Tea J. Bangladesh 2013

ISSN: 0253-5483
TEA JOURNAL OF BANGLADESH

Volume 42, 2013
BANGLADESH TEA RESEARCH INSTITUTE

SRIMANGAL-3210, MOULVIBAZAR
An organ of
BANGLADESH TEA BOARD
171-172, Baizid Bostami Road
Nasirabad, Chittagong
www.btri.gov.bd
Volume 42, 2013
Chief Editor Dr. Mainuddin Ahmed
EDITORIAL BOARD
Chairman
Major General Md. Abdus Salam Khan, afwc, psc
Chairman
Bangladesh Tea Board
Chittagong
Members
Mohammad Siddique
Member (R & D)
Chittagong
Dr. Mainuddin Ahmed

Director
Bangladesh Tea Research Institute
Srimangal, Moulvibazar
Md. Haroon-or-Rashid Sarker

Director
Project Development Unit
Srimangal, Moulvibazar
Volume 42, 2013
CONTENTS
Title
Page
Editorial vii
EFFECTS OF DIFFERENT PLUCKING SYSTEMS ON SHOOT GROWTH

DYNAMICS AND YIELD IN CONTRASTING TWO CULTIVARS OF TEA 1-9
(Camellia sinensis L.)
T. Ahmed, W.A.J.M. De Costa and M.A. Wijeratne
EVALUATION OF SOME INDIGENOUS PLANT EXTRACTS AGAINST

TEA MOSQUITO BUG, HELOPELTIS THEIVORA WATERHOUSE 10-20
(HEMIPTERA: MIRIDAE) INFESTING TEA
M.S.A. Mamun, M. Ahmed, S.K. Paul and R.S. Chowdhury
DEVELOPMENT OF IN VITRO MICRO PROPAGATION SYSTEM IN
TEA PLANT [CAMELLIASINENSIS (L.) O. KUNTZE] USING SHOOT 21-30
TIP AND NODAL SEGMENT EXPLANTS
S. Boonerjee, M. I. Hoque and R. H. Sarker
INTEGRATED MANAGEMENT OF GALL DISEASE IN TEA 31-36

M. Ali, M.S. Islam and M. Ahmed
EFFECT OF POULTRY MANURE ON THE YIELD OF MATURE TEA

VIS-À-VIS SOIL PROPERTIES 37-43
A. Q. Khan, A. Biswas, K. F. T.Zohora and A.K. Saha
EDITORIAL
The present issue (volume 42) of Tea Journal of Bangladesh contains five research
papers on different aspects of tea science.
The first paper deals with the effects of different plucking systems on shoot growth
dynamics and yield thereby in two cultivars of tea having some differing (opposing)
characteristics. The paper reveals that shoot growth dynamics of yield related
parameters for ultimate yield of tea are different for different plucking treatments
and vary from cultivar to cultivar. Though the original research work was carried out
In Sri Lanka, it also holds true for Bangladesh condition too. The work would have
some implications in upgrading the concept of plucking system in our tea.
The second article contains the evaluation of some indigenous plant extracts for
controlling Helopeltis commonly known as Tea Mosquito Bug. The work indicated
that most of the studied plants extracts were effective in controlling the target pest
Helopeltis. The order of their effectiveness was:
Bur weed>Lantana>Datura>Neem>Mahogani>Katamehedi.
All these plants are available in and around tea growing areas of Bangladesh. The
extracts are environment friendly and very effective. Those could be used safely both
in organic and inorganic tea fields.
The third paper is on the development of a suitable protocol for in vitro culture of tea
from shoot tip and nodal explants. The authors tried to establish a protocol for
micro-propagation of tea. It has been a long awaited technique to grab for the
commercialization of an easier, economic and suitable propagation method for our
tea clones.
The next paper is on the integrated management of Tea Galls - a minor disease in
tea. The work hints that Deep Skiff (DSK) operation followed by the application of
a systemic fungicide Carbendazim @ 750 gm per hectare is the most effective
method to minimize gall disease in tea.
The last paper is on the effect of poultry manure on the yield of mature tea as well as
on soil properties. It reveals that comparable yield can be achieved even after 75%
reduction of conventional chemical fertilizer (BTRI recommended dose) if poultry
manure is added in appropriate dose. Thus this widely available manure from poultry
droppings can minimize the use of chemical fertilizer and has a potential in reducing
the cost of inorganic fertilizer as well as ill effects on tea soil properties.
(Dr. Mainuddin Ahmed)

Chief Editor
Tea J. Bangladesh, 42: 1-9, 2013 ISSN: 0253-5483
EFFECTS OF DIFFERENT PLUCKING SYSTEMS ON SHOOT

GROWTH DYNAMICS AND YIELD IN CONTRASTING TWO
CULTIVARS OF TEA (CAMELLIA SINENSIS L.)
T. Ahmed1*, W.A.J.M. De Costa2 and M.A. Wijeratne3
Abstract
A field experiment was conducted to study the effects of different plucking
systems on shoot growth dynamics and yield of tea in two contrasting cultivars
(drought tolerant TRI2025 and moderately drought tolerant TRI2026). Depending
on the severity of plucking, four plucking systems were used as treatments namely,
mother-leaf plucking, fish-leaf plucking, janam-leaf plucking and estate practice.
Between the two cultivars, TRI2026 showed significantly higher yield because of
its greater number of harvested shoots m-2, higher average shoot fresh weight (g)
and less banji shoots (%) than the drought tolerant cultivar TRI2025. In TRI2025,
estate practice produced greater yield among the treatments, while TRI2026 gave
the highest yield in fish-leaf plucking system. It is concluded that shoot growth
dynamics, yield related parameters and yield of tea are affected by different
plucking treatments which vary due to cultivars.
Keywords: Tea cultivar, Severity of plucking, Shoot growth dynamics, Shoot
density, Yield
Introduction
The operation of harvesting crop in tea, termed plucking, is the removal of the
tender apical portions of secondary and higher order shoots which are subsequently
processed to give the commercial product. Shoot growth is the major physiological
process which determines the tea yield. The number of plucked shoots per unit land
area (shoot density) and the mean weight per shoot are the components of yield of
tea (Rahman, 1977). Of the two components mentioned, shoot density is the major
yield contributing factor which determines more than 80% of the variation in yield of
tea (Wijeratne, 2001). Although the number of harvested shoot has been identified as
the main factor responsible for the observed variation in yields between different
genotypes, variation in the rate of shoot growth is the main parameter that causes
season-to-season yield variation in a given genotype (Squire and Callander, 1981).
The number of harvested shoots per unit land area is determined by the rate of shoot
initiation whereas the mean weight per shoot is determined by the rate of shoot
expansion.
1SSO,Bangladesh Tea Research Institute, Srimangal–3210, Moulvibazar, Bangladesh

2Professor,
Department of Crop Science, Faculty of Agriculture, University of Peradeniya, Sri Lanka
3SRO, Tea Research Institute, Low-Country Station, Ratnapura, Sri Lanka
*Corresponding author’s email address: toufiqtea@yahoo.com

