BIBLIOGRAPHY DAO-INES, GABBY K. APRIL 2010....
BIBLIOGRAPHY
DAO-INES, GABBY K. APRIL 2010. Seed Germination and Seedling Growth of
Blueberry (Vacinium corymbossum) as Affected by Different Cold Storage Duration.
Benguet State University, La Trinidad, Benguet.
Adviser: Franklin G. Bawang, Msc.
ABSTRACT
The study was conducted at the Pomology Nursery of Benguet State University,
from November 2009 to March 2010 to find out the best durations of cold storage that
will effectively break the dormancy of blueberry seeds; and to determine the effect of
cold storage on seed germination and seedling growth of blueberry.
Results showed that the various durations of cold storage of blueberry seeds had
significantly affected seed germination and seedling emergence. Seedlings resulted from
the seeds subjected to one week cold storage had developed true leaf earlier; more seeds
emerged resulted to higher percentage of seedling emergence; and more normal seedlings
developed. Likewise, seeds subjected to one week cold storage resulted to seedlings
having longer roots.
TABLE OF CONTENTS
Page
Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
REVIEW OF LITERATURE
Description of the Plant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Seed Germination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Seed Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Methods of Breaking Dormancy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Effect of Cold Storage Duration
on Seed Germination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
RESULTS AND DISCUSSION
Number days from Sowing to
Emergence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Number of Days from Sowing
to First Appearance of Leaves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Percentage of Seedling Emergence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Percentage of Normal seedlings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Seedling Height . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Number of Leaves per Plant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Root Length . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
ii
Number of Roots . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Seedling Vigor Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
SAMMARY, CONCLUSION AND RECOMMENDATION
Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
APPENDICES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
iii
1
INTRODUCTION
Blueberries are flowering plants belonging to the genus Vaccinium ssp. The
species are native only to North America. The crop is considered as shrubs varying in
size from 10cm to 4m tall. The smaller species are known as “low bush blueberries”, and
the larger species are “high bush blueberries”. The leaves can be deciduous or evergreen,
ovate to lanceolate, and from1-8cm long and 0.5-3.5cm broad. The flowers are bell-
shaped, white, pale pink or red. Sometimes tinged greenish. The fruit is false berry 5-
16mm diameter with a flared “crown” at the end. They are pale greenish at first, then
reddish-purple and finally indigo on ripening. It has a sweet taste when mature, with
variable acidity. Blueberry bushes typically bear fruit from May to June in the Western
Hemisphere; “blueberry season’ peaks in July, which is National Blueberry Month in the
United States and Germany.
Blueberries grow wild in many parts of the world. However, the United States and
Canada supply about 95% of the blueberries used by the food industry. North American
farmers harvest about 103 million pounds (47 million kilograms) of blueberries annually.
For centuries, blue berries were gathered from the forest and bogs by the Native
Americans and these are usually consumed fresh and also it can be preserved. The
Northeast Native American tribes revered blueberries and much folklore developed
around them. The blossom ends of each berry, the calyx, forms a shaped of a perfect five-
. pointed star. Parts of the blueberry plant were also used as medicine, for coughs and
good for the circulation of the blood. Not only for medicine, the juice can also be made as
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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an excellent dye for baskets and cloth. It can also used in other food preparation.
Example, the dried berries were also crushed into a powder and into meat for flavor; it
can be processed such as jellies,jams,pies,muffins and snack foods.
For early colonist, blueberries were probably one of the first familiar foods
discovered on North American continent, since they were similar to other berry varieties
found in Europe. Their abundance, natural sweetness and visibility made them on
immediately welcome addition at a time when fresh food was in short supply, and sugar
was scarce and expensive. Blueberry were canned and shipped to union troops during
civil war soldiers in 1860’s.
Blueberries have gained increasing recognition from health professionals for their
very high antioxidant properties. Studies of the anthocyanin and pytochemicals found in
the blueberries have shown that they provide a range of benefits, including reducing
eyestrain, counteracting environmental carcinogens, promoting urinary tract health,
fighting sun damage and protecting against cardiovascular disease. These are low in
calories, low in fat and have no cholesterol, in addition to being a good source of dietary
fiber and is packed with other vitamins (A,C,E) and minerals (potassium, manganese and
magnesium).
`Blueberry can be potentially grown in forest or mountains. However, mass
production could become a problem under high demand because it takes time to produce
seedling from seeds. Therefore, it is imperative to conduct this study to be able to
determine the best duration of cold storage of blueberry seeds that can provides the
stimulus for earlier and uniform germination and reduce the longer duration and intervals
of waiting time leading to higher percentage survival of the seedling. In addition, to be
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
3
able to grow or to produce more blueberry in our community for economic, medicinal
and food purposes.
The results of this study will provide baseline information’s to researchers
interested to work on blueberry in the future in order to help in the establishment of
blueberry production industry and to encourage more farmers to produce more blueberry
that is considered as a medicinal and has great potential to market when it is processed.
The study was conducted at the Nursery of the Pomology Project, Benguet State
University, La Trinidad, Benguet second semester from November 15 to March 7, 2010
to find out the best duration of cold storage in breaking dormancy of blueberry seeds and
to determine the effect of cold storage on seed germination and seedling growth of
blueberry.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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REVIEW OF LITERATURE
Description of the Plant
Wild blueberries are perennial shrubs found in the undergrowth of forests in
northeastern United States and in the Atlantic provinces of Canada. Plants are propagated
from seeds or cuttings. They produced underground stems of rhizomes, which
periodically send up new shoots and developed roots. One original plant can thus cover
an area ranging from 75 to 250 square feet depending on the age of the plant. Originally
gathered by hand rakes although effective mechanical harvesters are being developed and
improved. Various species abound including low bush and high bush varieties, which
produced varying degrees of sweetness in the resulting fruit. For dried berries, the fruits
are infused with a sugar solution to create a uniformly sweet and appealing product for
used in baking and confectionery applications and granolos, as well as in shacks and trail
mixes (Kuepper and Diver, 2004).
Seed Germination
Seed germination is defined as the activation of the metabolic machinery of the
embryo leading to the emergence of new seedling plant (Hartman et. al., 1990). It is a
sequence of step beginning with the uptake of water leading to rupture of the seed coat
and germination usually determined visibly by observing the protrusion of the radicle or
shoot (Devlin, 1977). Germination of seed will occur only when the embryo development
is complete (Edmond et al., 1964)
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Seed Storage
Low temperature and low seed moisture are the most effective means of
maintaining seed quality in storage (Doije, 2001). Roberts (1985) broadly classified seeds
as orthodox and recalcitrant. Orthodox seeds can be dried to low moisture content and
stored longer at low temperature whereas recalcitrant seeds survive for a few week or
months in storage and are killed by desiccation below a certain high moisture content,
example are mango mangos teen, rambutan, jackfruit,durian,and avocado. In addition,
Hartman and Kester (1990) stated that seeds are usually in varying length of time after
harvest. Storage condition that maintain seed viability are the low content of the seed,
low storage temperature modification of the storage atmosphere, slow respiration and
other metabolic process without injuring the embryo.
Methods of Breaking Dormancy
Several methods of breaking seeds dormancy are available but the following will
just be mentioned:
Scarification. It is a treatment given to seeds having seed coat that is impermeable
to water or gases (Devlin, 1977). This mentioned of breaking dormancy decreases the
resistance of the seed coat to water absorption and embryo expansion by weakening the
seed coat.
Scarification is any process of breaking, scratching, mechanically altering or
softening the seed coverings to make them permeable to water and gases. Typical species
with hard seed coats include members of legume, geranium, morning glory and linden
families. The commonly used in scarification treatments were mechanical, chemical, and
hot water treatments (Hartmann et al., 1997).
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Moist chilling seeds: seeds have the specific temperature requirements for
germination and often contain inhibitors and promoters. Evidence to support their view
dormancy is controlled by inhibitor-promoter balance that is altered by exposing the
seeds to low temperature (scarification) while unimbibed are mentioned by Copeland and
McDonald (1985). Any seed treatment that may increase germination percentage would
improve the chances of escaping stress in the field (Antolin, 2001).
Stratification is an old method for germinating seeds of many species (Copeland
and McDonald 1985). Adriance and Brinson (1955) as cited by Paing (1980) reported
that stratification as a pre-plant treatment preserves viability and hastens the germination
of seeds by reducing the amount of the germination inhibitor with in the seeds there by
enhancing germination. In addition, stratification is a method of handling dormant seeds
in which the imbibed seeds are subjected to a period of chilling to after-ripen the embryo.
