BIBLIOGRAPHY URBAN, JONATHAN S. APRIL...
BIBLIOGRAPHY
URBAN, JONATHAN S. APRIL 2012. Effect of Cold Stratification period on the
Germination and Seedling growth of (Vaccinium corymbossum) seeds. Benguet State
University, La Trinidad, Benguet.
Adviser: Franklin G. Bawang, MSc.
ABSTRACT
The pre-sowing treatment and germination portion of blueberry seeds were
conducted at the Seed Laboratory and seedling emergence study was studied at the
Pomology Nursery of Benguet State University, from November 2011 to March 2012 to
assess the effect of stratification treatments, ascertain the best durations of stratification
treatment in breaking dormancy of blueberry seeds and to determine the different ways of
enhancing seed germination.
Results revealed that the various durations of cold storage of blueberry seeds had
significantly affected seed germination and seedling emergence. Seedlings resulting from
the seeds subjected to nine days cold storage had enhanced seed germination that resulted
to higher percentage of seed germination and development of more normal seedlings.
Likewise, seeds subjected to nine days cold storage resulted to taller seedlings.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
RESULTS AND DISCUSSION
A. Laboratory Germination Test
Figure 1. Counted seeds germinated and removed from the petri dishes
Percentage of Seeds Germinated
As shown in Table 1, there were highly significant differences on the percentage of
seedling emergence among the different treatments as affected by the various cold storage
durations. It was observed that blueberry seeds subjected to nine days cold storage attained
the highest percentage of seedling emergence with a mean of 96 % followed by seeds
subjected in cold storage duration for fifteen, six and twelve days having a mean of 93.33,
92 and 88.67% respectively, but were statistically comparable with each other. On the other
hand, seeds that were not subjected to cold storage had the lowest percentage of seedling
emergence with a mean of 69.33%.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 1. Percentage of seeds germinated
TREATMENT MEAN (%)
Control (no cold storage)
69.33b
Three days cold storage
74.67b
Six days cold storage
92a
Nine days cold storage
96a
Twelve days cold storage
88.67a
Fifteen days cold storage
93.33a
Means with the same letters are not significantly different at 5% level by DMRT
Median Germination Times.
Table 2 shows the median germination times (G50) of the stratified seeds. Statistical
results showed that seeds subjected in cold storage for nine days accelerated earlier
germination with a mean of 456.67 hours followed by the seeds subjected to cold storage
for fifteen and six days cold storage having a mean of 462.67 and 486.67. It was further
followed by the seeds subjected to cold storage for twelve days with a mean of 496 hours.
Seeds that were not subjected to cold storage had the longest duration to germination with
a mean of 536 hours but were comparable to the seeds subjected to three days cold storage
only.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 2. Median germination times.
TREATMENT MEAN (HOURS)
Control (no cold storage)
536a
Three days cold storage
535.33a
Six days cold storage
486.67bc
Nine days cold storage
456.67c
Twelve days cold storage
496b
Fifteen days cold storage
462.67bc
Means with the same letters are not significantly different at 5% level by DMRT
Median Spread of Germination Times.
The median spread of germination times (G90-G10) as affected by different durations
of cold storage is presented in Table 3. Apparently, seeds stored for cold storage durations
for fifteen days had 550.40 hours to show median spread of germination times followed by
the seeds subjected to cold storage durations for twelve, nine, six and three days showing
median spread of germination times with a mean of 537.07, 505.73, 488.03 and 449.33
hours, respectively having means that are highly significant with the other treatments
against the unstratified seeds.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 3. Median spread of germination times.
TREATMENT MEAN (HOURS)
Control (no cold storage)
332.68f
Three days cold storage
449.33e
Six days cold storage
488.03d
Nine days cold storage
505.73c
Twelve days cold storage
537.07b
Fifteen days cold storage
550.4a
Means with the same letters are not significantly different at 5% level by DMRT
B. Greenhouse Emergence Test
Figure 2. Overview of the experimental area
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Number of Days From Sowing to
Initial Seed Emergence
Seed emergence as affected by different cold storage durations was determined
based on the number of days from sowing to the time the seed had emerged on the soil
surface (Table 4). Statistical results showed that nine days cold storage enhanced earlier
emergence of blueberry with a mean of 53.67 days from sowing which was highly
significant as compared to the other treatments. It was followed by the seeds subjected to
six, twelve, fifteen and three days of cold storage with a mean of 55, 56, 56.67 and 59.33
days respectively. The seeds that were not subjected to cold storage were the last to emerge
with a mean of 68 days.
