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Effects of GA3 Pregerminative Treatment on Gentiana lutea L. var. aurantiaca Germination and Seedlings Morphology

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Gentiana lutea L. is widely used in bitter beverages and in medicine; Gentianae Radix is the pharmaceutical name of the root of G. lutea. These uses have generated a high demand. The wild populations of Gentiana lutea var. aurantiaca (M. Laínz) M. Laínz have been decimated; it is necessary to establish guidelines for its cultivation. Gentian as most alpine species has dormant seeds. Dormancy can be removed by cold and by means of a gibberellic acid (GA3) treatment. However, cold treatments produce low germination percentages and GA3 treatments may produce off-type seedlings. So, the objective was to determine, for the first time, the presowing treatments that allow high germination rate and good seedling morphology. The best pregerminative doses of GA3 to break seed dormancy were 100, 500, and 1000 ppm, while the best doses to optimize the seedling habit were 50 and 100 ppm. This study provides, for the first time, a 100 ppm GA3 dose that led to a high germination rate and good seedling morphology, as the starting point for gentian regular cultivation.
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Research Article
Effects of GA3Pregerminative Treatment on Gentiana lutea
L. var. aurantiaca Germination and Seedlings Morphology
Óscar González-López1and Pedro A. Casquero2
1Research Group of Engineering and Sustainable Agriculture, Natural Resources Institute, University of Le´
on,AvenuePortugal41,
24071 Le ´
on, Spain
2Research Group of Engineering and Sustainable Agriculture, Department of Agrarian Engineering and Sciences, University of Le´
on,
Ave n ue Po r tu g al 41 , 24 07 1 Le ´
on, Spain
Correspondence should be addressed to Pedro A. Casquero; pedro-casquero@unileon.es
Received  April ; Accepted  June ; Published  June 
Academic Editor: Antonio M. De Ron
Copyright ©  ´
O. Gonz´
alez-L´
opez and P. A. Casquero. is is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Gentiana lutea L. is widely used in bitter beverages and in medicine; Gentianae Radix is the pharmaceutical name of the root of
G. lutea. ese uses have generated a high demand. e wild populations of Gentiana lutea var. aurantiaca (M. La´
ınz) M. La´
ınz
have been decimated; it is necessary to establish guidelines for its cultivation. Gentian as most alpine species has dormant seeds.
Dormancy can be removed by cold and by means of a gibberellic acid (GA3) treatment. However, cold treatments produce low
germination percentages and GA3treatments may produce o-type seedlings. So, the objective was to determine, for the rst time,
the presowing treatments that allow high germination rate and good seedling morphology. e best pregerminative doses of GA3
to break seed dormancy were , , and  ppm, while the best doses to optimize the seedling habit were  and ppm.
is study provides, for the rst time, a  ppm GA3dose that led to a high germination rate and good seedling morphology, as
the starting point for gentian regular cultivation.
1. Introduction
Gentiana lutea L. (Gentianaceae) is a herbaceous perennial
plant native to the mountains of central and southern Europe
[,].Gentianrootsarewidelyusedinbitterbeverages,in
food products, and also in traditional medicine to stimulate
theappetiteandimprovedigestion[,]. ese uses have
generated a great demand, so that more than  tons of
gentian root is produced from  tons of wild stocks every
year around the world []. Due to the slow growth of this
species,thewildpopulationsofG. lutea have been decimated
and are now close to their disappearance and/or are dicult
to revive. e increasing demand has provoked alarm about
the extinction of the species, and for this reason gentian is
being protected throughout Europe by law.
In the Northwest part of Iberian Pen´
ınsula (Western
Cantabric Mountains), gentian root has been used as a tonic
for the stomach and to restore appetite []. In this area
Gentiana lutea L. owers show a colour ranging from orange
to almost red when compared to yellow owers of G. lutea
L. subsp. lutea. ese populations have been classied as
Gentiana lutea L. var. aurantiaca []. In the Le´
on province
some populations of aurantiaca variety populations show
signsofisolationwithlowlevelofgeneticdiversity[]
which indicates reduced reproductive tness and elevated
extinction risks [].
e main economic sources in European mountain areas
include mining (a decreasing activity), ranching (in crisis),
and tourism. Collection of gentian roots has traditionally
been a supplement to the family income. Due to the current
economic crisis, people have started to collect gentian again
in a furtive way for an extra income. Considering the harsh
climate conditions of the mountain regions, where both
horticulture and fruit growths are dicult to carry out, the
cultivation of gentian is an alternative economic activity for
large unused lands with agro climatic characteristics suitable
for its growth.