The common stratification temperature is 0 to 10°C for at higher temperatures; seeds
often sprout prematurely and lower temperatures just above freezing delay sprouting. The
time required for stratification defends on the kind of seed, and sometimes upon the
individual lot of seed as well. For seeds of most species, one to four months is sufficient
for low temperature stratification (Hartman et al., 1997).
Furthermore, (Antolin, 2001) shows that stratification should be done only on
fleshy extracted seeds. Outdoor stratification in other cold storage can be accomplished
if the temperature remains below 40 degree Fahrenheit for six to twelve weeks. In this
case, the seeds could be sown directly into the bed or growing medium.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Blueberries spoiled in NA with four weeks at 0-2°C and at preserves ranging from
10.7 to 26.7 kpa (80,200 mm hg) mold development limited their storage life to seven
weeks in low pressure (Burg, 1976).
Effect of Cold Storage Temperature on Seed Germination
Adriance and Brinson (1955) stated that cold storage with moisture and
temperature ranging from 1°C to 5°C is found to the effective in breaking the dormancy
period of seeds. Thus added further that seed coat under cold storage has a tendency to
soften, thus allowing moisture entrance into the seed resulting in rapid germination.
Dormancy is broken in seeds by nature when exposed to lower temperature during winter
(Daniel, 1980). Similarly, Halfacre (1979) mentioned that in seed stratification, seeds
must be placed in a moist medium at temperature ranging from 4 to 7°C for 1 to 3
months. His requirement is true for deciduous nut and and fruit trees buds. Furthermore,
Devlin (1977) and Leopold (1975) stressed that temperature near freezing point are
usually the most effective, however, 10°C is often low enough to break dormancy.
A stated by Klingman and Ashton (1975), refrigerator or stratification is
commonly used to shorten the period of dormancy. Many seeds especially grouse seeds,
require alteration of the temperature 20-20°C and exposing the seeds to a low
temperature for five days prior to germination test makes possible an accurate
determination of variability. Likewise, (Devlin, 1977) as cited by Agnaya (2004) stated
that the seeds of the plant that do not germinate immediately requires a period of after
ripening treatment either dry storage while others in moist and low temperature
condition. Furthermore, Janick (1972), Hartman and Kester (1975), mentioned that
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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subjecting the seeds to low temperature reduces the amount of germination inhibitor that
is present within the seed thereby hastening germination.
Paing (1980), in her experiment on garden pea observed that the cold stored seeds
for six weeks had the highest total yield, higher number of lateral, shoots, longer pods,
high percentage of germination, shorten period of germination and the highest percentage
of pod set.
Bucao (1983) reported that subjecting snap beans (Phaseolus vulgaris) seed in the
refrigerator for 42 days at 5°C had the highest percentage of germination, tallest plant,
earliest to flower and had the highest percentage of pod set.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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MATERIALS AND METHODS
The materials used in the study were blueberry seeds, refrigerator, tissue paper,
styrofoam plates, seedling tray, and measuring materials.
The blueberry seeds were extracted from the fresh fruit through pressing the fresh
fruit by using the hand. The seeds were placed in styrofoam plates with tissue paper and
it was added water just enough to moisten to facilitate seed germination.
The blueberry seeds were stored at different durations of cold treatments in the
refrigerator at 5°C. After subjecting to cold treatments, it was sown in the seedling tray
and it was covered thinly with sand to facilitate seed germination.
The experiment was laid out in a Complete Randomized Design (CRD) using 50
seeds per treatment replicated three times.
The treatments used were the following:
Code
Cold Stratification Period
T1
Control (no. cold storage)
T2
1 week cold storage
T3
2 weeks cold storage
T4
3 weeks cold storage
T5
4 weeks cold storage
T6
5 weeks cold storage
T7
6 weeks cold storage
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Data Gathered :
The data gathered were the following:
1. Number of days from sowing to seedling emergence. This was taken by
counting the days from sowing of seeds to seedling emergence.
2. Number of days to first appearance of leaves. This was taken by counting the
number of days from sowing to first appearance of leaves.
3. Percentage of seedling emergence. This was determined by using the formula:
Percentage of emergence (%) = Number of Seeds Germinant x 100
Total Number of Seeds Sown
4. Percentage of normal seedlings. Normal seedlings were those that have well
developed roots with straight hypocotyls and leaves are dark green. This was taken using
the formula:
Normal Seedlings (%) = Number of Normal Seedlings x 100
Total Number of Seeds Sown
5. Seedling height (mm). This was measured from the base up to the tip of the
leaves with 2 sample plants for every treatment using the ruler after 113 days from
sowing.
6. Number of leaves. The number of leaves per plant was taken and recorded.
This was done 113 days after sowing the seeds.
7. Root length (mm). This was taken by measuring 2 representatives sample roots
from the base to the tip.
8. Root number. This was taken by counting the roots 113 days after sowing the
seeds.
9. Seedling vigor index. This was taken using the following scale below.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Rating
Description
1
most vigorous - excellent growth with dark green
leaves
2
vigorous - good growth with green leaves
3
less vigorous – poor growth with yellow leaves
10. Documentation through pictures. Pictures were taken during planting stage
and at seedling stage. Figure 1 shows an overview of the experimental area.
Figure. 1. Overview of the experimental area
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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RESULTS AND DISCUSSION
Number of Days from Sowing to Emergence
Seedling emergence as affected by the different cold storage duration was
determined based on the number of days from sowing to the time the seedling had
emerged on the soil surface (Table 1). Statistical results show that one week cold storage
enhanced earlier emergence of blueberry with a mean of 14.67 days from sowing which
is highly significant compared to other treatments. It was followed by the seeds subjected
to three, five and four weeks cold storage with means of 17.67, 18.33 and 19.67 days
respectively. The seeds that were not subjected to cold storage were the last to emerge
with a mean of 22.33 days.
These findings corroborate with the earlier observations of Tocnang (2009),
Agnaya (2004) and Comnang (2005) that seeds subjected to cold storage prior to planting
had faster and uniform germination. Likewise, Paing (1980) found 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 are 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.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 1. Number of days from sowing to seedling emergence
TREATMENT
NUMBER OF DAYS
Control (no cold storage)
22.33a
One week cold storage
14.67d
Two weeks cold storage
22ab
Three weeks cold storage
17.67cd
Four weeks cold storage
19.67abc
Five weeks cold storage
18.33bcd
Six weeks cold storage
22ab
Means with the same letters are not significantly different at 5% level by DMRT
Number of Days from Sowing
to First Appearance of Leaves
Table 2 shows significant statistical differences among the different cold storage
duration affecting the number of days from sowing to first appearance of leaves. Results
shows that seeds stored for one week attained the shortest days for leaf development and
produced leaves within 20.33 days. This was followed by the seeds subjected to cold
duration for three, four, and five weeks having the means of 24, 25.33 and 27 days
respectively. Seeds subjected to two weeks cold storage and the control significantly had
shorter days to first appearance of leaves which was statistically comparable to the seeds
subjected to six weeks cold storage which were the last to develop leaves with a mean of
29 days.
Paing (1980) found in his experiment that garden pea refrigerated for a shorter
period of cold storage appeared to be the best, attaining the highest percentage of
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 2. Number of days from sowing to first appearance of leaves
TREATMENT
NUMBER OF DAYS
Control (no cold storage)
27.67ab
One week cold storage
20.33c
Two weeks cold storage
27.67ab
Three weeks cold storage
24bc
Four weeks cold storage
25.33ab
Five weeks cold storage
27ab
Six weeks cold storage
29 a
Means with the same letters are not significantly different at 5% level by DMRT
germination, complete emergence, percentage of normal seedlings, tallest seedlings and
had the shortest days for the true leaf to emerge. Figure 2 shows an overview of blueberry
seedlings during first appearance of leaves.
