BIBLIOGRAPHY ...
BIBLIOGRAPHY
MACLI-ING, JANETH P. APRIL 2012. Effect of Different Growing Media and
Frequency of Misting on the Acclimatization of Chrysanthemum Plantlets In Vitro. Benguet
State University, La Trinidad, Benguet.
Adviser: Leila Mary B. Alipio -Ayban MSc.
ABSTRACT
The study was conducted at the Ornamental Horticulture Area, Benguet State
University, La Trinidad, Benguet, from December 2011 to February 2012. The study was
conducted to adapt the most appropriate soil media to harden rooted chrysanthemum plantlets
before transplanting it to the field; to determine the frequency of misting that is appropriate
in the acclimatization of chrysanthemum plantlets; and to establish the best condition of soil
media and frequency of misting in the acclimatization of chrysanthemum plantlets.
Chrysanthemum plantlets grown in alnus compost + sand +burnt rice hull were the
best growing media to promote growth and development in acclimatizing the plantlets.
Likewise plantlets grown in alnus compost misted twice a day had the highest percentage
survival.
Planting chrysanthemum plantlets in a composition of 1:1:1 ratio of alnus compost+
sand+ burnt rice hull is recommended and the seedlings should be misted once a day for one
month to promote growth and development have a high percentage survival of the seedling.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
RESULTS AND DISCUSION
Plant Height
Effect of growing media. Highly significant differences were observed among the
different growing media used. Chrysanthemum plantlets grown in soil media containing
alnus compost + sand + burnt rice hull (1:1:1) markedly promoted plant growth faster
compared to other growing media.
Thompson and Troech (1978) added that the use of compost also resulted in humus
formation and promotes good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus (6%), calcium (13%), magnesium and organic matter content of 5.6%.
The presence of sand and burnt rice hull in the alnus compost media improved the
physical properties of the medium resulting to better drainage and aeration leading the better
plant growth performance (Dumaslan, M. 2006).
As previously mentioned the results collaborate with the findings of Gawadan (1999)
that a mixture of 1:1:1 alnus compost + rice hull + river sand had significantly promoted
longer roots.
Effect of frequency of misting. There were no significant differences on the height of
chrysanthemum plantlets as affected by misting (Table 1).
Interaction effect. Significant interaction effects were observed from the combination
of different growing media and frequency of misting on the height of plantlets, one month
from transplanting.
Plantlets grown in alnus compost + sand + burnt rice hull, misted once a day was the
tallest compared to other treatments but were comparable to those planted in alnus compost +
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
sand + burnt rice hull, misted twice a day; garden soil + alnus compost, misted twice a day
and alnus compost + sand + burnt rice hull, misted three times a day .
Table 1. Plant height
TREATMENT
MEAN (cm)
Growing Media
Alnus Compost + Garden Soil
13.62b
Alnus Compost
9.41c
Sand
5d
Burnt Rice Hull
3.20d
Alnus Compost + Sand + Burnt Rice Hull (1:1:1)
18.46a
Frequency of Misting
Once a day
9.0a
Twice a day
10.44a
Three times a day
10.29a
CV (%)
23.35
Means with a common letter are not significantly different at 5% level of DMRT
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Figure 1. Average plant height as affected by different growing media and frequency of
misting (Bars with the same letter are not significantly different at 5% level of DMRT)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Root Length
Effect of growing media. Plantlets grown in alnus compost + sand + burnt rice hull
significantly produced longer roots but it was not significantly different to those plantlets
grown in alnus compost and alnus compost + garden soil. Furthermore, plantlets grown in
sand and burnt rice hull alone produced the shortest roots.
In the Handbook of Texas Green house Management, burnt rice hulls have a light
weight but effective in improving drainage while sand do little to improve the physical
properties of growing media that may result in reduced drainage and aeration as a result of
compaction (Anon., 2006) as cited by Dumaslan M. (2006).
The above results agree with the earlier findings of Bisley (2008) that the seedlings
from seeds sown in 1:1 alnus compost + garden soil had the longest root length.
Effect frequency of misting. The effect of different growing media and frequency of
misting in the length of the roots in chrysanthemum plantlets was not significant.