Hindawi Publishing Corporation
e Scientific World Journal
Volume 2014, Article ID 751279, 6 pages
http://dx.doi.org/10.1155/2014/751279
e Scientic World Journal
G. lutea, as most alpine species, has dormant seeds,
exhibiting mainly physiological dormancy [,]. is
dormancy can be removed by cold treatment [,], by
cold stratication [], and by GA3treatment. GA3,alone
or together with other chemical or physical treatments, is
widely used to remove seed dormancy []ortoimprovethe
development of the seedlings, although diverse results have
been obtained []. Related to this, a study with G. lutea
seeds was carried out on germination rates in Petri dishes
using GA3and a stratication treatment []. However, some
of these treatments, such as gibberellic acid, may produce
weak and o-type seedlings in G. lutea [].
erefore, this study aims to establish which GA3dose,
applied as pregerminative treatment, improves the germina-
tion rate without aecting the development of seedlings, as
the rst step for gentian cultivation.
2. Materials and Methods
2.1. General Seed Trials. Seeds of Gentiana lutea L. var.
aurantiaca from population located in the western part of
the Cantabrian mountains (Spain) were collected (Figure ).
Seeds from mature capsules containing well-developed ripe
seeds were taken from  randomly selected plants. Aer this,
the seeds were manually cleaned, removing any damaged,
empty, or visually malformed seeds, kept in paper bags, and
then stored in a fridge at Cinsideadarkglassbottlewith
silica gel.
Stratication Treatment. Seeds were stratied for  months
over a silica sand base (Ø .–. mm). Both tray and silica
sand were sterilized by autoclaving at . kg cm−2 and C
for  h, on each of three consecutive days. Aer placing the
seeds well distributed over the tray surface, another layer
( cm in height) of silica sand was added covering the seeds
(Figure ). Finally, the sand was moistened with distilled
water, avoiding saturation and placing the tray inside the
refrigerator at C±C. e tray was checked every week
during the next  days in order to control the sand humidity.
Stratied seeds were cleaned using a metallic sieve with Ø
. mm.
GA3Tre atm ent. Seeds were soaked at Cforhours
in GA3(Sigma-Aldrich) water solution (, , , and
 ppm) and were cleaned with distilled water to remove
GA3residues before sowing.
2.2. Petri Dishes Germination Assay. Tw o t reat ment s were
applied to the gentian seeds: cold treatment (stratied or not)
and GA3dose(,,,andppm).Fivereplicates
of  seeds in each treatment were tested for germination
on top of a lter paper sheet with  ml distilled water in  cm
plastic Petri dishes (Figure (b)). Filter papers were rewetted
with distilled water as required. Dishes were checked twice
a week during a -day test period and germinated seeds
were counted and removed. It was established that seeds
were germinated when the length of the radicle exceeded
the seed coat with  mm (Figure (c)). e conditions during
Le ́
on
N
60 km Le ́
on province
F : Map showing the location of sampled population of
Gentiana lutea L. var. aurantiaca.
F : Addition of silica sand over gentian seeds for stratication
treatment.
this time were in a growth chamber (Figure (a)) with a
constant temperature of Candahlightphotoperiod
(provided by cool white uorescent tubes with an irradiance
of  𝜇molm−2s−1). At the end of the incubation period
( days), the nal germination percentage and the mean
germination time (MGT) were calculated according to Ellis
and Roberts [].
2.3. Forest Trays Assay. Five replicates of  seeds each were
tested with ve GA3dose (, , ,  and  ppm). All
seeds were stratied as specied in Section ..eforestry
seed trays had a volume of  cm3and a height of  cm. A
commercial substrate, n from Pindstrup Mosebrug Sae,
wasused.issubstratehasamixof%whitepeatand%
black peat (pH .–), with similar conditions to those of the
mountain soils. Pesticide seed treatment (Tiram % WP)
was applied according to Valenciano et al. []tocontrolthe
development of phytopathogenic fungi []. Every lot of 
seeds was sowed in a unique tray and every cell was covered
with  cm3of sterilized silica sand, which is equivalent to
cminheight.
 days aer sowing, the measurement of the plants
was carried out. Once the development of the plants was
observed, two dierent characteristics were measured: stem
length, that is, the length of the stem (mm) from the substrate
level to the apex of the primary stem, and leaf length, the
length (mm) of the most developed leaf in the third pair
e Scientic World Journal
(a)
(b)
(c)
F : Petri dishes germination assay of G. lutea L. var. aurantiaca: (a) growth chamber; (b) Petri dishes distribution; (c) germinated seeds.
of leaves starting from the base. Both measurements were
performed in  random seedlings per replication.
