds, which are leached out due to
the continuous flow of water in the natural swampy habitat. Now, due to anthropogenic
activity the natural habitats have been disturbed thus natural germination process is
affected. With regards to improved germination our results indicate that seed germination
can be enhanced up to 79 per cent using GA3@100ppm with partial removal of seed
coat. In the present scenario our results provide an alternative means to enhance the seed
germination, thus helps in conservation and restoration of a habitat specific, endemic
and endangered tree species of the Western Ghats, India. For the purpose of enrichment
planting, development of seedlings through seeds is known to be best method as it is
cheaper and also helps to maintain the natural genetic variability among the planting
material (Tilki, 2004).
Acknowledgements
We would like to thank Mr. P.G. Suraj, Dr. R. Vasudev, Mr. I. Sriram and students of
College of Forestry, Sirsi for the technical support during the fieldwork. We acknowledge
Karnataka Forest Department for the permission and British Petroleum (BP), UK for the
financial support.

5. 629
IMPROVED GERMINATION OF GYMNACRANTHERA CANARICA
F O R M A T T E D P R O O F
References
Agrawal, P.K. and Dadlani, M. (1995). Techniques seed science and technology. Second Edition. South Asian
publishers limited, India.
Anil Kumar, C., Babu, K.P. and Krishnan, P.N. (2002). Seed storage and viability of Myristica malabarica Lam.
an endemic species of Southern Western Ghats (India). Seed Science and Technology, 30, 651–657.
Bhat, R.P. and Kaveriappa, K.M. (2002). Polyembryony in Gymnocranthera farquhariana Hook. F. and
Thomson) Warb. Indian Forester, 6, 820–822.
Chandran, M.D.S., Divakar, K. Mesta and Manjunath, B. Naik (1999). Inventory and conservation of the
Myristica swamps of Uttara Kannada. Forest research and training institute Western Ghats forestry project,
Bangalore, (Final report).
Chandran, M.D.S. and Mesta, D.K. (2001). On the conservation of the Myristica swamps of the Western Ghats,
page no. 1-19. In Forest Genetic Resources: Status, Threats and Conservation Strategies (eds. Uma Shaanker,
R., K.N. Ganeshaiah and Kamaljit S. Bawa). Oxford and IBH Publishing Co. Pvt. Ltd. New Delhi.
Chatterjee, S.K. (1960). Effect of pre sowing treatment for tung (Aleurites fordii). Science and Culture, 26,
130-131.
Deno, N. (1993). Germination theory and practice, 2nd
edition, Lenor Drive, State college, Pennsylvania.
Evenari, M. (1949). Germination inhibitors. Botanical Review, 15,153-194.
Frankel, O.T., Brown, A.H.D. and Burdon, J.J. (1995). Conservation of Plant Biodiversity, Combridge University
Press, Pp 229.
Gamble, J.S. (1935). Flora of the Presidency of Madras, Vol. I to III. Adlard and Son, Limited, London.
IUCN (2002) Red listed plants of India.URL., www.iucn.com
Joshi, M. and Dhar, U. (2003). Effect of various treatment on seed germination of Heracleum condicans. Wall.
Ex DC.: A high value medicinal plant. Seed Science and Technology, 31, 737–743.
Karam, N.S. and Al-Salem, M.M. (2001). Breaking dormancy in Arbutus andrachne L. seeds by stratification
and Gibberlic acid. Seed Science and Technology, 29, 51–56.
Kattimani, K.N., Reddy, Y.N. and Rao, R.B. (1999). Effect of pre sowing treatments on germination, seedling
emergence, seedling vigour and root yield of Ashwagandha (Withania somnifera. Daunal). Seed Science and
Technology, 27, 483–488.
Kochankov, V.G., Grazesik, M., Chojnowski, M. and Nowak, J. (1998). Effect of temperature, growth regulators
and other chemicals on Echinacea purparea L. Moench seed germination and seedling survival. Seed Science
and Technology, 26, 547–554.
Mary, R.C. (1972). Tannins as gibberellins antagonists. Plant Physiology, 49, 323–330.
Myers, N., Mittermeier, R.A., Mittermeier, C.G., da Fonseca, G.A.B. and Kent, J., (2000). Biodiversity hot-spots
for conservation priorities. Nature, 403, 853–958.
Naidu, C.V., Rajendrudu, G. and Swamy, P.M. (2000). Effect of plant growth regulators on seed germination of
Sapindus trifoilatus Vahl. Seed Science and Technology, 28, 249–252.
Omran, A.F., El-Vakry, A.M. and Gawaish, R.A. (1980). Effect of soaking seeds in some growth regulator
solutions on the growth, Chemical constituents and yield of okra. Seed Science and Technology, 8, 161–
168.
Padma, V. Satyanarayana, G. and Muralimohan Reddy, B. (1996). Studies on pre-sowing seed treatments in
three species of Cassia. Seed Research, 24, 51–54.
Pandey, H., Nandi, S.K., Nadeem, M. and Palni, L.M.S. (2000). Chemical stimulation of seed germination in
Aconitum heterophyllum Well. and A. balfourii Stapf: important Himalayan species of medicinal value, Seed
Science and Technology, 28, 39–48.
Panse, V.S. and Sukhatme, P.V. (1967). Statstical Methods for Agricultural workers. Indian Council of
Agricultural Research Publication. New Delhi.
Pawshe, Y.H., Patil, B.N. and Patil, L.P. (1997). Effect of pre germination seed treatment on the germination
and vigour of seedlings in custard apple (Annona squamosa L.). Annals of Plant Physiology, 11, 150–154.
Purseglove, J.W., Brown, E.G., Green, C.L. and Robbins, S.R.J. (1981). Spice, Vol. I, Longman Scientific and
Technicals and Jihn Wiley and sons Inc. New York.
Subodh Airi, Rawar, R.S., Samant, S.S. and Dhar, U. (1998). Treatments to improve germination of four
multipurpose trees of central sub Himalaya. Seed Science and Technology, 26, 347–354.
Takashashi, N., Phinney, B. and MacMillan, J. (1991). Gibberellins, Springer Verlag, New york.

6. 630
B. TAMBAT, K. VISHWANATH, G.N. CHAITHRA AND T.S. HAREESH
F O R M A T T E D P R O O F
Tambat, B., Ravikanth, G., Nageswara Rao M. Chaithra G.N., Ramesh B.T. and R. Uma Shaanker, (2004).
Conservation of the Myristica Swamps - the highly threatened and unique ecosystem in the Western Ghats,
India. 2003 BP Conservation Programme, Silver award wining project, UK (Final Report).
Tigabu, M. and Oden, P. C. (2001). Effect of sacriication, gibberlic acid and temperature on seed germination of
two multipurpose Albizia species from Ethiopia. Seed Science and Technology, 29, 11–20.
Tilki, F. (2004). Improvement in Seed Germination of Arbutus unedo L. Pakistan Journal of Biological Sciences,
7, 1640–1642.
Yoganarasimhan, S.N. (2000). Medicinal Plants of India, Vol II, Published by Srinivasan V. and Kosal Ram K.
of cyber media, Bangalore.