These findings collaborate with the earlier observations of Paing (1980) that cold
storage shortened the period from sowing to germination of garden pea. Weaver (1974)
mentioned that pre-treated seeds in medium of carefully controlled temperature is
conditioned for rapid germination. However, many seeds placed in an environment
considered adequate for germination still do not germinate because of some internal
limitations and unfavorable environmental conditions (Devlin, 1977). Furthermore,
Daubenmire (1974) explained that the physiological explanation of this conditioning effect
is still unknown. In some seeds, low temperature may function to render the seed coat more
permeable to gasses and other substances.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 4. Number of days from sowing to initial seed emergence
TREATMENT MEAN (DAYS)
Control (no cold storage)
68a
Three days cold storage
59.33b
Six days cold storage
55bc
Nine days cold storage
53.67c
Twelve days cold storage
56.67bc
Fifteen days cold storage
56bc
Means with the same letters are not significantly different at 5% level by DMRT
Percentage Survival (%)
As shown in Table 5, there were highly significant statistical differences on the
percentage survival of seedlings among the different treatments as affected by various cold
storage durations. It was observed that blueberry seeds subjected to nine days storage
attained the highest percentage of seedling survival with a mean of 88 % which is
statistically different to the seeds that were not subjected to cold storage with a mean of
56.67 % which is also highly significantly different with the rest of the treatments.
It was observed further that some seeds did not emerge for reasons that cannot be
explained though it may be due to chilling injury that caused the death of the embryo in
the seeds during the long cold storage periods and may also be due to the not fully
decomposed coco coir dust added in the media that was used. Hartman et al. (1990) stated
that such result could be related to high concentration of soil solutes in the
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 5. Percentage survival
TREATMENT MEAN (%)
Control (no cold storage)
56.67d
Three days cold storage
66cd
Six days cold storage
78ab
Nine days cold storage
88a
Twelve days cold storage
70bc
Fifteen days cold storage
87a
Means with the same letters are not significantly different at 5% level by DMRT
upper layer of the germination media. Likewise, Handreck (1993) wrote that coco coir pith
has a carbon-nitrogen ratio of 104:1 which means that it would take decades to be
decomposed. The partly decomposed coco coir dust might have released some toxic
substance or gas that affected the emergence of seeds.
Number of Days From Sowing
to First Appearance of Leaves
Table 6 shows highly significant differences among the different cold storage
durations affecting the number of days from sowing to first appearance of leaves. Results
showed that seeds stored for nine days attained the shortest days for leaf development and
produced leaves within 57.33 days. This was followed by the seeds subjected to cold
storage for six, fifteen, twelve and two days having a mean of 60, 60.33, 61.33 and 64.67
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
days respectively. On the other hand, seeds that were not subjected to cold storage had the
longest days to leaf formation having a mean of 72 days.
Table 6. Number of days from sowing to first appearance of leaves
TREATMENT MEAN (DAYS)
Control (no cold storage)
72a
Three days cold storage
64.67b
Six days cold storage
60c
Nine days cold storage
57.33c
Twelve days cold storage
61.33bc
Fifteen days cold storage
60.33c
Means with the same letters are not significantly different at 5% level by DMRT
Dao-ines (2010) found in his experiment that blueberry seeds refrigerated for one
week cold storage appeared to be the best, as the seedlings attained the highest percentage
of germination, tallest seedlings, complete emergence, normal seedlings and included the
seedlings faster leaf to emergence.
Percentage of Normal Seedlings
Table 7 shows that there were highly significant differences observed among the
different cold storage durations in terms of percentage of normal seedlings. Based on the
results, blueberry seeds subjected to nine days cold storage had the highest percentage of
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
normal seedlings having a mean of 84.67 %. It was followed by the seeds subjected to cold
storage durations for fifteen, six, twelve and three days having a mean of 82.67, 74, 64.67
and 60.67% of normal seedlings respectively. Lastly, seeds that were not subjected to cold
storage had the lowest mean of 50% showing normal seedlings.
