Sugar apple (Annona squamosa L.
) seed germination
affected by the application of gibberellins
Germinación de semillas de anón (Annona squamosa L.)
afectada por la aplicación de giberelinas
Fabio Ernesto Martínez M.1, Diego Miranda L.2, and Stanislav Magnitskiy2
ABSTRACT
This research sought to establish the response of the germina-
tion percentage (PG), synchrony index (E), mean germination
time (MGT) and mean germination rate (MGR) of Annona
squamosa L. seeds from Apulo (province of Cundinamarca)
and Castilla (province of Tolima), Colombia, to treatments with
0, 50, 100, 200, 400, 600, or 800 mg L-1 of gibberellic acid (GA).
All of the treatments with GA increased the PG at each point of
time of seed incubation. The 600 mg L-1 GA treatment resulted
in higher PGs (92.3% at 16 days for Apulo and 95% at 24 days
for Castilla) and lower MGTs (8.75 and 5.38 days for Apulo and
Castilla, respectively) than those found with the concentration
of 0 mg L-1 GA (17.68 and 10.88 days for Apulo and Castilla,
respectively). Also, treating the seeds with 600 mg L-1 GA gen-
erated higher MGRs (0.18 and 0.12 germinated seeds/day for
Castilla and Apulo, respectively) than those obtained with 0
mg L-1 GA (Castilla = 0.09 and Apulo = 0.06 germinated seeds/
day). Likewise, the germination was synchronized with the
application of any concentration of GA. The results evidenced
a positive response to the GA application, which provided a
tool for the characterization of the phenomenon of dormancy
in the A. squamosa seeds.
Key words: germination percentage, mean germination rate,
synchrony index, mean germination time.
Introduction
The sugar apple or anon (Annona squamosa L.) is the
most widely distributed species of the Annona genus in
the world. In Colombia, it grows on the Atlantic Coast and
the dry areas of the valleys in the provinces of Valle, Cal-
das, Huila, Tolima, Cundinamarca, Meta, and Santander,
between 450 and 1,500 m a.s.l. (Buriticá and Cartagena,
2015). Although not yet officially exported, anon is a
promising fruit, with high potential for international
markets (Martínez, 2012).
Received for publication: 16 September, 2015. Accepted for publication: 28 March, 2016. Doi: 10.15446/agron.colomb.v34n1.53074
1
Tibaitata Research Center, Corporacion Colombiana de Investigacion Agropecuaria (Corpoica). Mosquera (Colombia). femartinez@corpoica.org.co
2
Department of Agronomy, Faculty of Agricultural Sciences, Universidad Nacional de Colombia. Bogota (Colombia)
RESUMEN
En esta investigación se buscó establecer la respuesta del
porcentaje de germinación (PG), índice de sincronía (E), el
tiempo medio de germinación (TMG) y la velocidad media de
germinación (MGR) de semillas de anón Annona squamosa
L. provenientes de Apulo (departamento de Cundinamarca) y
Castilla (departamento de Tolima), Colombia, a tratamientos
con 0, 50, 100 200, 400, 600 o 800 mg L-1 de ácido giberélico
(GA). Todos los tratamientos con GA incrementaron los PG
en cada punto del tiempo de incubación de las semillas. El
tratamiento 600 mg L-1 GA generó mayores PG (92,3%, a los
16 días para Apulo y 95% a los 24 días para Castilla), menores
TMG (8,75 y 5,38 días para Apulo y Castilla, respectivamente)
que los hallados con la concentración de 0 mg L-1 GA (17,68
días para Apulo y 10,88 días para Castilla) y mayores VMG
(0,18 y 0,12 semillas germinadas/día para Apulo y Castilla,
respectivamente) que las obtenidas con 0 mg L-1 GA (Castilla
= 0,09 y Apulo = 0,06 semillas germinadas/día). Así mismo, se
observó una germinación más sincronizada con la aplicación
de cualquier concentración de GA. Los resultados encontrados,
evidencian la respuesta positiva a la aplicación de GA, lo cual
brinda una herramienta en la caracterización del fenómeno de
latencia en semillas de A. squamosa.
Palabras clave: porcentaje de germinación, velocidad me-
dia de germinación, índice de sincronía, tiempo medio de
germinación.
The seeds of Annonaceae, within these A. squamosa, pos-
sess physical (impermeable seed coats) and morphological
dormancy (Lemos et al., 1988; Adeniji et al., 2014; Baskin
and Baskin, 2014). Morphological dormancy occurs when
a rudimentary embryo occupies less than 30% of the seed
volume at the time of seed dispersion, with an endosperm
(endospermic seeds) that fills most of the seed and a differ-
entiated embryo that needs specific conditions to complete
its growth before germination (Baskin and Baskin, 2014).
According to Pinto (2005), seed dormancy in Annonaceae
is due to the degree of seed maturity and, among other
Agronomía Colombiana 34(1), 17-24, 2016
factors, depends on climatic conditions during seed matura- Sciences, Universidad Nacional de Colombia, Bogota. The
tion on the mother plant and genetics of the mother plant. material (A. squamosa) was obtained from accessions col-
Therefore, difficulties in sexual propagation of the species of lected from two locations: Apulo (province of Cundina-
this family could be partially explained by seed dormancy. marca) and Castilla (province of Tolima) because plant
individuals are distributed spontaneously in agroecosys-
In A. squamosa, the seeds feature a rudimentary but dis-
tems and not cultivated. This study was carried with fresh
tinctly underdeveloped embryo. According to Hayat (1963),
seeds obtained from ripe and soft-to-the-touch fruits,
the A. squamosa embryo is embedded in an abundant en-
which were disinfected with 1% sodium hypochlorite im-
dosperm and has a slow growth rate, which could indicate
mersion for 9 min, washed with distilled water and 96%
the occurrence of morphological dormancy as specified by
ethanol three times, and, finally, rinsed three times with
Baskin and Baskin (2004). The erratic and slow germina-
distilled water to remove any ethanol residue that may have
tion of these seeds has been reported by several authors,
remained on the seeds.
who proved that this can take around 30 d (Morton, 1987;
Cruz, 2002), 50 d (Hernández, 1993) or 90 d (Hayat, 1963). To assess the effect of GA treatments on germination, the
Additionally, Stenzel et al. (2003) indicated that anon seeds seeds were soaked for 72 h in distilled water only (control,
have slow germination, with values that, after 2 months of 0 mg L-1 GA) and GA at 6 different concentrations: 50,
germination, reach 1 to 3.8%. 100, 200, 400, 600, or 800 mg L-1 in closed glass jars. The
Also anon seeds possess germination-inhibiting chemi- treatments with control were arranged in a completely
cal substances that cause dormancy; these substances randomized design with five replicates per treatment. After
together with durable and waterproof integuments are soaking, the seeds were removed to assess the germination.
antagonistic factors to rapid and uniform germination The seeds were sown in disinfected peat substrate Klas-
(Pawshe et al., 1997; Stenzel et al., 2003). The positive results mann® (Klasmann-Deilmann GmbH, Geeste, Germany)
of gibberellin (GA) treatments on the germination of A. without nutrients at a depth equal to twice the seed length
squamosa has been reported by several authors (Pawshe et and subsequently placed in a controlled environment cabi-
al., 1997; Stenzel et al., 2003; Sousa et al., 2005; Lima-Brito net (SGC066, Sanyo Gallenkamp, Leicester, UK) for 30 d
et al., 2006; Menegazzo et al., 2012) may also indicate the at a constant temperature of 35ºC in the absence of light
presence of morphophysiological dormancy in these seeds. because germination of seeds of A. squamosa is indifferent
Morphophysiological dormancy is characterised by an to light conditions (Ferreira et al., 2002). Observations were
underdeveloped embryo and a physiological mechanism made every 5 d for 30 d, since seeds that did not germi-
that inhibits seed germination (Baskin and Baskin, 2004; nate after 30 d could be classified as dormant (Baskin and
Baskin and Baskin, 2014). The embryo with this type of Baskin, 2014). Germination was recorded in those seeds
dormancy has a low potential or inhibition of growth due that had radicle protrusion.

