BIBLIOGRAPHY LAOYAN, JOLAFLOR C. MAY...
BIBLIOGRAPHY
LAOYAN, JOLAFLOR C. MAY 2012. Acclimatization of Carnation (Dianthus
caryophyllus var. Prodo Mint) Plantlets from in vitro. Benguet State University, La
Trinidad, Benguet.
Adviser: Araceli G. Ladilad PhD.
ABSTRACT
The study was conducted at the Ornamental Horticulture Area, Benguet State
University, La Trinidad, Benguet, from February 2012 to April 2012. The study was
conducted to establish the most appropriate growing media to harden rooted carnation
plantlets and to establish a protocol for the acclimatization of rooted carnation plantlets
grown from in vitro before transplanting them to the greenhouse benches.
Results shows that carnation plantlets grown in alnus compost + burnt rice hull 1:1
were the best growing media that promoted the production of longer roots, higher number
of developed leaves, and promoted faster plant growth. Likewise, plantlets grown in sandy
loam soil + burnt rice hull 1:1 produced higher number of roots. However, the different
soil media compositions did not significantly affect the percentage survival of the
acclimatized mint green carnation plantlets.
Soil media composition of alnus compost + burnt 1:1 or carbonized rice hull is
recommended for the acclimatization of carnation plantlets from in vitro for best
performance of the plantlets prior to in vivo transplanting in the greenhouse.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
RESULTS AND DISCUSSION
Root Length
Highly significant differences were observed among the different growing media
used. A mixture of alnus compost + burnt rice hull 1:1 promoted the longest roots with a
mean of 7.53 cm. Plantlets grown in sandy loam soil + sand 1:1 had the shortest length
of roots with a mean of 2.15 cm
According to Thompson and Troech (1978), the use of compost resulted in humus
formation and promotes good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus, calcium, magnesium and organic matter of 5.6 %.
On the other hand, Dumaslan (2006) stated that 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 to better plant growth and performance in strawberry
plantlets.
Table 1. Root length per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
3.18c
Alnus Compost + Sand (1:1)
3.02c
Alnus Compost + Burnt Rice Hull (1:1)
7.53a
Sandy Loam Soil + sand (1:1)
2.15c
Sandy Loam Soil + Burnt Rice Hull (1:1)
5.48b
Burnt rice Hull + sand (1:1)
6.51ab
Means followed by a common letter are not significantly different at 5% level (DMRT).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Number of Leaves
Results showed that plantlets grown in the different media had significantly
different number of leaves per plantlet after 45 days from transplanting. Plantlets grown in
alnus compost + burnt rice hull 1:1 produced the most number of leaves with a mean of
38.67 followed by those grown in burnt rice hull + sand 1:1 with a mean of 34.00,
respectively.
Thompson and Troech (1978) added that the use of compost resulted in humus
formation and promoted good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus, calcium, magnesium and organic matter of 5.6 %.
Length of Leaves
Plantlets grown in alnus compost + burnt rice hull 1:1 significantly produced
longer leaves but were not significantly different to the length of leaves of plantlet
grown in sandy loam soil + burnt rice hull 1:1 and in burnt rice hull + sand 1:1.
Table 2. Number of leaves per plantlet after 45 days from transplanting
TREATMENT
MEAN
Alnus Compost + Sandy Loam Soil (1:1)
19.33b
Alnus Compost + Sand (1:1)
16.00b
Alnus Compost + Burnt Rice Hull (1:1)
38.67a
Sandy Loam Soil + sand (1:1)
12.00b
Sandy Loam Soil + Burnt Rice Hull (1:1)
31.00a
Burnt rice Hull + sand (1:1)
34.00a
Means followed by a common letter are not significantly different at 5% level (DMRT).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 3. Length of leaves per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
1.51b
Alnus Compost + Sand (1:1)
1.41b
Alnus Compost + Burnt Rice Hull (1:1)
3.78a
Sandy Loam Soil + sand (1:1)
1.07b
Sandy Loam Soil + Burnt Rice Hull (1:1)
3.71a
Burnt rice Hull + sand (1:1)
3.33a
Means followed by a common letter are not significantly different at 5% level (DMRT).
Plantlets grown in sandy loam + sand 1:1 produced the shortest leaves with a mean
of 1.07 cm.
