COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

Rosa, S.D.V.F. da, Carvalho, A.M., McDonald, M.B., Von Pinho, E.R.V., Silva, A.P. and Veiga, A.D.
(2011), Seed Sci. & Technol., 39, 151-164

The effect of storage conditions on coffee seed and seedling

quality
S.D.V.F. DA ROSA1, A.M. CARVALHO2, M.B. McDONALD3, E.R.V. VON PINHO2, A.P. SILVA2 AND
A.D. VEIGA2
1

2
3

Embrapa, PqEB, Av. W3 Norte (final), Ed. Sede, CEP 70770-901, Braslia, DF, Brazil
(E-mail: sttelaveiga@dag.ufla.br)
Universidade Federal de Lavras, CEP 37200-000, Lavras, MG, Brazil
Seed Biology Program, Department of Horticulture and Crop Science, Ohio State University, 2021 Coffey Rd,
Columbus, OH 43210, USA

(Accepted October 2010)

Summary
Obtaining commercially useful coffee seedlings is hindered by slow, uneven germination and low tolerance to
desiccation as well as reduced coffee seed longevity. Coffee seeds have been considered recalcitrant, orthodox
and even intermediate with varying results. Current recommendations suggest that coffee seeds can be safely
stored between 10-11% f.w.b. at 15C. However, after drying and storage, coffee seeds lose vigour, and seeds
stored after drying cannot be used for producing seedlings. Coffee seedling producers usually sow seeds
immediately or lightly dry them after harvest for a short storage period, if necessary. It is highly desirable
that seeds are stored safely to optimize coffee seedling production at the appropriate time and season with
ideal climatic conditions for planting in the field. The objective of this study was to determine the quality
of coffee seedlings produced from seeds stored with high, medium and low moisture levels under hermetic
conditions at 10 and 20C. Seed and seedling quality were assessed before and after nine months storage. Only
the germination of seeds harvested at the cherry stage, evaluated before and after the storage at 10C, was not
affected by moisture content, but these seeds lost vigour and did not produce suitable seedlings for planting
when stored for nine months. Seedlings produced in the nursery from seeds with 47 and 12% moisture content
performed the same as those from greenish-yellow seeds, but they produced a leaf area that was five times
smaller, a stem height three times shorter, and had 1.7 times fewer pairs of true leaves than coffee seedlings
produced from fresh seeds. Storage at 20C was not suitable for coffee seeds, especially those at 18% moisture
content whose quality declined drastically. These results suggest that coffee seeds do not tolerate desiccation and
that their storage behaviour classification should be reviewed.

Introduction
Coffee is a significant agricultural product of economic and social importance for Brazil
and is important in world agribusiness. Coffee trees are propagated using seedlings derived
from seeds. However slow and uneven germination and poor storage potential slows
coffee seedling development at the time and season when ideal climatic conditions exist
for planting in the field in the principal Brazilian coffee producing regions. Therefore,
it is highly desirable that coffee seeds be stored safely to optimize coffee seedling
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S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

production. The intermediate storage category has been proposed for coffee seeds based
on the fact that coffee seeds survived storage for about 10 months at 15C at 10/11%
(f.w.b.) moisture content and decreased germination was obtained with progressive
reductions in seed moisture content and storage temperature (Ellis et al., 1990; 1991).
However, the ideal moisture content for coffee seed conservation is still being studied.
In a comprehensive review of literature from the 1920s to the present, more than
60 papers have provided contradictory information regarding storing wet or dry coffee
seeds. The results are inconsistent by showing that ideal storage moisture contents ranged
from wet (greater than 40% f.w.b.) to dry (10 to 12% f.w.b.), or even both conditions.
Studies conducted in the 1980s and 1990s have classified Coffea arabica L. seeds as
recalcitrant (King and Roberts, 1979), orthodox (Roberts et al., 1984) and intermediate
(Ellis et al., 1990). More recent studies have also shown inconsistent classifications
(Guimares et al., 2002; Dussert et al., 2006; Braghini and Fazuoli, 2007; Veiga et al.,
2007; Arajo et al., 2008).
For commercial coffee seedling production, Brazilian producers usually do not dry
the seeds to 10-11% moisture content. Instead, they immediately sow the seeds or only
lightly dry them after harvest to enable a short storage period. It is recommended that
coffee seeds be harvested at the cherry maturation stage. However, it has been reported
that coffee seeds acquire maximum germination capacity when the fruits are between
the yellow-green and cherry stages (Ellis et al., 1991; Guimares et al., 2002). Several
authors (Finch-Savage, 1996; Farrant et al., 1988; Pammenter and Berjak, 1999) have
suggested that completely mature seeds of species sensitive to desiccation may not possess
maximum vigour because the germination process may start at the end of the maturation
process.
The objective of this study was to evaluate the effects of seed moisture content and
storage temperature on coffee seed and seedling quality and to determine the seeding
production potential of seeds stored at high, intermediate and low moisture content.