Percentage of Seedling Emergence
As shown in Table 3, there were significant statistical 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 one
week cold storage attained the highest percentage of seedling emergence with a mean of
91.33 % which is not statistically different to the seeds that were not subjected to cold
storage with mean of 87.67 %. On the other hand, the seeds subjected to six weeks cold
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Figure 2. Overview of blueberry seedlings during first appearance of leaves
Table 3. Percentage of seedling emergence
TREATMENT
PERCENTAGE
Control (no cold storage)
87.67a
One week cold storage
91.33a
Two weeks cold storage
56.67bc
Three weeks cold storage
70.67b
Four weeks cold storage
67.33b
Five weeks cold storage
56.67bc
Six weeks cold storage
48.67c
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
16
storage had the lowest percentage of seedling emergence with a mean of 48.67% which is
significantly different with the rest of the treatments
It was observed however, that some seeds did not emerge for the reasons that can
not be precisely explained although it may be due to chilling injury that caused the death
of the embryo in the seeds during the long cold storage period and may also be due to the
not fully decomposed coco coil dust added in the media that was used. Hartman and
Kester (1990) stated that such results could be related this to high concentration of soil
solutes in the upper layer of the germination media. Likewise, Handreck (1993) wrote
that coir pith has a carbon-nitrogen ratio of 104:1 which means that it takes decades to
decompose. The containing partly decomposed coco coir dust migth have released some
toxic substance or gas that affected the emergence of seeds.
Percentage of Normal Seedlings
Table 4 shows that there were highly significant differences observed among the
different cold storage duration in terms of percentage of normal seedlings. Based on the
results, blueberry seeds subjected to one week cold storage had the highest percentage of
normal seedlings with a mean of 84.67 %. It was followed by the seedlings resulting from
seeds that were not subjected to cold storage with a mean of 78.67%. Seeds subjected to
cold storage durations for three, four, two and five weeks obtained means of 62, 58, 50.67
and 49.33% of normal seedlings. Lastly, six weeks cold storage duration of blueberry
seeds showed the lowest percentage of normal seedlings with a mean of 41.33%.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 4. Percentage of normal seedlings
TREATMENT
PERCENTAGE
Control (no cold storage)
78.67a
One week cold storage
84.67a
Two weeks cold storage
50.67bcd
Three weeks cold storage
62b
Four weeks cold storage
58bc
Five weeks cold storage
49.33cd
Six weeks cold storage
41.33d
Means with the same letters are not significantly different at level by DMRT
Salisbury and Ross (1992) explained that pre-chilling of seeds sometimes has a
strong effect on growth in addition to its dormancy breaking action. If the embryos of
peach seedlings are excised from their cotyledons, they will germinate without pre-
chilling, but the seedlings are frequently stunted and abnormal. However, when excised
embyos are treated with low temperature, they grow normal seedlings. Thus, it is pre-
chilling and not the presence of the cotyledons that ensures their normality.
Seedling Height
The seedling (Table 5) of blueberry was obtained by measuring the seedling from
the base up to the tip of its secondary leaves. Result shows that there were no significant
statistical differences among the various cold storage durations. However, numerical
results shows that seedlings from seeds subjected to one week cold storage had the tallest
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
18
Table 5. Seedling height
TREATMENT
HEIGHT (mm)
Control (no cold storage)
5a
One week cold storage
7.67a
Two weeks cold storage
5.17a
Three weeks cold storage
4.67a
Four weeks cold storage
5.50a
Five weeks cold storage
5.50a
Six weeks cold storage
5a
Means with the same letters are not significantly different at level by DMRT
seedlings with a mean of 7.67 mm. This was followed by the seedlings from the seeds
subjected to four and five weeks cold storage durations with a means of 5.50 mm.
However, seedlings from the seeds that were subjected to three weeks cold storage had
the shortest height with a mean of 4.67 mm.
Results showed that one week cold storage duration of seeds prior to sowing
enhanced growth of seedlings leading to taller seedlings. These findings corroborate with
the observation of Tocnang (2009) who stated that cold storage of guapple seeds for a
period of one week at 5 °C, will hasten seed germination, improve rate of seedling
emergence and promote the development of taller seedlings. Likewise, Banagen (2008)
reported that cold stratification of coffee Arabica seeds at 5 °C before planting stimulates
faster growth and the greatest effect can be obtained from cold stratification for one
week.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Number of Leaves per Plant
The result in Table 6 shows that there were no significant differences among the
different cold storage duration with regards to the number of leaves per plant. However,
numerical results revealed that seedlings from seeds subjected to one week cold storage
had the highest number of leaves per plant with a mean of 3.83. Whereas, seedlings from
seeds subjected to six weeks cold storage had the lowest number of leaves developed
with a mean of 2.83.
Root Length
Regarding the root length of blueberry, it can be observed in Table 7 that there
were significant statistical differences among the treatments as affected by the various
durations of cold storage used. Results shows that seeds subjected to one week cold
storage had the longest roots with a mean of 18.83 mm. Seedlings from the seeds
Table 6. Number of leaves per plant
TREATMENT
NUMBER OF LEAVES
Control (no cold storage)
3a
One week cold storage
3.83a
Two weeks cold storage
3a
Three weeks cold storage
3.33a
Four weeks cold storage
3.50a
Five weeks cold storage
3.17a
Six weeks cold storage
2.83a
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
20
Table 7. Root length
TREATMENT
ROOT LENGTH (mm)
Control (no cold storage)
14.50b
One week cold storage
18.83a
Two weeks cold storage
9c
Three weeks cold storage
9c
Four weeks cold storage
8.83c
Five weeks cold storage
10.17c
Six weeks cold storage
9c
Means with the same letters are not significantly different at level by DMRT
subjected to four weeks duration of cold storage had the shortest roots with a mean of
8.83 mm which is not statistically different with the root length of seedlings resulting
from seeds subjected to two, three, five and six weeks cold storage.
Results showed that subjecting seeds to shorter duration of cold storage for a
period of one week induce the production of roots. Meyer and Anderson (1952) explained
that optimum temperature is usually about midway between two extremes of temperature
at which germination will occur. It is not possible to designate any exact varies with the
other prevailing environmental conditions and also with the exact criterion selected as an
index of germination. The most favorable temperature for the elongation of the primary
roots for example does not always correspond to the most suitable temperature for the
development of the plumule.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
21
Number of Roots
Table 8 shows that there were no significant differences on the number of roots as
affected by different cold storage duration. Nevertheless, numerical figures suggest that
seedlings coming from seeds subjected to one week cold storage had the highest number
of roots with a mean of 4.17. Seedlings treated with four weeks duration of cold storage
had an average number of roots of 3.83 while seedlings subjected to two, three and five
weeks cold storage had the same means of 3.50. Seedlings subjected to six weeks
duration of cold storage produced the least number of roots with a mean of 2.83.
Figure 3 shows uprooted seedlings during gathering of data regarding root length
and root number 113 days from sowing.
Table 8. Number of roots
TREATMENT
NUMBER OF ROOTS
Control (no cold storage)
2.50a
One week cold storage
4.17a
Two weeks cold storage
3.50a
Three weeks cold storage
3.50a
Four weeks cold storage
3.83a
Five weeks cold storage
3.50a
Six weeks cold storage
2.83a
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
22
Figure 3. Uprooted seedlings of blueberry during gathering of data 113 days from sowing
Seedling Vigor Index
The results in Table 9 shows no significant statistical differences observed among
the different treatments used in the study affecting the seedling vigor of blueberry
seedlings. However, numerical results shows that seedlings coming from the seeds
subjected to one week cold storage promoted the excellent growth of seedlings.
Tocnang (2009) observed that guapple seed subjected to one week and two weeks
cold storage promoted excellent growth of seedlings. Likewise, Banagen (2008) noted
that storing coffee seeds under cold temperature influences 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. Figure 4 shows the overview of the experiment 113 days from
sowing the seeds.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
23
Table 9. Seedling vigor index
TREATMENT
SEEDLING VIGOR
Control (no cold storage)
2a
One week cold storage
1.67a
Two weeks cold storage
2.33a
Three weeks cold storage
2.33a
Four weeks cold storage
2.33a
Five weeks cold storage
2.33a
Six weeks cold storage
2.33a
Rating: 1= most vigorous; 2= vigorous; 3= less vigorous
Figure 4. Overview of the experiment 113 days from sowing seeds.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
24
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The germination and seedling growth of blueberry seeds as affected by different
cold storage duration was studied at the Pomology Project Nursery, Benguet State
University, La Trinidad, Benguet from November 15,2009 to March 7, 2010. the
blueberry seeds were subjected to low temperature with an average of 5 °C at various
periods from one to six weeks.
Result showed that there were significant statistical differences observed among
the various cold storage duration used. One week cold storage of blueberry seeds showed
the best performance among all treatments that enhanced earlier seedling emergence with
a mean of 14.67 days and shorter days to develop true leaves with a mean of 20.33 days.