Interaction effect. The interaction effect of the different growing media and frequency
of misting on the root length of chrysanthemum plantlets were highly significant.
Plantlets grown in alnus compost+ sand + burnt rice hull, misted in different
frequencies promoted the production of longer and more roots; but were not significantly
different from those grown in garden soil + alnus compost and alnus compost, misted once a
day; garden soil + alnus compost and alnus compost, misted twice a day; and garden soil +
alnus compost, alnus compost and sand, misted three times a day.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Table 2. Root length
TREATMENT
MEAN (cm)
Growing Media
Alnus Compost + Garden Soil
12.26a
Alnus Compost
12.80a
Sand
8.44b
Burnt Rice Hull
4.22c
Alnus Compost + Sand + Burnt Rice Hull 1:1:1)
14.06a
Frequency of Misting
Once a day
10.12a
Twice a day
10.12a
Three times a day
10.83a
CV (%)
22.45
Means with a common letter are not significantly different at 5% level of DMRT
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
16
a
a
ab
14
ab
ab
ab
a
12
ab
ab
ab
)
m
c
(
b
t
10
h
g
n
e
l
8
oot
R
e
g
c
r
a
6
e
cd
v
A
cd
4
2
d
0
Misted once a day
Misted twice a day
Misted three times a
day
Frequency of Misting
T1=alnus compost + garden soil
T2= alnus comost
T3= sand
T4= burn rice hull
T5= alnus compost + sand + burnt rice hull
Figure 2. Average root length as affected by different growing media and frequency of
misting (Bars with the same letter are not significantly different at 5% level of DMRT)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Number of Leaves
Effect of growing media. Plantlets grown in alnus compost + garden soil produced
significantly more leaves with a mean of 22.00 but were not significantly different from
plantlets grown in alnus compost and in alnus compost + sand + burnt rice hull.
Thompson and Troech (1978) added that the use of compost also resulted in humus
formation and promotes good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus (6%), calcium (13%), magnesium and organic matter content of 5.6%.
Effect of frequency of misting. There were no significant effects of the frequency of
misting in the number of leaves produced per seedling/ plantlets one month from
transplanting.
Interaction effect. The numbers of leaves of chrysanthemum plantlets were not
significantly affected by interaction effects between the different growing media and
frequency of misting.
Table 3. Number of leaves per plantlets
TREATMENT
MEAN
Growing Media
Alnus Compost + Garden Soil
22.00a
Alnus Compost
20.11ab
Sand
14.66bc
Burnt Rice Hull
11.11c
Alnus Compost + Sand + Burnt Rice Hull (1:1:1)
18.667ab
Frequency of Misting
Once a day
15.067a
Twice a day
17.66a
Three times a day
19.200a
CV (%)
32.87
Means with a common letter are not significantly different at 5% level of DMRT
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Occurrence of Insect Pest and Diseases
Insect Infestation
There was slight insect infestation observed during the first week of this study.
Results show that 25 % of the total plantlets grown in alnus compost + sand + burnt rice hull
were infested by army worms as compared to other growing media. Severe rotting of plantlet
roots was observed in chrysanthemum plantlets grown in burnt rice hull misted in different
frequencies. There was no disease infection observed during the conduct of the study.
Percent Survival
Effect of growing media. Highly significant effects were observed on the percentage
survival as affected by the different growing media. Results showed that plantlets grown in
alnus compost had the highest percentage survival but was not significantly different from
plantlets grown in alnus compost + sand + burnt rice hull; those grown in sand and alnus
compost + garden soil. On the other hand, plantlets grown in burnt rice hull had the lowest
percentage survival.
Effect of frequency of misting. Results revealed that the plantlets misted twice a day
had the highest percentage of survival with a mean of 97.13 % but was not significantly
different with plantlets misted three times a day.
Interaction effect. Significant interactions were observed between the different
growing media and frequencies of misting on the percentage survival.