These parameters largely depend on the quality of cultivar, plant density (spacing),
pruning and plucking policies, fertilizer management, pest and disease control as well
as climatic and soil conditions. Having the uniformity of tea bushes and high yield
potential, clonal tea produced higher yield in comparison to seedling tea
(Gunasekare, 2008). Tanton (1979) stated that semi-erect leaf possessing China-type
tea (var. sinensis) produced up to six times the number of shoots per unit area than
the horizontal Assam-type (var. assamica) and could be responsible mainly for yield
variation. Hadfield (1975) showed that China-type tea with a photosynthetically
favourable semi-erect leaf pose, produced four times the yield of Assam tea which
has a horizontal leaf pose, concluding that this difference was related to
photosynthetic efficiency.
The plucking of tea acts as a stimulus for further shoot production by temporarily
destroying apical dominance (Tubbs, 1937; Portsmouth and Rajiah, 1957). The
axillary bud immediately below the point of plucking is activated and develops into a
new shoot. At times more than one shoot is regenerated which may depend on
cultivar, environmental conditions and cultural practices. Severity of plucking
determines the post-harvesting point of a shoot. Different plucking systems are
followed in different agro-climatic regions due to difference in the rate of shoot
growth. Some follow mother-leaf plucking, while others adopt fish-leaf and janam-
leaf plucking. Mother-leaf plucking was the standard estate practice in Sri Lanka
(Portsmouth, 1957) but now the estate practice plucking system (a combination of
mother-leaf plucking and fish-leaf plucking) is popular while in North-East India
janam-leaf plucking is practiced due to high growth rate (Wettasinghe et al., 1981).
The appropriate plucking system may vary from cultivar to cultivar. In order to
maximize the crop production and for the sustainability of a bush, it is necessary to
find out the appropriate plucking system for a particular tea field. Therefore, this
study was undertaken to know the effects of different standard of plucking on shoot
growth dynamics and yield in two contrasting tea cultivars.
Materials and Methods
The experiment was conducted at the Tea Research Institute, Low Country Station,
Ratnapura (6040'N, 80025'E and 30 m amsl) and data were collected about a period
of one year (from March 2011 to February 2012). The experiment was consisted of a
two-factor factorial treatment structure. The treatment factors were a) two
contrasting tea cultivars and b) four plucking standards. The contrasting two tea
cultivars, drought tolerant TRI2025 (Assam-type) and moderately drought tolerant
TRI2026 (China-type) were used. The four different standard of plucking were
mother-leaf plucking (shoot is plucked leaving the oldest normal leaf), fish-leaf
plucking (plucking to the fish-leaf), janam-leaf plucking (plucking below the fish-leaf)
and estate practice. In the estate practice, about 60-75% of tea crop shoots were
plucked to the fish-leaf and the balance to the mother leaf (Wettasinghe et al., 1981).
There were four replicate plots for each cultivar x plucking standard treatment
combination. Altogether there were 32 experimental plots and each plot having 10
bushes were demarcated. So, the experimental area was laid in a randomized
complete block design. Tea bushes were 11 years old with double hedge spacing
(0.6m x 0.9m x 1.5m) and were in the third year of its pruning cycle. Application of
fertilizers and cultural practices were carried out according to the recommendations
of TRI, Sri Lanka.
Plucking was done in the morning on weekly rounds for estimation of the plot yield
(g plot-1) and it was used to extrapolate as tea yield in g m-2 and during the plucking
following data were collected e.g. harvested shoot population density (No. m -2), basal
shoot population density (No. m-2) and total shoot population density (No. m-2).
To know the shoot growth dynamics, five shoots each having three leaves and active
bud were randomly selected from the centre of the bush from each plot at two
months intervals. Selected shoots were plucked according to the treatment and
tagged for future identification. Tagged shoots were plucked when they reached at
the standard of two leaves with active bud and the following measurements were
made on these harvested shoots e.g. length of shoot replacement cycle (L SRC), fresh
weight per shoot at two leaves with active bud stage and number of shoot
regenerated from the plucking point. Collected data were statistically analyzed using
SAS computer package.
Results and Discussion
Shoot growth dynamics
The time taken by an axillary bud to grow into a harvestable shoot is termed as the
shoot replacement cycle. The LSRC was significantly longer for cultivar TRI2026 than
the TRI2025 (Table 1). In TRI2025, mother-leaf plucking showed the highest
duration for shoot replacement cycle (36.1 days) which was significantly different
with others. However in TRI2026, janam-leaf plucking had the highest LSRC (36.7
days) which was similar with mother-leaf plucking and fish-leaf plucking but
different with estate practice. Between two cultivars, greater fresh weight per shoot
was observed in TRI2026 (0.49 g) than TRI2025 (0.46 g). For both cultivars, mother-
leaf plucking showed greater fresh weight per shoot and the lowest was in janam-leaf
plucking (Table 1). In both cultivars, janam-leaf plucking regenerated greater number
of shoots which was significantly different and followed by fish-leaf plucking, estate
practice and mother-leaf plucking, respectively (Table 1).
Table 1. Effect of different plucking systems on length of shoot replacement cycle
(LSRC) and fresh weight per shoot at two leaves with active bud stage and
number of shoots regenerated
Plucking LSRC (Days) Shoot fresh weight (g) Number of shoot
system generations
TRI2025 TRI2026 TRI2025 TRI2026 TRI2025 TRI2026
Mother-leaf 36.1(±2.8)a 36.3(±2.4)a b 0.50(±0.04)a 0.50(±0.04)a 1.0(±0.04)b 1.0(±0.01)b
Fish-leaf 33.9(±2.2)b 35.8(±2.8)a b 0.45(±0.05)b c 0.49(±0.06)a b 1.1(±0.05)b 1.1(±0.02)b
Janam-leaf 34.2(±2.4)b 36.7(±3.2)a 0.42(±0.06)c 0.47(±0.04)b 1.4(±0.11)a 1.3(±0.09)a
Estate practice 34.8(±2.5)b 35.6(±3.4)b 0.48(±0.05)a b 0.49(±0.04)a b 1.1(±0.03)b 1.1(±0.02)b
Mean 34.7 B 36.1 A 0.46 B 0.49 A 1.2 1.1
Level of significance 0.05
LSD 1.01 1.10 0.03 0.03 0.09 0.08
CV (%) 10 11 25 22 27 27
The difference of shoot growth dynamics may be due to the variation of genetic
characteristics between two cultivars. Similar findings observed by Wijeratne (1994)
that shoot replacement cycle was shorter in drought tolerant TRI2025 for its early
bud break and faster rate of leaf appearance than the drought susceptible cultivar
TRI2023. Between two cultivars, greater fresh weight per shoot was observed in
TRI2026 than TRI2025. This may be due to the variation of L SRC or for genetic
characteristics resulting the variation of shoot weight between two cultivars most
probably caused by the inherited different characteristics of shoot internodes and
leaves as suggested by Wickramaratne (1981). The cultivar that takes longer period to
complete the shoot replacement cycle, naturally assimilates more photosynthates and
absorb more nutrients for its growth and the vice-versa. In agreement with present
findings, Wettasinghe et al. (1981) showed janam-leaf plucking resulted in a 26%
drop in average shoot weight. In both cultivars, mother-leaf plucking spent a longer
time period for LSRC in contrast to other systems. Number of shoot generations were
higher in janam-leaf plucking which is supported by Visser (1960), that harder
plucking systems influenced multiple shoot growth than the lighter system of
plucking. This would have resulted from the weak apical dominance over the second
axillary buds below the fish leaf in a region where internodes are very short.
Shoot density (Number m-2)
The difference in number of harvested shoot population density, basal shoot
population density and total shoot population density were highly significant for
cultivars, treatments and between their interaction effects. Greater number of shoots
m-2 were observed for all the above parameters in TRI2026 than TRI2025, presented
in the Table 2. In cultivar TRI2025, greater number of harvested shoots m -2 were
observed in estate practice (217) which was statistically similar with fish-leaf plucking
and mother-leaf plucking but significantly different with janam-leaf plucking.
However cultivar TRI2026 gave greater number of harvested shoots m -2 in fish-leaf
plucking (243) which was similar with janam-leaf plucking and estate practice but
significantly different with mother-leaf plucking.
Table 2. Effect of severity of plucking on harvested shoot density, basal shoot
density and total shoot density (No. m-2)
Plucking Harvested shoot density Basal shoot density Total shoot density
system
TRI2025 TRI2026 TRI2025 TRI2026 TRI2025 TRI2026
Mother-leaf 208(±4) a b 221(±6) b 190(±4) c 189(±4) c 398(±7) c 410(±9) b
Fish-leaf 215(±5) a b 243(±5) a 204(±4) b 224(±4) b 419(±7) b 467(±8) a
Janam-leaf 206(±5) b 242(±5) a 202(±5) b 231(±4) a 408(±9) b c 473(±9) a
Estate practice 217(±5) a 240(±6) a 222(±5) a 237(±5) a 438(±9) a 477(±11) a
Mean 212 B 237 A 205 B 220 A 417 B 457 A
LSD 9.3 10.1 7.1 7.4 13.1 13.7
CV (%) 20 19 16 15 14 14
From the above results, it is clearly indicated that density of harvested shoot
significantly varied due to cultivars. Similar findings observed by Stephens and Carr
(1994) that shoot population density of tea almost depended on its genetic makeup.
Although the numbers of shoots generated after plucking were greater in TRI2025,
at the time of plucking a greater number of shoots were being harvested in TRI2026.
This was due to the production of more than one secondary shoot generations which
became smaller in size than the generation of one shoot and often they do not reach
the plucking surface. This phenomenon happened especially in the drought tolerant
cultivar of TRI2025 and subsequently reduced the harvested shoot density, which is
supported by the findings of Wijeratne (1994). Plucking system also affected the
harvested shoot population density which varied from cultivar to cultivar. In
TRI2025, greater number of harvested shoot density observed in estate practice,
while in TRI2026 that was under fish-leaf plucking.
Banji shoots (%) in the harvested shoots
Formation of banji shoots (%) varied for cultivars. Between the two cultivars, higher
percentage of banji shoots observed in the drought tolerant TRI2025 than the
TRI2026 at most of the plucking dates. Severity of plucking also had significant
effect on the banji formation of shoots. In TRI2025, greater percentage of banji
shoots observed in janam-leaf plucking (about 63% in an average) which was
followed by fish-leaf plucking (61%), estate practice (59%) and mother-leaf plucking
(58%), respectively [Figure 1 (a)]. Similarly for the cultivar TRI2026, lower
percentage of banji shoots observed in mother-leaf plucking and estate practice
(about 55% in an average) which was followed by janam-leaf plucking (56%) and
fish-leaf plucking (57%) [Figure 1 (b)]. In the harvested leaf, percentage of banji
shoots were varied at different plucking dates for all of the four treatments in both
cultivars.
(a)
(b)
Fig. 1. Banji shoots (%) in the harvested leaf for the severity of plucking in (a)
TRI2025 and (b) TRI2026 at different plucking days of the year
Formation of banji reduces production of transpiring leaves and fast growing apices,
thus reducing the water requirement for growth. Hence, greater production of
dormant shoots can be considered as a useful mechanism of drought tolerance by
maintaining internal water balance of the bush (Odhiambo et al., 1993). Clonal
differences in banji shoot production also have been reported by Stephens and Carr
(1990) for Mufindi tea clones. Among the four treatments, mother-leaf plucking
produced less number of banji shoots than others in both cultivars. It happened
because results of this study showed that being the lightest system of plucking,
mother-leaf plucking took longer period for its shoot replacement cycle. And
consequently, banji percentage was greater in janam-leaf plucking.
Harvested yield records
Greater fresh weight and dry weight (g m-2) were recorded in TRI2026 than TRI2025
(Table 3). In TRI2025, estate practice produced higher harvested fresh leaf weight
and dry weight which was significantly different and followed by fish-leaf plucking,
mother-leaf plucking and janam-leaf plucking respectively. However for the cultivar
TRI2026, fish-leaf plucking produced greater harvested leaf fresh weight and dry
weight which was significantly different and followed by janam-leaf plucking, estate
practice and mother-leaf plucking, respectively.
Table 3. Effect of severity of plucking on harvested fresh leaf weight and dry
weight
Severity of Fresh leaf weight (g m-2) Dry leaf weight (g m-2)
plucking TRI2025 TRI2026 TRI2025 TRI2026
Mother-leaf 26.1(±0.8) b 25.9(±0.8) c
5.7(±0.2) c 5.4(±0.2) c
Fish-leaf 27.9(±0.9) a b 35.5(±1.3) a
6.4(±0.2) b 7.7(±0.3) a
Janam-leaf 22.6(±0.8) c 33.2(±1.0) b
5.4(±0.2) c 7.4(±0.2) a b
Estate practice 28.8(±1.0) a 31.1(±1.0) b
6.8(±0.2) a 7.0(±0.2) b
Mean 26.4 B 31.4 A 6.1 B 6.9 A
LSD 1.87 2.29 0.43 0.50
CV (%) 32 33 33 33
As already pointed out, the yield of tea depends on the average shoot weight and the
number of shoots harvested per unit area (harvested shoot density). The relative
importance of these components in the determination of tea yield has been discussed
by many researchers (Tanton, 1992; Odhiambo et al., 1993). Both the parameters are
significantly affected by the characteristics of the cultivar which is largely under
genetic control and also by the system of plucking. Having greater harvested shoot
density, less banji shoots and greater shoot weight, the cultivar TRI2026 produced
greater harvested leaf yield than TRI2025. For almost the similar reasons, greater
yield of tea was observed under estate practice in TRI2025 and that was in fish-leaf
plucking for TRI2026. In TRI2025, the lowest yield observed in janam-leaf plucking
which supported with the findings of Wettasinghe et al. (1981) who stated that
increasing severity of plucking increased the harvest index up to a certain period after
which it will be decreased due to aging of maintenance foliage. Moreover, present
study shows that in both cultivars, mother-leaf plucking producing lower yield and it
was more pronounce in TRI2026 than TRI2025. The main reason of low yield in
mother-leaf plucking was due to its lower number of harvested shoots per unit land
area which is considered as the major yield component of tea.
Conclusion
An improved vegetatively propagated tea cultivar has significant effect to increase
the yield due to its higher shoot density, greater average shoot weight and less
number of banji shoots. To ensure the highest productivity from a tea area, it is
essential to use the appropriate plucking policy which vary from cultivar to cultivar.
Acknowledgements
This work was supported by a grant from the National Agricultural Technology
Project (NATP), Bangladesh Agricultural Research Council (BARC) and constitutes
a part of a PhD training program. The authors are thankful to Mrs. P. Premathunga,
TRI, Low Country Station, Ratnapura for her assistance during the laboratory work.
References
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Wickramaratne, M.R.T. 1981. Variation in some leaf characteristics in tea (Camellia
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Wijeratne, M.A. 1994. Effect of climatic factors on the growth of tea (Camellia sinensis
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Wijeratne, M.A. 2001. Shoot growth and harvesting of tea. Tea Research Institute of
Sri Lanka, Talawakelle, Sri Lanka. pp. 9-19.
EVALUATION OF SOME INDIGENOUS PLANT EXTRACTS AGAINST
TEA MOSQUITO BUG, HELOPELTIS THEIVORA WATERHOUSE
(HEMIPTERA: MIRIDAE) INFESTING TEA
M.S.A. Mamun1*, M. Ahmed2, S.K. Paul3 and R.S. Chowdhury4
Abstract
An experiment was conducted to evaluate the toxicity of some indigenous plants
viz., Bur weed (Xanthium strumarium), Datura (Datura metel), Katamehedi (Duranta
erecta), Lantana (Lantana camara), Mahogani (Swietenia mahagoni) and Neem
(Azadirachta indica) at 5.0, 7.5 and 10.0% (w/v) concentrations against tea mosquito
bug, Helopeltis theivora Waterhouse under both laboratory and field conditions at
Bangladesh Tea Research Institute, Srimangal, Moulvibazar during 2010-2011. The
plant extracts were prepared with acetone and water solvents separately. Direct
toxicity test was done by topical application method in completely randomized
design under laboratory and spraying method in randomized complete block
design under field condition. Results indicated that all the plant extracts had
showed the toxic effect on Helopeltis under both laboratory and field conditions.
Under laboratory condition, the toxicity of Bur weed (52.86%) was found to be
highest whereas least action found in Katamehedi (27.24%). Among the solvents,
acetone extract (40.30%) was slightly more effective than water extract (37.01%).
The maximum average mortality (44.09%) was observed at the highest
concentration (10.0%) of plant extracts and the mortality percentage was directly
proportional to the level of concentration of plant extracts. In case of field
evaluation, the aqueous extract of Bur weed showed the highest percent reduction
in infestation (46.73%) over control and that of Katamehedi had the lowest action
(36.71%) at 10% concentration. The effectiveness of most of the plant extracts
were found to increase proportionally with the increase of doses and decreases
proportionally with increase of time. The order of toxicity of the plant extracts on
Helopeltis was: Bur weed>Lantana>Datura>Neem>Mahogani>Katamehedi. The
above mentioned plants are available throughout the tea growing areas and the
planters may use these plants in tea pest management especially for the control of
Helopeltis as safer phytopesticidal products in both organic and inorganic tea
estates.
Keywords: Tea, Plant extracts, Biopesticides, Toxicant, Tea mosquito bug, Helopeltis theivora
1SSO, Entomology Division, 2Director, 3SO, Entomology Division, Bangladesh Tea Research
Institute, Srimangal, Moulvibazar, Bangladesh; 4Assistant Professor, Department of Food Engineering
and Tea Technology, Shahjalal University of Science and Technology, Sylhet, Bangladesh
*Corresponding author’s email address: shameembtri@yahoo.com
Introduction
Tea is the most widely used non-alcoholic beverage all over the world, prepared
from the tender shoot of tea plant, Camellia sinensis (L) O. Kuntze. Tea plants are
subjected to the attack of several pests such as insects, mites and nematodes (Sana,
1989). In world tea, 1034 species of arthropods and 82 species of nematodes are
associated with tea plants (Chen and Chen, 1989). Among them 25 species of insects,
4 species of mites and 10 species of nematodes are recorded from Bangladesh
(Ahmed, 2005). Among the various insect pests that attack or cause severe damage
to tea plant, one is the Helopeltis theivora, commonly known as Tea mosquito bug
(Waterhouse, 1886). It is one the most serious pests of tea in almost all tea growing
countries like Sri Lanka, India, Indonesia and Bangladesh. The crop loss due to
Helopeltis to the tune of 10-15% is reported from Bangladesh (Ahmed, 2012). Both
nymphs and adults of Helopeltis suck the cell sap from the buds, young leaves and
tender stems. Generally infested shoots dry up which directly affect to crop loss. In a
severe attack, the plantation turns black and growth of flush i.e. two leaves and a
bud, is completely checked. In addition to direct crop loss, damage by Helopletis leads
to debilitation of bushes resulting in die-back with crows-feet and delayed or meager
flushing thereafter consequently almost the entire crop is lost.
To combat these problems different groups of pesticides have been used in the tea
fields with approval of BTRI since 1960 (Mamun et al., 2012). The large-scale use of
synthetic insecticides in tea leads to adverse effects such as development of pesticide
resistance, frequent pest outbreaks, emergence of new pests, environmental pollution
and health hazards. As tea is a consumable commodity, the effect of residue of
pesticides in made tea is harmful to human health. In this context, botanicals are
being considered as environmentally safe, selective, biodegradable, economical and
renewable alternatives for use in IPM programmes in tea (Mamun, 2011). Botanicals
are natural plant products that belong to the secondary metabolites, which include
alkaloids, terpenoids, phenolics, and minor secondary chemicals. It is estimated that
the plants may contain as many as 4000,000 secondary metabolites (Jacobson, 1989).
As many as 2121 plant species have been reported to posses’ pest control properties;
25 of these plants species posses the characteristics required for an ideal botanical
insecticide and are therefore more promising for use in organic pest control
programmes (Radhakrishnan, 2005). Botanicals like neem, ghora-neem, mahogani,
karanja, adathoda, sweet flag, tobacco, derris, annona, smart weed, bur weed, datura,
calotropis, bidens, lantana, chrysanthemum, artemisia, marigold, clerodendrum, wild
sunflower etc. are available surroundings of the tea garden or may be grown by the
planters with minimum expense and extracted by indigenous methods (Mamun and
Ahmed, 2011). These botanical materials can be used as an alternative to chemical
pesticides. This will be very helpful in minimizing the undesirable side effects of
synthetic pesticides.
Based on the foregoing, the present study was undertaken to evaluate the toxic effect
of six indigenous plant extracts namely Burweed (Xanthium strumarium), Datura
(Datura metel), Katamehedi (Duranta erecta), Lantana (Lantana camara), Mahogani
(Swietenia mahagoni) and Neem (Azadirachta indica) against tea mosquito bug, H.
theivora, a major insect pest of tea in Bangladesh.
An experiment was conducted both in the Entomology Laboratory and in the main
farm of Bangladesh Tea Research Institute (BTRI), Srimangal, Moulvibazar during
the period from January 2010 to December 2011. The Helopeltis was collected from
different sections of BTRI main farm and was reared in the Entomology Laboratory,
BTRI at 27 to 30°C temperature and 70-80% relative humidity on a susceptible tea
clone, BT3 by following the method of Muraleedharan (2006). The insects were
reared on tea shoots with three leaves and a bud in transparent glass jars (20 cm Х 15
cm) and mouths of transparent jars were covered with nylon mesh. Each jar was set
up with 20 pairs of adult insects and the newly emerged adults were utilized in the
subsequent experiments of the present study.
Preparation of plant extracts
Fresh leaves, succulent stems, flowers, seeds and whole plants of Bur weed
(Xanthium strumarium), Datura (Datura metel), Katamehedi (Duranta erecta), Lantana
(Lantana camara), Mahogani (Swietenia mahagoni) and Neem (Azadirachta indica) were
collected locally from nearby areas of BTRI main farm, Srimangal, Moulvibazar
(Table 1 & Plate 1). Each plant material was dried under shade and powdered by
using electric grinder and pass through a 20 mesh sieve and kept in a 1 kg capacity
polypropylene bag. 300 g of each powdered plant material were taken into a 2 litre
capacity conical flask and 1000 ml of distilled water and/or acetone was added to it
separately and shaken for 8 h in a mechanical shaker and then kept it for 24 h.
Acetone and distilled water were used as solvents. The extract was separated using
fine Muslin cloth and then filtered. The filtrate was collected in a 2 litre capacity
conical flask and volume was made up to 1000 ml. This was considered as stock
solution. Required concentrations (5.0, 7.5 and 10.0%) were prepared from the stock
solution.
Bur weed (Xanthium strumarium) Datura (Datura metel) Katamehedi (Duranta erecta)
Lantana (Lantana camara) Mahogani (Swietenia mahagoni) Neem (Azadirachta indica)
Tea mosquito bug (H. theivora) Rearing of H. theivora Bioassay of Plant extracts
Plate 1. Some indigenous plant extracts against tea mosquito bug