Likewise, the same duration of cold storage attained the highest percentage of seedling
emergence having a mean of 91.33 % and also with the percentage of normal seedlings
with a mean of 84.67 %. The seedlings coming from the seeds subjected to one week cold
storage had also produced longer root length with a mean of 18.83 mm which is highly
significant to the other treatments.
With regards to the seedling height, number of leaves, number of roots and
seedling vigor statistical results shows that there were no significant differences among
the different cold storage duration used however numerical results shows that seedlings
coming from the seeds stored for a period of one week had the tallest seedlings, more
number of leaves and roots, and had more vigorous seedlings as compared to other
treatments.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
25
Conclusion
Based on the results of the study, subjecting blueberry seeds to cold storage for
one week before sowing showed promising results. It was found out that it was the best
among the various cold storage durations studied in enhancing faster germination and
emergence of blueberry seeds.
Recommendation
From the preceding results, it is recommended that cold storage of blueberry
seeds for a period of one week at 5 °C prior to sowing should be done in order to
enhance faster seed germination and emergence as well as to promote longer root
development and enhanced faster and vigorous seedling growth. However, further study
along this line also recommended to verify these findings.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
26
LITERATURE CITED
ADRIANCE, G. W. and F. R. BRINSON. 1955. Propagation of Horticulture Plants: Mc
Grawhill Book Co., New York Inc. p. 82.
AGNAYA, J. S. 2004. Effect of cold stratification period on the germination of Benguet
“wild tea”. BS Thesis. BSU La Trinidad, Benguet.
ANTOLIN, N. C. 2001. Germination of Benguet “wild tea” as influence by
stratification periods. A Term Paper Submitted in Horticulture 315 (Advances in
plant Propagation and Management), BS Thesis. BSU, LA Trinidad, Benguet.
BANAGEN, M. L. 2008. Effect of cool storage duration on the germination and seedling
characteristics of coffee seeds. BS Thesis. BSU, LA Trinidad, Benguet.
BUCAO, N. J. 1983. Influence of storing seeds in cold storage on the growth and yield
of snap beans. BS Thesis. BSU, La Trinidad, Benguet.
BURG, S. P. 1976. Postharvest, Physiology and Hypobaric, Storage of fresh fruit
produce. USA: p. 383.
COMNANG, L. D. 2005. The effect of cold stratitification duration on the seedling
emergence of mountain tea “Gipas”. BS Thesis. BSU, La Trinidad, Benguet.
COPELAND, L. O. and M. B. Mc DONALD. 1985. Principles of seed Science and
Technology. Minnesota: Burgness Pub. p. 481.
DANIEL, T. W. 1980. Principles of Silviculture. 2nd edition.MCGraw Hill Book Co.,
New York. p. 379.
DAUBENMIRE, R. R. 1974. Plant and Environment. A textbook of Plant Autocology.
New York: John Welly and Sons. p.182.
DEVLIN, R. 1977. Plant Physiology. Third Edition Dvan Nestrand Co., New York. pp.
552-557.
DOIJE, S. D. 2001. Seed storage of Horticulture crops pp. 65-67.
EDMOND, J. B. T. C SEENN and P. S ANDREW. 1964. Fundamentals of Horticulture.
New Delhi, Tata McGraw hill Public. Co., p. 432
HALFACRE, R. G. 1979. Horticulture.MCGraw Hill Book Co., New York. p. 340.
HANDRECK, K. A. 1993. Properties of coco coir dust, and its use in the formulation of
soilless potting media. Community Soil and Plant Analysis. 24: 349-363.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
27
HARTMAN, H. T., D. E. KESTER, DAVIES, F. T. Jr., and R. L. GENEVE. 1997.
Plant Propagation: Principles and Practices.6th Ed. New Jersey: Prentice Hall.
pp. 218-220.
HARTMAN, H. T. and D. E. KESTER. 1990. Plant Propagation: Principles and
Practices 5th Ed. New Jersey: Prentice Hall. p. 98.
HARTMAN, H. T.and D. E. KESTER.1975. Plant Propagation, Principles and
Practices New Jersey: Prentice Hall of India. Inc. p. 120
JANICK, J. 1972. Horticulture Science Principle and Practices, Second Edition. San
Francisco.W.H. Freeman.Camp. p. 152.
KLINGMAN, G. C. and F. N. ASHTON.1975. Weed Science. Principles and Practices.
New York. A Welley Inter Science Publication. p. 52.
KUEPPER, G. L. and S. DIVER. 2004. Principle of Horticultural Crops. Horticulture
Publication Guide. p. 428.
LEOPOLD, 1975. Plant Growth and Development. New York. Judd Publishing Co. Inc.
p. 235.
MEYER, B. S. and D. B. ANDERSON. 1952. Plant Physiology. New Jersey: Van
Nostrand.
PAING, R. D. 1980. Effect of seed stratification on the growth and yield of the edible
podded pea. BS Thesis. BSU, La Trinidad, Benguet.
ROBERTS, E. H. 1985. Handbook of Seed Technology for Geneback Compedium of
Scientific Germination Information and Test Recommendations.
SALISBURY, F. B. and C. W. ROSS. 1992. Fourth Edititon. 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 and Development. Bombay, New Delhi: Tata Mc
Graw Hill, Publ., Co., Ltd. p. 37.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
28
APPENDICES
Appendix Table 1. Number of days from sowing to seedling emergence
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 22 24 21 67 22.33
T2 16 14 14 44 14.67
T3 23 19 24 66 22
T4 14 20 19 53 17.67
T5 19 19 21 59 19.67
T6 18 16 21 55 18.33
T7 21 22 23 66 22
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Between 6 145.905 24.317 5.738 2.85 4.46
Within 14 59.333 4.238
Total 20 205.238
**Highly significant
Coefficient of variation = 10.54%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
29
Appendix Table 2. Number of days from sowing to first appearance of leaves
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 27 30 26 83 27.67
T2 22 19 20 61 20.33
T3 28 25 30 83 27.67
T4 21 26 25 72 24
T5 24 24 28 76 25.33
T6 26 25 30 81 27
T7 27 29 31 87 29
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 155.905 25.984 5.006 2.85 4.4
Error 14 72.667 5.190
Total 20 228.571
**Highly significant
Coefficient of variation = 8.81%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
30
Appendix Table 3. Percentage of seedling emergence
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 96 76 92 264 88
T2 92 94 88 274 91.33
T3 64 48 58 170 56.67
T4 82 60 70 212 70.67
T5 72 74 56 202 67.33
T6 66 54 50 170 56,67
T7 54 50 42 146 48.67
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 4704.476 784.079 10.468 2.85 4.46
Error 14 1048.667 74.905
Total 20 228.571
**Highly significant
Coefficient of variation = 12.65%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
31
Appendix Table 4. Percentage of normal seedlings
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 78 76 82 236 78.67
T2 84 86 84 254 84.67
T3 56 44 52 152 50.67
T4 72 54 60 186 62
T5 62 64 48 174 58
T6 58 46 44 148 49.33
T7 46 40 38 124 41.33
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 4533.333 755.556 18.711 2.85 4.46
Error 14 565.333 40.381
Total 20 5098.667
**Highly significant
Coefficient of variation = 10.47%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
32
Appendix Table 5. Seedling height
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 4.5 4.5 6 15 5
T2 6 8 9 23 7.67
T3 5 5.5 5 15.5 5.17
T4 5 4.5 4.5 14 4.67
T5 4.5 7 5 16.5 5.5
T6 5.5 7 4 16.5 5.5
T7 5 6 4 15 5
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 2.143 0.357 2.55 2.85 4.46
Error 14 2.167 0.155
Total 20 4.310
Not significant
Coefficient of variation = 12.15%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
33
Appendix Table 6. Number of leaves per plant
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 3 3 3 9 3
T2 4 4 3.5 11.5 3.83
T3 3 3.5 2.5 9 3
T4 3.5 3 3.5 10 3.33
T5 4 3.5 3 10.5 3.5
T6 3.5 3.5 2.5 9.5 3.17
T7 5 3 2.5 8.5 2.83
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 18.000 3.000 2.55 2.85 4.46
Error 14 16.500 1.179
Total 20 5098.667
Not significant
Coefficient of variation = 19.74%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
34
Appendix Table 7. Root length
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 18.5 11.5 13.5 43.5 14.5
T2 15.5 18.5 22.5 56.5 18.83
T3 9 10 8 27 9
T4 10 8.5 8.5 27 9
T5 10.5 8 8 26.5 8.83
T6 9.5 11.5 9.5 30.5 10.17
T7 8 8.5 10.5 27 9
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 270.667 45.111 9.792 2.85 4.46
Error 14 64.500 4.607
Total 20 335.167
** Highly significant
Coefficient of variation = 18.94%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
35
Appendix Table 8 . Number of roots
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 2.5 2 3 7.5 2.5
T2 4.5 4 4 12.5 4.17
T3 4 3 3.5 10.5 3.5
T4 4 3 3.5 10.5 3.5
T5 2.5 4.5 4.5 11.5 3.83
T6 4.5 4 2 10.5 3.5
T7 3 3 2.5 8.5 2.83
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 5.810 0.968 1.69 2.85 4.46
Error 14 8.000 0.571
Total 20 13.810
Not significant
Coefficient of variation = 22.20%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
36
Appendix Table 9. Seedling vigor
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 2 2 2 6 2
T2 2 2 1 5 1.67
T3 3 2 2 7 2.33
T4 2 2 3 7 2.33
T5 3 2 2 7 2.33
T6 2 2 3 7 2.33
T7 2 2 3 7 2.33
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 1.238 0.206 0.72 2.85 4.46
Error 14 4.000 0.286
Total 20 5.238
Not significant
Coefficient of variation = 24.40%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
DAO-INES, GABBY K. APRIL 2010. Seed Germination and Seedling Growth of
Blueberry (Vacinium corymbossum) as Affected by Different Cold Storage Duration.