 

 
 
to a high ABA/GA ratio, which disables breaking through
the seed cover structures (Baskin and Baskin, 2014). With the sampling data, the following ! parameters were

  (GP) = !" ∗ 100 !!
evaluated: germination percentage (GP)
(GP) = !" = 100, where,
∗∗ 100
!" ∗ 100
Knowledge on seed germination is very important for the N = number of germinated seeds and N = total number of
! (MGT) = !!!! 𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡s; !!
sexual propagation of anon as a cultivated species. At the (GP)
seeds; mean= !" germination time (MGT)
∗ 100 (MGT)
(MGT)== !!! = !!!𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡 ;; mean
𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡
same time, most germination studies for this species were germination rate (MGR) (MGR)== !!! 𝑛𝑛𝑛𝑛 ∗ 𝑡𝑡𝑡𝑡  /   !!!!!! 𝑛𝑛𝑛𝑛 ,, where,
!
(MGR) == !
𝑛𝑛𝑛𝑛,,
ni = number!!!of germinated seeds(MGR)
(MGT) = !
𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡 ; !!!𝑛𝑛𝑛𝑛 𝑡𝑡𝑡𝑡  /   !!!!
𝑛𝑛𝑛𝑛 ∗∗ 𝑡𝑡𝑡𝑡  /   !!!𝑛𝑛𝑛𝑛
conducted in regions with very differented edaphoclimatic in the ith data !!! collection;
characteristics to those prevailing in the producing areas t(MGR) (E)ith=data
− !!! !
𝑓𝑓𝑓𝑓  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓, K  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓,
i = time=(in!days) of the ! collection
(E) == −
(E) − and
!! 𝑓𝑓𝑓𝑓 = time (in
!!! 𝑛𝑛𝑛𝑛 ∗ 𝑡𝑡𝑡𝑡  /   !!! 𝑛𝑛𝑛𝑛 , !!!𝑓𝑓𝑓𝑓  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓,
!!!
of Colombia (Menegazzo et al., 2012; Adeniji et al., 2014). days) duration of the germination ! test, and synchrony index
Also, there are contrasting reports on seed responses to the = , !
(E)== − !!! 𝑓𝑓𝑓𝑓  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓,, where, ni =
(E) ! !!!! (!!")
== number !
(!!"),
,of germinated
!!!!(!!")
!!!!
application of diverse hormones. Therefore, the aim of this seeds in the ith data collection 𝑡𝑡 =  
!"#
, 𝑦𝑦 and
=  
! f = relative frequency
i !"#
!"# !
study was to determine the effect of GA applications on !
= germination , !
𝑡𝑡𝑡𝑡!==  2006).
  ,, 𝑦𝑦𝑦𝑦 ==    !
of !!!! (!!") (Ranal and !"#
Santana, !! !!
the germination of A. squamosa seeds from two producing !"#
𝑡𝑡 = , !"#
!"#
areas in Colombia. 𝑡𝑡 =  assumptions
The , 𝑦𝑦 =  
!
of normality and 𝑡𝑡𝑡𝑡 =
!
,,
=homogeneity of vari-
! ! !!
27.2067
ance!"# were assessed using GAthe
0 mgSASL statistical
-1
𝑦𝑦 = package. The27.2067
R2=0.99398
27.2067 GA
𝑡𝑡 = , GA
GA 00 mg
mg −-1-19.361e(!!.!"#)
1LL 𝑦𝑦 =
2
RR2==
Materials and methods datas !howed no residual normality and were transformed 𝑦𝑦 =
1 − 9.361e
1 − 9.361e
(!!.!"#)
(!!.!"#)
92.5701
with arcosen function √(PG/100)
GA 0 mg L-1
GA 50 mg L-1 usually
27.2067 𝑦𝑦 = used
R2=0.99398
in(!!.!"#)
germi- R2=0.99341
92.5701
-1
92.5701
GA 9
LL-15.40eGA𝑦𝑦𝑦𝑦3 =200
= mg L (!!.!"#)
-1
𝑦𝑦 = (!!.!"#) GA GA 50 mg
50 1−
mg 𝑦𝑦 = RR22==
This research was conducted in the laboratories of Plant nation studies (Júnior Wagner et al., 2006). ANOVA 11was
1 − 9.361e 5.40e(!!.!"#) 1 − 0.
− 5.40e
−
90.199
Physiology and Genetic Resources, Faculty of Agricultural performed L-1 2 𝑦𝑦 =treatments 2
GA 50 mg Land the differences
-1 GA 100 mg among
92.5701 were90.199
R90.199
=0.99949
-1 GA9
𝑦𝑦 = R =0.99341
GA 100 1mg L-1-1 GA
− 1.321e 3 400 mg L
(!!.!"#)
𝑦𝑦 = R22=
1 − 5.40e(!!.!"#) GA 100 mg L 𝑦𝑦𝑦𝑦 = (!!.!"#) 1 −R0.=
=
11 −
− 1.321e
1.321e(!!.!"#)
18 Agron. 96.2871
𝑦𝑦 = Colomb. 34(1) 2016R =0.99413
-1 2
GA 800 mg L
90.199 96.2871
96.2871 9
GA 100 mg L-1 𝑦𝑦 = R2=0.99949
GA -1
1 − L1.56e
800 mg -1 GA
(!!.!"#) 600 -1
= mg L (!!.!"#) 𝑦𝑦 = RR22==
1 − 1.321e(!!.!"#) GA 800 mg L 𝑦𝑦𝑦𝑦3 =
11 − 1.56e(!!.!"#) 1 − 0.
− 1.56e
96.2871
GA 800 mg L-1 𝑦𝑦 = R2=0.99413
1 − 1.56e(!!.!"#)
70,348
 !
(GP) = !"
∗ 100