2.4. Statistical Analysis. e values of nal germination
percentages were arcsine transformed (untransformed data
appears in the gures). In each experiment, the data was ana-
lyzed using the general linear model (GLM procedure) and
whenever the analyses showed signicance least signicant
dierences (LSD) were computed at the . probability level
to compare means. All analyses were performed using the
SASversion..soware(SASInstituteInc.,,Cary,NC,
USA).
3. Results and Discussion
3.1. Germination. According to analysis of variance, stratied
seeds showed higher germination rates than nonstratied
seeds for each tested GA3dose. ere were no interactions
between the sources of variation used (cold treatments and
GA3treatments). e GA3treatment had an eect on germi-
nation time so a generalized early germination was observed
when  and  ppm doses were used (Figure ). In these
cases, aer  days, most of the seeds were germinated.
Medium germination time (MGT) for these doses was 19.82±
0.475 and 20.34 ± 0.545,respectively.Regardingppm,
a quick germination was also observed, but not as strong
(MGT = 23.77 ± 1.111), and newly germinated seeds were
ob s er ved until the  t h day. Fi nally, t h e   ppm d ose mea nt
a good nal germination rate, but the number of germinated
seeds increased until the th day at a gradual rate (MGT =
39.16 ± 2.186).
At the end of the  days of the germination test, none
of the sterilized water pretreated seeds were germinated.
All GA3treatments yielded signicantly higher germination
Germination (%)
c
b
ab
a
a
0
10
20
30
40
50
60
70
80
90
100
6 132027344249566370778490
Test da y
C
50 ppm
100 ppm
500 ppm
1000 ppm
F : Germination curves of Gentiana lutea var. aurantiaca
seeds using dierent doses of GA3(, , , and  ppm) and
control (C).
percentages compared to the control. e greater the hor-
monal dose used, the greater the percentage of germination
obtained. Seeds pretreated with  ppm of GA3resulted in
the highest germination percentage (%), not signicantly
dierent compared to the  ppm (%) or  ppm (%)
doses, but signicantly dierent to the  ppm dose (%)
(Figure ).
e results obtained in Petri dishes germination test are
in accordance with those obtained by P´
erez-Garc´
ıa et al. [],
which verify the high degree of success in the elimination of
seed dormancy using GA3, this eect being greater with high
doses (Figure ). e highest GA3doses ( and  ppm)
e Scientic World Journal
Germination (%)
c
b
ab aa
0
20
40
60
80
100
Control 50 100 500 1000
GA3(ppm)
F : Germination percentages of Gentiana lutea var. aurantiaca seeds using dierent GA3doses (, , , and  ppm) and control
at the end of the test of  days of germination. Bars with dierent letters are signicantly dierent (𝑃 = 0.05) according to LSD.
cc
b
a
0
5
10
15
20
25
30
50 100 500 1000
Length (mm)
GA3(ppm)
F : Stem length of Gentiana lutea var. aurantiaca seedlings using dierent GA3doses (, , , and ppm) on the th day
aer sowing. Bars with dierent letters are signicantly dierent (𝑃 = 0.05) according to LSD.
provided a quick and grouped germination. e eect of the
GA3dose of  ppm was that the germination was slower
during the rst part of the test compared to higher GA3doses,
but MGT and germination were similar for all three doses.
3.2. Seedlings Development. Analysis of variance shows sig-
nicant dierences between GA3treatments. Stem length
was greater when the GA3dose used was higher. e dose
of  ppm provided the longest (. mm) stem, being
signicantly dierent from the others, as well as the  ppm
dose (. mm). Signicant dierences were not found
between the remaining doses of  ppm and  ppm, both
providing the shortest stems of all the treatments (.mm
and . mm, resp.) (Figure ).
Depending on the GA3dose, gentian seedlings showed
dierent growth habits (Figure ). H: the aerial part of the
plant is formed by a basal rosette of well-developed leaves
and the stem is very short. H: the aerial part of the plant is
formed by a basal rosette of well-developed leaves; the stem
is formed and strong. H: the leaves are elongated; they are
arranged along a stem formed with short internodes. H:
small leaves are arranged on a very long and weak stem: long
internodes; light yellow-green color. H: poorly developed
and large leaves: liform appearance; long internodes; light
yellow-green color.