Table 1. Plants evaluated for insecticidal activities against Helopeltis
Common name Scientific name Family Plant parts used
Bur weed Xanthium strumarium Compositae Whole plant
Datura Datura metel Solanaceae Leaves and fruits
Katamehedi Duranta erecta Verbenaceae Leaves and flowers
Lantana Lantana camara Verbenaceae Leaves and flowers
Mahogani Swietenia mahagoni Meliaceae Seeds
Neem Azadirachta indica Meliaceae Seeds
Laboratory evaluation of toxicity of plant extracts against Helopletis
A laboratory test for direct toxicity by topical application method was conducted
according to the method of Talukder and Howse (1993) with slight modification.
Three different concentrations of each plant extracts (5.0, 7.50 and 10.0%) were
prepared with respective solvents (Acetone and Water). One microliter (μl) of
prepared solution was applied to the dorsal surface of the thorax of each Helopelits
using a micropipette. Ten insects per replication were treated and each treatment was
replicated thrice. In addition, the same numbers of insects were treated with solvent
only for control. After treatment, the insects were transferred into 9 cm diameter
petri dishes (10 insects/petri dish) containing tea shoots (two leaves and a bud).
Insect mortalities were recorded at 24, 48 and 72 hours after treatment (HAT).
Original data were corrected by Abbott’s (1987) formula:
P´-C
P = ----------------- x 100
100-C
Where,
P = Corrected mortality (%)
P´= Observed mortality (%)
C = Control mortality (%)
Field evaluation of toxicity of plant extracts against Helopeltis
Field evaluation of aqueous extracts of the above six plants was also conducted
against tea mosquito bug at the BTRI main farm in 2010-2011. No pesticide was
applied to the bushes prior to the start of the experiment. The experiment was laid
out in low skiffed sections following Randomized Block Design with three
replications each having plot size of 5m x 5m. The treatments were T1= Bur weed,
T2= Datura, T3= Katamehedi, T4= Lantana, T5= Mahogani, T6= Neem and T7=
control.
Plots with heavy infestation of H. theivora were chosen for this study. After selection
of the plots, pre treatment count was taken in the respective plots. The total quantity
of each aqueous extract for spraying in each treatment was calculated on the basis of
plot size. The plots were sprayed with 5.0, 7.5 and 10.0% concentrations of each of
the extracts separately. Spraying was done early in the morning with knapsack
sprayer where uniform coverage of each plot and bush was taken care of. Two
rounds of foliar spray were given at 15 days interval and post treatment observations
were taken in four consecutive weeks. One hundred shoots were randomly collected
from the harvested shoots of each plot and infested shoots were counted. A shoot
was considered as infested, if it contained even a single feeding spot. Effectiveness of
the plant extracts was calculated by using Henderson & Tilton’s (1955) formula:
cbxta
Percent effectiveness = (1- ) x 100
caxtb
Where,
cb = No. of insect population in control before treatment
ca = No. of insect population in control after treatment
tb = No. of insect population in treated before treatment
ta = No. of insect population in treated after treatment
Statistical Analysis
The experimental data were statistically analyzed by Completely Randomized Design
(factorial CRD) and Randomized Block Design in case of laboratory and field
experiment respectively using MSTAT statistical software in a microcomputer. The
results are expressed as Mean ± SE and data were statistically analyzed by one-way
ANOVA, with the level of significance set at p<0.05. The mean values adjusted by
Duncan’s Multiple Range Test (DMRT) (Duncan, 1951).
Laboratory evaluation of toxicity of plant extracts against Helopeltis
The effects of different of plant extracts of Bur weed, Datura, Katamehedi, Lantana,
Mahogani, Neem against tea mosquito bug, H. theivora are presented in Tables 2-5.
Average mortality percentage of tea mosquito bug at 24, 48 and 72 hours after
treatment indicated that Bur weed extract possessed the highest (52.86%) toxic
effect, whereas Katamehedi leaf extract possessed the lowest (27.24%) toxic effect
(Table 2). Among the solvents, acetone extract was found more toxic (40.30%) and it
was significantly different from water (37.01%) extract (Table 3). The maximum
average mortality (44.09%) was observed at higher concentration (10.0%) of plant
extract and the mortality percentage was directly proportional to the level of
concentration of plant extract (Table 4).
Table 2. Mean mortality percentage of tea mosquito bug, H. theivora treated with
different plant extracts (Interaction of plant extracts and time)
Mortality rate (%)
Name of the plant Mean (%)
24HAT 48HAT 72HAT
Bur weed 47.59 54.63 56.36 52.86a
Datura 35.18 44.02 47.59 42.26c
Katamehedi 22.59 27.96 31.48 27.24f
Lantana 38.70 45.68 49.26 44.55b
Mahogani 25.78 29.26 32.16 29.07e
Neem 32.13 35.69 39.70 35.84d
Sx 0.135 0.1051
Probability level NS 0.01
HAT= Hours after treatment
NS = Not Significant
Within column and row values followed by different letter(s) are significantly different by DMRT
Table 3. Effect of different solvents used in preparing various plant extracts on the
mortality of tea mosquito bug, H. theivora (Interaction of solvent and time)
Name of the Mortality rate (%)
Mean (%)
solvent 24HAT 48HAT 72HAT
Acetone 34.98 41.52 44.41 40.30a
Water 32.34 37.57 41.11 37.01b
Sx 0.0967 0.0821
Table 4. Effect of different doses of plant extracts on the mortality of tea mosquito
bug, H. theivora (Interaction of dose and time)
Mortality rate (%)
Dose (%) Mean (%)
24HAT 48HAT 72HAT
5.0 28.67 34.38 37.44 33.50c
7.5 34.26 37.75 43.12 38.38b
10.0 38.06 46.50 47.71 44.09a
Sx 0.1732 0.0958
70
60
50
% Mortality
40
30
20
10
0
Bur weed Datura Katamehedi Lantana Mahogani Neem
Plants
Fig. 1. Performance of plant extracts on the mortality of H. theivora