  (MGT) = !
!!! 𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡 ;

determined by the Tukey test (P≤0.05).! SAS (Statistical 8 d, the increases in the PG dwindled. The maximum PG
(MGR) = 𝑛𝑛𝑛𝑛 ∗ 𝑡𝑡𝑡𝑡  /   !!!! 𝑛𝑛𝑛𝑛 ,
Analysis System),
(GP) =
!
∗version
100 9.1 was used. !!! of 27.2% was reached after 30 d.
!"
(E) = − !
!!! 𝑓𝑓𝑓𝑓  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓,
PG response
(MGT) =to the ! application
!!! 𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡 ; of GA, for both areas was With the application of 200, 400, 600, and 800 mg L-1 GA,
!
adjusted to a logistic model = , where, K is the the better PG responses were observed during the incuba-
!!!! (!!")
maximum(MGR) = !!!
value
!
reached
!
𝑛𝑛𝑛𝑛 ∗ 𝑡𝑡𝑡𝑡  /  by !!! ,
the𝑛𝑛𝑛𝑛variable, in this case the tion period. Under these concentrations, the germination
!"# !
seed weight (g)! or the percentage 𝑡𝑡 =  of germination
, 𝑦𝑦 =  
! (%), A is patterns were similar, with values of PG much higher than
!
(E) = − 𝑓𝑓𝑓𝑓  𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓,
the product of!!! the initial value “a”
𝑡𝑡 = ,
!"#maximum “K”, B is a found for the treatment of 0 mg L-1 GA in the first 5 d of
scale parameter
=
!
,on the time t that influences the growth incubation (86.4, 85.6, 89.6, and 84.8%, respectively). The
!
(!!")
rate. On!!!!the logistic model it wasGAcalculated 0 mg L-1
an𝑦𝑦inflection
=
27.2067 shortest time to reach
R2=0.99398 GA3 200 the mgmaximum
L-1 𝑦𝑦 =
PG94.112
was obtained with
R2=0.99696
!"#
point 𝑡𝑡 =   ! , 𝑦𝑦 =   !! , in order to establish the change 1 −in the (!!.!"#)
9.361e -1
the application of 600 mg L at 16 d GA (PG = 92.3%), 1 − 0.419e(!!.!"#)
 
concavity !"# of the curve in order GA to determine
50 mg L-1 the
𝑦𝑦 =
growth92.5701 followed
R2=0.99341by 800GA mg L-1 GA-1 to 𝑦𝑦17.7
3 400 mg L = d (PG(!!.!"#)
94.0994= 96.3%). The
R2=0.99809
𝑡𝑡 = , 1 − 5.40e(!!.!"#)
dynamics !of variation represented by the percentage of treatments of 200 and 400 mg L GA obtained maximum -1 1 − 0.535e
!
germination (Hsu, 1984). (GP) = !" ∗ 100 90.199
PG values at 2294.112
dGAof3incubation 92.2718
-1 GA 100 mg L2-1
27.2067 𝑦𝑦 = R2=0.99949 600 mg L2-1 (PG 𝑦𝑦 = = 94.1%). For the same
R2=0.92104
GA 0 mg L 𝑦𝑦 = R =0.993981 − 1.321e GA3 200 mg L-1
(!!.!"#)
𝑦𝑦 = R =0.996961 − 0.379e(!!.!"#)
1 − 9.361e (!!.!"#)
concentrations, geometric
1 − 0.419e (!!.!"#)
progression of PG or acceler-
(MGT) = !!!! 𝑛𝑛𝑛𝑛/𝑡𝑡𝑡𝑡 96.2871 ;
ResultsGAand 50 mgdiscussion
L-1
GA 800 mg L2-1
92.5701 𝑦𝑦 =
R =0.99341 1 − GA
ated-1Rgrowth
2
=0.99413of94.0994
this variable was presented during 1st d of
R2=0.99809
3 400 mg L
1.56e (!!.!"#)
𝑦𝑦 = 𝑦𝑦 =
(MGR) = !!!! 𝑛𝑛𝑛𝑛 ∗ 𝑡𝑡𝑡𝑡  /   incubation.
1 − 5.40e (!!.!"#) 1 − 0.535e (!!.!"#)
!
!!! 𝑛𝑛𝑛𝑛 ,
The Fig.GA 1 shows
100 mg the L-1 effect 𝑦𝑦 = of GA (!!.!"#) application on PG of GAA.
90.199 92.2718
R2=0.99949 3 600 mg L
-1
𝑦𝑦 = R2=0.92104
(E) =Tolima − !!!! 𝑓𝑓𝑓𝑓 1 − 0.379e(!!.!"#)
squamosa seeds obtained1 from − 1.321e
Castilla, and its Without the
 𝐿𝐿𝐿𝐿𝐿𝐿2  𝑓𝑓𝑓𝑓, application of phytoregulator (0 mg L-1 GA),
70,348 87.6346
variationGAduring
800 mg30 L-1d of𝑦𝑦germination.
=
GA 0 mg Without
96.2871 L-1 R2=0.99413
!
𝑦𝑦 the
= applica- theR2germination
=0.99272 GAof3 200themgseeds
L-1 from𝑦𝑦 = Apulo, was lower than
R2=0.9756
1 − 14.05e(!!,!"#) 1 − 2.273e(!!.!"#)
1 − 1.56e
tion of phytoregulator (0 mg L GA) the germination -1
(!!.!"#)
= , was those values achieved with any dose of the hormone. In
!!!! (!!")
78.7976 100.423
lower than the one achieved with GA any50 mg L-1 of!"#
dose the
𝑦𝑦 = hormone;
! R2=0.97243
(!!.!"#)this case, at 5 d of
GA3 400 mg L-1
incubation 𝑦𝑦PG
= was 9.6% R2=0.99292
and increased
𝑡𝑡 =   , 𝑦𝑦 1=−   7.220e 1 − 1.412e(!!.!"#)
after 5 d of incubation 8.8% PG was obtained. !
According
!
to a value of 59.3% at 30 d of evaluation. By analyzing the
-1 !"# 103.45 95.138
GA 100 mg L R2=0.98739inflection mg L2-1 it𝑦𝑦could
GA3 600 point, R2that
=0.98012
to theGA inflection point 𝑦𝑦 = calculated for 𝑡𝑡R==0.99272 , during GA3the mgcalculated be observed the
-1 70,348 2 𝑦𝑦 = 87.6346 =
0 mg L (!!,!"#) ! 1 − 1.508e 200 (!!.!"#) L-1 𝑦𝑦 = (!!.!"#)
R =0.9756 1 − 2.389e (!!.!"#)
1 − 14.05e 1 − 2.273e
first 8 d of incubation, the value of PG grew in geometric accelerated growth of PG occured until 18.9 d, where it
27.2067 94.112
progression, that
L-1 is, the 𝑦𝑦 = PG presented high growth = L rates 𝑦𝑦to= mgreaches 𝑦𝑦 =half the maximum 2 load
3 200 value
mg L-1 (36.6%), after which R2=0.99696
78.7976 2GA 0𝑦𝑦mg -1 88.4138 100.423
R2=0.99398
GA 50 mg GA 800 mg R-1
L =0.97243 GA3 400 L-1R2=0.97761 RGA
=0.99292 𝑦𝑦 =
1 − 7.220e(!!.!"#) 1 − 1.202e(!!.!"#) 1 − 9.361e(!!.!"#) 1 − 1.412e(!!.!"#) 1 − 0.419e(!!.!"#)
reach half of the maximum value of its load (13.6%). After the rate of PG growth slowed down (Fig. 2).
103.45 -1 92.5701 95.138 94.0994
GA 100 mg L-1 𝑦𝑦 = R2GA 50 mg L GA3 600
=0.98739 𝑦𝑦 =mg L-1 𝑦𝑦 = R2=0.99341 RGA
2
3 400 mg L
=0.98012
-1
𝑦𝑦 = R2=0.99809
1 − 1.508e(!!.!"#) 1 − 5.40e(!!.!"#)
1 − 2.389e(!!.!"#) 1 − 0.535e(!!.!"#)
120
-1 90.199 0 mg L-1 92.2718
R2=0.99949 GA3 600 mg L-1 R2=0.92104
(𝑡𝑡 = ) R2GA 100 mg L
!"# 𝑦𝑦 = 𝑦𝑦 =
-1 88.4138 1 − 1.321e(!!.!"#) 1 − 0.379e(!!.!"#)
GA 800 mg L 𝑦𝑦100
= ! =0.97761
1 − 1.202e(!!.!"#) 50 mg L -1