Benguet State University, La Trinidad, Benguet.
Adviser: Franklin G. Bawang, Msc.
ABSTRACT
The study was conducted at the Pomology Nursery of Benguet State University,
from November 2009 to March 2010 to find out the best durations of cold storage that
will effectively break the dormancy of blueberry seeds; and to determine the effect of
cold storage on seed germination and seedling growth of blueberry.
Results showed that the various durations of cold storage of blueberry seeds had
significantly affected seed germination and seedling emergence. Seedlings resulted from
the seeds subjected to one week cold storage had developed true leaf earlier; more seeds
emerged resulted to higher percentage of seedling emergence; and more normal seedlings
developed. Likewise, seeds subjected to one week cold storage resulted to seedlings
having longer roots.
TABLE OF CONTENTS
Page
Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
REVIEW OF LITERATURE
Description of the Plant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Seed Germination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Seed Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Methods of Breaking Dormancy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Effect of Cold Storage Duration
on Seed Germination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
RESULTS AND DISCUSSION
Number days from Sowing to
Emergence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Number of Days from Sowing
to First Appearance of Leaves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Percentage of Seedling Emergence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Percentage of Normal seedlings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Seedling Height . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Number of Leaves per Plant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Root Length . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
ii
Number of Roots . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Seedling Vigor Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
SAMMARY, CONCLUSION AND RECOMMENDATION
Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
APPENDICES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
iii
1
INTRODUCTION
Blueberries are flowering plants belonging to the genus Vaccinium ssp. The
species are native only to North America. The crop is considered as shrubs varying in
size from 10cm to 4m tall. The smaller species are known as “low bush blueberries”, and
the larger species are “high bush blueberries”. The leaves can be deciduous or evergreen,
ovate to lanceolate, and from1-8cm long and 0.5-3.5cm broad. The flowers are bell-
shaped, white, pale pink or red. Sometimes tinged greenish. The fruit is false berry 5-
16mm diameter with a flared “crown” at the end. They are pale greenish at first, then
reddish-purple and finally indigo on ripening. It has a sweet taste when mature, with
variable acidity. Blueberry bushes typically bear fruit from May to June in the Western
Hemisphere; “blueberry season’ peaks in July, which is National Blueberry Month in the
United States and Germany.
Blueberries grow wild in many parts of the world. However, the United States and
Canada supply about 95% of the blueberries used by the food industry. North American
farmers harvest about 103 million pounds (47 million kilograms) of blueberries annually.
For centuries, blue berries were gathered from the forest and bogs by the Native
Americans and these are usually consumed fresh and also it can be preserved. The
Northeast Native American tribes revered blueberries and much folklore developed
around them. The blossom ends of each berry, the calyx, forms a shaped of a perfect five-
. pointed star. Parts of the blueberry plant were also used as medicine, for coughs and
good for the circulation of the blood. Not only for medicine, the juice can also be made as
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
2
an excellent dye for baskets and cloth. It can also used in other food preparation.
Example, the dried berries were also crushed into a powder and into meat for flavor; it
can be processed such as jellies,jams,pies,muffins and snack foods.
For early colonist, blueberries were probably one of the first familiar foods
discovered on North American continent, since they were similar to other berry varieties
found in Europe. Their abundance, natural sweetness and visibility made them on
immediately welcome addition at a time when fresh food was in short supply, and sugar
was scarce and expensive. Blueberry were canned and shipped to union troops during
civil war soldiers in 1860’s.
Blueberries have gained increasing recognition from health professionals for their
very high antioxidant properties. Studies of the anthocyanin and pytochemicals found in
the blueberries have shown that they provide a range of benefits, including reducing
eyestrain, counteracting environmental carcinogens, promoting urinary tract health,
fighting sun damage and protecting against cardiovascular disease. These are low in
calories, low in fat and have no cholesterol, in addition to being a good source of dietary
fiber and is packed with other vitamins (A,C,E) and minerals (potassium, manganese and
magnesium).
`Blueberry can be potentially grown in forest or mountains. However, mass
production could become a problem under high demand because it takes time to produce
seedling from seeds. Therefore, it is imperative to conduct this study to be able to
determine the best duration of cold storage of blueberry seeds that can provides the
stimulus for earlier and uniform germination and reduce the longer duration and intervals
of waiting time leading to higher percentage survival of the seedling. In addition, to be
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
3
able to grow or to produce more blueberry in our community for economic, medicinal
and food purposes.
The results of this study will provide baseline information’s to researchers
interested to work on blueberry in the future in order to help in the establishment of
blueberry production industry and to encourage more farmers to produce more blueberry
that is considered as a medicinal and has great potential to market when it is processed.
The study was conducted at the Nursery of the Pomology Project, Benguet State
University, La Trinidad, Benguet second semester from November 15 to March 7, 2010
to find out the best duration of cold storage in breaking dormancy of blueberry seeds and
to determine the effect of cold storage on seed germination and seedling growth of
blueberry.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
4
REVIEW OF LITERATURE
Description of the Plant
Wild blueberries are perennial shrubs found in the undergrowth of forests in
northeastern United States and in the Atlantic provinces of Canada. Plants are propagated
from seeds or cuttings. They produced underground stems of rhizomes, which
periodically send up new shoots and developed roots. One original plant can thus cover
an area ranging from 75 to 250 square feet depending on the age of the plant. Originally
gathered by hand rakes although effective mechanical harvesters are being developed and
improved. Various species abound including low bush and high bush varieties, which
produced varying degrees of sweetness in the resulting fruit. For dried berries, the fruits
are infused with a sugar solution to create a uniformly sweet and appealing product for
used in baking and confectionery applications and granolos, as well as in shacks and trail
mixes (Kuepper and Diver, 2004).
Seed Germination
Seed germination is defined as the activation of the metabolic machinery of the
embryo leading to the emergence of new seedling plant (Hartman et. al., 1990). It is a
sequence of step beginning with the uptake of water leading to rupture of the seed coat
and germination usually determined visibly by observing the protrusion of the radicle or
shoot (Devlin, 1977). Germination of seed will occur only when the embryo development
is complete (Edmond et al., 1964)
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Seed Storage
Low temperature and low seed moisture are the most effective means of
maintaining seed quality in storage (Doije, 2001). Roberts (1985) broadly classified seeds
as orthodox and recalcitrant. Orthodox seeds can be dried to low moisture content and
stored longer at low temperature whereas recalcitrant seeds survive for a few week or
months in storage and are killed by desiccation below a certain high moisture content,
example are mango mangos teen, rambutan, jackfruit,durian,and avocado. In addition,
Hartman and Kester (1990) stated that seeds are usually in varying length of time after
harvest. Storage condition that maintain seed viability are the low content of the seed,
low storage temperature modification of the storage atmosphere, slow respiration and
other metabolic process without injuring the embryo.
