Horticulturae 08 00561
Horticulturae 08 00561
Horticulturae 08 00561
Article
Seaweed Extracts as Substitutes of Synthetic Hormones for
Rooting Promotion in Rose Cuttings
Silvia Traversari 1,2 , Sonia Cacini 2, * and Beatrice Nesi 2
1 Research Institute on Terrestrial Ecosystems (IRET), National Research Council (CNR), Via Moruzzi 1,
56124 Pisa, Italy; [email protected]
2 CREA Research Centre for Vegetable and Ornamental Crops, Council for Agricultural Research and
Economics, Via dei Fiori 8, 51012 Pescia, Italy; [email protected]
* Correspondence: [email protected]
Abstract: In the horticultural sector, the achievement of an efficient and eco-friendly sustainable
production of plants is nowadays challenging. Indeed, in plant vegetative propagation of woody
ornamentals, the substitution of chemical products used to promote rooting of cuttings with natural
extracts would be a desirable goal. Thus, the aim of this work was to test the replacement of synthetic
phytoregulators, such as auxins and brassinosteroids, with biostimulants, such as seaweed extracts,
for the rooting promotion of rose cuttings. The rooting rate and biometric parameters of control
cuttings treated with distilled water were compared with those of cuttings treated with synthetic
hormones, i.e., auxins or 22(S),23(S)-homobrassinolide, or two commercial products based on low
temperature seaweed extracts, i.e., Kelpak® and Phylgreen. Two scented hybrid tea rose cultivars
were used to assess possible genotype-dependent effects, i.e., ‘Michelangelo® ’ and ‘Cosmos® ’. Auxins
confirmed their role in root growth enhancement in ornamental plant cuttings. Like these phytoreg-
ulators, Kelpak® improved the survival rate and root biometric parameters of both rose cuttings,
Citation: Traversari, S.; Cacini, S.; highlighting its suitability for the replacement of synthetic products used for rooting promotion in
Nesi, B. Seaweed Extracts as
rose propagation. Brassinosteroids showed a species-dependent effect, increasing the root biomass
Substitutes of Synthetic Hormones
in ‘Cosmos® ’ while it resulted as distilled water in ‘Michelangelo® ’. Phylgreen did not improve the
for Rooting Promotion in Rose
rooting of both rose cuttings, highlighting the necessity of evaluating the applicability and methodol-
Cuttings. Horticulturae 2022, 8, 561.
ogy for this product before its use. In conclusions, our results highlighted the possibility to replace
https://doi.org/10.3390/
horticulturae8070561
chemical products in rose cutting production.
Academic Editors: Jean Carlos Keywords: auxins; homobrassinolide; Kelpak® ; ornamental plant propagation; Phylgreen
Cardoso, Wagner Vendrame and
Chao Ma
root length as well as the total soluble sugars in stem cuttings of two cultivars of barberry
ornamental plants [10]. However, the small amount and fast metabolization of BRs in plant
tissues make the production costs of natural compounds too high, and therefore structural
and functional analogues are commonly used, such as the 24-epibrassinolide [11].
In modern agriculture, the substitution of chemical inputs, such as phytoregulators,
with alternative natural eco-friendly products presents a key challenge [12] also to meet the
transition towards agroecological production systems and organic farming principles [13].
In this context, seaweed extracts constitute a promising alternative solution to the use
of synthetic products in the promotion of plant yield and health [14]. Different seaweed
species have shown their activity as biostimulants, probably related to the presence of
bioactive ingredients such as phytohormones as well as carbohydrates, proteins, and min-
eral elements [15,16]. Moreover, seaweed extracts have been shown to be not only naturally
enriched in phytohormones but also able to promote the endogenous biosynthesis of aux-
ins, cytokinin, and gibberellins [17]. Beyond their effect as growth promoters, seaweed
extracts have also disease suppressing effects [18], potentially reducing the application of
phytochemicals as well. However, the molecular mechanisms behind their function are
still little known. Moreover, new sustainable extraction procedures have been developed
to avoid the use of chemicals and preserve the seaweed’s original quality, increasing their
environmentally friendly aspects [18]. Despite these promising features, to our knowl-
edge, only a few seaweed extracts have been tested for rooting promotion in rose cutting
propagation, prevalently containing the Ascophyllum nodosum extract [6].
Thus, the aim of this work was to test the replacement of synthetic phytoregulators
with two seaweed extracts, i.e., Kelpak® and Phylgreen, for promoting the rooting of rose
cuttings. The two seaweed products were evaluated in comparison with the absence of
treatment (only distilled water), or two different synthetic phytoregulators, i.e., auxins or
BRs. The extracts were tested on two rose cultivars to assess possible differences related to
genotype-dependent responses.
3 min for 3 weeks, while after this period they were sprayed 3 times per day for 3 min.