96.2871
GA 800 mg L-1 𝑦𝑦 = R2=0.99413
Germination (%)

80 1 − 1.56e(!!.!"#) 100 mg L-1

60 200 mg L-1
  (𝑡𝑡 = !"#
)
!
40 400 mg L-1
70,348 87.6346
    GA 0 mg L-1 𝑦𝑦 = R2=0.99272 GA3 200 mg L-1 𝑦𝑦 = R2=0.9756
1 − 14.05e(!!,!"#) 1 − 2.273e(!!.!"#)
600 mg L-1
20
    78.7976 100.423
27.2067 GA2 50 mg L-1 R2=0.97243
94.112 GA23 400 L-1L-1
mgmg R2=0.99292
  GA 0 mg L
-1
  𝑦𝑦 =01 − 9.361e(!!.!"#) R =0.99398 GA𝑦𝑦3=200
1 −mg L-1 (!!.!"#)
7.220e 𝑦𝑦 = 800
R =0.99696 𝑦𝑦 =
1 − 1.412e(!!.!"#)
1 − 0.419e(!!.!"#)
1  
0 27.2067 5 10 2 27.2067
15 20 2 25 103.45 30 35294.112 95.138
GA 0 mg L-1 GA -1
𝑦𝑦 = 0 mg L (!!.!"#) 𝑦𝑦 = GA 100 mg L-1 GA𝑦𝑦3R=
R =0.99398 200
=0.99398 𝑦𝑦 =3 200R mg
L-1 (!!.!"#) =0.98739 𝑦𝑦 = GA
L-1(!!.!"#) 600
R23=0.99696mg L-1
94.112 𝑦𝑦R=2 R2=0.98012
1 − 9.361e
92.5701 1 − 9.361eTime (d)
(!!.!"#) 1 −mg
1.508e GA
1 − 0.419e
94.0994 1 − 0.419e (!!.!"#)
=0.99696
1 − 2.389e(!!.!"#)
GA 50 mg L-1 𝑦𝑦 =   R2=0.99341 GA3 400 mg L-1 𝑦𝑦 = R2=0.99809
1 −27.2067
5.40e (!!.!"#) (!!.!"#)
-1 2 27.2067 2 1 − 0.535e
94.112 -1 94.112
GA 0 mg L-1 𝑦𝑦GA = 0 mg L (!!.!"#) 𝑦𝑦 = R =0.99398 -1
(!!.!"#)
GA 3R 200=0.99398
mg L -1
88.4138 GA
𝑦𝑦 = 3 200 mg
2
L 𝑦𝑦 = R 2
=0.99696 (!!.!"#)
R2=0.99696
1 −22 9.361e (!!.!"#) 1 −22 0.419e
-1 L-1-1
27.2067
1 − 9.361e
92.5701
-1-1 GA 800 mg
27.2067
92.5701
R =0.99341 L 𝑦𝑦 =2 -1 R
1 − -1 =0.97761
94.112
0.419e
94.0994-1 94.112
94.0994 R22=0.99809
GA 0 mg GA 0 mg
mgLL (!!.!"#) =0.99398 GA3R3R400
2200
=0.99398
mg LGA(!!.!"#)
-1 GA 200 mg LL (!!.!"#)
-1 R =0.99809
=0.99696 =0.99696
2
GA
GA0 mg
50 Lmg L 𝑦𝑦𝑦𝑦 =
GA =50 𝑦𝑦𝑦𝑦 =
= R R(!!.!"#)
=0.99398
GA =0.99341
1 −mg L
1.202e 3 200
GA =mg
𝑦𝑦𝑦𝑦 = L mg
33 400 𝑦𝑦𝑦𝑦 =
= R (!!.!"#) R R2=0.99696
11 −
− 9.361e
5.40e(!!.!"#) 11 − − 9.361e
5.40e(!!.!"#) 11 −
− 0.535e
0.419e(!!.!"#) 11 − − 0.535e
0.419e(!!.!"#)
1  
-1 90.199
92.5701
-1 22 92.5701 2 -1-1 92.2718
-194.0994-1 22 94.0994
GA
GAGA 100
5050
mg mg L-1L
L-1mg GA
𝑦𝑦 = 50 mg L (!!.!"#) 𝑦𝑦
(!!.!"#) = RR =0.99949
=0.99341 R 2 GA
GA
=0.99341
(!!.!"#) 3R
3 600
400 mg
mg LL
=0.99341 GA 3 GA
400
𝑦𝑦
𝑦𝑦 ==mg
3 400
L mg L 𝑦𝑦
(!!.!"#) = RR =0.92104
=0.99809 R2=0.99809
R2=0.99809
 GA 50 mg L-1 1.321e
92.5701
1 − 5.40e -1 1− 2 5.40e
92.5701 2 11−−0.535e
0.379e(!!.!"#)
94.0994 1 −2 0.535e
94.0994 (!!.!"#)
𝑦𝑦 =50 mg L (!!.!"#)𝑦𝑦 = R =0.99341
GA (!!.!"#)
GA3R400 mg L-1 GA
=0.99341 -1
𝑦𝑦 =3 400 mg L (!!.!"#) 𝑦𝑦 = R =0.99809 (!!.!"#)
R2=0.99809
1 − 5.40e
90.199 1 − 5.40e
90.199 1 − 0.535e
92.2718 1 − 0.535e
92.2718
GAGA
100100
mg Lmg
-1
L-1 GA -1
𝑦𝑦 = 100 mg L (!!.!"#) R2=
𝑦𝑦 = (𝑡𝑡 !"# R2=0.99949
=0.99949
) (!!.!"#)
2
GA3R600 =0.99949
mg LGA-1
GA
3 600 3 600
𝑦𝑦 =mg
-1
L-1 mg L (!!.!"#)
2
𝑦𝑦 = R =0.92104 (!!.!"#)
R2=0.92104
R2=0.92104
1 − 1.321e 1 − 1.321e! 1 − 0.379e 1 − 0.379e
90.199
96.2871
-1 22 90.1992 2 92.2718
-1 2 92.2718
GAGA
800100
GA mg mg
800 -1 LL-1
𝑦𝑦GA
Lmg
-1
𝑦𝑦==100 mg L (!!.!"#) 𝑦𝑦 = RR=0.99949
=0.99413 GA3R600
R(!!.!"#)
=0.99413 mg L-1 𝑦𝑦GA
=0.99949 = 3 600 mg L(!!.!"#)
𝑦𝑦 = R =0.92104 (!!.!"#)
R2=0.92104
1 1−−1.321e
1.56e (!!.!"#)
90.199 1 −2 1.321e
90.199 92.2718
1 − 0.379e 1 −2 0.379e
92.2718
GA 100 mg L-1GA
𝑦𝑦 = 100 mg L -1
𝑦𝑦 = R =0.99949
(!!.!"#) 1 − 1.321e(!!.!"#)
GA R
3
2
600
=0.99949
mg L -1
GA
𝑦𝑦 =3 600 mg L -1