Methods of Breaking Dormancy
Several methods of breaking seeds dormancy are available but the following will
just be mentioned:
Scarification. It is a treatment given to seeds having seed coat that is impermeable
to water or gases (Devlin, 1977). This mentioned of breaking dormancy decreases the
resistance of the seed coat to water absorption and embryo expansion by weakening the
seed coat.
Scarification is any process of breaking, scratching, mechanically altering or
softening the seed coverings to make them permeable to water and gases. Typical species
with hard seed coats include members of legume, geranium, morning glory and linden
families. The commonly used in scarification treatments were mechanical, chemical, and
hot water treatments (Hartmann et al., 1997).
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Moist chilling seeds: seeds have the specific temperature requirements for
germination and often contain inhibitors and promoters. Evidence to support their view
dormancy is controlled by inhibitor-promoter balance that is altered by exposing the
seeds to low temperature (scarification) while unimbibed are mentioned by Copeland and
McDonald (1985). Any seed treatment that may increase germination percentage would
improve the chances of escaping stress in the field (Antolin, 2001).
Stratification is an old method for germinating seeds of many species (Copeland
and McDonald 1985). Adriance and Brinson (1955) as cited by Paing (1980) reported
that stratification as a pre-plant treatment preserves viability and hastens the germination
of seeds by reducing the amount of the germination inhibitor with in the seeds there by
enhancing germination. In addition, stratification is a method of handling dormant seeds
in which the imbibed seeds are subjected to a period of chilling to after-ripen the embryo.
The common stratification temperature is 0 to 10°C for at higher temperatures; seeds
often sprout prematurely and lower temperatures just above freezing delay sprouting. The
time required for stratification defends on the kind of seed, and sometimes upon the
individual lot of seed as well. For seeds of most species, one to four months is sufficient
for low temperature stratification (Hartman et al., 1997).
Furthermore, (Antolin, 2001) shows that stratification should be done only on
fleshy extracted seeds. Outdoor stratification in other cold storage can be accomplished
if the temperature remains below 40 degree Fahrenheit for six to twelve weeks. In this
case, the seeds could be sown directly into the bed or growing medium.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Blueberries spoiled in NA with four weeks at 0-2°C and at preserves ranging from
10.7 to 26.7 kpa (80,200 mm hg) mold development limited their storage life to seven
weeks in low pressure (Burg, 1976).
Effect of Cold Storage Temperature on Seed Germination
Adriance and Brinson (1955) stated that cold storage with moisture and
temperature ranging from 1°C to 5°C is found to the effective in breaking the dormancy
period of seeds. Thus added further that seed coat under cold storage has a tendency to
soften, thus allowing moisture entrance into the seed resulting in rapid germination.
Dormancy is broken in seeds by nature when exposed to lower temperature during winter
(Daniel, 1980). Similarly, Halfacre (1979) mentioned that in seed stratification, seeds
must be placed in a moist medium at temperature ranging from 4 to 7°C for 1 to 3
months. His requirement is true for deciduous nut and and fruit trees buds. Furthermore,
Devlin (1977) and Leopold (1975) stressed that temperature near freezing point are
usually the most effective, however, 10°C is often low enough to break dormancy.
A stated by Klingman and Ashton (1975), refrigerator or stratification is
commonly used to shorten the period of dormancy. Many seeds especially grouse seeds,
require alteration of the temperature 20-20°C and exposing the seeds to a low
temperature for five days prior to germination test makes possible an accurate
determination of variability. Likewise, (Devlin, 1977) as cited by Agnaya (2004) stated
that the seeds of the plant that do not germinate immediately requires a period of after
ripening treatment either dry storage while others in moist and low temperature
condition. Furthermore, Janick (1972), Hartman and Kester (1975), mentioned that
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
8
subjecting the seeds to low temperature reduces the amount of germination inhibitor that
is present within the seed thereby hastening germination.
Paing (1980), in her experiment on garden pea observed that the cold stored seeds
for six weeks had the highest total yield, higher number of lateral, shoots, longer pods,
high percentage of germination, shorten period of germination and the highest percentage
of pod set.
Bucao (1983) reported that subjecting snap beans (Phaseolus vulgaris) seed in the
refrigerator for 42 days at 5°C had the highest percentage of germination, tallest plant,
earliest to flower and had the highest percentage of pod set.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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MATERIALS AND METHODS
The materials used in the study were blueberry seeds, refrigerator, tissue paper,
styrofoam plates, seedling tray, and measuring materials.
The blueberry seeds were extracted from the fresh fruit through pressing the fresh
fruit by using the hand. The seeds were placed in styrofoam plates with tissue paper and
it was added water just enough to moisten to facilitate seed germination.
The blueberry seeds were stored at different durations of cold treatments in the
refrigerator at 5°C. After subjecting to cold treatments, it was sown in the seedling tray
and it was covered thinly with sand to facilitate seed germination.
The experiment was laid out in a Complete Randomized Design (CRD) using 50
seeds per treatment replicated three times.
The treatments used were the following:
Code
Cold Stratification Period
T1
Control (no. cold storage)
T2
1 week cold storage
T3
2 weeks cold storage
T4
3 weeks cold storage
T5
4 weeks cold storage
T6
5 weeks cold storage
T7
6 weeks cold storage
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Data Gathered :
The data gathered were the following:
1. Number of days from sowing to seedling emergence. This was taken by
counting the days from sowing of seeds to seedling emergence.
2. Number of days to first appearance of leaves. This was taken by counting the
number of days from sowing to first appearance of leaves.
3. Percentage of seedling emergence. This was determined by using the formula:
Percentage of emergence (%) = Number of Seeds Germinant x 100
Total Number of Seeds Sown
4. Percentage of normal seedlings. Normal seedlings were those that have well
developed roots with straight hypocotyls and leaves are dark green. This was taken using
the formula:
Normal Seedlings (%) = Number of Normal Seedlings x 100
Total Number of Seeds Sown
5. Seedling height (mm). This was measured from the base up to the tip of the
leaves with 2 sample plants for every treatment using the ruler after 113 days from
sowing.
6. Number of leaves. The number of leaves per plant was taken and recorded.
This was done 113 days after sowing the seeds.
7. Root length (mm). This was taken by measuring 2 representatives sample roots
from the base to the tip.
8. Root number. This was taken by counting the roots 113 days after sowing the
seeds.
9. Seedling vigor index. This was taken using the following scale below.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Rating
Description
1
most vigorous - excellent growth with dark green
leaves
2
vigorous - good growth with green leaves
3
less vigorous – poor growth with yellow leaves
10. Documentation through pictures. Pictures were taken during planting stage
and at seedling stage. Figure 1 shows an overview of the experimental area.
Figure. 1. Overview of the experimental area
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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RESULTS AND DISCUSSION
Number of Days from Sowing to Emergence
Seedling emergence as affected by the different cold storage duration was
determined based on the number of days from sowing to the time the seedling had
emerged on the soil surface (Table 1). Statistical results show that one week cold storage
enhanced earlier emergence of blueberry with a mean of 14.67 days from sowing which
is highly significant compared to other treatments. It was followed by the seeds subjected
to three, five and four weeks cold storage with means of 17.67, 18.33 and 19.67 days
respectively. The seeds that were not subjected to cold storage were the last to emerge
with a mean of 22.33 days.
These findings corroborate with the earlier observations of Tocnang (2009),
Agnaya (2004) and Comnang (2005) that seeds subjected to cold storage prior to planting
had faster and uniform germination. Likewise, Paing (1980) found 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 are 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.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 1. Number of days from sowing to seedling emergence
TREATMENT
NUMBER OF DAYS
Control (no cold storage)
22.33a
One week cold storage
14.67d
Two weeks cold storage
22ab
Three weeks cold storage
17.67cd
Four weeks cold storage
19.67abc
Five weeks cold storage
18.33bcd
Six weeks cold storage
22ab
Means with the same letters are not significantly different at 5% level by DMRT
Number of Days from Sowing
to First Appearance of Leaves
Table 2 shows significant statistical differences among the different cold storage
duration affecting the number of days from sowing to first appearance of leaves. Results
shows that seeds stored for one week attained the shortest days for leaf development and
produced leaves within 20.33 days. This was followed by the seeds subjected to cold
duration for three, four, and five weeks having the means of 24, 25.33 and 27 days
respectively. Seeds subjected to two weeks cold storage and the control significantly had
shorter days to first appearance of leaves which was statistically comparable to the seeds
subjected to six weeks cold storage which were the last to develop leaves with a mean of
29 days.