Table 7. Percentage of normal seedlings
TREATMENT MEAN (%)
Control (no cold storage)
50e
Three days cold storage
60.67d
Six days cold storage
74bc
Nine days cold storage
84a
Twelve days cold storage
64.67cd
Fifteen days cold storage
82.67ab
Means with the same letters are not significantly different at 5% level by DMRT
Salisbury and Ross (1992) explained that pre-chilling of seeds sometimes has a
strong effect on growth in addition to its breaking dormancy action. If the embryos of peach
seed are exercised from their cotyledons, they will germinate without pre-chilling, but the
seedlings are frequently stunted and abnormal. However, when the exercised embryos are
treated with low temperature, they grow into normal seedlings. Thus, it is pre-chilling and
not the presence of the cotyledons that ensures their normality.
Seedling Height (mm)
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
The seedling height (Table 8) of blueberry was obtained by measuring the seedlings
from the base up to the tip of its secondary leaves. Results showed that seeds subjected to
cold storage for nine days had the tallest seedlings having a mean of 9 mm followed by the
seeds subjected to cold storage durations for six, fifteen, twelve and three days have a mean
of 8.33, 7.67, 7 and 6.67 mm respectively. However, blueberry seeds that were not
subjected to cold storage had the shortest height with a mean of 5.33 mm.
Table 8. Seedling height (mm)
TREATMENT MEAN
Control (no cold storage)
5.33d
Three days cold storage
6.67cd
Six days cold storage
8.33ab
Nine days cold storage
9a
Twelve days cold storage
7bc
Fifteen days cold storage
7.76abc
Means with the same letters are not significantly different at 5% level by DMRT
Results showed that nine days cold storage duration of seeds prior to sowing
enhanced growth of seedlings leading to taller seedlings. These findings collaborated with
the findings of Dao-ines (2010) who stated that cold storage of blueberry seeds for a period
of one week at 5 oC hastened seed germination, improved rate of seed emergence and
promoted the development of taller seedlings.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Number of Leaves Per Plant
Regarding the number of leaves of blueberry seedlings per plant, it was shown in
Table 9 that there were no significant statistical differences among the different cold
storage durations with regards to the number of leaves per plant. However, numerical
results revealed that seedlings from the seeds subjected to nine days cold storage had the
highest number of leaves with a mean of 3. Whereas, seeds that was not subjected to cold
storage had the lowest number of leaves developed with a mean of 2.
Table 9. Number of leaves per plant
TREATMENT MEAN
Control (no cold storage)
2a
Three days cold storage
2.33a
Six days cold storage
2.67a
Nine days cold storage
3a
Twelve days cold storage
2.33a
Fifteen days cold storage
2.67a
Means with the same letters are not significantly different at 5% level by DMRT
Seedling Vigor Index
The results in Table 10 showed that there were no significant statistical differences
observed among the different treatments used in the study affecting the seedling vigor of
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
blueberry. However, numerical results showed that seedlings from the seeds subjected to
nine days cold storage exhibited excellent growth.
Tocnang (2009) observed that guapple subjected to nine days cold storage
promoted excellent growth of seedlings. Likewise, Banagen (2008) noted that storing
coffee seeds under cold temperature influenced seedling vigor. Cold stratification enhanced
faster growth which may explain the performance of coffee seeds subjected to cool
treatments as having seedlings that are more vigorous as compared to the unstratified seeds.
Table 10. Seedling vigor rating
TREATMENT MEAN
Control (no cold storage)
3a
Three days cold storage
3.33a
Six days cold storage
3.33a
Nine days cold storage
3.67a
Twelve days cold storage
3.33a
Fifteen days cold storage
3.33a
Means with the same letters are not significantly different at 5% level by DMRT
Rating
Description Remarks
4
Most vigorous
excellent growth with dark green leaves
3
Vigorous
good growth with green leaves
2
Less vigorous
slightly growth with light green leaves
1
Poor
poor growth with yellow leaves
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The laboratory germination test of blueberry seeds as affected by cold storage
durations were conducted at Seed Laboratory and seedling emergence and seedling growth
of blueberry seeds was done at the Pomology Project Nursery, Benguet State University,
La Trinidad, Benguet from November 30, 2011 to March 11, 2012. The blueberry seeds
were subjected to low temperature with an average of 5 oC at various periods from three
days to fifteen days.