Material and methods

Seed harvesting and processing
Coffea arabica seeds, cultivar Rubi, were harvested from fruits at the cherry and
greenish-yellow maturation stages in an experimental field at the Federal University of
Lavras, MG, Brazil, in the 2006/2007 growing season. Fruits were pulped in a hand pulp
separator and the mucilage removed by natural fermentation in water at 30C for 24 h,
washed in running water, and left on an absorbent paper towel to remove excess surface
water before drying. Seeds were dried in a small-scale stationary dryer (Navratil and
Burris, 1982) with 12.5m3 min-1 ton-1 air flow at 30C (i.e. slow drying; 0.15%. h-1) to
obtain three levels of moisture (47, 18 and 12% [f.w.b.]). Based on previous studies,
these moisture contents were optimum for determining the effects of high, intermediate
and low moisture contents on seed and seedling quality. Seed moisture content was
determined before storage on seeds without parchments (i.e after removal of the endocarp,
the remnant of the fruit) in a chamber at 105C 3C according to the Rules for Seed
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COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

Analysis (Brazil, 1992). Seeds were stored with their parchment in waterproof packages
at 10C and 20C for nine months to test the potential of the seeds to produce coffee
seedlings in a favorable climatic season for planting in the field since six months are
necessary for coffee tree formation.
Seed quality evaluation
Seed physiological quality was assessed before and after the storage period when the
standard germination test was used for determining the percentage radicle protrusion,
standard germination (normal seedlings), and seedlings with unfolded cotyledonary
leaves. The standard germination test was carried out with four subsamples of 50 seeds
each without the outer skin, spread on moist germination test paper with a quantity of
water equivalent to two and a half times the weight of the dry substrate, at 30C with
fluorescent white light at 750lux. Standard germination percentage was assessed 30
days after sowing following the Rules for Seed Analysis (Brazil, 1992). Physiological
germination was assessed at 15 days after the start of the germination test when seeds
with at least 2.0 mm root protrusion were counted and the results expressed in percentage.
Unfolded cotyledonary leaves (jaguar ear stage) were assessed 45 days after the start of
the germination test and the results expressed in percentage.
Seedling production
After the seed storage period, coffee seedling production was tested in a nursery under
ambient environmental conditions. Seeds were sown with attached parchments as in
commercial seedling production in 11 22 cm perforated polyethylene bags with standard
substrate (CFSMG, 1999) consisting of soil, manure, simple super phosphate, and
potassium chloride at a ratio of 70, 30, 0.5 and 0.1%, respectively. After purging the
substrate with methyl bromide (Guimares and Mendes, 1998) and preparing the bags,
two seeds were sown directly per bag and covered with 2 cm substrate. Each experimental
plot consisted of 20 seedlings that were watered daily and appropriate crop treatments and
weeding were carried out whenever necessary.
Seedling quality evaluation
The quality of coffee seedlings produced from the stored seed was assessed in the nursery
by the following: emergence percentage (final stand), emergence speed index, and seedling
developmental parameters (number of pairs of true leaves, stem diameter, seedling height,
leaf area, and the root and canopy dry matter). The total number of emerged seedlings at
120 days after sowing in each experimental plot of 20 coffee seedlings at the jaguar ear
stage was expressed as a percentage. The speed of emergence index was calculated using
data from the daily count of emerged seedlings according to the formula by Edmond
and Drapala (1958). Coffee seedling developmental parameters were determined for
six coffee seedlings in each experimental plot and the following measurements taken
180 days after sowing. The pairs of true leaves were calculated and the mean pair of
true leaves determined. Stem diameters were measured in the plant culm region using
a pachymeter and the results expressed in millimeters per plant. Seedling height was
obtained by measuring the region between the culm and the point of the terminal sprouts
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S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

of the coffee seedlings on the orthotropic branch. Seedling height means were calculated
and the results expressed in centimeters per seedling. Length and width of one leaf of
each pair of true leaves were measured and the leaf area of each pair of leaves obtained
multiplying the width length 0.667 2 (pair of leaves) proposed by Barros et al.
(1973). Coffee seedling leaf area was obtained by the sum of the area of the pairs of
leaves of each coffee seedling. Average leaf area was obtained from the leaf areas values
of the coffee seedlings in the experimental plot and the results expressed in cm2. Root and
canopy dry weight determinations for the coffee seedlings were obtained by removing the
coffee seedlings from the bags, washing the root system in running water and the roots
were separated from the canopy by cutting the stem at the culm; the weights of the root
system and canopy were obtained after drying in a forced air chamber at 60C to constant
weight and the mean results expressed in g per coffee seedling.
Statistical design
A complete randomized block design was used to assess laboratory seed quality with four
replications and a randomized block design was used with three replications to assess
nursery coffee seedling quality. A two (maturation stages) three (seed moisture content)
two (storage temperature) factorial scheme was used for a total of 12 treatments.
The mean results of the assessments were compared by the Scott and Knott test at 5%
probability by transforming the percentage data using [(arc sen X100-1/2) + 0.5]. The
analyses were carried out using the Sisvar program (Ferreira, 2000).