Considering these ve growth habits the number of plants
showing each of the habits was counted according to the
dierent GA3doses (Figure ). e habit shown by the
seedlings at low doses was similar, H being the predominant.
Meanwhile, for high GA3doses,  and  ppm, the pre-
dominant growth habit was H, and also H was abundant. In
both,  ppm and  ppm, none of the seedlings showed
the H habit, which is considered the best for the gentian
seedlings.
e negative eect on the development of the seedlings
was observed only at  and  ppm doses, whereas a
high percentage of seedlings showed o-type growth habits
with long stems. Using these doses, seedlings were very weak,
poorly developed, and with a threadlike appearance, there-
fore, more susceptible to low temperatures, insect attacks,
physical breakage, or drying out. While at low doses ( ppm
andppm)thestemswereveryshortandthepredominant
seedlings morphology was very similar to that observed in
nature, where the seedlings are formed by a basal rosette, like
other perennial mountainous plants, protecting themselves
from the hard conditions of the mountain climate.
4. Conclusion
e best GA3pregerminative doses to break seed dormancy
of Gentiana lutea L. var. aurantiaca were , , and
 ppm. Doses of  and ppm GA3not only allow
obtaining a high and uniform germination, but also produce
high percentages of weak and o-type seedlings. e best
doses to optimize seedling habit were  and  ppm. is
studyprovides,forthersttime,appmGA
3doses that
e Scientic World Journal
H1H2H3H4H5
F : Growth habits (H–H) of Gentiana lutea var. aurantiaca seedlings on the th day aer sowing treated seeds with GA3(, ,
, and  ppm).
Seedlings (%)
aa
ee
bb
a
b
bc
b
a
b
dc
b
b
dd
c
0
10
20
30
40
50
60
70
80
90
100
50 100 500 1000
GA3(ppm)
H1
H2
H3
H4
H5
F : Percentage of Gentiana lutea var. aurantiaca seedlings
showing dierent growth habits for each of the GA3doses (, ,
, and  ppm). Bars with dierent letters for each GA3doses
are signicantly dierent (𝑃 = 0.05) according to LSD.
led to a high germination rate and good seedling morphology,
as the starting point for gentian regular cultivation.
Conflict of Interests
e authors declare that there is no conict of interests
regarding the publication of this paper.
Acknowledgments
e authors acknowledge the Regional Ministry of Education
of the Junta de Castilla y Le´
on and the European Social
Fund PIRTU Grants (Orden EDU//), the Regional
Ministry of the Environment of the Junta de Castilla y Le´
on
Project no. //, the sta from the Forest and
Range Management Research Institute (CRA Agricultural
Research Council-Trento), and Asociaci´
on Promotora de la
Genciana y otras plantas de inter´
es de la monta˜
na occidental
leonesa for collaborating to collect gentian material.
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... Cold stratification and pre-treatment with gibberellic acid (GA 3 ) may help to overcome dormancy in several Gentiana species and improve seed germination (Struwe and Albert 2002), even though high concentrations may have inhibitory effects (Mikula and Rybczyńsky 2001). In particular, Pérez-García et al. (2012) and González-López and Casquero (2014) reported that GA 3 may efficiently increase germination in seeds of G. lutea under different temperatures (from 15 to 20°C). Seed storage is considered amongst the most important approaches to preserve plant germplasm (Bacchetta et al. 2006, Smith et al. 2011, and seed germination is a key technique to better understand plant species biology and ensure effective ex situ conservation (Carta et al. 2014, Ludewig et al. 2014. ...
... symphyandra (Mt Grande population). These results are consistent with those obtained in previous studies with comparable protocols (Pérez-García et al. 2012, González-López andCasquero 2014), supporting the reliability of the method with regard to GA 3 concentration and germination temperature, and confirming its effectiveness as a successful treatment to enhance seed germination. As in most mountain plants, G. lutea seeds exhibit dormancy which can be overcome by means of cold treatments or gibberellic acid application. ...