Plant Height
Table 4 shows that there were highly significant differences observed on the
height of plantlets grown in different soil media combinations. The prodo mint carnation
plantlets grown in alnus compost + burnt rice hull 1:1 were the tallest among the treatments
with a mean of 3.15 cm, followed by plantlets grown in burnt rice hull + river sand 1:1,
sandy loam + burnt rice hull 1:1 with means of 2.83 cm and 2.28 cm, respectively. It was
also observed that plantlets acclimatized in sandy loam soil + sand
1:1 have the shortest height with a mean of 1.45 cm and was comparable with plantlets
grown in alnus compost + sand 1:1 with means of 1.51 cm and 1.63 cm, respectively.
Macli-ing (2012) found that a media of alnus compost +sand + burnt rice hull
1:1:1 promoted faster plant growth for the acclimatization of chrysanthemum plantlets.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 4. Plant height per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
1.51c
Alnus Compost + Sand (1:1)
1.63c
Alnus Compost + Burnt Rice Hull (1:1)
3.15a
Sandy Loam Soil + sand (1:1)
1.45c
Sandy Loam Soil + Burnt Rice Hull (1:1)
2.28b
Burnt rice Hull + sand (1:1)
2.83ab
Means followed by a common letter are not significantly different at 5% level (DMRT).
Number of Roots
Based on the results presented in Table 5, sandy loam soil + burnt rice hull
1:1 enhanced the production of more roots, followed by soil media composition: alnus
compost + burnt rice hull 1:1, burnt rice hull + sand 1:1, and alnus compost + sandy loam
soil 1:1 with means of 24.55, 23.33, 13.00 and 11.00 roots per plantlet, respectively, while,
plantlets grown in alnus compost + sand 1:1 had the lowest number of roots per plantlet
after 45 days from transplanting with means of 9.11 roots per plantlet.
In the handbook of Texas Greenhouse Management, burnt rice hull 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 as cited by Dumaslan (2006).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 5. Number of roots per plantlet after 45 days from transplanting
TREATMENT
MEAN
Alnus Compost + Sandy Loam Soil (1:1)
13.00bc
Alnus Compost + Sand (1:1)
9.11c
Alnus Compost + Burnt Rice Hull (1:1)
24.55ab
Sandy Loam Soil + sand (1:1)
11.00bc
Sandy Loam Soil + Burnt Rice Hull (1:1)
32.45a
Burnt rice Hull + sand (1:1)
23.33abc
Means followed by a common letter are not significantly different at 5% level (DMRT).
Percentage Survival
Based on the results in Table 6, there were no significant effects of the different
growing media on the percentage survival of the prodo mint carnation plantlets. However,
it was observed that plantlets grown on alnus compost + sandy 1:1, alnus compost+ burnt
rice hull 1:1 and burnt rice hull + sand 1:1 have 100 percent survival, while those that
were grown in sandy loam soil+ burnt rice hull 1:1 and sandy loam soil + sand 1:1 have
88.89 percent plant survival. Among the treatments, plantlets grown in alnus compost +
sandy loam soil 1:1 had the least number of established plants.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 6. Percentage survival
TREATMENT
MEAN
(%)
Alnus Compost + Sandy Loam Soil (1:1)
77.78
Alnus Compost + Sand (1:1)
100.00
Alnus Compost + Burnt Rice Hull (1:1)
100.00
Sandy Loam Soil + sand (1:1)
88.89
Sandy Loam Soil + Burnt Rice Hull (1:1)
88.89
Burnt rice Hull + sand (1:1)
100.00
Means followed by a common letter are not significantly different at 5% level (DMRT)
Occurrence of Insect Pests and Diseases
Insect occurrence was observed during the study. Results show that 50% of the
total plantlets grown in sandy loam soil + sand 1:1 were infested by cabbage butterfly as
compared to the other growing media however disease occurrence was not observed during
the study.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Figure1. Overview of the experimental area
Figure 2. Carnation plantlets from culture bottles before transplanting in the different
growing or acclimatization media
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Alnus compost +
a
Alnus compost + sand b
sandy loam soil
Alnus compost + burnt
c
Sandy loam soil + sand
d
rice hull
Sandy loam soil +
e
Burnt rice hull + sand
f
burnt rice hull
Figure 3. Plantletys grown in 1:1 a) alnus compost + sandy loam soil; b) alnus compost
sand; c) alnus compost + burnt rice hull; d) sandy loam soil + sand; e) sandy loam soil +
burnt rice hull; and f) burnt rice hull + sand 45 days after transplanting.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
SUMMARY, CONCLUSION AND RECOMMENDATIONS
Summary
The study was conducted to determine the most appropriate soil media to harden
rooted carnation and to establish a protocol for the acclimatization of rooted carnation from
in vitro.