Results
Seed quality evaluation results
After harvest and drying, the maturation stage had a significant effect on the standard
germination; no significant effects were observed for physiological germination, moisture
content or interaction among these factors. The effect of maturation stage on the percentage
of standard germination was significant for the 18 and 47% moisture contents (figure 1),
but not for the 12% moisture content.
After nine months storage, however, the interaction between maturation stage, seed
moisture content, and storage temperature was significant for all seed physiological
quality assessment variables. With respect to fruit maturation stage (figure 2), significant
differences were detected in all assessments, except for the physiological germination
(radicle protrusion) values of 47% moisture content and open cotyledonary leaves at 18
and 12% moisture content. Seeds at the greenish-yellow maturation stage had poorer
physiological performances than those at the cherry maturation stage and the greatest
reductions in seed quality were observed in seeds stored at 18% moisture content at 20C.
Considering storage temperature, seeds of the same maturity level stored at 12%
moisture content had the same physiological performance (table 1). The physiological
performance of seeds stored at 18% moisture content decreased when stored at 20C
at both maturation stages. Among seeds stored at 47% moisture content, there were no
differences in greenish-yellow seed quality following storage at the two temperatures, but
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COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

PG
100

PG

SG
A

PG

SG

SG
a

90

80

(%)

70
60
50
40
30
20
10
0

12

18

47

Water Content (%)

Figure 1. Comparisons between cherry ( ) and greenish-yellow ( ) maturity stages within each moisture
contents. Seed physiological quality was assessed by physiological germination ( ) and standard germination
( ), after drying of the seeds to different moisture contents (12, 18 and 47%). The results are from one-way
ANOVAs. Lower cases are used for comparing physiological germination (PG) values and upper ones for
standard germination (SG). Averages with equal case letters are not significantly different at P = 0.05 according
to the Scott Knott test. Embrapa/UFLA/OSU, 2008.

seeds at the cherry maturation stage had lower standard germination and percentage open
cotyledonary leaves when stored at 20C. Coffee seed performance was affected very
little by moisture content when seeds were stored at 10C, regardless of the maturation
stage (table 1). However, at 20C, both ripe (cherry) and non-ripe (greenish-yellow)
seeds showed marked reductions in physiological quality when stored at 18% moisture
content.
Table 1. Assessment results of the Coffea arabica L. seeds obtained from cherry fruits and greenish-yellow
fruits dried to different moisture contents, and stored at different temperatures during nine months. The results
are from three-way ANOVAs. Averages showing the same letters were not significantly different at P = 0.05
according to the Scott Knott test. UFLA, Lavras, MG, 2007. Embrapa / UFLA / OSU, 2008.
Cherry

Moisture content

Greenish-Yellow

10C

20C

10C

20C

93 Aa
92 Aa
88 Aa

90 Aa
36 Bb
91 Aa

81 Ba
77 Ba
86 Aa

81 Ba
6 Cb
91 Aa

86 Aa
90 Aa
91 Aa

90 Aa
40 Cb
58 Bb

81 Aa
77 Aa
77 Aa

76 Ba
11 Cb
87 Aa

69 Ba
67 Ba
81 Aa

60 Aa
5 Cb
48 Bb

72 Aa
72 Aa
67 Aa

63 Aa
2 Bb
72 Aa

Physiological germination
12
18
47
Standard germination
12
18
47
Open Cotyledonary Leaves
12
18
47

Capital letters are used for comparisons within columns and lower cases letters within lines.

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Radicle Protrusion (%)

S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

10oC

120
100

20oC

10oC

20oC

10oC

80

20oC

60

40
20

12

18

47

Water Content (%)

20oC

120

10oC

Viability (%)

100

10oC

10oC
a

20oC

a
b

80

20oC
60

40

20
0

12

18

47

Op en Co tyl ed o n ary L eaves (% )

Water Content (%)

100
80

10oC
a

20 C

a
A

10oC

10oC
a
a

20oC

a
b

60

40

20oC
20

12

18

47

-20

Water Content (%)

Figure 2. Comparisons between cherry ( , ) and greenish-yellow ( , ) maturity stages within each moisture
contents (12, 18 and 47%) and storage temperature (10 and 20C). Seed physiological quality was assessed after
nine months of storage, and the results are from three-way ANOVAs. Lower cases are used for comparing
within 10C and upper ones within 20C. Averages with equal case letters are not significantly different at
P = 0.05 according to the Scott Knott test. Embrapa/UFLA/OSU, 2008.

Generally, when seeds were dried to 12% moisture content, the maturation stage and
storage temperature had more influence on physiological quality. However, when seeds
were stored at 47% moisture content at 20C, reductions in seed quality were observed
only for the cherry maturation stage.
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COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

Seedling quality assessment

Seeds harvested at the cherry and greenish-yellow maturation stages and dried to different
moisture contents followed by storage at 10C and 20C were tested for their capacity
to produce coffee seedlings that were capable of propagating coffee plants. Generally,
seed moisture content followed by storage temperature, were the most important factors
that influenced coffee seedling quality whereas the fruit maturation stage least influenced
quality. There was a triple interaction of factors that influenced final stand and speed of
emergence index of seedlings produced in the nursery, and also an interaction between
seed maturity and storage temperature for the other variables in the assessment of the
quality of coffee seedlings.
The best performance as assessed by the smallest emergence speed index values
calculated according to the formula by Edmond and Drapala (1958) were from coffee
seedlings derived from greenish-yellow seed at all the storage moisture contents and
temperatures (figure 3). Forty days after sowing, coffee seedlings derived from seeds at
20oC

Final Emergency (%)