𝑦𝑦 = R =0.92104 R2=0.92104
1 − 1.321e
96.2871 1 − 0.379e(!!.!"#) 1 − 0.379e(!!.!"#)
-1 -1 2 96.2871 2
GA 800 mg L GA𝑦𝑦 =800 mg L (!!.!"#)𝑦𝑦 = R =0.99413 R =0.99413
FIGURE 1. Effect of imbibition 1in−the1.56e
water and GA 1.56e(!!.!"#)
1 −solutions on the accumulated germination of seeds of A. squamosa from Castilla (province of
Tolima,GA
Colombia), 96.2871
800 mg Lfor
-1 30 d of incubation
-1 in moist2 peat
96.2871 at 35ºC temperature and 60% relative air humidity.
GA
𝑦𝑦 = 800 mg L 𝑦𝑦 = R =0.99413 R2=0.99413 (!!.!"#) (!!.!"#)
96.2871
1 − 1.56e 1 − 1.56e
96.2871
GA 800 mg L-1 GA mg L-1(!!.!"#)𝑦𝑦 = R2=0.99413
𝑦𝑦 =80070,348 R2=0.99413 87.6346
1 − 1.56e 21 − 1.56e(!!.!"#)
Martínez
GAM.,0 mg L-1 L., and𝑦𝑦 Magnitskiy:
Miranda = Sugar apple (Annona
(!!,!"#)
R squamosa GA
=0.99272 L.) seed
3 200 mg L-1 affected
germination 𝑦𝑦 = by the application
(!!.!"#)
2
ofRgibberellins
=0.9756 19
1 − 14.05e 1 − 2.273e
70,348 2 70,348 87.6346 87.6346
GA 0 mg L-1 GA
𝑦𝑦 = 0 mg L
-1
𝑦𝑦 = R =0.99272
(!!,!"#) 1 − 14.05e(!!,!"#)
GAR 2
3 200 mg L-1
=0.99272 GA -1
𝑦𝑦 =3 200 mg L (!!.!"#)
2
𝑦𝑦 = R =0.9756 R2=0.9756
1 − 14.05e
78.7976 1 − 2.273e
100.423 1 −2 2.273e(!!.!"#) 1  
GA 50 mg L-1 𝑦𝑦 = (!!.!"#)
R =0.97243 2
GA3 400 mg L -1
𝑦𝑦 = R =0.99292
1 − 7.220e
70,348
-1 70,348 2 1.412e-1(!!.!"#)
1 − 87.6346 2 87.6346
GA 0 mg L-1 𝑦𝑦GA= 0 mg L (!!,!"#)
2
𝑦𝑦 = R =0.99272 GA3R200
(!!,!"#)
mg L-1
=0.99272 GA
𝑦𝑦 =3 200 mg L(!!.!"#) 𝑦𝑦 = R =0.9756(!!.!"#) R2=0.9756
 27.2067
-1 27.2067 94.112 94.112
GA 0 mg L-1 GA
𝑦𝑦 =0 mg L 𝑦𝑦 = R2=0.99398 R2=0.99398-1
(!!.!"#) GA3 200 mg L
GA
𝑦𝑦 =3 200 mg L
-1
𝑦𝑦 = R2=0.99696 R2=0.99696
(!!.!"#)
1 − 9.361e 1 − 9.361e 1 − 0.419e(!!.!"#) 1 − 0.419e(!!.!"#)
  GA 27.2067
-1 27.2067
R2=0.99398-1 94.112
-1 94.112
R2=0.99696
GA 0 mg L-1 𝑦𝑦 =0 mg L 𝑦𝑦 = R2=0.99398 (!!.!"#) GA3 200 mg L
27.2067
(!!.!"#) 1 − 9.361e
GA
𝑦𝑦 =3 200 mg L (!!.!"#) 𝑦𝑦 = R2=0.99696 (!!.!"#)
94.112
1 − 27.2067
9.361e
-1 1 − 94.112
0.419e 1 − 0.419e
GA 0 mg -1 GA 0 mg LL-1
mg92.5701 𝑦𝑦𝑦𝑦== R 2 92.5701 R3 22200
=0.99398
mg L-1 GA =33 200 mg L -1
= R 2 94.0994 R 2
R2=0.99696
mgLL-1 2=0.99398
(!!.!"#) GAR
GA
𝑦𝑦𝑦𝑦==50 =0.99341-1 GA
𝑦𝑦 = 400 94.0994
mg L -1 𝑦𝑦 = 2=0.99696 =0.99809
GA 50 (!!.!"#)
1 − 9.361e(!!.!"#) 1R =0.99341
− 9.361e
1 − 5.40e(!!.!"#) GA3 400 mg L 𝑦𝑦 1−− 0.535e 𝑦𝑦
(!!.!"#) 1 R
− =0.99809
0.419e(!!.!"#) 1 − 0.535e(!!.!"#)
0.419e (!!.!"#)
1 − 5.40e 1
100 92.5701 0 mg L-1
94.0994
GA 92.5701
-1
R2=0.99341-1 94.0994
-1
R2=0.99809
=50 mg L (!!.!"#) GA
𝑦𝑦 =3 400 mg L (!!.!"#)
-1
GA 50 mg L 𝑦𝑦90 𝑦𝑦 = R2=0.99341(!!.!"#) GA3 400 mg L 𝑦𝑦 = R2=0.99809
1− 92.5701
5.40e
-1 92.5701
1 −2 5.40e 2 94.0994
1 − 0.535e-1 1 − 0.535e (!!.!"#)
94.0994
GA 50 mgLL-1-1 GA 50 mg L
27.2067 𝑦𝑦 = RR =0.99341
2 GAR3 200
=0.99341
400 mg L -1
L-1 GA 3 400 mg L
94.112 𝑦𝑦 = R R22=0.99696
=0.99809 R2=0.99809
GA 0 mg 𝑦𝑦𝑦𝑦80
== 1 − 5.40e(!!.!"#) 1 −=0.99398
5.40e (!!.!"#) GA 3 mg 𝑦𝑦
𝑦𝑦 =
= 1 − 0.535e(!!.!"#) 1 − 50 mg(!!.!"#)
0.535e L-1
1 − 90.199
9.361e -1 (!!.!"#) 90.199 2 92.2718
1 − 0.419e-1(!!.!"#) 92.2718
GA 100 mg L-1 GA
𝑦𝑦 =100 mg L (!!.!"#)
2
𝑦𝑦 = R =0.99949 GAR3 600
=0.99949
mg L-1 GA
𝑦𝑦 =3 600 mg L (!!.!"#) 𝑦𝑦 = R2=0.92104 R2=0.92104
70 1 − 1.321e 1 − 1.321e(!!.!"#) 1 − 0.379e 1 − 0.379e(!!.!"#)
100 mg L