... (a)  Mt Grande and (b)  Mt Nanos population. drastically enhances seed germination in G. lutea (Pérez-García et al. 2012, González-López andCasquero 2014), hence our results obtained from GA 3 addition do not reflect the natural conditions. However, the small-scale complexity of landscape in natural habitats may enhance the heterogeneity of several environmental factors (Scherrer andKorner 2011, Opedal et al. 2015); the spatial variability of environmental conditions within and among sites may therefore reduce the probability of obtaining homogeneous germination conditions in the field. ...
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Gentiana lutea L. (yellow gentian, Gentianaceae) is a protected orophyte of central and southern Europe, mainly threatened by the uncontrolled collection of its rhizome, used in traditional medicine and for liquor production. The species is self-compatible, but outcrossing mediated by pollinators is needed to obtain a viable progeny. In this study, we considered five natural populations belonging to the four subspecies of G. lutea. We performed controlled pollinations in the field (self- versus cross-pollination) followed by seed germination tests in laboratory conditions, adding a solution of gibberellic acid, in order to evaluate the seed performance. A cumulative index of inbreeding depression was calculated considering maternal reproductive output as well as seed performance traits. Seed weight and seed germination performance was similar between seeds resulting from naturally pollinated and pollen-augmented flowers and higher compared to selfed flowers, highlighting a disadvantage of selfing and the importance of cross-pollen transfer in natural conditions. However, in the small and isolated population of G. lutea subsp. symphyandra on Mt Grande we found a general reduction in seed germination rate, likely due to increased selfing or mating among close relatives as a consequence of a severe bottleneck. We discuss our results with regards to implications for conservation practices.
... In a previous study, dormancy breaking and germination were improved by applying GA3 to Gentiana species; this has also been observed in numerous other plant species [22,[43][44][45][46]. Similarly, in this study, the final germination was enhanced by the application of 100 and 1000 mg•L −1 exogenous GA3 at 90.0 ± 2.0% and 84.0 ± 4.3%, respectively ( Figure 6A). ...
... In a previous study, GA3 application was more effective at 1000 mg•L −1 than at 50 and 100 mg•L −1 and positively affected the percentage and speed of germination in G. lutea L. var. aurantiaca seeds [43]. In addition, 250 mg•L −1 GA3 remarkably influenced the germination percentage compared to the results of 3-month cold stratification in G. lutea L. subsp. ...
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This study investigated the kind of seed dormancy and seed germination of Gentiana triflora var. japonica (Kusn.) H. Hara for developing a seed propagation method. The seeds were collected in October 2020 from plants at Mt. Sobaeksan, Korea. In a water imbibition experiment, seed weights increased by > 101.9% of their initial masses over 12 h. Effects of incubation temperature (5, 15, 20, 25, 15/6, or 25/15 °C), cold stratification period (5 °C; 0, 4, 8, or 12 weeks), and gibberellic acid (GA3; 0, 10, 100, or 1000 mg∙L−1) and potassium nitrate treatment (KNO3; 0, 1000, 2000, or 4000 mg∙L−1) on seed germination were investigated to characterize seed dormancy. These seeds exhibited underdeveloped embryos during seed dispersal. The seeds failed to reach the final germination of 15.0% after treatment at 5, 15, 20, 25, 15/6, or 25/15 °C. After cold stratification for 8 weeks, the germination increased dramatically by > 90.0% compared to that at 0 weeks. After the GA3 treatment, the germination reached > 80.0% within 5 days. The final germination was 90.0% in the 100 mg∙L−1 GA3 treatment group. However, the KNO3 treatment had no effect on seed germination. Therefore, the G. triflora var. japonica seeds exhibited non-deep simple morphophysiological dormancy.
... GA 3 pretreatment is known to promote the germination of seeds with non-deep PD, but this is not the case for all species with intermediate PD [16,53]. GA 3 enhances non-dormant seed germination and germination maintenance, it has also been observed in numerous other plant species [54][55][56][57]. The GR of L. coreana increased from 14.0% (control) to 82.0% at 25/15 • C with 250 mg·L −1 GA 3 treatment, and improved 76.0% and 66.0% at 20/10 and 25 • C with 250 mg·L −1 GA 3 treatment , respectively. ...