The order of toxicity of the plant extracts on tea mosquito bug, H. theivora was: Bur
weed>Lantana>Datura>Neem>Mahogani>Katamehedi (Fig. 1). Among the test
plants, Bur weed extracts showed the highest toxic effect, whereas Katamehedi
showed the lowest toxic effect. Control of Helopeltis with some indigenous plant
extracts were reported by Gogoi et al. (2003), Deka et al. (1998), Sarmah and Bhola
(2008). The present results are in agreement with the findings of those workers.
Field evaluation of toxicity of plant extracts
The data for two years experiment at BTRI main farm reveal that at 5% aqueous
extracts of the plants gave 28.30% reduction in infestation compared to control and
those of 33.26% and 41.34% at 7.5 and 10% respectively (Fig. 2). Higher
concentration of aqueous extract of all the tested plants showed higher percent
reduction in infestation. The highest percent reduction in infestation was found
46.73% at 10% of concentration of aqueous extract of Bur weed (Table 5). Another
plant, Lantana occupied the second position in effectiveness by reducing infestation
which was 43.90% over untreated control at the same concentration and followed by
Datura (41.82%), Neem (40.39%), Mahogani (38.50%) and Katamehedi (36.71%).
Fig. 2. Effect of doses of plant extracts on Helopeltis
Table 5. Field evaluation of different plant extracts against Helopeltis theivora
Treatments Dose Pre Post treatment observations
(%) treatment (% reduction of infestation over control)
(% shoot 1st spray 2nd spray Average
infestation) 7 days 14 days 21 days 28 days
Bur weed 5.0 30 39.56 31.17 33.11 23.08 31.73i
7.5 35 47.62 29.82 39.13 28.57 36.29f
10.0 40 48.41 41.52 56.52 40.48 46.73a
Datura 5.0 28 35.06 23.44 32.81 25.32 29.16j
7.5 32 38.78 32.33 37.89 28.57 34.39g
10.0 36 46.43 39.47 45.65 35.71 41.82c
Katamehedi 5.0 38 28.57 21.05 29.35 19.64 24.65l
7.5 43 36.51 23.98 37.20 20.63 29.58j
10.0 34 46.43 29.82 42.03 28.57 36.71f
Lantana 5.0 42 38.10 26.32 33.33 23.81 30.39i
7.5 37 40.48 32.75 39.61 28.57 35.35g
10.0 40 54.55 35.41 50.59 35.06 43.90b
Mahogani 5.0 44 34.52 21.05 27.54 19.64 25.69k
7.5 39 41.96 24.34 37.50 21.88 31.42i
10.0 36 48.98 32.33 44.10 28.57 38.50e
Neem 5.0 34 38.10 22.81 30.43 21.43 28.19j
7.5 30 39.29 28.95 36.96 25.00 32.55h
10.0 37 48.05 35.41 44.66 33.44 40.39d
Control - 29 36 42 49 57 -
CV% - - 8.77 6.45 7.08 5.36 -
Mean of three observations
The results of the studies showed that aqueous extracts of the test plants were
effective in reducing the mosquito bug infestation. However, variations existed in
their efficacy. Similar results were reported by Sing et al. (2012), Anonymous (2010),
Roy et al. (2009), Sarmah et al. (2006) and Gogoi et al. (2003).
Conclusion
It has been observed from the result that the botanicals used in this experiment had
direct toxic effect on tea mosquito bug, H. theivora. The tested indigenous plants are
available throughout the tea growing areas and the planters may use these plants in
tea pest management especially for the control of tea mosquito bug, H. theivora.
However, before releasing it as a new technology further investigation is needed to
confirm the result. Furthermore, natural products are notoriously variable, and
therefore consistency of the final product will be much harder to achieve. Also, it
may be necessary to determine both the shelf stability of the active ingredient and its
fate in the environment or in animals. Such studies can be complicated enough when
only a single compound is being tracked, but the effort required to track several
putative active principles in a single product should not be underestimated. These
insecticidal activities of tested plants can therefore be incorporated into other insect
control techniques in the strategy of integrated pest management (IPM) in tea.
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DEVELOPMENT OF IN VITRO MICRO PROPAGATION SYSTEM IN

TEA PLANT [CAMELLIA SINENSIS (L.) O. KUNTZE] USING SHOOT
TIP AND NODAL SEGMENT EXPLANTS
S. Boonerjee1*, M.I. Hoque2 and R.H. Sarker3
Abstract
A successful method of in vitro propagation system for tea plant [Camellia sinensis
(L.) O. Kuntze] was developed. Shoot tip and nodal segment explants of elite tea
clone BT2 were used for direct shoot regeneration. The experiments were carried
out following the method of shoot initiation, multiple shoot proliferation, root
induction and transfer of the plantlets to the soil after proper hardening. Among
the different hormone supplements examined, combined application of BAP and
IBA was found to be effective for shoot initiation and multiple shoot
proliferation. The best shoot initiations (48% for shoot tip and 52% for nodal
segment explants) were observed and the highest mean number and mean length
of shoots/explant for shoot tip (13.2 and 4.5cm.) and nodal segment (15.1 and 5.0
cm.) explants were achieved on MS medium supplemented with 2.0 mg/l BAP
and 0.1 mg/l IBA. Addition of 0.5 mg/l GA3 on the shoot multiplication medium
showed optimum results on shoot elongation. Moreover 85% rooting was
achieved when the isolated microshoots were pretreated with high concentration
of IBA (500 mg/l; 30 min) and transferred to the soil directly. After proper
hardening rooted plantlets were transplanted to the pot and 80% plantlets were
found to survive.
Keywords: BAP-6-benzyleaminopurine, IBA- indole-3-butyric acid, MS- Murashige
& Skoog (1962) medium, micropropagation, explants, plantlet.
Introduction
Tea (Camellia sinensis (L.) O. Kuntze) is one of the most popular mild stimulating, caffeine
containing drink used all over the world. Medicinal values have increased its popularity as a
health drink also. In Bangladesh it has been habituated as a daily consumption. Because of
increasing demand world wide and in domestic level tea is now an important agricultural
product in our country. Bangladesh is now facing the challenging competition in the
international market as the demand of world tea consumers are on good quality. Considering
tea as an important commercial crop, Bangladesh has to give emphasis on its improved
production and quality as well. Intensive plantation with elite clones and seed stocks is one
of the options to achieve the goal for the tea industries of Bangladesh.
1SO, Botany Division, Bangladesh Tea Research Institute, Srimangal, Moulvibazar, 2&3Professor,
Department of Botany, University of Dhaka, Bangladesh.