A. Laboratory Seed Germination Test.
Results showed that there were highly significant statistical differences observed
among the various cold storage durations used. Nine days cold storage of blueberry seeds
showed the best performance among all the treatments that enhanced highest seed
emergence with a mean of 96 % compared to the unstratified seeds having the lowest
percentage of seed germination with a mean of 69.33%. Concerning median germination
times (G50), storing blueberry seeds at nine days considerably accelerated germination
compared with the rest of the other treatments and the unstratified seeds resulted with
highly significant differences to the other treatments. As for the median spread of
germination times (G90- G10), it was observed that cold storage of blueberry seeds for nine
days markedly improved uniformity of seeds germination due to the statistically similar
responses with that of the unstratified seeds.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
B. Greenhouse Seed Emergence Test.
Results showed that there were highly significant differences observed among the
various cold storage durations used. Nine days cold storage of blueberry showed the best
performance among all treatments that enhanced earlier seed emergence with a mean of
53.67 days and shorter days to develop true leaves with a mean of 57 .33 days. Furthermore,
seeds subjected to cold storage for nine days had the tallest seedlings having a mean of 9
mm. Likewise, the same duration of cold storage attained the highest percentage of seed
emergence having a mean of 88 % and also had the highest percentage of normal seedlings
with a mean of 84.67 % which is highly significant with the other treatments.
With regards to the number of leaves and seedling vigor, statistical results showed
that there were no significant differences among the different cold storage durations used.
However, numerical results showed that seedlings from seeds stored for nine days had
more number of leaves and more vigorous seedlings as compared to other treatments.
Conclusion
Based on the results of the study, subjecting blueberry seeds to cold storage for nine
days before sowing hastened germination (G50) and showed the best results. It was also
found that cold storage of blueberry seeds for a period of nine days at 5oC markedly
improved uniformity of germination (G90-G10) and enhanced faster germination and seed
emergence as well as higher percentage of normal seedlings and vigorous seedling growth.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Recommendation
Based on the results, it is recommended that cold storage of blueberry seeds for a
period of nine days at 5 oC prior to sowing should be done in order to enhance faster and
uniform seed germination and seed emergence as well as to promote taller seedlings,
development of more leaves and vigorous seedling growth.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
LITERATURE CITED
ADRIANCE, G. W. and F. R. BRISON. 1955. Propagation of Horticulture Plants: Mc Graw Hill
Book Co., New York Inc. P. 82.
AGNAYA, J. S. 2004. Effect of cold stratification period on germination of Benguet “wild tea”.
BS Thesis. BSU, La Trinidad, Benguet.
ANTOLIN, N. C. 2001. Germination of Benguet Tea as influenced by stratification periods. A
term paper submitted in Horticulture b315 (Advances in Plant Propagation and
Management). BSU, La Trinidad, Benguet. Pp 1-8.
BANAGEN, M. L. 2008. Effect of cool storage seeds in cold duration on the germination and
seedling characteristics of coffee seeds. BS Thesis. BSU, La Trinidad, Benguet.
COPELAND, L. O. and M. B. MCDONALD. 1985. Principles of Seed Science and Technology.
Burgess Publ., Co., Minnesota. Pp. 111-113.
DAO-INES, G. K. 2010. Seed germination and seedling growth of Blueberry as affected by
different cold storage. BS Thesis. BSU, La Trinidad, Benguet.
DAUBENMIRE, R. R. 1974. Plant and Environment. A textbook of Plant Autocology. New York:
John Welly and Sons. P. 82.
DELA CRUS, M. P. 2005. Germination and Seedling Emergence behavior of Chive seeds
pretreated with sea water. BS. Thesis. BSU, La Trinidad, Benguet.
DEVLIN, R. 1977. Plant Physiology. AVI Publ., Co., New York. Pp. 552-557.
HANDRECK, K. A. 1993. Properties of coco coir dust, and its use in the formulation of soilless
potting media. Community Soil and Plant Analysis. Pp. 349-363.