Paing (1980) found in his experiment that garden pea refrigerated for a shorter
period of cold storage appeared to be the best, attaining the highest percentage of
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 2. Number of days from sowing to first appearance of leaves
TREATMENT
NUMBER OF DAYS
Control (no cold storage)
27.67ab
One week cold storage
20.33c
Two weeks cold storage
27.67ab
Three weeks cold storage
24bc
Four weeks cold storage
25.33ab
Five weeks cold storage
27ab
Six weeks cold storage
29 a
Means with the same letters are not significantly different at 5% level by DMRT
germination, complete emergence, percentage of normal seedlings, tallest seedlings and
had the shortest days for the true leaf to emerge. Figure 2 shows an overview of blueberry
seedlings during first appearance of leaves.
Percentage of Seedling Emergence
As shown in Table 3, there were significant statistical 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 one
week cold storage attained the highest percentage of seedling emergence with a mean of
91.33 % which is not statistically different to the seeds that were not subjected to cold
storage with mean of 87.67 %. On the other hand, the seeds subjected to six weeks cold
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Figure 2. Overview of blueberry seedlings during first appearance of leaves
Table 3. Percentage of seedling emergence
TREATMENT
PERCENTAGE
Control (no cold storage)
87.67a
One week cold storage
91.33a
Two weeks cold storage
56.67bc
Three weeks cold storage
70.67b
Four weeks cold storage
67.33b
Five weeks cold storage
56.67bc
Six weeks cold storage
48.67c
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
16
storage had the lowest percentage of seedling emergence with a mean of 48.67% which is
significantly different with the rest of the treatments
It was observed however, that some seeds did not emerge for the reasons that can
not be precisely explained although it may be due to chilling injury that caused the death
of the embryo in the seeds during the long cold storage period and may also be due to the
not fully decomposed coco coil dust added in the media that was used. Hartman and
Kester (1990) stated that such results could be related this to high concentration of soil
solutes in the upper layer of the germination media. Likewise, Handreck (1993) wrote
that coir pith has a carbon-nitrogen ratio of 104:1 which means that it takes decades to
decompose. The containing partly decomposed coco coir dust migth have released some
toxic substance or gas that affected the emergence of seeds.
Percentage of Normal Seedlings
Table 4 shows that there were highly significant differences observed among the
different cold storage duration in terms of percentage of normal seedlings. Based on the
results, blueberry seeds subjected to one week cold storage had the highest percentage of
normal seedlings with a mean of 84.67 %. It was followed by the seedlings resulting from
seeds that were not subjected to cold storage with a mean of 78.67%. Seeds subjected to
cold storage durations for three, four, two and five weeks obtained means of 62, 58, 50.67
and 49.33% of normal seedlings. Lastly, six weeks cold storage duration of blueberry
seeds showed the lowest percentage of normal seedlings with a mean of 41.33%.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 4. Percentage of normal seedlings
TREATMENT
PERCENTAGE
Control (no cold storage)
78.67a
One week cold storage
84.67a
Two weeks cold storage
50.67bcd
Three weeks cold storage
62b
Four weeks cold storage
58bc
Five weeks cold storage
49.33cd
Six weeks cold storage
41.33d
Means with the same letters are not significantly different at level by DMRT
Salisbury and Ross (1992) explained that pre-chilling of seeds sometimes has a
strong effect on growth in addition to its dormancy breaking action. If the embryos of
peach seedlings are excised from their cotyledons, they will germinate without pre-
chilling, but the seedlings are frequently stunted and abnormal. However, when excised
embyos are treated with low temperature, they grow normal seedlings. Thus, it is pre-
chilling and not the presence of the cotyledons that ensures their normality.
Seedling Height
The seedling (Table 5) of blueberry was obtained by measuring the seedling from
the base up to the tip of its secondary leaves. Result shows that there were no significant
statistical differences among the various cold storage durations. However, numerical
results shows that seedlings from seeds subjected to one week cold storage had the tallest
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 5. Seedling height
TREATMENT
HEIGHT (mm)
Control (no cold storage)
5a
One week cold storage
7.67a
Two weeks cold storage
5.17a
Three weeks cold storage
4.67a
Four weeks cold storage
5.50a
Five weeks cold storage
5.50a
Six weeks cold storage
5a
Means with the same letters are not significantly different at level by DMRT
seedlings with a mean of 7.67 mm. This was followed by the seedlings from the seeds
subjected to four and five weeks cold storage durations with a means of 5.50 mm.
However, seedlings from the seeds that were subjected to three weeks cold storage had
the shortest height with a mean of 4.67 mm.
Results showed that one week cold storage duration of seeds prior to sowing
enhanced growth of seedlings leading to taller seedlings. These findings corroborate with
the observation of Tocnang (2009) who stated that cold storage of guapple seeds for a
period of one week at 5 °C, will hasten seed germination, improve rate of seedling
emergence and promote the development of taller seedlings. Likewise, Banagen (2008)
reported that cold stratification of coffee Arabica seeds at 5 °C before planting stimulates
faster growth and the greatest effect can be obtained from cold stratification for one
week.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
19
Number of Leaves per Plant
The result in Table 6 shows that there were no significant differences among the
different cold storage duration with regards to the number of leaves per plant. However,
numerical results revealed that seedlings from seeds subjected to one week cold storage
had the highest number of leaves per plant with a mean of 3.83. Whereas, seedlings from
seeds subjected to six weeks cold storage had the lowest number of leaves developed
with a mean of 2.83.
Root Length
Regarding the root length of blueberry, it can be observed in Table 7 that there
were significant statistical differences among the treatments as affected by the various
durations of cold storage used. Results shows that seeds subjected to one week cold
storage had the longest roots with a mean of 18.83 mm. Seedlings from the seeds
Table 6. Number of leaves per plant
TREATMENT
NUMBER OF LEAVES
Control (no cold storage)
3a
One week cold storage
3.83a
Two weeks cold storage
3a
Three weeks cold storage
3.33a
Four weeks cold storage
3.50a
Five weeks cold storage
3.17a
Six weeks cold storage
2.83a
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
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Table 7. Root length
TREATMENT
ROOT LENGTH (mm)
Control (no cold storage)
14.50b
One week cold storage
18.83a
Two weeks cold storage
9c
Three weeks cold storage
9c
Four weeks cold storage
8.83c
Five weeks cold storage
10.17c
Six weeks cold storage
9c
Means with the same letters are not significantly different at level by DMRT
subjected to four weeks duration of cold storage had the shortest roots with a mean of
8.83 mm which is not statistically different with the root length of seedlings resulting
from seeds subjected to two, three, five and six weeks cold storage.
Results showed that subjecting seeds to shorter duration of cold storage for a
period of one week induce the production of roots. Meyer and Anderson (1952) explained
that optimum temperature is usually about midway between two extremes of temperature
at which germination will occur. It is not possible to designate any exact varies with the
other prevailing environmental conditions and also with the exact criterion selected as an
index of germination. The most favorable temperature for the elongation of the primary
roots for example does not always correspond to the most suitable temperature for the
development of the plumule.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
21
Number of Roots
Table 8 shows that there were no significant differences on the number of roots as
affected by different cold storage duration. Nevertheless, numerical figures suggest that
seedlings coming from seeds subjected to one week cold storage had the highest number
of roots with a mean of 4.17. Seedlings treated with four weeks duration of cold storage
had an average number of roots of 3.83 while seedlings subjected to two, three and five
weeks cold storage had the same means of 3.50. Seedlings subjected to six weeks
duration of cold storage produced the least number of roots with a mean of 2.83.
Figure 3 shows uprooted seedlings during gathering of data regarding root length
and root number 113 days from sowing.
Table 8. Number of roots
TREATMENT
NUMBER OF ROOTS
Control (no cold storage)
2.50a
One week cold storage
4.17a
Two weeks cold storage
3.50a
Three weeks cold storage
3.50a
Four weeks cold storage
3.83a
Five weeks cold storage
3.50a
Six weeks cold storage
2.83a
Means with the same letters are not significantly different at level by DMRT
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
22
Figure 3. Uprooted seedlings of blueberry during gathering of data 113 days from sowing
Seedling Vigor Index
The results in Table 9 shows no significant statistical differences observed among
the different treatments used in the study affecting the seedling vigor of blueberry
seedlings. However, numerical results shows that seedlings coming from the seeds
subjected to one week cold storage promoted the excellent growth of seedlings.