Soil media composition of alnus compost + burnt rice hull 1:1 significantly
promoted faster plant growth, longer root length, produced more leaves and longer length
of leaves. Moreover, plantlets grown in sandy loan soil + burnt rice hull 1:1 produced more
number of roots. However the different soil media compositions have no significant effects
on the percentage survival of plantlets grown from in vitro.
Conclusion
Based on the results of the study the use of alnus compost + burnt rice hull 1:1
media composition promoted the best root and growth performance of acclimatized
carnation plantlets.
Recommendations
From the preceding results, acclimatizing carnation plantlets is recommended to be
done on alnus compost + burnt rice hull 1:1 for best plantlets root and shoot establishment.
The following protocol is recommended in acclimatization of carnation plantlets from in
vitro:
1. To harden the tissue cultured plantlets, the bottled carnation plantlets will be first
exposed to partial shade conditions and ambient temperature;
2. After one week, the plantlets will be removed from the bottle and washed with
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
tap water to remove the gelling agent;
3. The plantlets will be soaked in Previcur fungicide (2ml/L) for five minutes then
transplanted into alnus compost + burnt rice hull 1:1 growing media;
4. Before planting, the alnus compost + burnt rice hull 1:1 will be drenched with
Previcur fungicide solution (2ml/L) to avoid unnecessary infection. The plantlets will be
planted under partial shade and will be misted once a day to prevent excessive drying; and,
5. The plantlets will be maintained for 45 days before transplanting in the
greenhouse benches.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Recommended Acclimatization Protocol of Carnation Plantlets
Removed plantlet from the
bottle
Washing of the agar
medium from the roots
Bottled carnation
plantlets from in
vitro
Plantlets are soaked in
fungicide solution for 5
minutes
Preparation: 1:1 alnus
compost + burned rice
Successfully acclimatized
hull
plantlets 60 days after
transplanting
Drenching of the soil media
Transplanting of the
with fungicide solution
plantlets in the sterilize
soil media
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 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
BUTTATI, M. 1983. The use of Cell and Tissue Culture for Mutation Breeding. Science
for Plant Breeding. Europe Congress, Gottinger. Pp. 295- 296.
CASSELLS,A.C. And C. WALSH. 1994. The Influence of Gas Permeability of the Culture
Lid on Calcium Uptake AndStomatal Function In Dianthus Microplants. Plant Cell
Tissue Organ Culture 37:171-178.
DE FOSSARD, R.A. 1981. Tissue Culture Micropropagation, Notes on
TissueCulture.
DUMASLAN, M. R. 2006. Growth Performance of Hardened Tissue Cultured Strawberry
Plantlets Under Greenhouse Condition. MS Thesis, BSU La Trinidad, Benguet.Pp
7
LARSON, R. R. 1980. Introduction to Floriculture. New York : Academic Press, Inc. Pp.
73-78.
MORGAN ED R., SEELVY, JHON F., and BURGE GARRY K., 2003. Acclimatizing
Tissue Culture Plants: Reducing The Shock. New Zealand Institute for Crop and
Food Res. Lt, Prvt Bag 11 600, Palmerston North, New Zealand.
MURASHIGE ,T. and F. SKOOG. 1962. A Revised Medium for Rapid Growth and Bio
Assays with Tobacco Tissue Culture. PhysiologicaPlantarium 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 F. R. TROECH. 1978. Soils and Soil Fertility. New York.
McGraw Hillbook Co. Inc. Pp. 108-111
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
LAOYAN, JOLAFLOR C. MAY 2012. Acclimatization of Carnation (Dianthus
caryophyllus var. Prodo Mint) Plantlets from in vitro. Benguet State University, La
Trinidad, Benguet.