Germination (%)
-1

90.199 90.199 92.2718 92.2718

mgLL-1-1 𝑦𝑦𝑦𝑦60
GA ==100 mg L-1 𝑦𝑦 = R22=0.99949 R2=0.99949 GA 3 600 mg L-1 𝑦𝑦 = R22=0.92104 R2=0.92104
92.5701 -1 94.0994
GA
GA 100
50 mg (!!.!"#) 1 R (!!.!"#) GA
GA33 2600
400 mg
mg L
L-1 𝑦𝑦
𝑦𝑦 = (!!.!"#) 1 R
11−−mg90.199
1.321e-1(!!.!"#) −2=0.99341
90.199
1.321e = 1 − 0.379e
92.2718 − =0.99809
0.379e (!!.!"#)
92.2718
200 mg L-1
GA 100 mg L-1 GA
𝑦𝑦 50
= 100 L
5.40e 𝑦𝑦 = R =0.99949 R =0.99949
(!!.!"#) GA3 600 mg L
-1 GA
𝑦𝑦 =3 600
1 − mg L-1(!!.!"#)
0.535e 𝑦𝑦 = R2=0.92104(!!.!"#)
R2=0.92104
1 − 1.321e(!!.!"#) 1 − 1.321e 1 − 0.379e(!!.!"#) 1 − 0.379e
96.2871
-1 96.2871
GA 800 mg L-1 𝑦𝑦40
GA =800 mg L 𝑦𝑦 = R2=0.99413 R2=0.99413 400 mg L-1
1− 1.56e(!!.!"#) 1 − 1.56e(!!.!"#)
30
96.2871 96.2871
GA 100
GA 800 mg
mg L -1
L-1 GA
𝑦𝑦𝑦𝑦20
800 mg L-1
90.199
== 1 − 1.56e R22=0.99949
𝑦𝑦 = R =0.99413 R2=0.99413-1
(!!.!"#) GA3 600 mg L 𝑦𝑦 =
92.2718 600 mg L-1
R2=0.92104
(!!.!"#)
96.2871
-1 1 − 96.2871
1.56e
GA 800 mg L-1 GA
𝑦𝑦 =800
1 − mg L
1.321e (!!.!"#) 2
𝑦𝑦 = R =0.99413 R2=0.99413 1 − 0.379e(!!.!"#)
10 1 − 1.56e(!!.!"#) 1 − 1.56e(!!.!"#) 800 mg L-1
0
GA 0 mg0 L70,348
-1 5 102 70,34815 20 25 -1
R2=0.99272 30 200 mg
GA 35 L-1
87.6346 87.6346
R2=0.9756
GA 0 mg L-1 96.2871𝑦𝑦 = R =0.99272 (!!,!"#) GA3 200 mg L
𝑦𝑦 = R2=0.9756
GA 800 mg L-1 𝑦𝑦 =𝑦𝑦1=− 14.05e(!!,!"#) 𝑦𝑦 =
3
(!!.!"#)
R2=0.99413
1 − 14.05e Time (d) (!!.!"#)
1 − 2.273e (!!.!"#)
1 − 2.273e
1 − 1.56e
70,348
-1 70,348 87.6346 87.6346
GA 0 mg L-1 GA
𝑦𝑦 =0 mg L 𝑦𝑦 = R2=0.99272 R2=0.99272-1 GA
(!!,!"#) GA3 200 mg L 3 200 mg L
𝑦𝑦 =
-1
𝑦𝑦 = R2=0.9756 (!!.!"#)
R2=0.9756
1 − 70,348
14.05e
-1 70,348
(!!,!"#) 1 − 14.05e 2 1 − 2.273e 87.6346
-1(!!.!"#) 1 −22.273e
87.6346 2
GA -1 GA 0 mg L R 2 78.7976 R =0.99272 -1 GA 200-1mg L R 100.423 R
R2=0.9756
GA0 050mg
L-1LL-1
GA mg 78.7976
-1 𝑦𝑦 = =0.99272
R2 GA
=0.99272 2200 mg LGA 200 mg L 100.423
-1 𝑦𝑦 = 2=0.9756 R2=0.9756
GA
𝑦𝑦 =50
𝑦𝑦 = mg L (!!,!"#) GAR3 400
=0.97243 3 GA
𝑦𝑦 3 400 mg L (!!.!"#)
= =0.99292
2 -1 3
mg 𝑦𝑦
(!!,!"#)
1 − 14.05e(!!.!"#) = 1 R
− =0.97243
14.05e (!!.!"#)
3 mg L 𝑦𝑦 = 1 − 2.273e(!!.!"#)𝑦𝑦 = 1 R
− =0.99292
2.273e (!!.!"#)
(!!.!"#)
1 − 7.220e 1 − 7.220e 1 − 1.412e 1 − 1.412e
78.7976
-1 78.79762 2 100.423
-1 100.423
GA
GA 5050
mgmg
L L -1 -1 GA
𝑦𝑦 =50 mg L (!!.!"#)
2
𝑦𝑦 = R =0.97243 GAR3 400
R =0.97243
(!!.!"#)
=0.97243 -1 GA
mg LGA3 400 400
3mg
𝑦𝑦 = L mg L (!!.!"#)
-1
𝑦𝑦 = R2=0.99292 (!!.!"#)
R2=0.99292
R2=0.99292
1− 78.7976
7.220e
-1 78.7976
1 −27.220e 2 100.423
1 − 1.412e -1 100.423
1 −21.412e
mgLL-1-1
GA 050mg GA
𝑦𝑦 50 mg L
70,348 𝑦𝑦 = R =0.99272
2 =0.97243 R =0.97243
GA3 200
400 mg
mg L
L -1
-1 GA 400 mg L
87.6346 𝑦𝑦 = R =0.9756
2 =0.99292 R2=0.99292
𝑦𝑦 = 𝑦𝑦
𝑦𝑦 =
3
GA (!!.!"#) 1 R
= 1 − 7.220e(!!,!"#) − 7.220e (!!.!"#) GA 3 (!!.!"#) 1 R
= 1 − 1.412e(!!.!"#) − 1.412e (!!.!"#)
103.45
1 − 14.05e -1 103.45 2 95.138
1 −-12.273e 95.138
GAGA
100100
mgmg
L-1 L
-1 GA
𝑦𝑦 =100 mg L (!!.!"#) 𝑦𝑦 = R2=0.98739 R2 GAR3 600
=0.98739
(!!.!"#)
=0.98739 -1
mg LGA GA
600 600
3mg
𝑦𝑦 = L mg L-1 𝑦𝑦 = R2=0.98012 R2=0.98012
R2=0.98012
1 − 1.508e 1 − 1.508e 3
1 − 2.389e(!!.!"#) 1 − 2.389e(!!.!"#)
103.45 103.45 95.138 2 95.138
-1 GA =100 mg L-1
78.7976 𝑦𝑦 = R22=0.98739 R2=0.98739 -1 GA 3 600 mg L-1
100.423 R2=0.98012
𝑦𝑦 = R
GAGA
800100
GA 50mgmg
mgL-1LL-1-1 𝑦𝑦 =
𝑦𝑦 1−−mg 103.45
1.508e (!!.!"#) 1 R
-1 (!!.!"#) =0.97243
−21.508e
GA
GA332600
R2=0.97761
(!!.!"#)
103.45 400 mg
mg L
L-1 𝑦𝑦
𝑦𝑦 =
= 1 − 2.389e
95.138 R2=0.98012
=0.99292
(!!.!"#) 1 −22.389e
-1(!!.!"#)
(!!.!"#)
95.138
GA 100 mg L GA
𝑦𝑦 =100
1 L
7.220e 𝑦𝑦 = R =0.98739 R =0.98739
(!!.!"#) GA3 600 mg L
-1 GA
𝑦𝑦 =
3 600
1 − mg L
1.412e R2=0.98012
𝑦𝑦 = R =0.98012
1 − 1.508e 88.4138
-1 (!!.!"#) 1 −21.508e
88.4138 2 1 − 2.389e (!!.!"#) 1 − 2.389e(!!.!"#)
GA 800 mg L -1 GA
𝑦𝑦 =800 mg L (!!.!"#) 𝑦𝑦 = R =0.97761 R =0.97761
1 − 1.202e 1 − 1.202e(!!.!"#)
FIGURE 2. Effect of imbibition in the water and GA solutionson
88.4138
the accumulated germination of seeds of A. squamosa from Apulo (province of Cun-
88.4138
-1 2
GA 800
dinamarca, mg L-1
Colombia),-1
GA
𝑦𝑦 =80030
during mgd Lof-1 incubation
103.45 𝑦𝑦 = R22in
=0.97761
moist peat at R
35ºC=0.97761
temperature
-1 and 60% relative air humidity.
95.138
GA 100 mg L-1 GA 𝑦𝑦 = 1 − 88.4138
1.202e (!!.!"#) 1R
−21.202e (!!.!"#)
88.4138
=0.98739 GA R 2600 mg L 𝑦𝑦 = R2=0.98012
𝑦𝑦 =800
1 −mg L (!!.!"#) =0.97761
𝑦𝑦 = R =0.97761 3
GA 800 mg L 1.508e (!!.!"#) 1 − 2.389e(!!.!"#)
1 − 1.202e(!!.!"#) 1 − 1.202e
-1
The seeds!"# treated with GA !"# at a concentration of 600 mg L In the present study, there were observed differences in
(𝑡𝑡 =highest
had the (𝑡𝑡 = )
) values of!PG during the incubation time. the patterns of seed germination of Castilla, Tolima and
!
-1 88.4138
The PG
GA maximum
800
!"# L
mg of
𝑦𝑦 =95%
!"# was obtained R2=0.97761
with the shortest Apulo, Cundinamarca affected by the application of GA.
(𝑡𝑡 = !"#) (𝑡𝑡 =1!"# )
− 1.202e (!!.!"#)
!
incubation
(𝑡𝑡 = time
!
) (between
(𝑡𝑡 = )24 and 28 d) compared to the At all GA concentrations, the seeds from Castilla, Tolima
! !
other concentrations. The period of accelerated growth had higher PG values (> 70%) at 5 d of incubation than the
ended on 3.5 d of incubation and at 5 d of germination seeds from Apulo (PG > 50%), and a shorter time to obtain
!"#
yielded
(𝑡𝑡 =a 56.8%
!
) PG. The curve reflects that the seed treat- the maximum germination value (maximum PG “K”) and
ment with 800 mg L-1 GA presented a dynamics similar to shorter periods (of PG accelerated growth, indicating that,
that one found with treatment with 600 mg L-1 GA although in seeds from Tolima, stabilization occured in less time.
the time to reach the maximum value of PG (88.4%) was
obtained according to the projection model and param- GA effect on germination variables
eters, after the last day of the incubation. The period of In seeds from Apulo, only the concentration of 600 mg L-1
accelerated growth was 1.22 d and generated a PG of 56.8% GA was effective in promoting a synchronized germination
during first 5 d of incubation. to generate a synchronization 1  
index significantly
1   lower
(0.34) than the control (0.41). In the seeds from Castilla, a
There   are few studies
  that describe patterns of germination trend was observed for a decreased
1   synchrony
1   index with
in seeds of Annonaceae species. Stenzel et al. (2003) found 1  
1   Again, it was
increasing concentrations of GA. possible to
that the  
  seed germination of the cultivars Atemoya Gefner
    demonstrate differences in the response among the two
and Anon PR-3, when treated with 50-100 mg L-1 GA, oc- locations, with the seeds of Tolima more influenced by the
curred mainly between 14 and 28 d. Seed germination of application of GA compared to1  the seeds of Cundinamarca
cultivar PR-1 of Atemoya was presented mainly between 21 (Fig. 3A).
 