HARTMANN, H. T. and D. E. KESTER. 1990. Plant Propagation: Princilpes and Practices 5th Ed.
New Jersey: Prentice Hall of India. Inc. p. 120.
HARTMANN, H. T. and D. E. KESTER. 1972. Plant Propagation: Principles and Practices.
Prentice Hall of India, New Delhi. Pp. 126-127.
HILL, M. J. 1987. Seed Development, Maturity and Ripeness. Lecture Handouts in Certificate in
Seed Technology Course (February 9- May 5, 1987). Seed Technology Center, massey
University, New Zealand.
KLINGMAN, G. C. and F. N. ASHTON. 1975. Weed Science. Principles and Practices. New
York. A Welley Inter Science Publication. P. 52.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
MABESA, R .C. 1980. Seed Maturation. Paper presented during the International Training
Programme on the Seed Technology for Vegetable Crops (August 31 Nov. 17, 1980). UP
at Los Banos, Laguna, Philippines.
PAING, R. D. 1980. Effect of seed stratification on the growth and yield of the edible podded pea.
BS Thesis. BSU, La Trinidad, Benguet.
SALISBURY, F. B. and C. W. ROSS. 1992. Fourth Edition. Belmont, California, Wadworth
Publishing Co. Inc. p. 497.
TOCNANG, R. G. 2009. Effect of cold storage duration on the germination and seedling
characteristics of guapple seeds. BS Thesis. BSU, La Trinidad, Benguet.
WEAVER, M. B. 1974. Plant Growth Development, New Delhi: Tata Mc Graw Hill. Publ. Co.
Inc. P. 37.
WELLS, J. S. 1955. Plant Propagation Practices. The Macmillan Company. New York. P. 61.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
URBAN, JONATHAN S. APRIL 2012. Effect of Cold Stratification period on the
Germination and Seedling growth of (Vaccinium corymbossum) seeds. Benguet State
University, La Trinidad, Benguet.
Adviser: Franklin G. Bawang, MSc.
ABSTRACT
The pre-sowing treatment and germination portion of blueberry seeds were
conducted at the Seed Laboratory and seedling emergence study was studied at the
Pomology Nursery of Benguet State University, from November 2011 to March 2012 to
assess the effect of stratification treatments, ascertain the best durations of stratification
treatment in breaking dormancy of blueberry seeds and to determine the different ways of
enhancing seed germination.
Results revealed that the various durations of cold storage of blueberry seeds had
significantly affected seed germination and seedling emergence. Seedlings resulting from
the seeds subjected to nine days cold storage had enhanced seed germination that resulted
to higher percentage of seed germination and development of more normal seedlings.
Likewise, seeds subjected to nine days cold storage resulted to taller seedlings.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
RESULTS AND DISCUSSION
A. Laboratory Germination Test
Figure 1. Counted seeds germinated and removed from the petri dishes
Percentage of Seeds Germinated
As shown in Table 1, there were highly significant differences on the percentage of
seedling emergence among the different treatments as affected by the various cold storage
durations. It was observed that blueberry seeds subjected to nine days cold storage attained
the highest percentage of seedling emergence with a mean of 96 % followed by seeds
subjected in cold storage duration for fifteen, six and twelve days having a mean of 93.33,
92 and 88.67% respectively, but were statistically comparable with each other. On the other
hand, seeds that were not subjected to cold storage had the lowest percentage of seedling
emergence with a mean of 69.33%.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 1. Percentage of seeds germinated
TREATMENT MEAN (%)
Control (no cold storage)
69.33b
Three days cold storage
74.67b
Six days cold storage
92a
Nine days cold storage
96a
Twelve days cold storage
88.67a
Fifteen days cold storage
93.33a
Means with the same letters are not significantly different at 5% level by DMRT
Median Germination Times.
Table 2 shows the median germination times (G50) of the stratified seeds. Statistical
results showed that seeds subjected in cold storage for nine days accelerated earlier
germination with a mean of 456.67 hours followed by the seeds subjected to cold storage
for fifteen and six days cold storage having a mean of 462.67 and 486.67. It was further
followed by the seeds subjected to cold storage for twelve days with a mean of 496 hours.