Average temperature, humidity, and lighting during the experimental period were 21 ◦ C,
70%, and 322 Watt m−2 . Data were acquired by an EnviroMonitor weather station (Davis
Instruments, Hayward, CA, USA).
2.4. Statistics
Data were tested for normal distribution using the Shapiro–Wilk normality test and
then analyzed by a two-way ANOVA (treatment and cultivar as variables) to assess differ-
ences in treatment between the cultivars and then with a one-way ANOVA, followed by a
Tukey’s post-hoc test, to highlight significant differences between treatments (p ≤ 0.05, 0.01,
and 0.001). The statistical analyses and graphs were performed with Prism 9 (GraphPad
Software, Inc., La Jolla, CA, USA).
3. Results
3.1. Survival Percentage and Biometric Measures
Control cuttings of both cultivars treated with distilled H2 O showed a rooting percent-
age on average of 65% (Figure 1). Cuttings treated with BRA and PHY had a slightly lower
rooting rate than control cuttings in cv. ‘Michelangelo® ’. The highest rooting percentage
in this cultivar was measured under KEL treatment (77%, + 19% in comparison to control
cuttings). On the contrary, all treatments increased the rooting percentage in cv. ‘Cosmos® ’,
particularly the cuttings treated with AUX, which had a survival rate 22% higher than the
control ones.
After 16 days from the treatments, callus formation was mostly present in the lowest
part of cuttings treated with AUX, BRA, and KEL (Figure 2). Callus was absent from
cuttings treated with PHY, while it was a small portion in those treated with H2 O.
Every cutting produced one single sprout and sprout length was not significantly
different between treatments in both cultivars (Figure 3).
Treatments had a different effect on the root biometric parameters of the two rose
cultivars (Figure 4). The root FW (Figure 4A) was higher under AUX and KEL treatments in
cv. ‘Michelangelo® ’ (+99 and 62%, respectively) and under BRA treatment in cv. ‘Cosmos® ’
(+82%). The same trend was shown in root DW (Figure 4B) by cv. ‘Cosmos® ’. Root length
(Figure 4C) was higher under AUX and KEL treatments in cv. ‘Michelangelo® ’ (+103 and
75%, respectively) and under BRA and PHY treatments in cv. ‘Cosmos® ’ (+76 and 71%,
respectively). Root area (Figure 4D) was higher under AUX, KEL, and PHY treatments in
cv. ‘Michelangelo® ’ and under AUX, BRA, and PHY treatments in cv. ‘Cosmos® ’.
‘Cosmos
‘Cosmos
®’ (+ 82%). The same trend was shown in root DW (Figure 4B) by cv. ‘Cosmos®’.
®’ (+ 82%). The same trend was shown in root DW (Figure 4B) by cv. ‘Cosmos®’.
Root length (Figure
Root length (Figure 4C) was 4C)higher
was higher
underunder AUXKEL
AUX and andtreatments
KEL treatments
in cv. in cv. ‘Michelangelo
‘Michelangelo ®’
®’
(+ 103(+and
10375%,
and respectively)
75%, respectively) and under BRAPHY
and treatments
PHY treatments
in cv. in cv. ‘Cosmos
®’ (+ 76’ (+ 76
®
and under BRA and ‘Cosmos
and 71%, respectively). Root area (Figure 4D) was higher under AUX, KEL, and PHY PHY
and 71%, respectively). Root area (Figure 4D) was higher under AUX, KEL, and
Horticulturae 2022, 8, 561 treatments
treatments in cv. in cv. ‘Michelangelo
‘Michelangelo ®’ and ’under
® and under AUX, BRA, and PHY treatments in cv. ‘Cos-
AUX, BRA, and PHY treatments in cv. ‘Cos- 4 of 9
mos®’.mos ’.
®
H2O H2O AUX AUX BRA BRA KEL KEL PHY PHY
100 100
77 78 78
80 80 77
71
7070 7170
40 40
20 20
0 0
cv. 'Michelangelo
®' ®' cv. 'Cosmos
®' ®'
cv. 'Michelangelo cv. 'Cosmos
® ’‘Cosmos ® ’ cuttings.
FigureFigure 1.1.Rooting rate (%) of cv.
of ‘Michelangelo
Rooting ®’ and ®’ cuttings. Values were calculated on
Figure
1. Rooting rate (%) ofrate
cv. (%) cv. ‘Michelangelo
‘Michelangelo ®’ and ‘Cosmos and ‘Cosmos
®’ cuttings. Values Values
were calculatedwere
on calcu-
the 900
the 900 lated total cuttings.
on the 900
total cuttings. Cuttings
total cuttings.