and 42 d when 50 mg L-1 GA was applied. Toll-Jubes et al.
(1975) found a higher percentage of germination between The germination percentage (PG) was influenced by the
7 and 38 d in cherimoya (A. cherimola) seeds scarified and external application of GA as could be seen in Fig. 3B. In
imbibed in a GA solution and water. Pawshe et al. (1997) both locations, all concentrations significantly increased
found a higher percentage of germination performed at 22 the final PG (30 d incubation), as compared to the control
and 26 d in seeds of A. squamosa immersed in GA at 50 (0 mg L-1). However, among the tested concentrations (50,
and 100 mg L-1. 100, 200, 400, 600, and 800 mg L-1), statistically significant

20 Agron. Colomb. 34(1) 2016

differences were not found. In seeds from Apulo, higher
PG values were obtained in the treatments with 400 and
600 mg L-1 GA (96.8 and 97.6%, respectively), as compared
to the control (59.2%), with an increase of 63 and 64.9% of
PG. In the seeds from Castilla, one can see a trend in which
the PG increaseed with increasing concentrations of GA.
All concentrations contributed to a significant increase of
PG (>200%) and, although these differences were minimal,
the application of 800 mg L-1 GA resulted in higher values
of PG (96%), which represented an increase of 252% over
the control (PG = 27.2%).
Our results, obtained with Colombian assesions of A. squa-
mosa, are comparable to those found by several authors in
other tropical regions, where the PG increased after the
application of GA at different concentrations. Pawshe et
al. (1997), Stenzel et al. (2003), and Menegazzo et al. (2012)
with low concentrations of GA (from 50 to 100 mg L-1) ob-
tained the best PG (around 75%); the latter study reported
durations of seed imbibitions in GA solutions from 5 to 24
h as sufficient ones. Sousa et al. (2005) obtained 90% PG
after imbibition of A. squamosa seeds for 12 hat 400 mg L-1
GA, indicating that doses of 50 to 750 mg L-1 GA also pro-
duced positive results. Ferreira et al. (1998, 2002) reported
that the application of 200 and 250 mg L-1 GA significantly
promoted germination (up to 77%) of A. squamosa seeds
in germination chamber at alternating temperatures of 20
to 30ºC. Likewise, Lima-Brito et al. (2006) found that the
use of GA between 250 and 1000 mg L-1 increased the PG
and IVG of the seeds.
In other Annonaceae species, the same positive effects
were reported. De Smet et al. (1999) reported, in cherimoya
seeds treated with 500; 1,000; 5,000, or 10,000 mg L-1 GA,
a germination of 58.5, 65.5, 69.5, or 74.5%, respectively.
Meanwhile, González-Esquinca et al. (1997), indicated
that seeds of A. diversifolia have dormancy that could be
overcomed with the use of GA. Gimenez et al. (2014) in
Annona emarginata obtained 91% germination with the
use of 250 mg L-1 GA3. Ferreira et al. (2002) showed, in
cultivars Gefner and Thompson of Atemoya, significant
differences in germination between the concentrations
500, 750, and 1,000 mg L-1 GA (for cultivar Thompson)
and 1,000 mg L-1 GA (for cultivar Gefner) compared with
the control. At the same time, Stenzel et al. (2003) reported
no differences between the control and GA concentrations
(50 and 100 mg L-1) used for three cultivars of Atemoya
(Gefner, PR-1, and PR-3).
Shorter germination times were recorded for treatments
with GA for seeds from both locations, where a trend of
Martínez M., Miranda L., and Magnitskiy: Sugar apple (Annona squamosa L.) seed germination affected by the application of gibberellins
a decreasing MGT with increasing concentrations of GA
could be observed (Fig. 3C). In seeds from Cundinamarca,
no statistical differences were found between the GA treat-
ments during the time required for maximum germination
(MGT), but the treatment with 600 mg L-1 GA gave the best
values for MGT that decreased from 17.68 d (control data
at 0 mg L-1) down to 8.75 d. In Castilla, Cundinamarca, the
different GA concentrations were not statistically different
in the time required for maximum germination (MGT);
however, concentrations of 200, 400, 600, and 800 mgL-1 GA
led to lower values of MGT of 5.52, 5.55, 5.38, and 5.82 d,
respectively, compared to the control with MGT of 10.88 d.
Compared to the control (0 mg L-1 GA), the treatments of
seeds of A.squamosa with GA at all concentrations were
effective in promoting a greater MGR (Fig. 3D). It could
be seen from the resultsfor the two locaions that the seeds
tended to increase for the MGR with increasing concentra-
tions from 0 to 600 mg L-1 GA. The concentration of 600
mg L-1 GA produced the highest MGR values for both seeds
from Castilla, Tolima and Apulo, Cundinamarca (0.18 and
0.12 germinated seeds/d, respectively), compared with
the values of the respective controls (Castilla = 0.09 and
Apulo = 0.06 germinated seeds/d). However, the seeds from
Castilla had significantly higher values of MGR relative to
the control than the seeds from Apulo at each tested GA
concentration, indicating a greater influence of the GA on
the MGR of the seeds from Castilla, Tolima.
Increases in the speed of germination of the seeds of
A.squamosa were also found with the application of 50
mg L-1 (Toll-Jubes et al., 1975), 100 mg L-1 (Stenzel et al.,
2003) and 250 mg L-1 GA (Ferreira et al., 1999). Our results
are also consistent with those of Sousa et al. (2008), who
found that the highest rates of germination were obtained
with GA treatments. Stenzel et al. (2003) in atemoya seeds
indicated that GA increased the germination rate, where
IVGs of 0.48 were obtained for cultivar Gefner and of 0.46
for cultivar PR-3.
The results showed a significant effect of the exogenous
application of GA on seed germination, this being very
effective treatment for breaking dormancy in seeds of A.
squamosa. It is possible to observe a great sensitivity of the
seeds to application of this phytoregulator since with any
tested dose of GA, the responses were improved, indicating
that the phenomenon of dormancy in these seeds might
depend on a balance between hormonal promoters and
inhibitors of growth (Weaver, 1987) and its breakage could
be affected by the change in hormonal balance, in which
GA acts to promote germination (Kigel and Galili, 1995).
21
 120 0.40