Seeds that were not subjected to cold storage had the longest duration to germination with
a mean of 536 hours but were comparable to the seeds subjected to three days cold storage
only.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 2. Median germination times.
TREATMENT MEAN (HOURS)
Control (no cold storage)
536a
Three days cold storage
535.33a
Six days cold storage
486.67bc
Nine days cold storage
456.67c
Twelve days cold storage
496b
Fifteen days cold storage
462.67bc
Means with the same letters are not significantly different at 5% level by DMRT
Median Spread of Germination Times.
The median spread of germination times (G90-G10) as affected by different durations
of cold storage is presented in Table 3. Apparently, seeds stored for cold storage durations
for fifteen days had 550.40 hours to show median spread of germination times followed by
the seeds subjected to cold storage durations for twelve, nine, six and three days showing
median spread of germination times with a mean of 537.07, 505.73, 488.03 and 449.33
hours, respectively having means that are highly significant with the other treatments
against the unstratified seeds.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 3. Median spread of germination times.
TREATMENT MEAN (HOURS)
Control (no cold storage)
332.68f
Three days cold storage
449.33e
Six days cold storage
488.03d
Nine days cold storage
505.73c
Twelve days cold storage
537.07b
Fifteen days cold storage
550.4a
Means with the same letters are not significantly different at 5% level by DMRT
B. Greenhouse Emergence Test
Figure 2. Overview of the experimental area
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Number of Days From Sowing to
Initial Seed Emergence
Seed emergence as affected by different cold storage durations was determined
based on the number of days from sowing to the time the seed had emerged on the soil
surface (Table 4). Statistical results showed that nine days cold storage enhanced earlier
emergence of blueberry with a mean of 53.67 days from sowing which was highly
significant as compared to the other treatments. It was followed by the seeds subjected to
six, twelve, fifteen and three days of cold storage with a mean of 55, 56, 56.67 and 59.33
days respectively. The seeds that were not subjected to cold storage were the last to emerge
with a mean of 68 days.
These findings collaborate with the earlier observations of Paing (1980) that cold
storage shortened the period from sowing to germination of garden pea. Weaver (1974)
mentioned that pre-treated seeds in medium of carefully controlled temperature is
conditioned for rapid germination. However, many seeds placed in an environment
considered adequate for germination still do not germinate because of some internal
limitations and unfavorable environmental conditions (Devlin, 1977). Furthermore,
Daubenmire (1974) explained that the physiological explanation of this conditioning effect
is still unknown. In some seeds, low temperature may function to render the seed coat more
permeable to gasses and other substances.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 4. Number of days from sowing to initial seed emergence
TREATMENT MEAN (DAYS)
Control (no cold storage)
68a
Three days cold storage
59.33b
Six days cold storage
55bc
Nine days cold storage
53.67c
Twelve days cold storage
56.67bc
Fifteen days cold storage
56bc
Means with the same letters are not significantly different at 5% level by DMRT
Percentage Survival (%)
As shown in Table 5, there were highly significant statistical differences on the
percentage survival of seedlings among the different treatments as affected by various cold
storage durations. It was observed that blueberry seeds subjected to nine days storage
attained the highest percentage of seedling survival with a mean of 88 % which is
statistically different to the seeds that were not subjected to cold storage with a mean of
56.67 % which is also highly significantly different with the rest of the treatments.
It was observed further that some seeds did not emerge for reasons that cannot be
explained though it may be due to chilling injury that caused the death of the embryo in
the seeds during the long cold storage periods and may also be due to the not fully
decomposed coco coir dust added in the media that was used. Hartman et al. (1990) stated
that such result could be related to high concentration of soil solutes in the
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Table 5. Percentage survival
TREATMENT MEAN (%)
Control (no cold storage)
56.67d
Three days cold storage
66cd
Six days cold storage
78ab
Nine days cold storage
88a
Twelve days cold storage
70bc
Fifteen days cold storage
87a
Means with the same letters are not significantly different at 5% level by DMRT
upper layer of the germination media. Likewise, Handreck (1993) wrote that coco coir pith
has a carbon-nitrogen ratio of 104:1 which means that it would take decades to be
decomposed. The partly decomposed coco coir dust might have released some toxic
substance or gas that affected the emergence of seeds.