*Corresponding author’s email address: sboonerjee55@yahoo.com
But conventional propagation system of tea plant is very slow and has some
unavoidable limitations. Sufficient planting materials are not available throughout the
year due to winter dormancy and season depending rooting ability of the cuttings
results the poor survival rate at the nursery level. Besides, production of large
number of cuttings is impossible immediately after releasing of an elite clone from
the experimental plot as the number of mother plants are limited (80-100). At
present 15000-18000 plants/ha is recommended to get optimum production and
10% more saplings are required for infilling. In this situation in vitro
micropropagation can be considered as useful attempt to solve unavailability of
sufficient planting materials for tea plantation. This technique can be an ideal choice
for the circumvention of conventional multiplication system (Mondal et. al., 2004)
and commercially, it can be effectively utilized for introducing improved clones in a
relatively short period of time (Sharma et. al., 1999).
Several reviews on micropropagation of tea and related species have been reported in
past (Creze 1980, Kato 1989, Phukan and Mitra 1984, Sharma et. al., 1999, Das 2001,
Mondal 2003). Haque et. al. (2002) tried to initiate shoot regeneration from shoot tip
culture of some BT (Bangladesh Tea) clones as the initial work of BTRI. The present
study is aimed to develop the complete regeneration protocol for the rapid
propagation of desired clones of tea plant using shoot tip and nodal segment
explants.
Shoot tips and nodal segments were collected from new shoots of the first flush of
mature tea field of BT2 clone (growing in the Nucleus Clone Plot of Bangladesh Tea
Research Institute). The explants were thoroughly cleaned and surface sterilized with
70% alcohol for 2 minutes followed by washing with 0.2% HgCl2 for 17 minutes. All
traces of mercuric chloride were then washed off with sterile distilled water prior to
cutting into shoot tip (0.5-1.0 cm.) and nodal segment (1.0-2.0 cm.) explants. The
responsive explants were incubated for shoot initiation and multiplication on the
basal medium of MS (Murashige and Skoog 1962) inorganic salts supplemented with
3% sucrose and 0.8% agar. After 30-35 days of culture in the multiplication media,
individual microshoots (2.5-4 cm.) were isolated and used for rooting experiments.
The final pH of MS basal medium was adjusted to 5.8 before autoclaving for 20
minutes at 121º C. The incubation was made in a growth room under 16 hour
photoperiod regime. The temperature and humidity of the culture room were
maintained at 25±2º C and 65% respectively. Weekly visual observation was made
and the data on shoot initiation, proliferation and root induction were used to
calculate the percentage of cultures responding per treatment.
Different experiments were conducted to establish the direct regeneration system for
tea plant using shoot tip and nodal segment explants. The findings of the
experiments show that both the explants were responded towards regeneration on
MS medium and different hormone compositions have significant impact on their
regeneration system. In hormone free MS medium the response towards growth of
incubated explants were found very slow even when kept maintained on subculture.
2.0 mg/l BAP showed best response towards regeneration for both the explants
when various concentrations of cytokinins (Kn, BAP, 2ip and TDZ) alone were
examined. Nakamura (1988) reported that the addition of cytokinins is essential for
the growth of shoot tips of tea plant and he found more accelerated shoots with
large leaves in MS medium with 3.0 mg/l BAP. Multiple shoot formation was not
occurred when BAP was tried alone during this study. It indicates that cytokinin
alone is not suitable for multiple shoot proliferation from both the explants. Similar
observation was reported by George and Sherrington (1984). They stated that the
application of just a cytokinin has been commonly effective but optimum rates of
shoot initiation generally occur with the combinations of auxin and cytokinin. The
results of combined application of different concentrations of cytokinins and auxins
on multiple shoot proliferation are presented in Table 1. It reveals from the table
that highest proliferation of shoots was recorded on the MS medium supplemented
with different concentrations of BAP and IBA. Shoot initiation was observed in all
the treatments of BAP and IBA although the percentages of shoot initiation and
multiplication varied in different combinations. The best shoot initiation for shoot
tip (48%) and nodal segment (52%) explants were observed within 30-40 days on MS
medium supplemented with 2.0 mg/l BAP and 0.1 mg/l IBA. Shoot growth and
newly developed shoot buds were observed to initiate on the same medium
combination after each subculture (Fig. 1, 2, 3, 4, 5 & 6). The highest mean number
and mean length of shoots/explant for shoot tip (13.2 and 4.5cm.) and nodal
segment (15.1 and 5.0 cm.) explants were achieved on this medium. Jha and Sen
(1992) achieved multiple shoots using 5.0 mg/l BAP and 0.1 mg/l IBA.
Combinations of BAP and IBA were also suggested in several works for multiple
shoot proliferation in tea micropropagation (Kato 1985, Kuranuki and Shibata 1992,
Nakamura 1990). Table 1 reveals that multiple shoot proliferation was also observed
from both the explants in the combination of TDZ and NAA but lower than that of
BAP and IBA. Rest combinations did not showed significant impact on shoot
multiplication.
Table 1. Response of shoot tip and nodal segment explants to shoot initiation and
multiple shoot formation using different concentrations of cytokinins and
auxins
No. of % of Days required Average Mean no. of
Concentration explants responsive for shoot length of shoots/explan
(mg/l)
inoculated explants initiation shoot (cm.) t after 4
month
TDZ NAA ST NS ST NS ST NS ST NS
2.0 0.1 25 22 20 40-43 40-43 1.5 2.2 4.3 4.6

2.0 0.5 25 28 26 33-37 38-40 2.6 2.6 4.3 4.8
2.0 1.0 25 29 28 32-36 36-41 3.3 3.4 6.0 5.8
2.0 1.5 25 32 33 32-33 34-37 4.0 4.3 8.0 8.6
2.0 2.0 25 28 27 33-38 36-40 3.4 3.6 6.5 7.0
2.0 2.5 25 24 23 35-37 35-38 2.5 3.0 4.7 4.8
BAP Kn
2.0 0.1 25 08 12 33-40 36-40 1.5 2.4 1.00 1.35
2.0 0.5 25 16 20 35-38 34-38 3.8 4.3 2.02 1.88
2.0 1.0 25 24 27 32-37 31-36 3.64 3.87 4.3 4.8
2.0 1.5 25 12 18 33-37 34-37 2.4 3.8 2.52 3.07
2.0 2.0 25 16 16 36-38 34-40 3.0 2.7 1.96 2.60
2.0 2.5 25 10 14 34-39 35-37 2.6 2.5 1.72 2.35
BAP IBA
2.0 0.01 25 32 36 30-40 32-40 3.4 3.6 4.70 5.70
2.0 0.05 25 36 32 28-38 33-38 3.8 3.8 9.80 8.80
2.0 0.10 25 48 52 28-33 28-32 4.5 5.0 13.2 15.1
2.0 0.50 25 36 40 30-35 30-33 4.3 4.2 10.3 10.5
2.0 1.00 25 28 32 30-33 32-35 3.2 4.3 8.50 7.50
2.0 1.5 25 28 36 30-35 32-35 2.2 3.2 5.30 6.30
BAP NAA
2.0 0.1 25 18 18 42-45 40-45 2.5 3.4 0.8 1.2
2.0 0.5 25 24 24 40-43 40-42 3.6 3.6 1.8 2.0
2.0 1.0 25 27 30 37-40 35-38 3.0 3.4 4.6 5.0
2.0 1.5 25 24 26 38-41 38-40 4.0 4.3 2.4 2.8
2.0 2.0 25 22 23 38-43 39-42 3.8 4.0 2.2 2.5
2.0 2.5 25 19 20 38-44 38-42 3.5 3.5 2.0 1.9
Effects of GA3 on shoot development and elongation of multiple shoots derived
from shoot tip and nodal segment explants were also examined (Table 2). When
GA3 added in the multiplication media the shoot growth was found stimulated. It
was observed that the mean length of the longest shoots of shoot tip and nodal
segment explants were 5.4 cm. and 5.8 cm. respectively after using 0.5 mg/l GA3 on
MS medium supplemented with 2.0 mg/l BAP and 0.1 mg/l IBA (Fig. 7 & 8).
Similar results were found by Jacqueline and Torres (1986). The suitability of GA3
for the proliferation and shoot growth were also reported by Arulpragasum and
Latiff (1986) and Jain et. al. (1993). Findings of these workers are in agreement with
the present findings on multiple shoot proliferation and growth.
Table 2. Effect of GA3 on elongation of multiple shoots derived from shoot tip and
nodal segment explants
Name of the Hormone concentrations (mg/l) Average Average length of
explants length of shoots after using
shoots before GA3 (cm)
BAP IBA GA3
using GA3
(cm)
Shoot tip 2.0 0.1 0.5 4.5 5.4
Nodal 2.0 0.1 0.5 5.0 5.8
segment
After successful development of microshoots, induction of healthy roots is an
essential part for the establishment of plantlets in hardening chamber. Several
researchers reported a significant increase in rooting percentage of micropropagated
shoots of tea plant from the effect of high concentration of IBA (0.5-1.0 g/l)
treatment (Bidarigh and Azarpour 2011, Sandal et. al., 2001, Mondal et. al., 2004). Jain
et. al. (1993) reported that ex vitro rooting was better than all types of in vitro rooting
methods. In the present study root induction was found when microshoots (2.5-4.0
cm) were treated with 500 mg/l IBA for 30 minutes followed by direct transplanting
into the soil mix (Table 3). Root development was started to initiate within 45 days
and after three months 85% rooting was achieved (Fig. 9, 10 & 11). Similar findings
were reported by Sharma et. al. (1999). They got maximum 71.6% rooting after
treating the microshoots in 500 mg/l IBA for 30 minutes prior to transfer into the
soil. Ranaweera et. al. (2013) developed low cost method of micropropagation and
achieved 100% rooting and survivability of plantlets using ex vitro rooting. They
stated that ex vitro rooting of tea microshoots reduce the cost of production of
micropropagated plants by 71% compared to that of micropropagation with an in
vitro rooting step.
Table 3. Effect of microshoots treated with high concentrations of IBA (500 mg/l)
on root formation
Dipping No. of % of Days Mean no. of Mean length

time in shoots roots/shoot of
responsive taken to
500 mg/l transferred shoots in (cm.) after 3 root/shoot
root
IBA(min) to the soil rooting months (cm.)
initiation
after 3
5 10 - - - -
10 10 20 65 3.5 2.6
20 10 48 60 4.2 3.2
30 10 85 45 8.5 5.4
40 10 30 68 2.0 1.0
50 10 nacrosis - - -
Success of survivality of tissue culture derived plants depends upon proper