Adviser: Araceli G. Ladilad PhD.
ABSTRACT
The study was conducted at the Ornamental Horticulture Area, Benguet State
University, La Trinidad, Benguet, from February 2012 to April 2012. The study was
conducted to establish the most appropriate growing media to harden rooted carnation
plantlets and to establish a protocol for the acclimatization of rooted carnation plantlets
grown from in vitro before transplanting them to the greenhouse benches.
Results shows that carnation plantlets grown in alnus compost + burnt rice hull 1:1
were the best growing media that promoted the production of longer roots, higher number
of developed leaves, and promoted faster plant growth. Likewise, plantlets grown in sandy
loam soil + burnt rice hull 1:1 produced higher number of roots. However, the different
soil media compositions did not significantly affect the percentage survival of the
acclimatized mint green carnation plantlets.
Soil media composition of alnus compost + burnt 1:1 or carbonized rice hull is
recommended for the acclimatization of carnation plantlets from in vitro for best
performance of the plantlets prior to in vivo transplanting in the greenhouse.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
RESULTS AND DISCUSSION
Root Length
Highly significant differences were observed among the different growing media
used. A mixture of alnus compost + burnt rice hull 1:1 promoted the longest roots with a
mean of 7.53 cm. Plantlets grown in sandy loam soil + sand 1:1 had the shortest length
of roots with a mean of 2.15 cm
According to Thompson and Troech (1978), the use of compost resulted in humus
formation and promotes good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus, calcium, magnesium and organic matter of 5.6 %.
On the other hand, Dumaslan (2006) stated that 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 to better plant growth and performance in strawberry
plantlets.
Table 1. Root length per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
3.18c
Alnus Compost + Sand (1:1)
3.02c
Alnus Compost + Burnt Rice Hull (1:1)
7.53a
Sandy Loam Soil + sand (1:1)
2.15c
Sandy Loam Soil + Burnt Rice Hull (1:1)
5.48b
Burnt rice Hull + sand (1:1)
6.51ab
Means followed by a common letter are not significantly different at 5% level (DMRT).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Number of Leaves
Results showed that plantlets grown in the different media had significantly
different number of leaves per plantlet after 45 days from transplanting. Plantlets grown in
alnus compost + burnt rice hull 1:1 produced the most number of leaves with a mean of
38.67 followed by those grown in burnt rice hull + sand 1:1 with a mean of 34.00,
respectively.
Thompson and Troech (1978) added that the use of compost resulted in humus
formation and promoted good soil structure. Compost also supplies nutrients such as
nitrogen, phosphorus, calcium, magnesium and organic matter of 5.6 %.
Length of Leaves
Plantlets grown in alnus compost + burnt rice hull 1:1 significantly produced
longer leaves but were not significantly different to the length of leaves of plantlet
grown in sandy loam soil + burnt rice hull 1:1 and in burnt rice hull + sand 1:1.
Table 2. Number of leaves per plantlet after 45 days from transplanting
TREATMENT
MEAN
Alnus Compost + Sandy Loam Soil (1:1)
19.33b
Alnus Compost + Sand (1:1)
16.00b
Alnus Compost + Burnt Rice Hull (1:1)
38.67a
Sandy Loam Soil + sand (1:1)
12.00b
Sandy Loam Soil + Burnt Rice Hull (1:1)
31.00a
Burnt rice Hull + sand (1:1)
34.00a
Means followed by a common letter are not significantly different at 5% level (DMRT).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 3. Length of leaves per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
1.51b
Alnus Compost + Sand (1:1)
1.41b
Alnus Compost + Burnt Rice Hull (1:1)
3.78a
Sandy Loam Soil + sand (1:1)
1.07b
Sandy Loam Soil + Burnt Rice Hull (1:1)
3.71a
Burnt rice Hull + sand (1:1)
3.33a
Means followed by a common letter are not significantly different at 5% level (DMRT).
Plantlets grown in sandy loam + sand 1:1 produced the shortest leaves with a mean
of 1.07 cm.