Number of Days From Sowing
to First Appearance of Leaves
Table 6 shows highly significant differences among the different cold storage
durations affecting the number of days from sowing to first appearance of leaves. Results
showed that seeds stored for nine days attained the shortest days for leaf development and
produced leaves within 57.33 days. This was followed by the seeds subjected to cold
storage for six, fifteen, twelve and two days having a mean of 60, 60.33, 61.33 and 64.67
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
days respectively. On the other hand, seeds that were not subjected to cold storage had the
longest days to leaf formation having a mean of 72 days.
Table 6. Number of days from sowing to first appearance of leaves
TREATMENT MEAN (DAYS)
Control (no cold storage)
72a
Three days cold storage
64.67b
Six days cold storage
60c
Nine days cold storage
57.33c
Twelve days cold storage
61.33bc
Fifteen days cold storage
60.33c
Means with the same letters are not significantly different at 5% level by DMRT
Dao-ines (2010) found in his experiment that blueberry seeds refrigerated for one
week cold storage appeared to be the best, as the seedlings attained the highest percentage
of germination, tallest seedlings, complete emergence, normal seedlings and included the
seedlings faster leaf to emergence.
Percentage of Normal Seedlings
Table 7 shows that there were highly significant differences observed among the
different cold storage durations in terms of percentage of normal seedlings. Based on the
results, blueberry seeds subjected to nine days cold storage had the highest percentage of
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
normal seedlings having a mean of 84.67 %. It was followed by the seeds subjected to cold
storage durations for fifteen, six, twelve and three days having a mean of 82.67, 74, 64.67
and 60.67% of normal seedlings respectively. Lastly, seeds that were not subjected to cold
storage had the lowest mean of 50% showing normal seedlings.
Table 7. Percentage of normal seedlings
TREATMENT MEAN (%)
Control (no cold storage)
50e
Three days cold storage
60.67d
Six days cold storage
74bc
Nine days cold storage
84a
Twelve days cold storage
64.67cd
Fifteen days cold storage
82.67ab
Means with the same letters are not significantly different at 5% level by DMRT
Salisbury and Ross (1992) explained that pre-chilling of seeds sometimes has a
strong effect on growth in addition to its breaking dormancy action. If the embryos of peach
seed are exercised from their cotyledons, they will germinate without pre-chilling, but the
seedlings are frequently stunted and abnormal. However, when the exercised embryos are
treated with low temperature, they grow into normal seedlings. Thus, it is pre-chilling and
not the presence of the cotyledons that ensures their normality.
Seedling Height (mm)
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
The seedling height (Table 8) of blueberry was obtained by measuring the seedlings
from the base up to the tip of its secondary leaves. Results showed that seeds subjected to
cold storage for nine days had the tallest seedlings having a mean of 9 mm followed by the
seeds subjected to cold storage durations for six, fifteen, twelve and three days have a mean
of 8.33, 7.67, 7 and 6.67 mm respectively. However, blueberry seeds that were not
subjected to cold storage had the shortest height with a mean of 5.33 mm.
Table 8. Seedling height (mm)
TREATMENT MEAN
Control (no cold storage)
5.33d
Three days cold storage
6.67cd
Six days cold storage
8.33ab
Nine days cold storage
9a
Twelve days cold storage
7bc
Fifteen days cold storage
7.76abc
Means with the same letters are not significantly different at 5% level by DMRT
Results showed that nine days cold storage duration of seeds prior to sowing
enhanced growth of seedlings leading to taller seedlings. These findings collaborated with
the findings of Dao-ines (2010) who stated that cold storage of blueberry seeds for a period
of one week at 5 oC hastened seed germination, improved rate of seed emergence and
promoted the development of taller seedlings.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Number of Leaves Per Plant
Regarding the number of leaves of blueberry seedlings per plant, it was shown in
Table 9 that there were no significant statistical differences among the different cold
storage durations with regards to the number of leaves per plant. However, numerical
results revealed that seedlings from the seeds subjected to nine days cold storage had the
highest number of leaves with a mean of 3. Whereas, seeds that was not subjected to cold
storage had the lowest number of leaves developed with a mean of 2.