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This study aimed to determine the type of seed dormancy and to identify a suitable method of dormancy-breaking for an efficient seed viability test of Lysimachia coreana Nakai. To confirm the effect of gibberellic acid (GA3) on seed germination at different temperatures, germination tests were conducted at 5, 15, 20, 25, 20/10, and 25/15 °C (12/12 h, light/dark), using 1% agar with 100, 250, and 500 mg·L−1 GA3. Seeds were also stratified at 5 and 25/15 °C for 6 and 9 weeks, respectively, and then germinated at the same temperature. Seeds treated with GA3 demonstrated an increased germination rate (GR) at all temperatures except 5 °C. The highest GR was 82.0% at 25/15 °C and 250 mg·L−1 GA3 (4.8 times higher than the control (14.0%)). Additionally, GR increased after cold stratification, whereas seeds did not germinate after warm stratification at all temperatures. After cold stratification, the highest GR was 56.0% at 25/15 °C, which was lower than the GR observed after GA3 treatment. We hypothesized that L. coreana seeds have a non-deep physiological dormancy and concluded that 250 mg·L−1 GA3 treatment is more effective than cold stratification (9 weeks) for L. coreana seed-dormancy-breaking.
... The plant is herbaceous perennial and grows in Europe and Asia. The plant has slow growth habit and produces dormant seed ( González-López and Casquero 2014). The species is assessed as least concern (LC) in the IUCN European Red List ( Bilz et al. 2011). ...
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The twentieth century witnessed deterioration of biodiversity and loss of natural habitats of many plant species. However, it was also an era of significant progress in tissue culture technology which opened further vistas for multiplication and conservation of plant species. Synthetic seed technology is one such method which involves selection of a suitable explant and encapsulating it in an apposite matrix for successful germination and conversion into a healthy plantlet. The underlying basis of synthetic seed technology is to imitate natural plant development that occurs through seed germination. This method has been successfully employed for propagation and storage of various forest, medicinal, and vegetable plant species. The technique is of pivotal importance for species which produce non-viable seeds, recalcitrant seeds, or have limited and rare seed production. Threatened and endangered plant species are one such category which has several bottlenecks in seed development, reproduction, and establishment in their natural environments that have undergone disruptive changes. This review aims to explore and assess the potential of synthetic seed technology as an effective approach to support conservation strategies for endangered plant species.
... Sustainable use of natural resources can only be achieved with the introduction of plants, as well as cultivation, which is an important way to protect endangered plant species (Menges, 2000). Therefore, in recent decades of the last and at the beginning of this century, abroad, but also in Serbia and Slovenia great efforts had been made to develop a technology for gentian cultivation, especially in rural mountainous areas (Kušar and Baričević, 2006;Radanović et al., 2007b;2007c;Radanović, 2008;Franz, 2012;Radanović et al., 2013;Balijagić, 2013;González-López et al., 2014;Sand, 2015). ...
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Malvella sherardiana (L.) Jaub & Spach (Malvaceae) is a perennial herbaceous plant listed as Vulnerable in the Red List of threatened plant species for Spain. The germination characteristics under controlled conditions of light and temperature of M. sherardiana seeds were studied. Mechanical scarification, cold stratification, liquid nitrogen, freezing, hot water, dry heat, sulphuric acid, soaking in distilled water and soaking in gibberellic acid were used as pre-sowing treatments applied for enhancing germination. The untreated seeds showed a high dormancy at all temperature regimens tested and sulphuric acid scarification drastically improved final germination percentage and germination rate. Germination of seeds soaked in concentrated sulphuric acid arithmetically increased as soaking time increased (from 5 min to 3 h), but seeds soaked in acid for more than 4 h failed to germinate. The application of a gibberellic acid solution (1000 mg l−1) increased the germination percentages for all tested soaking times in sulphuric acid. However, it was only significantly effective for the soaking time of 3 h. The other pre-sowing treatments resulted in some germination, but none resulted in greater germination than sulphuric acid. As embryos are fully developed and the seed coat is water permeable, we conclude that M. sherardiana seeds present physiological dormancy. The main conclusion of the study is that dormancy of M. sherardiana seeds is attributed to mechanical restriction of the embryo caused by the tough seed coat. This is the first report on germination requirements of the Malvella genus.