a a A A a 0.35
100 A

MGR (germinated seeds/d)

A a a 0.30
a A A
Germination (%)

80
0.25
B a
60 0.20 ab ab
ab b ab
0.15 A A A
40
c AB A A
0.10
20 b B
0.05
0 0
0 50 100 200 400 600 800 0 50 100 200 400 600 800

20 1.00
A
18 0.90
16 0.80
14 0.70
B
12 a B B 0.60
MGT (d)

B B 0.50 a

E
10 B AB
0.40 AB AB AB B AB
8 b b
b b b b A
6 0.30 ab ab b
4 0.20 b
b b
2 0.10
0 0
0 50 100 200 400 600 800 0 50 100 200 400 600 800
GA (mg L-1) GA (mg L-1)

Tolima Cundinamarca

FIGURE 3. Effect of different concentrations of GA on germination percentage (PG), mean germination time (MGT), mean germination rate (MGR)
and synchronization (E) in seeds of A. squamosa during incubation for 30 d in moist peat. Means with different letters on each location indicate
significant differences according to Tukey test (P≤0.05).

With respect to the patterns of germination, GA treatments
of seeds increased PG at each time point of incubation as
the effect of large increases ininitial germination rates and
shorter times to reach maximum PG, which was more
pronounced in seeds from Tolima.
Although all of the GA concentrations, applied by soaking,
enhanced the potential for seed germination, compared to
the non-application of GA, it is possible to highlight the
performance of the seeds treated with 600 mg L-1 GA. With
this concentration, it took less time for the seeds to reach
the maximum PG, also it generated a shorter time of ac-
celerated growth and, therefore, the seeds had the highest
PG in the first 5 d of incubation.
As mentioned, one of the characteristics of Annonaceae
seeds is the presence of an inmature, slow growing em-
bryo that is not yet developed when the fruit is riped. In
this state, the embryo remains even after the seeds are
removed from the fruits (Sanewski, 1991; De Smet et al.,
1999). The positive response of the GA application on
the seed germination was due to its effect on the embryo
growth (Bewley, 1997). Additionally, the GA is required
for the activation of embryonic growth and weakening of
tissues (endosperm, perisperm, seed coats) surrounding the
embryo and restricting its growth (Bewley, 1997; Bradford
et al., 2000; Hegashi et al., 2002). During the germination,
the embryo synthesizes and releases GA that promotes the
production of various hydrolytic enzymes, including the
hemicellulases, involved in the reserve solubilization of
seeds. In seeds of Annonaceae, hemicellulases digest the
reserves of galactomannans held in the endosperm (Baskin
and Baskin, 2014), forming sugars that are further trans-
ported to embryo growth, stimulating cell elongation and
promoting the rupture of seed coats by the radicle, thus,
improving greater uniformity of germination (Hopkins and
Hüner, 2009). Likewise, many of the enzymes involved in
the process of germination, such as lipases, proteases, phos-
phatases, and other hydrolases, are controlled or activated
by plant regulators, such as GA (Arteca, 1995).

22 Agron. Colomb. 34(1) 2016

The difference between the seeds of two locations to
treatment responses may be because plant responses to
phytoregulators depend on many factors; among them
are genetic and environmental ones that influence the
level of endogenous hormones and substances antagonistic
to germination (Agustí and Almela, 1991). Additionally,
the depth of primary dormancy is highly related to the
nutritional status of the mother plant (Geneve, 2003;
El-Keblawy and Al-Rawai, 2006) and the environmental
conditions during seed development (Finch-Savage and
Leubner-Metzger, 2006); these conditions strongly influ-
ence the ABA/GA ratio in seeds at the final phases of seed
maturation (Gutierrez et al., 2007) and this induced seed
dormancy. The seeds from Apulo, Cundinamarca were col-
lected at an altitude of 635 m a.s.l., an average temperature
of 24.7ºC, a monthly average sunshine of 118.8 h and an
annual rainfall of 1,146 mm, while seeds from Tolima were
collected in a lower region, with an altitude of 447 m a.s.l.,
annual average temperature of 28.2ºC, average monthly
sunshine of 183.23 h, and average annual rainfall of 1,732
mm. The habitat differences of the plant material could
influence the seed responses of the same species to treat-
ments (Nikolaeva, 2004).
Conclusions
The exogenous application of GA increased the unifor-
mity of the germination and germination potential of the
seeds of A. squamosa and generated large increases in the
initial germination rates and germination rates during the
incubation time.
There was agreat sensitivity of seeds to exogenously applied
GA, indicating that the embryo may present low growth
potential, which could explain the poor development and
rudimentary characteristic.
Positive responses to the application of GA are valuable
evidence that allows demonstrating a physiological com-
ponent of dormancy in the seeds of A. squamosa.
Acknowledgements
The authors express their gratitude to the Graduate School
of the Faculty of Agricultural Sciences of the Universidad
Nacional de Colombia, Bogota, for help in developing this
study.
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