Results show that plantlets grown in alnus compost, sand, alnus compost + sand +
burnt rice hull misted in different frequencies and plantlets grown in garden soil + alnus
compost misted twice and three times day exhibited the highest plantlet survival.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Table 4. Percentage survival
TREATMENT
MEAN (%)
Growing Media
Alnus Compost + Garden Soil
96.80a
Alnus Compost
100a
Sand
98.40a
Burnt Rice Hull
63.40b
Alnus Compost + Sand + Burnt Rice Hull (1:1:1)
98.40a
Frequency of Misting
Once a day
83.80b
Twice a day
97.10a
Three times a day
93.30a
CV (%)
12.99
Means with a common letter are not significantly different at 5% level of DMRT
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
120
a a
a a a
a
a a
a
100
a
a
ab
l
ab
a
v
i
v
r
80
u
b
S
e
g
a
t
n
60
e
c
r
e
P
e
g
40
c
a
r
e
v
A
20
0
Misted once a day
Misted twice a day
Misted three times a
day
Frequency of Misting
T1= alnus compost + garden soil
T2= alnus compost
T3= sand
T4= burnt rice hull
T5= alnus comost + sand+ burnt rice hull
Figure 3. Average percentage survival of different growing media and frequency of misting
(Bars with the same letter are not significantly different at 5% level of DMRT)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Other Observations
It is observed that the plantlets grown in burnt rice hull produced brownish roots; while
plantlets grown in sand alone were stunted and exhibited nutrient deficiency symptoms.
It was observed that the roots of plantlets grown in burnt rice hull produced brownish
color and this could be the effect of the toxic component present in the growing medium.
Burnt rice hull is ash which contains potentially toxic levels of silicon which could
plasmolized the roots of the acclimatized seedlings when planted in a longer period that lead
to the rotting of the roots and exhibiting leaf yellowing. (Dumaslan, M. 2006).
Moreover, plantlets grown in sand alone had stunted growth that indicated nutrient
deficiency symptom.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
Plate 1. Overview of the experimental area
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
F1B1R3
F1B1R2
F1B1R1
F2B1R1
F2B1R2
F2B1R3
a
b
F3B1R2
F3B1R1
F3B1R3
F4B1R1
F4B1R2
c
d
F5B1R1
F5B1R2
F5B1R3
e
Plate 2. Uprooted plantlets in different growing media misted once a day (a= alnus compost
+ garden soil, b= alnus compost, c= sand, d= burnt rice hull, e= alnus compost + sand + burnt
rice hull)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
F2B2R1
F2B2R2
F1B2R2
F2B2R
F1B2R1
F1B2R3
3
a
b
F3B2R2
F4B2R3
F3B2R1
F3B2R3
F4B2R1
F4B2R2
c
d
F5B2R2
F5B2R1
F5B2R3
e
Plate 3. Uprooted plantlets in different growing media misted twice a day (a= alnus compost
+ garden soil, b= alnus compost, c= sand, d= burnt rice hull, e= alnus compost + sand + burnt
rice hull)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
F1B3R2
F1B3R1
F1B3R3
F2B3R2
F2B3R3
F2B3R1
a
b
F3B3R1
F4B3R1
F3B3R3
F3B3R2
F4B3R2
F4B3R3
c
d
F5B3R3
F5B3R3
F5B3R3
e
Plate 4. Uprooted plantlets from different growing media misted three times a day
(a= alnus compost + garden soil, b= alnus compost, c= sand, d= burnt rice hull, e= alnus
compost + sand + burnt rice hull)
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The study was conducted to determine the most appropriate soil media to harden
rooted chrysanthemum plantlets before transplanting to the field; to determine the frequency
of misting that is appropriate in the acclimatization of chrysanthemum plantlets; and to
establish the best condition of soil media and frequency of misting in the acclimatization of
chrysanthemum plantlets.
A medium of alnus compost+ sand+ burnt rice hull, significantly promoted faster
plantlet growth that remarkably produced longer roots. Moreover, plantlets grown in alnus
compost + garden soil significant produced more leaves; plantlets grown in alnus compost
alone had the highest percentage of plantlets survival.
Plantlets misted twice a day revealed a highest percentage of survival. There were no
significant effects in the frequency of misting noted in plant height, root length and number
of leaves produced.
Plantlets grown in alnus compost+ sand+ burnt rice hull, misted in different
frequencies had the best plantlet growth and longer fibrous roots. Moreover, plantlets misted
twice a day grown in alnus compost had the highest percentage of survival. There were no
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
significant interaction effects noted between the different growing media and frequency of
misting on the number of leaves.