hardening. In the present study ex vitro rooted plantlets were transferred into small
pots containing soil mix and kept deeply covered with transparent polythene. To
reduce sudden shock, the pots were kept in growth room for three weeks. These
plantlets were exposed to environment for one hour daily and then again placed in
growth room for another week. When the regenerated plants were fully established
in the small pots, they were kept inside a polyshaded chamber with intermittent
watering. 80% plantlets were observed to survive. Sharma et. al. (1999) reported that
survival percentage depends on maintaining a special designed hardening chamber
with controlled soil pH, CO2 enrichment, light conditions and relative humidity. In
this investigation rooted plants were kept inside a polyhouse with continuous
maintaining of high humidity for further growth.
1 2 3
4 5
6
7 8
Figs. 1-6. Different stages of direct organogenesis from shoot tip and nodal segment
explants on MS medium supplemented with 2.0 mg/l BAP and 0.1 mg/l IBA. 1, 2 &
3. Shoot initiation multiple shoot formation and shoot elongation from shoot tip
explants. 4, 5 & 6. Shoot initiation multiple shoot formation and shoot elongation
from nodal segment explants.
Figs. 7 & 8. Shoot elongation from nodal segment and shoot tip explants on MS
medium supplemented with 2.0 mg/l BAP, 0.1 mg/l IBA and 0.5 mg/l GA3.
9
10
11
Figs. 9-10. Different stages of ex vitro rooting. 9. Transfer the microshoot onto the
soil in covered jar after 30 min 500 mg/l IBA treatment, 10 rooted plants after 4
months and 11. Transfer to the pot.
Conclusion
In conclusion, the present study clearly defines the establishment of a functional
protocol for in vitro micropropagation of desired tea clones. Ex vitro rooting of
microshoots is cost effective, profitable and time saving method for the
multiplication of newly developed tea clones. Tissue culture technique provides the
significant application for rapid clonal propagation as well as the tool of modern
technology for crop improvement. Through this work for the first time a suitable
protocol has been developed for in vitro regeneration of tea clones of Bangladesh.
Acknowledgement
This work is a part of NATP Phase-1 Ph.D. research program funded by Bangladesh
Agricultural Research Council. Experiments were carried out in the Plant Breeding
and Biotechnology Laboratory of the Department of Botany, University of Dhaka.
The kind help and encouragement of Dr. Mainuddin Ahmed, Director, Bangladesh
Tea Research Institute is gratefully acknowledged by the authors during the
preparation of the manuscript.
References
Arulpragasum, P.V. and Latiff, R. 1986. Studies on the tissue culture on tea (Camellia
sinensis (L.) O. Kuntze)1. Development of a culture method for the
multiplication of shoots. Sri Lanka J. Tea Sci., 55: 44-47
Bidarigh, S. and Azarpour, E. 2011. The study effect of cytokinin hormone types on
length shoot in vitro culture of tea (Camellia sinensis L.). World Applied Sciences
Journal. IDOSI Publications, 13 (7): 1726-1729.
Creze, J. 1980. Camellia cultivation in vitro. Int. Camellia J., 12: 31-34.
Das, S.C. 2001. Tea. In: Parthasarathy V.A., Bose T. K., Deka P.C., Das P., Mitra
S.K. & Mondals S. (eds) Biotechnology of Horticultural Crops. Vol.1. Naya
Prakash, India, pp. 526-546.
George, E.F. and Sherrington, P.D. 1984. Plant propagation by tissue culture.
Exegetics Ltd. Eversiey, England.
Haque, S.K.L., Samad, M.A. and Alam, A.F.M.B. 2002. Varietal improvement and
multiplication of tea plant through tissue culture technique. Final report of
contract research project of Bangladesh Tea Research Institute with Bangladesh
Agricultural Research Council.
Jacqueline, A.C. and Torres, K.C. 1986. In vitro proliferation of Camellia “Purple
Dawn”. Hort Science, 21(2): 314.
Jain, S.M., Das, S.C. and Barman, T.S. 1993. Enhancement of root induction from in
vitro regenerated shoots of tea (Camellia sinensis L. ) Proc. Ind. Nati. Sci. Acad.,
59: 623-628.
Jha, T.B. and Sen, S.K. 1992. Micropropagation of an elite Darjeeling tea clone. Plant
Cell Rep., 11: 101-104.
Kato, M. 1985. Regeneration of plantlets from tea stem culture. Jpn. J. Breed., 35:
317-322.
Kato, M. 1989. Camellia sinsensis L. (Tea): in vitro regeneration. In: Bajaj YPS (ed.)
Biotechnology in Agriculture and Forestry, Vol. 7. Medicinal and Aromatic
Plants II. Springer-Verlag, Berlin Heidelberg, pp. 8-98
Kuranuki, Y. and Shibata, M. 1992. Effect of concentration of plant growth
regulators on the shoot apex culture of tea plant. Bull. Shizuoka Tea Expt.
Station., 16: 1-6.
Mondal, T.K. 2003. Micropropagation of tea (Camellia sinensis) In: Jain SM & Ishii K
(eds) Micropropagation of woody trees and fruits (pp. 671-720). Kluwer
Academic Publishers. The Naderlands.
Mondal, T.K., Bhattacharya, A., Laxmikumaran, M. and Ahuja, P.S. 2004. Recent
advances of tea (Camellia sinensis L.) biotechnology. Review of Plant
Biotechnology and Applied genetics. Plant cell, Tissue and Organ Culture, 76:
195-254.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassay
with tobacco tissue culture. Physiol. Plant, 15: 474-49.
Nakamura, Y. 1988. Effective methods of in vitro propagation of tea plant.
Proceedings of the International Symposium on Recent Development of Tea
Production Taiwan Tea Experiment Station, Yangmei, Taiwan, pp. 63-75.
Nakamura, Y. 1990. Effect of sugar on formation of adventitious buds and growth
of auxiliary buds in tissue culture of tea. Bull. Shizuoka Tea Expt. Station., 15: 1-5.
Phukan, M.K. and Mitra, G.C. 1984. Regeneration of tea shoots from nodal explants
in tissue culture. Curr. Sci., 53: 874-876.
Ranaweera, K.K., Gunasekara, M.T.K. and Eeswara, J.P. 2013. Ex vitro rooting: A
low cost micropropagation technique of Tea (Camellia sinensis (L.) O. Kuntze)
hybrids. Scientia Horticulturae, 155: 8-14
Sandal, I., Bhattacharya, A. and Ahuja, P.S. 2001. An efficient liquid culture system
for tea shoot proliferation. Plant Cell Tiss. Org. Cult., 65: 75-80.
Sharma, M., Sood, A., Nagar, P.K., Prakash, O. and Ahuja, P.S. 1999. Direct rooting
and hardening of tea microshoots in the field. Plant Cell Tiss. Org. Cult., 58:
111-118.
INTEGRATED MANAGEMENT OF GALL DISEASE IN TEA

M. Ali1, M.S. Islam2 and M. Ahmed3
Abstract
An experiment was conducted at Bilashcherra Experimental Farm of
Bangladesh Tea Research Institute during 2006- 2013 to find out the
potential control methods of Gall disease in tea. The experiment was laid
out in a RCBD design with ten treatments and three replications. The
treatments consist of different pruning operations and followed by foliar
application of Carbendazim based fungicide @ 750g ha-1. All the treatments
were applied during pruning operation. Repeated application of the said
fungicide was applied after first rainfall of the year. Yield data and disease
severity were recorded at seven days and thirty days intervals respectively.
The disease severity was assessed using a 0-5 scale. All treatments caused
significant reduction (P=0.05) in severity of the disease in terms of PDI and
increase of yield of made tea as compared to control. The highest yield
(2392.45 kg/ha) and highest reduction (65.17%) in PDI was obtained from
T5 followed by T7, T4 and T3. The highest marginal rate of return (1,792%)
was also found in same treatment. Deep Skiff (DSK) operation and
following application of systemic fungicide Carbendazim @ 750 g /ha gave
the better result to minimize the Gall disease as well as to achieve higher
yield over other treatments.
Keywords: Gall disease, Integrated, Carbendazim, Deep Skiff
Introduction
Tea, the most popular beverages in the world, obtained from the flush shoots of the
plant Camellia sinensis (L). O. Kuntze. It has been cultivated in more than 30 countries
over the world. Most of the tea world production is produced in Asia (81.4%),
second is in Africa (15.5%), which is being followed by Latin America (2.0%) (FAO,
1999). It is one of the largest agro-based industries in Bangladesh. There are about
55.25 thousand hectares (Anon., 2013) of land under tea plantation producing about
61.93 million kg of made tea (Anon, 2013). The average yield per hectare of tea in
Bangladesh is 1,144 kg/ ha (Anon., 2013) which is quite low as compared to some
other tea growing countries of the world. Many factors are associated with the low
yield of our tea. Crop loss due to diseases is one of the key factors. Tea is a crop that
1CSO Pest Management Department, 2SSO Plant Pathology Division, 3Director Bangladesh Tea
Research institute, Srimangal-3210, Moulvibazar.
*Corresponding author’s email address: ali63bd@yahoo.com
is grown extensively under monoculture conditions, remote from its natural
environment. The tea bush like any other living plant is susceptible to attacks by
diseases, more so as it has been forced to grow, under varying climatic and soil
conditions. Disease attack may be in a sporadic form or in an epidemic form. When
the disease occurs in an epidemic form, it can cause colossal crop loss and can
damage the national economy of a country. The majority of the diseases in tea are of
fungal origin. More than 400 pathogens cause various diseases in tea (Chen and
Chen, 1990) viz., foliage, stem and root all over the world. The disease spectrum of
Bangladesh tea consists of one algal disease, 18 fungal diseases and a few epiphytes
(Sana, 1989). The total crop loss in Bangladesh tea, though not yet precisely
ascertained, a conservative estimate to the tune of 10-15% may be reckoned annually
(Sana, 1989). Among the diseases, Gall is the major one. Several isolations were
made in the laboratory to identify the causal agent of the disease but no other
organisms were observed except Fusarium oxysporum Schlecht. The disease was first
noticed in the year 1998 at Bilashcherra Experimental Farm of Bangladesh Tea
Research Institute on a tea clone BT1. The disease was found to be confined on the
upper canopy of bushes mostly within 5 cm of the plants. Galls were found to form
at the base of leaf petioles, leaf scars, on pruning cuts, at the base of twigs, in
between nodes particularly at the axial of branches. In some cases galls were found
to develop on the edges of thick cut surfaces of branches. The disease causes
massive gall formation on the upper canopy of the tea bushes resulting scanty and
weak flushes (shoots). With the advancement of time infected upper twigs die and
falloff. Severe gall formation restricts emergence of new shoots, consequently
substantial crop loss may occur if not controlled at the initial stage.
Integrated Disease Management (IDM) is an approach that attempts to use all
available methods of control of a disease or of all the diseases and pests of a crop
plant for best control results with the least cost and the least damage to the
environment. Cultural, Physical, Biological and Chemical control are the key
components of Integrated Disease Management. Available literatures in relation to
control of the disease in tea cultivation are very limited. Different chemical groups
like carbendazim, copper oxychloride, copper hydroxide, hexaconazole,
propiconazole are recommended for controlling the disease in Bangladesh (Huq et al.
2007). Effective, economically viable, environment friendly and acceptable
management practices are yet to be developed. So, a research work has been under
taken to establish potential control measures of the leading disease.
A field experiment was conducted at the Bilashcherra Experimental Farm (BEF) of
Bangladesh Tea Research Institute (BTRI) during 2006 to 2013 to establish potential
control measures of the disease. The experiment was laid out in a RCBD design with
ten treatments each of replicated thrice. The treatments were T0= Untouched
(Control), T1= T0+ Carbendazim @ 750 gm ha-1, T2= LP, T3= LP+ Carbendazim @
750 gm ha-1, T4= DSK, T5= DSK+ Carbendazim @ 750 gm ha-1, T6= MSK, T7=
MSK+ Carbendazim @ 750 gm ha-1, T8= LSK and T9= LSK+ Carbendazim @ 750
gm ha-1. All the treatments were applied during pruning operation. After pruning and
cleaning of litters Carbendazim based fungicide was applied @ 750 gm ha-1.
Repeated application of the said fungicide was applied after first rainfall of the year.
Data were collected on the disease severity at monthly interval and on yield of green
leaves at weekly interval against each treatment. The disease severity was assessed
using a 0-5 scale; 0= no infection, 1= 1-20% infection, 2= 21-40% infection, 3= 41-
60% infection, 4= 61-80% infection and 5= 81-100% infection of the disease
(Rashid et al. 1987). Data were converted as production of made tea (kg ha-1) and
Percent Disease Index (PDI). Percent Disease Index (PDI) was computed following
standard formula as described by Singh (2000).
Sum of (disease score × number of plants within that score)