10oC
100
90

10oC

10oC
a
a

80

20oC
A

20oC

70
60

50
40

30
20
10
0

12

18

47

Emergence speed index

Water Content (%)

80

60
50

20oC

10oC

70

A
b

20oC
A

10oC
a

20oC

10oC

a
b
b

40

30
20
10
0

12

18

47

Water Content (%)

Figure. 3 Comparisons between cherry ( ,
) and greenish-yellow ( ,
) maturity stages within each
moisture contents (12, 18 and 47%) and storage temperature (10 and 20C). Seedlings were produced in the
nursery from seeds stored for nine months. The results of seedling physiological quality are from three-way
ANOVAs. Lower cases are used for comparing within 10C and upper ones within 20C. Averages with equal
case letters are not significantly different at P = 0.05 according to the Scott Knott test. Embrapa/UFLA/OSU,
2008.

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S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

the greenish-yellow maturation stage and stored at 47% moisture content had produced
open cotyledonary leaves (data not shown) that normally occurs at 90 days after sowing.
Although these seedlings had higher emergence speed indices at the end of planting period,
coffee seedlings derived from greenish-yellow seeds had final emergence percentages
similar to those derived from cherry stage seeds (figure 3). There was a significant
difference only in coffee seedlings derived from greenish-yellow seeds stored at 18%
moisture content at 20C that resulted in the poorest performance.
Coffee seedlings produced from cherry or greenish-yellow fruit seeds stored at 18%
moisture content at 20C produced the poorest quality seedlings and this finding was
consistent with the seed quality assessments in the laboratory.
There was a significant interaction between seed moisture content and storage
temperature for all other variables for coffee seedling quality assessment, but no
significant effect was observed for fruit maturation stage. Seed moisture content and
storage temperature had less impact on coffee seedling production from seed stored at
18% moisture content at 20C (table 2). In contrast, coffee seedlings produced from seeds
harvested at the greenish-yellow or cherry stage stored at 12 or 47% moisture content did
not have differences in seed quality following storage at either 10C or 20C.
Poorer physiological quality was observed in most of the treatments in coffee seedlings
stored at 18% moisture content. Seeds stored at 18% moisture content at 20C for both
maturation stages consistently gave poor results. These results are further supported at 85
days, when the final stand had already been established and most seedlings already had
unfolded cotyledonary leaves (data not presented). The best seedling performance was
from seedlings derived from seeds at the cherry maturation stage at 47% moisture content
and stored at 10C.

Discussion
Slow and uneven germination of coffee seeds greatly hinders obtaining high quality
coffee seedlings to form a coffee plantation. Furthermore, in the main Brazilian coffee
producing regions, temperatures are in the 15 to 19C range during the seed germination
and seedling emergence period causing complete seedling emergence to take up to 120
days (Guimares and Mendes, 1998). The ideal temperature for coffee seed germination
is 30C (Brazil, 2009). However, this temperature results in coffee seedling damage that
is already a slow process, taking at least six months to be concluded. Thus, in addition
to germination capacity, it is highly desirable that coffee seeds have a high vigour index
that leads to a better performance during germination and coffee seedling formation in
the nursery, especially under adverse conditions, so that more vigorous seedlings are
obtained. Under these conditions, vigorous coffee seedlings have faster establishment in a
productive plantation and a faster return for the producer in invested capital.
The results presented here show that coffee seeds can germinate when they are dried
to 12% moisture content, an intermediate moisture level among the levels tolerated by
recalcitrant and orthodox seeds. After nine months storage they still have the ability to
germinate. Although the fruit maturation stage and seed moisture content after drying
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Table 2 Assessment results of the Coffea arabica L. seedlings obtained from fruits dried to different moisture
contents, and stored at different temperatures during nine months. The results are from two-way ANOVAs.
Averages showing the same letters were not significantly different at P = 0.05 according to the Scott Knott test.
Data are the average SE from six replications. UFLA, Lavras, MG, 2007. Embrapa / UFLA / OSU, 2008.
Storage
Temperature
(C)

12

18

10C

2.6 Aa

Pairs of true leaves

2.5 Aa

20C

2.8 Aa

10C

Water content (%)

P
47

2.9 Aa

0.5752

0.8 Bb

2.8 Aa

<0.0000

1.57 Aa

Stem diameter
1.59 Aa

1.50 Aa

0.6220

20C

1.59 Aa

0.70 Bb

1.67 Aa

<0.0000

10C

5.40 Aa

Seedling height
5.35 Aa

5.21 Aa

0.9329

20C

5.40 Aa

2.10 Bb

5.24 Aa

<0.0000

10C

0.98 Aa

Root dry matter

0.88 Aa

1.00 Aa

0.6885

20C

1.10 Aa

0.18 Bb

0.98 Aa

<0.0000

10C

2.23 Aa

Canopy dry matter

2.10 Aa

2.13 Aa

0.9688

20C

2.46 Aa

0.64 Bb

2.32 Aa

0.0027

10C

44.11 Aa

Leafy area
45.58 Aa

44.37 Aa

0.9889

20C

51.55 Aa

12.36 Bb

49.69 Aa

0.0010

Lower cases are used for comparisons within lines and capital letters within columns.