Plant Height
Table 4 shows that there were highly significant differences observed on the
height of plantlets grown in different soil media combinations. The prodo mint carnation
plantlets grown in alnus compost + burnt rice hull 1:1 were the tallest among the treatments
with a mean of 3.15 cm, followed by plantlets grown in burnt rice hull + river sand 1:1,
sandy loam + burnt rice hull 1:1 with means of 2.83 cm and 2.28 cm, respectively. It was
also observed that plantlets acclimatized in sandy loam soil + sand
1:1 have the shortest height with a mean of 1.45 cm and was comparable with plantlets
grown in alnus compost + sand 1:1 with means of 1.51 cm and 1.63 cm, respectively.
Macli-ing (2012) found that a media of alnus compost +sand + burnt rice hull
1:1:1 promoted faster plant growth for the acclimatization of chrysanthemum plantlets.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 4. Plant height per plantlet after 45 days from transplanting
TREATMENT
MEAN
(cm)
Alnus Compost + Sandy Loam Soil (1:1)
1.51c
Alnus Compost + Sand (1:1)
1.63c
Alnus Compost + Burnt Rice Hull (1:1)
3.15a
Sandy Loam Soil + sand (1:1)
1.45c
Sandy Loam Soil + Burnt Rice Hull (1:1)
2.28b
Burnt rice Hull + sand (1:1)
2.83ab
Means followed by a common letter are not significantly different at 5% level (DMRT).
Number of Roots
Based on the results presented in Table 5, sandy loam soil + burnt rice hull
1:1 enhanced the production of more roots, followed by soil media composition: alnus
compost + burnt rice hull 1:1, burnt rice hull + sand 1:1, and alnus compost + sandy loam
soil 1:1 with means of 24.55, 23.33, 13.00 and 11.00 roots per plantlet, respectively, while,
plantlets grown in alnus compost + sand 1:1 had the lowest number of roots per plantlet
after 45 days from transplanting with means of 9.11 roots per plantlet.
In the handbook of Texas Greenhouse Management, burnt rice hull 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 as cited by Dumaslan (2006).
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 5. Number of roots per plantlet after 45 days from transplanting
TREATMENT
MEAN
Alnus Compost + Sandy Loam Soil (1:1)
13.00bc
Alnus Compost + Sand (1:1)
9.11c
Alnus Compost + Burnt Rice Hull (1:1)
24.55ab
Sandy Loam Soil + sand (1:1)
11.00bc
Sandy Loam Soil + Burnt Rice Hull (1:1)
32.45a
Burnt rice Hull + sand (1:1)
23.33abc
Means followed by a common letter are not significantly different at 5% level (DMRT).
Percentage Survival
Based on the results in Table 6, there were no significant effects of the different
growing media on the percentage survival of the prodo mint carnation plantlets. However,
it was observed that plantlets grown on alnus compost + sandy 1:1, alnus compost+ burnt
rice hull 1:1 and burnt rice hull + sand 1:1 have 100 percent survival, while those that
were grown in sandy loam soil+ burnt rice hull 1:1 and sandy loam soil + sand 1:1 have
88.89 percent plant survival. Among the treatments, plantlets grown in alnus compost +
sandy loam soil 1:1 had the least number of established plants.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Table 6. Percentage survival
TREATMENT
MEAN
(%)
Alnus Compost + Sandy Loam Soil (1:1)
77.78
Alnus Compost + Sand (1:1)
100.00
Alnus Compost + Burnt Rice Hull (1:1)
100.00
Sandy Loam Soil + sand (1:1)
88.89
Sandy Loam Soil + Burnt Rice Hull (1:1)
88.89
Burnt rice Hull + sand (1:1)
100.00
Means followed by a common letter are not significantly different at 5% level (DMRT)
Occurrence of Insect Pests and Diseases
Insect occurrence was observed during the study. Results show that 50% of the
total plantlets grown in sandy loam soil + sand 1:1 were infested by cabbage butterfly as
compared to the other growing media however disease occurrence was not observed during
the study.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Figure1. Overview of the experimental area
Figure 2. Carnation plantlets from culture bottles before transplanting in the different
growing or acclimatization media
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Alnus compost +
a
Alnus compost + sand b
sandy loam soil
Alnus compost + burnt
c
Sandy loam soil + sand
d
rice hull
Sandy loam soil +
e
Burnt rice hull + sand
f
burnt rice hull
Figure 3. Plantletys grown in 1:1 a) alnus compost + sandy loam soil; b) alnus compost
sand; c) alnus compost + burnt rice hull; d) sandy loam soil + sand; e) sandy loam soil +
burnt rice hull; and f) burnt rice hull + sand 45 days after transplanting.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
SUMMARY, CONCLUSION AND RECOMMENDATIONS
Summary
The study was conducted to determine the most appropriate soil media to harden
rooted carnation and to establish a protocol for the acclimatization of rooted carnation from
in vitro.