PDI= ---------------------------------------------------------------------------- × 100
Highest score × total number of plants checked
Data were analyzed using MSTAT C computer programme. Mean separation was
done using Duncan’s Multiple Range Test (DMRT).
All treatments with pruning and following foliar application of Carbendazim @ 750
gm ha-1 caused significant reduction in severity of Gall disease in terms of PDI and
increase of yield of made tea as compared to control, where no pruning and
fungicide was applied. Among the treatments the highest reduction in PDI was
obtained with T5, which is followed by T7 and T3. They caused 65.17, 58.95 and
43.53% reduction in PDI over control, respectively. The least effective treatment was
T1, which gave only 9.76% reduction in PDI (Figure 2). The highest yield of tea was
achieved with the T5, which was followed by T7, T4 and T3. The yield was 2392.45,
2204.02, 2094.32 and 2080.62 kg ha-1 respectively. Treatments T4 and T3 are
statistically similar. They gave 63.33, 50.47, 42.98 and 42.04% increase yield of made
tea, respectively (Table 1). The yield of made tea was linearly and inversely correlated
with severity of Gall disease in terms of PDI. There relation could be expressed by
the regression equation Y = 40.792x + 1680.2, where Y represented yield of tea and
x represented slop of the equation. The R2 value indicated that their relationship may
be expressed by 17.45% (Figure 1). The highest marginal rate of return (1,792%) was
obtained in the treatment T5 receiving DSK operation and following application of
Carbendazim @ 750 gm ha-1 (Table 3). Results of the experiment revealed that foliar
spray with Carbendazim @ 750 gm ha-1 after pruning reduced severity of the disease
and increased yield of made tea considerably.
Carbendazim is a systemic fungicide that is absorbed through the foliage and is
translocated within the plant. After removal the infected plant debris or pruning
litters, the latent inocula within the plant may not be aggravated further due to fungi
toxiccity (Pandey, 2001)
Table 1. Production of made tea (Kg ha-1) and Percent Disease Index (PDI) against
different treatments during (2006-2013)
Treatment Made tea (Kg/ha) % Disease
(Mean of 3 replications) Index
T0= Untouched (Control) 1,464.72 f 22.83 a
T1= T0+ Carbendazim @ 750 gm ha-1 1,538.02 f 20.60 ab
T2= LP 1,809.74 de 16.60 bcd
T3= LP+ Carbendazim @ 750 gm ha-1 2,080.62 bc 12.89 de
T4= DSK 2,094.32 bc 14.38 cd
T5= DSK+ Carbendazim @ 750 gm ha-1 2,392.45 a 7.95 f
T6= MSK 1,859.16 cde 14.96 cd
T7= MSK+ Carbendazim @ 750 gm ha-1 2,204.02 ab 9.37 ef
T8= LSK 1,675.14 ef 18.38 bc
T9= LSK+ Carbendazim @ 750 gm ha-1 1,927.57 cd 15.95 cd
3000
2,392.45
2500
2,204.02
2,080.62
2,094.32
2000 1,809.74
Production (kg/ha)
1,859.16 1,927.57
1500 1,538.02 1,675.14
1,464.72
2
y = 40.792x + 1680.2 R = 0.1745
1000
500
20.6
22.83 16.6 12.89 14.38 7.95 14.96 9.37 18.38 15.95
0
1 2 3 4 5 6 7 8 9 10
% Disease Index
Fig.1. Effect of disease incidence on the production of made tea (kg/ha)

65.17
70
58.95
% Reduction of PDI 60
50 43.53
40 30.13
30 37.01 34.47
27.28
20
19.49
10 0 9.76
0
T0 T1 T2 T3 T4 T5 T6 T7 T8 T9
Treatments
Fig. 2. Showing Reduction of PDI over the control

Table 2. Partial budget of different treatments used for controlling Gall disease in tea
Treatments Yield Variable cost Gross Return Gross Margin
(Kg/ha) (Tk/ha) (Tk/ha) (Tk/ha)
T0 1,464.72 0 2,93,000 2,96,000
T1 1,538.02 1,565 3,07,600 3,06,035
T2 1,809.74 7,752 3,62,000 3,54,248
T3 2,080.62 9,317 4,16,200 4,06,883
T4 2,094.32 5,574 4,18,800 4,13,226
T5 2,392.45 7,036 4,78,400 4,71,364
T6 1,859.16 3,484 3,71,800 3,68,316
T7 2,204.02 5,049 4,40,800 4,35,751
T8 1,675.14 3,049 3,35,000 3,31,951
T9 1,927.57 4,613 3,85,600 3,80,987
Table 3. Marginal analysis of different treatments used for controlling Gall disease in
tea
Treatment Gross Variable Margin Gross Margin Margin
Margin cost Margin(Tk/ha) Variable Rate of
(Tk/ha) (Tk/ha) cost Return
(Tk/ha) (%)
T5 4,71,364 7,036 35,613 1,987 1,792
T7 4,35,751 5,049 22,525 (-) 525 -
T4 4,13,226 5,574 - - -
Conclusion
It can be concluded from the experiment that Deep Skiff (DSK) operation and
following application of systemic fungicide Carbendazim @ 750 gm ha-1 gave better
result to minimize the Gall disease as well as to achieve higher yield over other
treatments.
References
Anonymous. 2013. Annual Report of 2011 & 2012. Bangladesh Tea Association, PP:
167-261.
Chen, Z.M. and X.F. Chen, 1990. The diagnosis of tea diseases and their control (in
Chinese) Shanghai. Shangahai Scientific and Technical Publishers, 9: 73-88.
FAO, 1999. FAO year book of production Vol 52, 1998. FAO Statistic Series No.
148. Food and Agriculture Organization of the United Nations, Rome: P. 170
Huq, M., M. Ali and M.S. Islam. 2007. Approved Fungicides and Weedicides for
Bangladesh Tea. Cireular No. 119. Bangladesh Tea Research Institute. Srimangal, PP.
1-6
Pandey, B.P. 2001. Principles of Plant Disease control; 6th edition. 121 pp.
Rashid, A.Q.M.B, M.B. Meah and M. Jalaluddin. 1987. Effect of leaf blight caused by
Drechslera sorokiniana on some yield components of wheat. Crop Protect. 6 (4):
256-260
Sana, D.L. 1989. Tea Science. Ashrafia Boi Char. 36. Bangla Bazar, Dhaka, pp. 96-99.
Singh, R.S. 2000. Assessment of disease incidence and loss. Introduction to
principles of pathology. 3rd edition: 328 pp.
EFFECT OF POULTRY MANURE ON THE YIELD OF MATURE TEA

VIS-À-VIS SOIL PROPERTIES
A.Q. Khan , A. Biswas2, K.F.T. Zohora2 and A.K. Saha3
1
Abstract
A trial was conducted at BTRI farm during January 2012 to December 2013 to
evaluate the performance of poultry manure on the yield of mature tea and on soil
properties. Poultry manure was applied in different rates (0, 0.25, 0.50, 0.75
ton/ha) with 100%, 75%, 50% and 25% of the BTRI recommended fertilizer
doses. Poultry manure was applied once while BTRI recommended fertilizer was
applied in two splits in a year. Yield of tea was recorded at seven days interval
during the harvesting season. Soil samples were collected before starting the
experiment as well as after completion of the experiment and analyzed
accordingly. Highest yield (1442 kg made tea/ha) was recorded in T4 where 0.75
ton/ha poultry manure was applied along with 25% of BTRI recommended
chemical fertilizer. The yield increase was 21.69% over the control. A slight
increase of the macro and micro nutrients were found. But the changes of the
nutrient status due to the application of poultry manure were insignificant.
Keywords: Poultry manure, mature tea, soil properties
Introduction
Tea (Camellia sinensis) is an important cash crop as well as long established plantation
crop of enormous economic importance to Bangladesh that can meet the domestic
demand of this cheapest health beverage. It is an intensively managed perennial
monoculture crop cultivated on large and small scale plantation.
Fertilizer is one of the major agro-inputs contributing to the productivity in tea
plantation. For proper maintenance of the health of tea bushes and to obtain high
yield, a well-balanced fertilization is necessary at certain intervals throughout the year
(Sarwar et al, 2007). But the use of inorganic fertilizers has been observed to cause
the destruction of soil texture and structure which often leads to soil erosion, acidity
as a result of leaching effects of mineral nutrients (Ojeniyi, 2000). All these gave rise
to reduced crop yields as a result of soil degradation and nutrient imbalance (Ojeniyi,
2000).
1 & 3SSO, 2SO, Soil Science Division, BTRI, Sreemangal-3210, Moulvibazar

*Corresponding author’s email address: scientist08@yahoo.com
The sustainable crop production needs proper management of nutrient resources
and conservation of soil fertility. A combined application of balance inorganic
fertilizer and supplementary dose of organic matter sustains the crop productivity
and maintain the soil fertility. Integrated supply of nutrients to plants through
organic and inorganic sources is becoming an increasingly important aspect of
environmentally sound agriculture. Nutrients contained in organic manure are
released slowly and are stored for a longer time in the soil, thereby ensuring a longer
residual effect and persistence of nutrient availability (Singh et al, 2011).
Improvement in environmental conditions with respect to public health has been
observed as some of the major reasons for the need to adopt organic farming by
farmers in the world (Eifediyi and Remison, 2010). Organic manures sustain
cropping systems through better nutrient recycling and improvement in soil physical,
chemical and biological properties (Ojeniyi, 2000). It increases water holding capacity
of soil and nutrients will be made more readily available to crops where organic
manures will be added to the soil (Dada and Fayinminnu, 2010).
Poultry manure is one of the main sources of organic plant nutrient has long been
used by ancient farmers as a source of nutrition. It has a great potential for soil
fertility maintenance. It has been used to amendment soil in order to provide
nutrients for plant growth, increase organic matter in the soil, improve microbial
populations in the soil, and to control plant pests such as nematodes and weeds
(Shepherd and Withers, 2009). It supplies chelating agents that aid in maintaining the
solubility of micronutrients (Sinnadurai, 1992).
If poultry manure is added in combination with chemical fertilizer, it will supplement

all nutrients to crop and increase the productivity of the crop (Edward and Daniel,
1992). This study was therefore designed to investigate the effect of poultry manure
on the yield of mature tea and on soil properties.
Materials & Methods