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Gentians are alpine plants and two endemic species, Gentiana triflora and G. scabra, are cultivated as ornamentals in Japan. Because most of the Japanese cultivars are F1 hybrids, seed germination rate is an important factor for maintaining these cultivars. However, germination rates frequently differ among cultivars. Poor germination is a severe problem for gentian production, although there is almost no difference in their germination period and growth rate. In this study, we compared seed germination rates among three cultivars of G. triflora. A high rate of seed germination was observed in ‘Majel’ (MJ) and ‘Iwate’ (IW), but a low rate was recorded in ‘Iwate-Yumeminori’ (YM). We conducted a targeted metabolome analysis using these cultivars to elucidate the potential cause of inhibited seed germination. Multivariate analysis revealed that the aberrant accumulation of specific amino acids and a decrease in energy metabolites were observed in YM seed. Furthermore, protein concentrations and proteolytic activities in YM seed were lower than in MJ seed. These results imply that energy depletion was the main reason for the decreased rate of germination and that this depletion inhibited processes involved in seed germination, including de novo synthesis of the proteins necessary for germination.
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Gentiana lutea L. (yellow gentian, Gentianaceae) is an important medicinal plant under protection as endangered species in most European countries. The aim of this work was to evaluate variation in seed mass, seed water content, and seed germination among 56 wild accessions of G. lutea. The effect of gibberellic acid (GA3), putrescine, moist chilling, and level of ripeness of seeds on subsequent germination was also investigated. Seeds of G. lutea showed physiological dormancy (final germination percentages ranged from 0% to 11%, depending on the accession) and GA3 enhanced seed germination drastically in all the accessions. The highest germination (99%) of GA3-treated seeds was reached at 15 °C. Final germination percentage and germination rate (as expressed by mean germination time), as well as seed mass and seed water content, varied significantly among accessions. In general, 1 year moist chilling did not significantly enhance G. lutea seed germination. For most accessions, no significant differences were found between fully ripe seeds and less ripe seeds for seed water content, seed mass, and seed germination. Applications of GA3 were always most effective than those of putrescine for increasing seed germination.
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Gentiana lutea is a high mountain wild plant whose roots have been traditionally used for medicinal purposes in the northwest mountains of the Iberian Peninsula, mainly as a stomach tonic or to solve a lack of appetite. These medicinal properties are mainly due to bitter compounds such as amarogentin, gentiopicrin and swertiamarin, which are mainly found in the outer layers of the roots. The aim of the project was to establish the relationship between soil characteristics and the development of gentian root system in adult plants. Soil and underground part samples have been collected from twelve populations. The underground parts, conveniently called ‘roots’, has been separated in rhizome, principal and secondary (or lateral) roots. The diameter and cortex thickness of the roots were measured. The results show that there was a positive significant correlation between Mn and K content in the soil and the ratio between cortex thickness and the total thickness of the roots. A significant positive correlation was also found for altitude and cortex thickness in both primary and secondary roots. In conclusion, high altitude and soils with elevated levels of Mn and K improved the growth and the quality of the roots.
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Gentiana lutea is a high mountain wild plant whose roots have been traditionally used for medicinal purposes in the northwest mountains of the Iberian Peninsula, mainly as a stomach tonic or to solve a lack of appetite. These medicinal properties are mainly due to bitter compounds such as amarogentin, gentiopicrin and swertiamarin, which are mainly found in the outer layers of the roots. The aim of the project was to establish the relationship between soil characteristics and the development of gentian root system in adult plants. Soil and underground part samples have been collected from twelve populations. The underground parts, conveniently called 'roots', has been separated in rhizome, principal and secondary (or lateral) roots. The diameter and cortex thickness of the roots were measured. The results show that there was a positive significant correlation between Mn and K content in the soil and the ratio between cortex thickness and the total thickness of the roots. A significant positive correlation was also found for altitude and cortex thickness in both primary and secondary roots. In conclusion, high altitude and soils with elevated levels of Mn and K improved the growth and the quality of the roots.
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Kelussia odoratissima Mozaff. is one of the endogenous plant species of Iran which is exposed to extinction during the recent decades. Seeds of this plant have dormancy that causes reduction of seed germination. Experiment was performed as factorial with complete randomized design with 3 factors: polyethylene glycol (PEG) priming, gibberellic acid (GA(3)) treatment, GA(3) application time. This research was performed to evaluate priming effect alone and in combination with GA(3), on seed germination and seedling growth parameters. Results show that seed priming with PEG alone had negative effect and GA(3) application solely did not have any significant effect on the seed germination percentage in comparison with the control. Seedling growth responses to PEG priming was similar (in -1 MPa) or lesser (in -1.5 MPa) than control, GA(3) significantly improved seedling growth as compared to the control. Application of GA(3) after PEG was better than using of GA(3) before PEG or co-application of GA(3) and PEG. These results suggest that probably, GA(3) compensated negative effect on long time priming.