Soil media composition of alnus compost + burnt rice hull 1:1 significantly
promoted faster plant growth, longer root length, produced more leaves and longer length
of leaves. Moreover, plantlets grown in sandy loan soil + burnt rice hull 1:1 produced more
number of roots. However the different soil media compositions have no significant effects
on the percentage survival of plantlets grown from in vitro.
Conclusion
Based on the results of the study the use of alnus compost + burnt rice hull 1:1
media composition promoted the best root and growth performance of acclimatized
carnation plantlets.
Recommendations
From the preceding results, acclimatizing carnation plantlets is recommended to be
done on alnus compost + burnt rice hull 1:1 for best plantlets root and shoot establishment.
The following protocol is recommended in acclimatization of carnation plantlets from in
vitro:
1. To harden the tissue cultured plantlets, the bottled carnation plantlets will be first
exposed to partial shade conditions and ambient temperature;
2. After one week, the plantlets will be removed from the bottle and washed with
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
tap water to remove the gelling agent;
3. The plantlets will be soaked in Previcur fungicide (2ml/L) for five minutes then
transplanted into alnus compost + burnt rice hull 1:1 growing media;
4. Before planting, the alnus compost + burnt rice hull 1:1 will be drenched with
Previcur fungicide solution (2ml/L) to avoid unnecessary infection. The plantlets will be
planted under partial shade and will be misted once a day to prevent excessive drying; and,
5. The plantlets will be maintained for 45 days before transplanting in the
greenhouse benches.
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
Recommended Acclimatization Protocol of Carnation Plantlets
Removed plantlet from the
bottle
Washing of the agar
medium from the roots
Bottled carnation
plantlets from in
vitro
Plantlets are soaked in
fungicide solution for 5
minutes
Preparation: 1:1 alnus
compost + burned rice
Successfully acclimatized
hull
plantlets 60 days after
transplanting
Drenching of the soil media
Transplanting of the
with fungicide solution
plantlets in the sterilize
soil media
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012
LITERATURE CITED
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Beneficial Effect of Arbuscular Mycorrhizason Acclimatization of
Micropropagated Cassava Plantlets. Scientia Hort. 72:63-71
BUTTATI, M. 1983. The use of Cell and Tissue Culture for Mutation Breeding. Science
for Plant Breeding. Europe Congress, Gottinger. Pp. 295- 296.
CASSELLS,A.C. And C. WALSH. 1994. The Influence of Gas Permeability of the Culture
Lid on Calcium Uptake AndStomatal Function In Dianthus Microplants. Plant Cell
Tissue Organ Culture 37:171-178.
DE FOSSARD, R.A. 1981. Tissue Culture Micropropagation, Notes on
TissueCulture.
DUMASLAN, M. R. 2006. Growth Performance of Hardened Tissue Cultured Strawberry
Plantlets Under Greenhouse Condition. MS Thesis, BSU La Trinidad, Benguet.Pp
7
LARSON, R. R. 1980. Introduction to Floriculture. New York : Academic Press, Inc. Pp.
73-78.
MORGAN ED R., SEELVY, JHON F., and BURGE GARRY K., 2003. Acclimatizing
Tissue Culture Plants: Reducing The Shock. New Zealand Institute for Crop and
Food Res. Lt, Prvt Bag 11 600, Palmerston North, New Zealand.
MURASHIGE ,T. and F. SKOOG. 1962. A Revised Medium for Rapid Growth and Bio
Assays with Tobacco Tissue Culture. PhysiologicaPlantarium 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 F. R. TROECH. 1978. Soils and Soil Fertility. New York.
McGraw Hillbook Co. Inc. Pp. 108-111
Acclimatization of Carnation (Dianthus caryophyllus var. Prodo Mint) Plantlets from in
vitro | LAOYAN, JOLAFLOR C. MAY 2012