had significant effects on seed quality, seeds submitted to the different treatments
had physiological germination (radicle protrusion) and standard germination (normal
seedlings) values similar to each other and greater than 80%. Seeds harvested at the cherry
maturation stage had statistically equal germination percentage at all moisture contents,
indicating that coffee seeds tolerate drying to 12% moisture content, supporting the results
reported by others (Ellis et al., 1990 and 1991; Hong and Ellis, 1992 and 1995; Dussert
et al., 1999; Eira et al., 1999; Dussert et al., 2006). However, it is important to note that
in all the former studies to define coffee seed response following drying and storage,
seeds were tested without the parchment layer under laboratory conditions. These results
failed to determine the potential of dry stored seeds for producing seedlings under nursery
conditions as done in this study.
In addition to slow and uneven coffee seed germination characteristics, in commercial
practice, coffee seeds are planted with the parchment because there is no safe method
for its removal without adversely affecting seed quality. That is the reason why coffee
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S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

seeds were planted in this study with the parchment attached to evaluate their potential
for producing seedlings in a nursery. The outer skin is a mechanical barrier to seed
germination that makes coffee seedling development even more difficult (Vlio, 1976;
Rena and Maestri, 1986; Guimares, 1995; Rosa et al. 2002 or 2005). Ellis et al. (1990)
also indicated that not all planted seeds are able to emerge from the soil surface and grow,
and some have just their radicles emerged but remain under the soil.
Although seeds germinated after drying and after nine months storage, they had low
vigour index values that reduced the capacity to form coffee seedlings. Coffee seeds with
high vigour and storage capacity for at least nine or ten months are highly desirable because
the germination and emergence processes can start under more favorable temperature
conditions and successful coffee seedlings can be obtained in a climate season more
suitable for planting from September to November. Obtaining coffee seedlings at the start
of October, as long as they have commercialization standards, with three to seven pairs
of true leaves, increases the likelihood of successful coffee seedling production because
the climatic conditions are more favorable to plant the coffee seedling in the field during
this period.
Unfortunately, storing coffee seeds at intermediate moisture content (18%) reduced
physiological quality, especially when the seeds were stored at 20C. Respiration damage
from free radicals can occur at an intermediate moisture content level (between -15 and
-5 MPa), but the injury repair process is not activated. Seeds stored or submitted to
slow drying so they remain at an intermediate moisture content level for a long period
of time deteriorate in a similar fashion to the physiological mechanisms associated with
the accelerated aging test (Walters et al., 2001; Leprince et al., 2000). Dussert et al.
(2006) showed that, although slow drying did not directly influence coffee seed viability,
it induced oxidative injury and deterioration processes in the seed because the seeds
remained at an intermediate moisture content level.
All the treatments, except for the seeds with 18% moisture content stored at 20C,
produced coffee seedlings with statistically equal developmental parameters. Coffee
seedlings produced in the present study, including those derived from seeds processed
and stored according to the recommendations by Hong and Ellis (1992, 1995), i.e., 1011% (f.w.b.) moisture content and 10-15C storage temperature, did not produce coffee
seedlings suitable for planting. Although these coffee seedlings were produced during
the summer under higher temperatures, they had characteristics very different from
seedlings produced during the winter, the usual season for commercial coffee seedling
production. Coffee seedlings produced in this study had, on average, a leaf area five
times smaller, stem height three times shorter, and 1.7 times fewer pairs of true leaves
than those produced in the winter (Rosa et al., 2007). These results show that even though
coffee seeds germinated after drying, their vigour decreased during storage. These results
confirm that the ideal approach to processing and storing coffee seeds in the wet or dry
condition still remains to be determined.
Desiccation-tolerant seeds whose longevity is enhanced by reduced moisture content,
temperature, and decreased relative humidity have desiccation tolerance mechanisms such
as late embryonic abundant protein synthesis and accumulation (lea or lea like) (Walters
et al., 1997; Wolkers et al., 2001) and sugars such as sucrose and some oligosaccharides
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COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