Conclusion
Based on the results of the study alnus compost + sand + burnt rice hull (1:1:1), once,
twice and three times misting a day was the best combination in acclimatizing
chrysanthemum plantlets.
Recommendation
From the preceding, results and discussion, acclimatizing chrysanthemum plantlets in
a composition of 1:1:1 ratio of alnus compost+sand+ burnt rice hull and garden soil + alnus
compost, misted once a day is recommended.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
LITERATURE CITED
AZCON- AGUILAR, C., M. Cantos, A. Trancoso and J. M Barea. 1997. Beneficial Effect
of Arbuscular Mycorrhizason Acclimatization of Micropropagated Cassava
Plantlets. Scientia Hort. 72:63-71
BISLEY, M. B. 2008. Germination of Papaya (Carica Papaya) Seeds and Seedling
Characteristics as Affected by Different Growing Media in Camp 3 Tuba, Benguet.
Under Grad. Thesis BSU La Trinidad, Benguet Pp 25
CARVALHO, L. C., M.L. OSARIO, M. M. CHAVES, and S. AMACIO. 2001.
Chlorophyll Fluorescence as Indicator of Photosynthetic Functioning of In Vitro
Grapevine and Chestnut Plantlets Under Ex Vitro Acclimatization. Plant cell
Tissue Culture and Organ Culture.
CASSELLS, A.C. And C. WALSH. 1994. The Influence of Gas Permeability of the Culture
Lid on Calcium Uptake And Stomatal Function In Dianthus Microplants. Plant Cell
Tissue Organ Culture 37:171-178.
DE FOSSARD, R.A. 1981. Tissue Culture Micropropagation, Notes on Tissue
Culture.
DE KLERK, G-J. 2000. Rooting Treatment and Ex Vitrum Performance of Micropropagated
Plants. ActaHortic. 530:277-228.
DIAZ-PEREZ, J.C., K.A. SHAKEL and E.G. SUTTER. 1995a. Effects of In Vitro Formed
Roots and Acclimatization on Water Status And Gas Exchange of Tissue Cultured
Apple Shoots. J. AMER. Soc.Hort.Sc.120:435-440
DUMASLAN, M. R. 2006. Growth Performance of Hardened Tissue Cultured Strawberry
Plantlets Under Greenhouse Condition. MS Thesis, BSU La Trinidad, Benguet. Pp 7
GAMBORG, O. L. 2002. Plant Tissue Culture. Biotechnology Milestones.In Vitro Cell
Development Biology Plant.
GEORGE, E. F, and P. D. SHERINGTON. 1984. Plant Propagation by Tissue Culture,
Handbook and Directory of Commercial Laboratories.
JANNA, O. A., OHIDIN, R., MAZIAH, M. 2005. Glass House Acclimatization of Tissue-
Cultured Melastoma Malabathricum Plantlets. Tropical Sci. 45(1):45-49
MANTEL, S.H. and H. SMITH.1983, Plant Biotechnology. Cambridge University Press,
Cambridge.
MORGAN ED R., SEELVY, JHON F., and BURGE GARRY K., 2003. Acclimatizing
Tissue Culture Plants: Reducing The Shock. New Zealand Institute for Crop & Food
Res. Lt, Prvt Bag 11 600, Palmerston North, New Zealand.
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012
MURASHIGE ,T. and F. SKOOG. 1962. A Revised Medium for Rapid Growth and Bio
Assays with Tobacco Tissue Culture. Physiologica Plantarium 15:473-49
SLUIS, C.J. 2005. Protocols Greenhouse Planting Of Tissue Culture Transplants. In Plant
Tissue Culture Engineering (Gupta,S.D. And Ibaraki,Y.Eds)
THOMPSON, I.M. and TROECH, F.R. 1978. Soils and Soil Fertility, New York. Mc Graw
Hillbook Co. Inc. Pp. 108-111
Effect of Different Growing Media and Frequency of Misting on the Acclimatization of Chrysanthemum
Plantlets In Vitro | MACLI-ING, JANETH P. APRIL 2012