Table 9. Number of leaves per plant
TREATMENT MEAN
Control (no cold storage)
2a
Three days cold storage
2.33a
Six days cold storage
2.67a
Nine days cold storage
3a
Twelve days cold storage
2.33a
Fifteen days cold storage
2.67a
Means with the same letters are not significantly different at 5% level by DMRT
Seedling Vigor Index
The results in Table 10 showed that there were no significant statistical differences
observed among the different treatments used in the study affecting the seedling vigor of
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
blueberry. However, numerical results showed that seedlings from the seeds subjected to
nine days cold storage exhibited excellent growth.
Tocnang (2009) observed that guapple subjected to nine days cold storage
promoted excellent growth of seedlings. Likewise, Banagen (2008) noted that storing
coffee seeds under cold temperature influenced seedling vigor. Cold stratification enhanced
faster growth which may explain the performance of coffee seeds subjected to cool
treatments as having seedlings that are more vigorous as compared to the unstratified seeds.
Table 10. Seedling vigor rating
TREATMENT MEAN
Control (no cold storage)
3a
Three days cold storage
3.33a
Six days cold storage
3.33a
Nine days cold storage
3.67a
Twelve days cold storage
3.33a
Fifteen days cold storage
3.33a
Means with the same letters are not significantly different at 5% level by DMRT
Rating
Description Remarks
4
Most vigorous
excellent growth with dark green leaves
3
Vigorous
good growth with green leaves
2
Less vigorous
slightly growth with light green leaves
1
Poor
poor growth with yellow leaves
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The laboratory germination test of blueberry seeds as affected by cold storage
durations were conducted at Seed Laboratory and seedling emergence and seedling growth
of blueberry seeds was done at the Pomology Project Nursery, Benguet State University,
La Trinidad, Benguet from November 30, 2011 to March 11, 2012. The blueberry seeds
were subjected to low temperature with an average of 5 oC at various periods from three
days to fifteen days.
A. Laboratory Seed Germination Test.
Results showed that there were highly significant statistical differences observed
among the various cold storage durations used. Nine days cold storage of blueberry seeds
showed the best performance among all the treatments that enhanced highest seed
emergence with a mean of 96 % compared to the unstratified seeds having the lowest
percentage of seed germination with a mean of 69.33%. Concerning median germination
times (G50), storing blueberry seeds at nine days considerably accelerated germination
compared with the rest of the other treatments and the unstratified seeds resulted with
highly significant differences to the other treatments. As for the median spread of
germination times (G90- G10), it was observed that cold storage of blueberry seeds for nine
days markedly improved uniformity of seeds germination due to the statistically similar
responses with that of the unstratified seeds.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
B. Greenhouse Seed Emergence Test.
Results showed that there were highly significant differences observed among the
various cold storage durations used. Nine days cold storage of blueberry showed the best
performance among all treatments that enhanced earlier seed emergence with a mean of
53.67 days and shorter days to develop true leaves with a mean of 57 .33 days. Furthermore,
seeds subjected to cold storage for nine days had the tallest seedlings having a mean of 9
mm. Likewise, the same duration of cold storage attained the highest percentage of seed
emergence having a mean of 88 % and also had the highest percentage of normal seedlings
with a mean of 84.67 % which is highly significant with the other treatments.
With regards to the number of leaves and seedling vigor, statistical results showed
that there were no significant differences among the different cold storage durations used.
However, numerical results showed that seedlings from seeds stored for nine days had
more number of leaves and more vigorous seedlings as compared to other treatments.
Conclusion
Based on the results of the study, subjecting blueberry seeds to cold storage for nine
days before sowing hastened germination (G50) and showed the best results. It was also
found that cold storage of blueberry seeds for a period of nine days at 5oC markedly
improved uniformity of germination (G90-G10) and enhanced faster germination and seed
emergence as well as higher percentage of normal seedlings and vigorous seedling growth.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
Recommendation
Based on the results, it is recommended that cold storage of blueberry seeds for a
period of nine days at 5 oC prior to sowing should be done in order to enhance faster and
uniform seed germination and seed emergence as well as to promote taller seedlings,
development of more leaves and vigorous seedling growth.
Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012
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Effect of Cold Stratification period on the Germination and Seedling growth of (Vaccinium
corymbossum) seeds | URBAN, JONATHAN S. APRIL 2012