(Obendorf, 1997; Buitink et al., 2000; Oliver et al., 2001; Buitink et al., 2002; Buitink
and Leprince, 2004) involved in the formation of aqueous glasses, as well as the presence
of enzymatic and non-enzymatic anti-oxidant systems (Berjak, 2006). These mechanisms
should be completely expressed and activated in the locations where they are required to
protect against free radical attack (Berjak, 2006). Sugar accumulation (Chabrillange et al.,
2000; Guimares et al., 2002) and cytoplasmatic glass formation (data being prepared for
publication) do not appear to exist in coffee seeds following drying and storage. These
data indicate that desiccation tolerance mechanisms in coffee may not associated with
water removal as occurs in desiccation-tolerant seeds (Bewley and Black, 1994; Leprince
et al., 1993).
It has been suggested that the reduction in orthodox seed moisture content during
maturation drying elicits a switch from a developmental to a germinative mode as shown
in changes in genome activity resulting in the synthesis of germination-specific enzymes
(Evans et al., 1975; Dasgupta and Bewley, 1982; Armstrong et al., 1982; Dasgupta et al.,
1982; Kermode and Bewley, 1985; Misra et al., 1985; Kermode et al., 1989; Bewley et
al., 1989; Oishi and Bewley, 1990; Burris et al., 1997).
Seeds of other species, which develop within a fleshy fruit and frequently do not
dry out prior to dispersal, can germinate when removed from the fruit without any
requirement for drying. Thus,removal of the seeds from the constraints of the fruit is
sufficient for germination (Bewley and Black, 1994). In this study, coffee seeds harvested
in the greenish-yellow maturation stage, germinated as they were removed from the fruits
without any drying treatment indicating that drying is not a requirement for germination.
Recent research concerning the effects of post-harvesting processing of coffee fruits
(Selmar et al., 2004; Selmar et al., 2006; Bytof et al., 2007) show that in the course
of processing, after the fruit flesh is removed, the seeds have the germination process
initiated on the basis of the expression of the germination-specific isocitrate lyase and an
increase in the abundance of -tubulin, a maker for cell division or elongation.
Other studies on coffee seed performance during drying and storage and in wet and
dry conditions have not shown an interaction among physiological and pathological
effects on seed quality. Because coffee seeds germinate slowly and have low storage
potential, it is important to consider their state of health, because interactions among
the physiological and pathological parameters can lead to erroneous interpretations and
contradictory results. It is known that this interaction can influence the assessments of
seed performance sensitive to desiccation during storage, especially when they are stored
under wet conditions (Mycock and Berjak, 1995). The influence of this interaction on the
assessment of seed quality was observed in this study. Coffee seeds derived from fruits at
the greenish-yellow stage after nine months storage performed better than those extracted
at the cherry stage. Figure 3 shows enhanced speed of emergence indices and final stand
percentage for greenish-yellow seeds compared to cherry seeds. In other assessments
of coffee seedling performance, differences were not observed between the greenishyellow and cherry seeds, and only the effects of moisture content and storage temperature
prevailed. This better performance of seedlings produced from greenish-yellow seeds
is also observed for seedlings from seeds stored at 47% moisture content at 20C. The
pathological quality of the seeds was not determined, but greenish-yellow seeds were
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S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

visibly less infected by fungi than seeds at the cherry stage and this may explain their
enhanced performance when stored at 20C due to the better pathological health quality.
Thus, according to the data presented and discussed here, dry stored coffee seeds do not
produce suitable seedlings for planting and the statement they are tolerant to desiccation
to 10-11% moisture content should be reviewed.

References
Armstrong, C., Black, M., Chapman, J.M., Norman, H.E. and Angold, R. (1982). The induction of sensivity to
gibberellin in aleuronic tissue of developing wheat grains. I. The effect of dehydration. Planta, 154, 573577.
Barros, R.S., Maestri, M. and Braga Filho, L.J. (1973). Determinao da rea de folhas de caf (Coffea arabica
L., cv. Bourbon amarelo). Revista Ceres, 20, 44-53.
Berjak, P. (2006). Unifying perspectives of some mechanisms basic to desiccation tolerance across life forms.
Seed Science Research, 16, 1-15.
Bewley, J.D. and Black, M. (1994). Seeds: physiology of development and germination. New York: London
Plenum Press, 367 p.
Bewley, J.D., Kermode, A.R. and Misra, S. (1989). Desiccation and minimal drying treatments of seeds of
castor bean and Phaseolus vulgaris which terminate development and promote germination cause changes in
protein and messenger RNA syntheses. Annals of Botany, 63, 3-17.
Brasil (1992). Ministrio da Agricultura e Reforma Agrria. Secretaria Nacional de Defesa Agropecuria. Regras
para Anlise de Sementes. Braslia, DF. 365 p.
Buitink, J. and Leprince, O. (2004). Glass formation in plant anyhdrobiotes: survival in the dry state. Cryobiology,
48, 215-228.
Buitink, J., Hemminga, M.A. and Hoekstra, F.A. (2000). Is there a role for oligosaccharides in seed longevity?
An assessment of intracellular glass stability. Plant Physiology, 122, 1217-1224.
Buitink, J., Hoekstra, F.A. and Leprince, O. (2002). Biochemistry and biophysics of tolerance systems. pp.
293-318 in Black, M.; Pritchard, H.W. (Eds) Desiccation and survival in plants: Drying without dying.
Wallingford, CABI Publishing.
Braghini, M.T. and Fazuoli, L.C. (2007). Armazenamento de sementes de caf. O Agronmico, 59, 44-45.
Burris, J.S., Peterson, J.M., Perdomo, A.J. and Feng, D.S. (1997). Morphological and physiological changes
associated with desiccation in maize embryos. In: Ellis, R.H.; Black, M.; Murdoch, A.J. and Hong, T.D.
Basic and applied aspects of seed biology. Dordrecht: Kluwer Academic Publishers 1997. 103-111.
Bytof, G., Knopp, S., Kramer, D., Breitenstein, B., Bergervoet, J.H., Groot, S.P.C. and Selmar, D. (2007).
Transient occurrence of seed germination processes during coffee post-treatment. Annals of Botany, 100,
61-66.
CFSMG-Comisso de Fertilidade do Solo do Estado de Minas Gerais (1999). Recomendaes para o uso de
corretivos e fertilizantes em Minas Gerais, 5 aproximao. Lavras, 1999. 359p.
Chabrillange, N., Dussert, S., Engelmann, F., Douldeau, S. and Hamon, S. (2000). Desiccation tolerance in
relation to soluble sugar contents in seeds of ten coffee (Coffea L.) species. Seed Science Research, 10,
393-396.
Dasgupta, J. and Bewley, J.D. (1982). Desiccation of axes of Phaseolus vulgasis during development causes
a switch from a developmental pattern of protein synthesis to a germination pattern. Plant Physiology, 70,
1224-1227.
Dasgupta, J., Bewley, J.D. and Yeung, E.C. (1982). Desiccation-tolerant and desiccation-intolerant stages during
the development and germination of Phaseolus vulgaris seeds. Journal of Experimental Botany, 33, 10451057.
Dussert, S., Chabrillange, N., Engelmann, F. and Hamon, S. (1999). Quantitative estimation of seed desiccation
sensitivity using a quantal response model: application to nine species of the genus Coffea L. Seed Science
Research, 9, 135-144.