Cuttings were treated
Cuttings
were treated with:
were
with: H
H2Otreated
2 O = distilled
with:water;
= distilled water;
H2 O AUX AUX
= distilled = auxins;
water;
= auxins; BRAAUXBRA = ho-
= auxins;
= ho-
mobrassinolide;
KEL = KEL
BRA = homobrassinolide;
mobrassinolide; Kelpak KEL;==
= Kelpak
®; PHY
® PHY = Phylgreen.
Kelpak® ; PHY = Phylgreen.
Phylgreen.
Figure 2. Images of cuttings 16 days after the start of treatments. Orange arrows indicate the
callus formation.
Horticulturae 2022, 8, x FOR PEER REVIEW 5 of 9
Horticulturae 2022, 8, x FOR PEER REVIEW 5 of 9
Horticulturae 2022, 8, 561 Figure 2. Images of cuttings 16 days after the start of treatments. Orange arrows indicate the callus
5 of 9
Figure 2. Images of cuttings 16 days after the start of treatments. Orange arrows indicate the callus
formation.
formation.
(cm)
lenght(cm)
5
5
Sproutlenght 4
4
3
3
Sprout
2
2
1
1
0
0
cv. ® ®
cv. 'Michelangelo
'Michelangelo®'' cv.
cv. 'Cosmos
'Cosmos®''
Cultivar ×× Treatment
Cultivar Treatment == ns
ns
Figure
Figure3. 3.
Sprout length
Sprout of cv.
length of ‘Michelangelo
cv. ‘Michelangelo ® ’‘Cosmos
®’ and
and ‘Cosmos ® ’ cuttings,
®’ cuttings, 14 weeks
14 after
weeks theafter
treatments.
the treat-
Figure 3. Sprout length of cv. ‘Michelangelo®’ and ‘Cosmos®’ cuttings, 14 weeks after the treatments.
Cuttings
ments. were treated
Cuttings werewith: H2O
treated = distilled
with: H 2 O = water;
distilledAUX =
water;auxins;
AUX = BRA = homobrassinolide;
auxins; BRA = KEL =
homobrassinolide;
Cuttings were treated with: H2O = distilled water; AUX = auxins; BRA = homobrassinolide; KEL =
Kelpak ®; PHY =®Phylgreen. Two-way and one-way ANOVA p-values are reported in the figure (ns,
p-values
Kelpak ; PHY = Phylgreen. Two-way and one-way ANOVA p-values are reported in the figurein(ns,
KEL =® Kelpak ; PHY = Phylgreen. Two-way and one-way ANOVA are reported the
not significant).
figure
not (ns, not significant).
significant).
Figure 4. Root fresh weight (A), dry weight (B), length (C), and area (D) of cv. ‘Michelangelo ® ’ and
Figure 4. Root
® fresh weight (A), dry weight (B), length (C), and area (D) of cv. ‘Michelangelo®®’ and
Figure 4. Root fresh weight (A), dry weight (B), length (C), and area (D) of cv. ‘Michelangelo
‘Cosmos ’ cuttings, after 14 weeks from the treatments. Cuttings were treated with: H2 O = distilled ’ and
‘Cosmos®®’ cuttings, after 14 weeks from the treatments. Cuttings were treated with: H2O = distilled
‘Cosmos
water;AUX ’ cuttings,
AUX= =auxins; after
auxins;BRA 14 weeks from the
BRA= =homobrassinolide; treatments.
homobrassinolide;KEL Cuttings
KEL= =Kelpak
Kelpak ®were treated
; PHY= = with: H O
Phylgreen.Two-way= distilled
Two-wayand
2 and
water; ®; PHY Phylgreen.
water; AUX
one-way = auxins;
ANOVA BRA =and
p-values homobrassinolide;
Tukey’s post hoc KEL = Kelpak®are
comparisons ; PHY = Phylgreen.
reported in the Two-way
figure (* p < and
0.05,
one-way ANOVA p-values and Tukey’s post hoc comparisons are reported in the figure (* p < 0.05,
one-way ANOVA p-values and Tukey’s post hoc comparisons are reported in the figure (* p < 0.05,
****pp<<0.01;
0.01; *** pp<<0.001;
0.001;ns,ns,
notnot
significant). Letters
significant). in thein
Letters figure indicateindicate
the figure the levelthe
of difference significance.
level of difference
** p < 0.01; *** p < 0.001; ns, not significant). Letters in the figure indicate the level of difference
significance.
significance.
3.2. Soluble Sugars
Total soluble sugars at the beginning of the trial and after 4 and 16 days from the
treatments are reported in Table 1. At the beginning of the trial, the amount of soluble
sugar was the highest measured, and it was similar between the two cultivars. Four days
after the treatments, the soluble sugar content was lower in both cultivars, and it was
Horticulturae 2022, 8, 561 6 of 9
not significantly different between the treatments. After 16 days from the treatments, the
soluble sugar content was lower than the previous days in all treatments and, moreover, it
was lower under PHY treatment than in control cuttings in cv. ‘Michelangelo® ’.