A trial was conducted at BTRI farm during January 2012 to December 2013 to
evaluate the performance of poultry manure on the yield & soil properties of mature
tea in a randomized block design with four treatments and three replications. The
unit plot size was 36.36 m2. The treatment combinations were T1 = Control (No
poultry manure) + BTRI recommended fertilizer dose (N130 P35 K75 Zn10 kg/ha); T2
= Poultry manure (0.25 ton/ha) + 75% of BTRI recommended fertilizer dose; T3 =
Poultry manure (0.50 ton/ha) + 50% of BTRI recommended fertilizer dose; T4 =
Poultry manure (0.75 ton/ha) + 25% of BTRI recommended fertilizer dose. Urea
(N) and Muriate of Potash (K) fertilizer were applied in two splits. First dose of
fertilizer was applied in the month of April when soil contained sufficient moisture
& the second dose was applied in the first week of August in the year 2012 and 2013
respectively. Poultry manure was applied once in a year.
Soil samples were collected from a depth of 0-23 cm before starting the experiment
as well as after completion of the experiment. Yield of tea i.e. the weight of green
leaves was recorded at seven days interval. Irrigation, pruning, pest control and other
intercultural operations were done as and when necessary. Data on yield were
recorded and analyzed statistically. Texture, pH, organic carbon (%), total nitrogen
(%), available phosphorus (ppm), potassium (ppm), calcium (ppm), magnesium
(ppm), sulphur (ppm), boron (ppm), copper (ppm), iron (ppm), manganese (ppm)
and zinc (ppm) of the soil samples were determined before as well as after
completion of the experiment. Soil texture was determined by hydrometer method.
However, pH was determined by using pH meter (soil : distilled water = 1 : 2.5).
Walkley and black wet oxidation method was applied for soil organic carbon
determination (Imamul Huq and Alam, 2005). Total nitrogen was determined by
Micro kjeldahl steam distillation method (Imamul Huq and Alam, 2005).
Colorimetric estimation of available phosphorus was done by Bray-II ascorbic acid
method (Imamul Huq and Alam, 2005). Available potassium, calcium and
magnesium were extracted with 77% ammonium acetate solution. Available
potassium was determined by flame photometer while calcium and magnesium was
determined by AAS (atomic absorption spectrophotometer). Available zinc,
manganese, copper and iron were extracted with 1M ammonium acetate (pH 4.8)
and determined by AAS. Colorimetric determination of available boron was done by
UV-spectrophotometer.
Chemical analysis of poultry manure was done. It was digested with nitric-perchloric
acid mixture. After digestion of the poultry manure, phosphorus & sulfur was
analyzed by UV-spectrophotometer while potassium was analyzed through flame
photometer. Determination of zinc, copper, chromium, cadmium, lead and nickel
was done by AAS. For nitrogen analysis poultry manure was digested with sulphuric-
perchloric acid mixture. Then nitrogen was determined through micro-kjeldahl
method (Imamul Huq and Alam, 2005). Organic carbon content of poultry manure
was determined by dry combustion method.
Tea may be grown on soils of diverse geological origins. In Bangladesh most soils are
quaternary and recent alluvia. Tea soils are highly weathered, extremely acidic and
low of fertility status. Furthermore these soils do not receive deposits of fertile silt by
flooding; rather they suffer from erosion (Alam, 1999). Table 1 shows analytical
results of the soil samples which were collected from the experimental plots before
setting up the experiment as well as after completion of the experiment. The soil was
light textured with highly acidic in nature. After completion of the experiment a
slight increase of all the macro and micro nutrient was found except copper. But
from the statistical analysis (t-test) it is found that the changes of nutrient content in
soil due to the application of poultry manure is insignificant.
Table 1. Soil analytical results
Soil parameters Before experiment (Initial) After experiment
(Final)
Texture Sandy loam Sandy loam
pH 4.2 4.5
Organic carbon (%) 1.13 1.29
Total Nitrogen (%) 0.110 0.120
Available P (ppm) 3.36 4.22
Av. K (ppm) 28 32
Av. Ca (ppm) 76 93
Av. Mg (ppm) 12 19
Av. S (ppm) 11 15.6
Av. B (ppm) 0.46 0.47
Av. Cu (ppm) 7.60 6.66
Av. Fe (ppm) 198 201.3
Av. Mn (ppm) 35.6 37.8
Av. Zn (ppm) 1.16 1.19
Sana (1989) reported that the most suitable tea soil is thought to be light, friable and
well drained having soil pH from 4.5 to 5.8. So, before setting up the experiment pH
of the soil was below the critical limit while after the completion of the experiment
pH increased slightly which was within the critical limit. The critical values in
Bangladesh condition have been fixed at 0.1% for nitrogen and 1% for organic
matter (Alam, 1999). Content of organic carbon and total nitrogen of the soil
samples collected before as well as after the experiment was above the critical limit.
Available phosphorus, potassium, calcium and magnesium content were below the
critical limit in the soil samples which were collected before setting up the
experiment. But after completion of the experiment only calcium content increased
and it was above the critical limit. Alam(1999) reported that the critical limit of
nutrient status of tea soil is 10 µg/g for P, 80 µg/g K, 25 µg/g for Mg and 90 µg/g
for Ca.
The maximum permissible limits of micronutrients in soils are 2 to 250 mg/kg for
copper and 10 to 300 mg/kg for zinc respectively (katabata-Pendias and Pendias,
1984). Before and after the experiment copper content was within the permissible
limit but zinc content was below the permissible limit. Ma. et al., (2003) reported that
critical limit of available boron was 0.50 mg/kg from the analysis of tea growing soils
of China. The boron content was found 0.46 ppm & 0.47 ppm in the soil samples
collected before and after the experiment respectively. The critical limit of available
sulphur is 40 ppm and the soil test values indicate lower sulphur content than the
critical limit. Iron is the predominant metal with the concentration of 198 and 201.3
ppm in the soil whereas Mn content is 35.6 and 37.8 ppm.
Table 2. Chemical analytical results of poultry manure
Parameters Poultry manure Govt. Standard (critical limits)
Color Grey Dark grey to black
Physical condition Non-granular Non-granular
Odor Absence of foul Absence of foul odor
odor
Moisture 22.1% 15-20%
pH 8.5 6.0-8.5%
Organic carbon 19.7% 10-25%
Total nitrogen 2.52% 0.5-4.0%
C:N 7.8 : 1 20 : 1
Phosphorus (P) 1.9% 0.5-3.0%
Potassium (K) 1.14% 0.5-3.0%
Sulphur (S) 0.33% 0.1-0.5%
Zinc (Zn) 0.02% 0.1%
Copper (Cu) 0.003% 0.05%
Chromium (Cr) 0.99 ppm 50 ppm (max)
Cadmium (Cd) 0.59 ppm 5 ppm (max)
Lead (Pb) 0.99 ppm 30 ppm (max)
Nickel (Ni) 1.02 ppm 30 ppm (max)
Inert material (max) 1% 1%
Analytical results of the poultry manure is presented in Table 2 which was applied
during the experiment. From the phsico-chemical analysis of poultry manure it is
found that all the physical properties as well as chemical constituents are within the
critical limits. The content of toxic heavy metals such as chromium, cadmium, lead
and nickel in the poultry manure which was applied in the experimental plots were
0.99, 0.59, 0.99, 1.02 ppm respectively. The permissible limits of chromium,
cadmium, lead and nickel are 50, 5, 30, 30 ppm respectively (Fertilizer
Recommendation Guide, 2012). So the presences of toxic heavy metals in the
poultry manure were below the critical limit. Supplied poultry manure was a very
good source of organic carbon (19.7%), total nitrogen (2.52%), phosphorus (1.9%),
potassium (1.14%) and sulphur (0.33%) etc.
Table 3. Effect of poultry manure on the yield of tea
Treatment Average yield Average yield Made tea Increase over
(kg/plot) (kg/ha) (kg/ha) control (%)
T1 18.75 5156 1185 -
T2 20.75 5704 1312 10.72
T3 21.26 5846 1344 13.42
T4 22.81 6272 1442 21.69
The effect of poultry manure on the yield of tea is presented in Table 3. Yield
differences between the treatments were statistically insignificant. That means similar
yield can be achieved even after 75% reduction of chemical fertilizer (BTRI
recommended dose) if poultry manure is added in appropriate dose. Though not
statistically significant, increased yield was recorded in all the treatments when
poultry manure was applied. Highest yield was recorded in T4 (1442 kg made tea/ha)
which was 21.69% over the control. In case of T2 and T3, yield increase was 10.72%
and 13.42% respectively.
Conclusion
Both organic and inorganic fertilization are essential to increase tea yield. Tea
planters could minimize the use of NPK fertilizers with poultry manure to reduce
the cost of inorganic fertilizers as well as to reduce their damaging effects on the soil.
This trial clearly indicated that production of tea can be enhanced by the application
of poultry manure. It may be recommended as organic fertilizer for improving the
tea soil health as well as the production of mature tea. The highest rate of poultry
manure (0.75 ton/ha) with 25% of the BTRI recommended fertilizer doses gave the
highest yield (1442 kg/ha) of made tea which was 21.69% higher than the control.
A long term experiment on integrated nutrient management in combinations of
poultry manure and NPK could also be undertaken to support general fertilizer
application.
References
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Advances in Tea Science (Edited by N. K. Jain), Aravali Books International (P).
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regimes on period of weed control in okra. Not. Bot. Hort. Agrobot. Cluj. 38(1)
149-154.
Edward, D.R. and Daniel, T.C. 1992. A review on poultry manure.Bioresour.
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Eifediyi, E.K. and Remison, S.U. 2010. Growth and yield of cucumber (Cucumis
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(BARC), Farmgate, Dhaka.
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Press, Boca Raton, Fla
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tea gardens in zhejiang province and its affecting factors. Scientia Agric. Sinica
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ISSN: 0253-5483

TEA JOURNAL OF BANGLADESH

BANGLADESH TEA RESEARCH INSTITUTE

Chief Editor Dr. Mainuddin Ahmed

Dr. Mainuddin Ahmed

Md. Haroon-or-Rashid Sarker

EFFECTS OF DIFFERENT PLUCKING SYSTEMS ON SHOOT GROWTH

EVALUATION OF SOME INDIGENOUS PLANT EXTRACTS AGAINST

INTEGRATED MANAGEMENT OF GALL DISEASE IN TEA 31-36

EFFECT OF POULTRY MANURE ON THE YIELD OF MATURE TEA

(Dr. Mainuddin Ahmed)

EFFECTS OF DIFFERENT PLUCKING SYSTEMS ON SHOOT

1SSO,Bangladesh Tea Research Institute, Srimangal–3210, Moulvibazar, Bangladesh

*Corresponding author’s email address: toufiqtea@yahoo.com

Lantana (Lantana camara) Mahogani (Swietenia mahagoni) Neem (Azadirachta indica)

Plate 1. Some indigenous plant extracts against tea mosquito bug

Field evaluation of toxicity of plant extracts against Helopeltis

Fig. 1. Performance of plant extracts on the mortality of H. theivora

DEVELOPMENT OF IN VITRO MICRO PROPAGATION SYSTEM IN

Department of Botany, University of Dhaka, Bangladesh.

2.0 0.1 25 22 20 40-43 40-43 1.5 2.2 4.3 4.6

Dipping No. of % of Days Mean no. of Mean length

Success of survivality of tissue culture derived plants depends upon proper

INTEGRATED MANAGEMENT OF GALL DISEASE IN TEA

Sum of (disease score × number of plants within that score)

Fig.1. Effect of disease incidence on the production of made tea (kg/ha)

Fig. 2. Showing Reduction of PDI over the control

EFFECT OF POULTRY MANURE ON THE YIELD OF MATURE TEA

1 & 3SSO, 2SO, Soil Science Division, BTRI, Sreemangal-3210, Moulvibazar

If poultry manure is added in combination with chemical fertilizer, it will supplement

Materials & Methods

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