162

COFFEE SEEDLING PRODUCTION FROM STORED SEEDS

Dussert, S., Davey, M.W., Laffargue, A., Doulbeau, S., Swennen, R. and Etienne, H. (2006). Oxidative stress,
phospholipids loss and lipid hydrolysis during drying and storage of intermediate seeds. Physiologia
Plantarum, 127, 192-204.
Edmond, J.B. and Drapala, W.S. (1958). The effects of temperature, sand and acerone on germination of okra
seed. Proceedings of American Society for Horticultural Science, 71, 428-434, June.
Eira, M.T.S., Walters, C., Caldas, L.S., Fazuoli, L.C., Sampaio, J.B. and Dias, M.C.L.L. (1999). Tolerance of
coffea spp. seeds to desiccation and low temperature. Revista Brasileira de Fisiologia Vegetal., 11, 97-105.
Ellis, R.H., Hong, T.D. and Roberts, E.H. (1990). An intermediate category of seed storage behavior? I. Coffee.
Journal of Experimental Botany, 41, 167-1174, Sept.
Ellis, R.H., Hong, T.D. and Roberts, E.H. (1991). An intermediate category of seed storage behaviour? II.
Effects if provenance, immaturity and imbibition on desiccation tolerance in coffee. Journal of Experimental
Botany, 42, 653-657, May.
Evans, M., Black, M. and Chapman, J.M. (1975). Induction of hormone sensivity by dehydration is one positive
role for drying in cereal seed. Nature, 258, 144-145.
Farrant, J.M., Pammenter, N.W. and Berjak, P. (1988). Recalcitrance a current assessment. Seed Science and
Technology, 16, 155-166.
Ferreira, D.F. (2000). Sistema de anlise estatstica- SISVAR. Lavras: DCE, UFLA.
Finch-Savage, W.E. (1996). The role of developmental studies in research on recalcitrant and intermediate seeds.
In: Workshop on Improved Methods for Handling and Storage of Intermediate/Recalcitrant Tropical Forest
Tree Seeds, 1995, Humlebaek, Denmark. Proceedings... Humlebaek, Denmark, 1996. p.83-97.
Guimares, R.J. (1995). Formao de mudas de cafeeiro: (Coffea arabida L.): Efeitos de reguladores de
crescimento e remoo do pergaminho na germinao de sementes e do uso de N e K em cobertura,
no desenvolvimento de mudas. 133p. Tese (Doutorado em Fitotecnia) Universidade Federal de Lavras.
Lavras.
Guimares, R.J. and Mendes, A.N.G. (1998). Produo de mudas de cafeeiro. In: Mendes, ANG, Guimares, RG
(eds). Cafeicultura empresarial - produtividade e qualidade. Lavras: UFLA/FAEPE, p.1-60.
Guimares, R.M., Vieira, M.G.G.C., Fraga, A.D., Von Pinho, E.V.R. and Ferraz, V.P. (2002). Tolerncia
dessecao em sementes de cafeeiro (Coffea arabica L.). Ciencia e Agrotecnologia, 26, 128-139.
Hong, T.D. and Ellis, R.H. (1992). Optimum air-dry seed storage environments for arabica coffee. Seed Science
and Technology, 20, 547-560.
Hong, T.D. and Ellis, R.H. (1995). Interespecific variation in seed storage behaviour within two genera Coffea
and Citrus. Seed Science and Technology, 23, 165-181.
Hong, T.D. and Ellis, R.H. (2002). Optimum moisture status for the exceptional survival of seeds of arabica
coffee (Coffea arabica L.) in medium-term storage at -20C. Seed Science Technology, 30, 131-136.
Kermode, A.R. and Bewley, J.D. (1985). The role of maturation drying in the transition from seed development
to germination. I. Acquisition of desiccation-tolerance and germinability during development of Ricinus
communis L. seeds. Journal of Experimental Botany, 36, 1906-1915.
Kermode, A.R., Oishi, M.Y. and Bewley, J.D. (1989). Regulatory roles for desiccation and abscisic acid in seed
development: a comparison of the evidence from whole seeds and isolated embryos. In: Stanwood, P. C. and
Mcdonald, M.B. (eds.). Seed Moisture. Madison: CSSA, 1989. p.23-50. (Special Publication, 14).
King, M.W. and Roberts, E.H. (1979). The storage of recalcitrant seeds: achievements and possible approaches.
International Board for Plant Genetic Resources, Rome.
Leprince O., Harren, F.J.M., Buitink J., Alberda, M. and Hoekstra, F.A. (2000). Metabolic dysfunction and
unabated respiration of germinating radicles. Plant Physiology, 112, 597-608.
Leprince, O., Hendry, G.A.F. and McKersie, B.D. (1993). The mechanims of deseccation tolerance in developing
seeds. Seed Science Research, 3, 231-246.
Misra, S., Kermode, A. and Bewley, J.D. (1985). Molecular form and function of the plant genome. New York:
Plenum, 1985. (NATO Advanced Studies Institute Proceedings). Maturation drying as the switch that
terminates seed development and promotes germination, p.113-128.
Mycock, D.J. and Berjak P. (1995). The implications of Seed-Associated Mycoflora During Storage. In: Kigel,
J. and Galili, G., ed. Seed Development and Germination. Boca Raton, NY: CRC Press, 747-766.
Navratil, R.J. and Burris, J.S. (1982). Small-scale dryer designer. Agronomy Journal, 74, 159-161.
Obendorf, R.L. (1997). Oligosaccharides and galactosyl cyclitols in seed desiccation tolerance. Seed Science
Research, 7, 63-74.