Table 1. Soluble sugars within the bark and the emerging callus of cv. ‘Michelangelo® ’ (M)
and ‘Cosmos® ’ (C) cuttings, 0, 4, and 16 days after the treatments. Cuttings were treated with:
H2 O = distilled water; AUX = auxins; BRA = homobrassinolide; KEL = Kelpak® ; PHY = Phylgreen.
Two-way and one-way ANOVA p-values and Tukey’s post hoc comparisons are reported in the table
(* p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant).
4. Discussion
The rose is one of the most important and valuable ornamental shrubs worldwide [20].
Its propagation is usually achieved by cuttings with a rooting efficiency varying from 0 to
100% [4]. The cvs. ‘Cosmos® ’ and ‘Michelangelo® ’ showed a rooting percentage of about
65%, highlighting a medium rooting ability. The possible failures in root formation are
usually overcome by producers with the application of plant growth regulators [3]. In
particular, the rooting promotion of cuttings using auxins is widely used in commercial
plant propagation [7]. Indeed, AUX treatment showed an increase in survival percentage,
callus formation, and root biometric parameters in both cultivar cuttings in comparison
with the control conditions, i.e., only distilled water. Other authors reported an increase
from 33 to 65% in rooting of R. centifolia medial cuttings in the same substrate using
3500 ppm of auxins [21] or from 67 to 75% or 81% in rooting of R. damascena cuttings in
sand using 200 mg dm−3 IAA or 25 mg dm−3 of NAA, respectively [22]. Thus, the aim of
this work was to find alternative substances to auxins for the promotion of cutting rooting
suitable for rose propagation.
BRs have already shown their role in rooting promotion of ornamental plant cut-
tings [10], but commercial synthetic products are still based principally on auxins and, to
our knowledge, they have never been tested on rose cuttings. BRs have shown contrasting
effects on rooting in woody cuttings of different plant genotypes [23,24]. In addition, in
our experimental conditions, 22(S),23(S)-homobrassinolide showed opposite effects on the
two rose cultivars. The cuttings of cv. ‘Michelangelo® ’ treated with BRA had a similar
behavior to those treated with only distilled water, while in cv. ‘Cosmos® ’ this treatment
showed the best results in terms of root biometric parameters. Thus, BRs were found to be
unsuitable for propagation in operational conditions since testing on every specific rose
genotype is required. The worst performance in rooting under BRA treatment retrieved
in cv. ‘Michelangelo® ’ matched with a low amount in soluble sugars within the emerging
callus 16 days after the treatment. Since a positive correlation between root number and
soluble sugar concentration was reported in other rose cultivar cuttings [19], this result
might support a less susceptibility of cv. ‘Michelangelo® ’ to rooting under BRA treatment.
Indeed, other authors also highlighted that a depletion of carbohydrates reduced callus and
root formation in single-node leafy stem cuttings [25]. However, a constitutional decrease
in soluble sugars during the first 15 days has already been reported within the bark of
cuttings of Zizyphus jujuba Mill. [26] and could explain the lower level of soluble sugars
found in all rose cuttings in comparison with the beginning of the trial.
Horticulturae 2022, 8, 561 7 of 9
5. Conclusions
In conclusion, Kelpak® was shown to improve both rooting percentage and root
biometric parameters of both rose cultivar cuttings, highlighting its suitability for the
replacement of synthetic products used for rooting promotion. BRs showed a species-
dependent effect, highlighting the necessity of evaluating their applicability before their use
as alternative synthetic phytoregulators to auxins. Phylgreen did not improve the rooting
performance of rose cuttings, indicating that this natural product cannot be suitable for rose
propagation using this methodology. Since the use of authorized hormonal agrochemicals
is very limited in some countries and is getting even lower every year, this study offers a
new possible product also available for organic systems. Further efforts must be conducted
to clarify the biochemical and molecular mechanisms of action of this seaweed extract.
Author Contributions: Conceptualization and investigation, S.T., S.C. and B.N.; trial set up and
sampling, S.T., S.C. and B.N.; formal analysis and data curation, S.T.; funding acquisition, S.C. and
B.N.; Writing—original draft, S.T.; Writing—review & editing, S.T., S.C. and B.N. All authors have
read and agreed to the published version of the manuscript.
Funding: This research was funded by the Italian Ministry of Agriculture, Food and Forestry Policies,
sub-project ‘Tecnologie digitali integrate per il rafforzamento sostenibile di produzioni e trasfor-
mazioni agroalimentari (AgroFiliere)’, AgriDigit program (DM 36503.7305.2018 of 20 December 2018).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Horticulturae 2022, 8, 561 8 of 9
Acknowledgments: The authors would like to thank Monica Michelotti and Paolo Bini for their
technical support during the trial.
Conflicts of Interest: The authors declare no conflict of interest.
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