Tocnang (2009) observed that guapple seed subjected to one week and two weeks
cold storage promoted excellent growth of seedlings. Likewise, Banagen (2008) noted
that storing coffee seeds under cold temperature influences 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. Figure 4 shows the overview of the experiment 113 days from
sowing the seeds.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
23
Table 9. Seedling vigor index
TREATMENT
SEEDLING VIGOR
Control (no cold storage)
2a
One week cold storage
1.67a
Two weeks cold storage
2.33a
Three weeks cold storage
2.33a
Four weeks cold storage
2.33a
Five weeks cold storage
2.33a
Six weeks cold storage
2.33a
Rating: 1= most vigorous; 2= vigorous; 3= less vigorous
Figure 4. Overview of the experiment 113 days from sowing seeds.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
24
SUMMARY, CONCLUSION AND RECOMMENDATION
Summary
The germination and seedling growth of blueberry seeds as affected by different
cold storage duration was studied at the Pomology Project Nursery, Benguet State
University, La Trinidad, Benguet from November 15,2009 to March 7, 2010. the
blueberry seeds were subjected to low temperature with an average of 5 °C at various
periods from one to six weeks.
Result showed that there were significant statistical differences observed among
the various cold storage duration used. One week cold storage of blueberry seeds showed
the best performance among all treatments that enhanced earlier seedling emergence with
a mean of 14.67 days and shorter days to develop true leaves with a mean of 20.33 days.
Likewise, the same duration of cold storage attained the highest percentage of seedling
emergence having a mean of 91.33 % and also with the percentage of normal seedlings
with a mean of 84.67 %. The seedlings coming from the seeds subjected to one week cold
storage had also produced longer root length with a mean of 18.83 mm which is highly
significant to the other treatments.
With regards to the seedling height, number of leaves, number of roots and
seedling vigor statistical results shows that there were no significant differences among
the different cold storage duration used however numerical results shows that seedlings
coming from the seeds stored for a period of one week had the tallest seedlings, more
number of leaves and roots, and had more vigorous seedlings as compared to other
treatments.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
25
Conclusion
Based on the results of the study, subjecting blueberry seeds to cold storage for
one week before sowing showed promising results. It was found out that it was the best
among the various cold storage durations studied in enhancing faster germination and
emergence of blueberry seeds.
Recommendation
From the preceding results, it is recommended that cold storage of blueberry
seeds for a period of one week at 5 °C prior to sowing should be done in order to
enhance faster seed germination and emergence as well as to promote longer root
development and enhanced faster and vigorous seedling growth. However, further study
along this line also recommended to verify these findings.
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
26
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Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
28
APPENDICES
Appendix Table 1. Number of days from sowing to seedling emergence
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 22 24 21 67 22.33
T2 16 14 14 44 14.67
T3 23 19 24 66 22
T4 14 20 19 53 17.67
T5 19 19 21 59 19.67
T6 18 16 21 55 18.33
T7 21 22 23 66 22
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Between 6 145.905 24.317 5.738 2.85 4.46
Within 14 59.333 4.238
Total 20 205.238
**Highly significant
Coefficient of variation = 10.54%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
29
Appendix Table 2. Number of days from sowing to first appearance of leaves
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 27 30 26 83 27.67
T2 22 19 20 61 20.33
T3 28 25 30 83 27.67
T4 21 26 25 72 24
T5 24 24 28 76 25.33
T6 26 25 30 81 27
T7 27 29 31 87 29
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 155.905 25.984 5.006 2.85 4.4
Error 14 72.667 5.190
Total 20 228.571
**Highly significant
Coefficient of variation = 8.81%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
30
Appendix Table 3. Percentage of seedling emergence
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 96 76 92 264 88
T2 92 94 88 274 91.33
T3 64 48 58 170 56.67
T4 82 60 70 212 70.67
T5 72 74 56 202 67.33
T6 66 54 50 170 56,67
T7 54 50 42 146 48.67
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 4704.476 784.079 10.468 2.85 4.46
Error 14 1048.667 74.905
Total 20 228.571
**Highly significant
Coefficient of variation = 12.65%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
31
Appendix Table 4. Percentage of normal seedlings
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 78 76 82 236 78.67
T2 84 86 84 254 84.67
T3 56 44 52 152 50.67
T4 72 54 60 186 62
T5 62 64 48 174 58
T6 58 46 44 148 49.33
T7 46 40 38 124 41.33
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 4533.333 755.556 18.711 2.85 4.46
Error 14 565.333 40.381
Total 20 5098.667
**Highly significant
Coefficient of variation = 10.47%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
32
Appendix Table 5. Seedling height
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 4.5 4.5 6 15 5
T2 6 8 9 23 7.67
T3 5 5.5 5 15.5 5.17
T4 5 4.5 4.5 14 4.67
T5 4.5 7 5 16.5 5.5
T6 5.5 7 4 16.5 5.5
T7 5 6 4 15 5
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 2.143 0.357 2.55 2.85 4.46
Error 14 2.167 0.155
Total 20 4.310
Not significant
Coefficient of variation = 12.15%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
33
Appendix Table 6. Number of leaves per plant
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 3 3 3 9 3
T2 4 4 3.5 11.5 3.83
T3 3 3.5 2.5 9 3
T4 3.5 3 3.5 10 3.33
T5 4 3.5 3 10.5 3.5
T6 3.5 3.5 2.5 9.5 3.17
T7 5 3 2.5 8.5 2.83
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 18.000 3.000 2.55 2.85 4.46
Error 14 16.500 1.179
Total 20 5098.667
Not significant
Coefficient of variation = 19.74%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
34
Appendix Table 7. Root length
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 18.5 11.5 13.5 43.5 14.5
T2 15.5 18.5 22.5 56.5 18.83
T3 9 10 8 27 9
T4 10 8.5 8.5 27 9
T5 10.5 8 8 26.5 8.83
T6 9.5 11.5 9.5 30.5 10.17
T7 8 8.5 10.5 27 9
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 270.667 45.111 9.792 2.85 4.46
Error 14 64.500 4.607
Total 20 335.167
** Highly significant
Coefficient of variation = 18.94%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
35
Appendix Table 8 . Number of roots
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 2.5 2 3 7.5 2.5
T2 4.5 4 4 12.5 4.17
T3 4 3 3.5 10.5 3.5
T4 4 3 3.5 10.5 3.5
T5 2.5 4.5 4.5 11.5 3.83
T6 4.5 4 2 10.5 3.5
T7 3 3 2.5 8.5 2.83
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 5.810 0.968 1.69 2.85 4.46
Error 14 8.000 0.571
Total 20 13.810
Not significant
Coefficient of variation = 22.20%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
36
Appendix Table 9. Seedling vigor
REPLICATION
TREATMENT I II III TOTAL MEAN
T1 2 2 2 6 2
T2 2 2 1 5 1.67
T3 3 2 2 7 2.33
T4 2 2 3 7 2.33
T5 3 2 2 7 2.33
T6 2 2 3 7 2.33
T7 2 2 3 7 2.33
ANALYSIS OF VARIANCE
SOURCE DEGREES SUM OF MEAN COMPUTED TABULAR F
OF OF SQUARES SQUARE F 0.05 0.01
VARIANCE FREEDOM
Treatment 6 1.238 0.206 0.72 2.85 4.46
Error 14 4.000 0.286
Total 20 5.238
Not significant
Coefficient of variation = 24.40%
Seed Germination and Seedling Growth of Blueberry (Vacinium corymbossum) as Affected by
Different Cold Storage Duration / Gabby K. Dao-ines. 2010
Document Outline
- Seed Germination and Seedling Growth ofBlueberry (Vacinium corymbossum) as Affected by Different Cold Storage Duration.
- BIBLIOGRAPHY
- ABSTRACT
- TABLE OF CONTENTS
- INTRODUCTION
- REVIEW OF LITERATURE
- Description of the Plant
- Seed Germination
- Seed Storage
- Methods of Breaking Dormancy
- Effect of Cold Storage Temperature on Seed Germination
- MATERIALS AND METHODS
- RESULTS AND DISCUSSION
- Number of Days from Sowing to Emergence
- Number of Days from Sowingto First Appearance of Leaves
- Percentage of Seedling Emergence
- Percentage of Normal Seedlings
- Seedling Height
- Number of Leaves per Plant
- Root Length
- Number of Roots
- Seedling Vigor Index
- SUMMARY, CONCLUSION AND RECOMMENDATION
- Summary
- Conclusion
- Recommendation
- LITERATURE CITED
- APPENDICES