163

S.D.V.F. DA ROSA, A.M. CARVALHO, M.B. McDONALD, E.R.V. VON PINHO, A.P. SILVA AND A.D. VEIGA

Oishi, M.Y. and Bewley, J.D. (1990). Distinction between the responses of developing maize kernels to fluridone
and desiccation in relation to germinability, -amylase activity, and abscisic acid content. Plant Physiology,
94, 592-598.
Oliver , A.E., Leprince, O., Wolkers, W.F., Hincha, D.K., Heyer, A.G. and Crowe, J.H. (2001). Non-disaccharidebased mechanisms of protection during drying. Cryobiology, 44, 151-167.
Pammenter, N.W. and Berjak, P.A. (1999). Review of recalcitrant seed physiology in relation to dessication
tolerance mechanisms. Seed Science Research, 9, 13-37.
Rena, A.B. and Maestri, M. (1986). Fisiologia do Cafeeiro. In: Simpsio sobre fatores que afetam a produtividade
do cafeeiro, Poos de Caldas, 1986. Anais... Piracicaba: POTAFS, 13-85.
Roberts, E.H. (1973). Predicting the storage life of seeds. Seed Science and Technology, 1, 499-514.
Roberts, E.H., King, M.W. and Ellis, R.H. (1984). Recalcitrant seeds: their recognition and storage. In: Crop
genetic resources: conservation and evaluation. Eds. J.H.W. Holden and J.T. Williams. George Allen and
Unwim, London, pp.38-52.
Rosa, S.D.V.F., Brando Junior, D.S., Von Pinho, E.V.R., Veiga, A.D. and Silva, L.H.C. (2005). Effects of
different drying rates on the physiological quality of Coffea canephora Pierre seeds. Brazilian Journal Plant
Physiology, 17, 199-205.
Rosa, S.D.V.F., Melo, L.Q., Veiga, A.D., Oliveira, S., Souza, C.A.S. and Aguiar, V.A. (2007). Formao de
mudas de Coffea arabica L., cv Rubi, utilizando sementes e frutos em diferentes estdios de desenvolvimento.
Cincia e Agrotecnologia, 31, 349-356.
Selmar , D., Bytof, G., Knopp, S.E., Bradbury, A., Wilkens, J. and Becker, R. (2004) Biochemical insights into
coffee processing: quality and nature of green coffee are interconnected with an active seed metabolism. In:
20th International Conference in Coffee Science. Bangalore 2004, ASIC, India cd-room.
Selmar, D.Bytof, G., Knopp, S.-E. and Breitenstein, B. (2006). Germination of coffee seeds and its significance
for coffee quality. Plant Biology, 8, 260-264.
Valio, I.F.M. (1976). Germination of coffee seeds (Coffea arabica L., cv. Mundo Novo). Journal of Experimental
Botany, 27, 983-991.
Veiga, A.D., Guimares, R.M., Rosa, S.D.V.F. da, Von Pinho, E.V. de R, Silva, L.H. de C. e, Veiga, A.D.
(2007). Armazenabilidade de sementes de cafeeiro colhidas em diferentes estdios de maturao e submetidas
a diferentes mtodos de secagem. Revista Brasileira de Sementes, 29, 83-91.
Walters, C., Ried, J.L. and Walker-Simmons, M.K. (1997). Heat-soluble proteins extracted from wheat embryos
have tightly bound sugars and hydration properties. Seed Science Research, 7, 125-134.
Walters, W.F., McCready, S., Brandt, W.F., Lindsey, G. and Hoekstra, F.A. (2001). Isolation and characterization
of a D-7 LEA protein from pollen that stabilizes glasses in vitro. Biochimica et Biophysica Acta, 1544.
Wolkers, W.F., McCready, S., Brandt, W.F., Lindsey, G.C. and Hoekstra, F.A. (2001). Isolation and characterization of a D-7 LEA protein from pollen that stabilizes glasses in vitro. Biochimica et Biophysica Acta, 1544,
196-206.

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