Journal Journal

of Applied
Appl Journal of Applied Horticulture, 22(2): 159-163, 2020 Horticulture
DOI: 10.37855/jah.2020.v22i02.29 ISSN: 0972-1045

Effect of exogenous hormone and rooting medium on cutting

propagation of golden Camellia (Camellia impressinervis)

Tran Van Do1*, Tran Duc Manh1, Dao Trung Duc1, Mai Thi Linh1, Nguyen Toan Thang1, Dang
Van Thuyet1, Ly Thi Thanh Huyen1, Nguyen Van Tuan1, Phung Dinh Trung1, Nguyen Thi Thu
Phuong1, Ninh Viet Khuong1, Dang Thi Hai Ha1, Tran Cao Nguyen1, Tran Hoang Quy1, Pham
Dinh Sam1, Vu Tien Lam1, Nguyen Huu Thinh1, Hoang Thanh Son1, Trinh Ngoc Bon1, Ho
Trung Luong1, Tran Anh Hai1, Duong Quang Trung1, Nguyen Quang Hung1, Tran Hong Van1,
Nguyen Thi Hoai Anh1, Dinh Hai Dang1, Vu Van Thuan2 and Do Anh Tuan3
Silviculture Research Institute, Vietnamese Academy of Forest Sciences, Hanoi, Vietnam. 2Centre for Applied Silviculture
1

Research and Extension, Silviculture Research Institute, Vietnamese Academy of Forest Sciences, Hanoi, Vietnam. 3Vietnam
National University of Forestry, Xuan Mai, Hanoi, Vietnam. *E-mail: [email protected]

Abstract
Vegetative propagation method such as cutting, provides uniform plant materials from mother plants. The effect of medium and
exogenous hormone pretreatment on rooting were studied for golden camellia - Camellia impressinervis, a tree species which is used
for healthcare. The results showed that different concentrations of exogenous hormones IBA (Indole-3-butyric acid), IAA (Indole-3-
acetic acid), significantly affected rooting efficiency. Cuttings treated with IBA and IAA had significantly higher rooting percentage
than control (54.2 %). Higher rooting efficiency was obtained with IBA, which resulted in highest rooting percentage of 91.4 % at
4 months of growth by using concentration of 0.5 % (by weight). Hormone types and their concentrations did not affect mean root
number (MRN) and mean root length (MRL). Rooting media significantly affected rooting efficiency, MRN per cutting, and MRL.
Using medium of 100 % sand resulted in highest rooting percentage ( 91.4 %), roots per cutting (4.2) and MRL (5.4 cm) at 4 months
of growth. It is concluded that to produce seedlings for C. impressinervis by cuttings, rooting medium of 100 % sand should be used
and cuttings should be pretreated by 0.5 % IBA. In addition, cutting management is also important for rooting efficiency. It must be
controlled with humidity of > 95 % and temperature of < 27 oC.
Key words: Auxin, environment condition, healthcare value, indole-3-butyric acid, rooting efficiency

Introduction species (Wheeler and Rivers, 2015). Leaves and flowers of C.

impressinervis have been used to make tea by soaking in hot water
The vegetative propagation of superior genotypes is the basis (Tran, 2018). Several researches indicated that the extracts from
to create intensive plantations (Seth and Panigrahi, 2019). golden camellias have antioxidant activities, superoxide anions,
Cutting propagation has been given priority in vegetative and hydroxyl free radicals scavenging assays (Wan et al., 2011;
propagation, as it is easy to apply and usually results in high Wei et al., 2015). Golden camellias can be used to treat sore
rooting percentage for many plants (Eed and Burgoyne, 2014). throat, diarrhea, irregular menstruation, and cancer prevention
The physiological conditions of the stock plants (Mitchell et (Guangxi Institute of Botany, 1991). Currently, market price (600-
al., 2004), time and place of cutting collection (Crawford et 700 US$/ 1 kg) of dry flowers and leaves of golden camellias is
al., 2016), and environmental conditions significantly affect quite high (Tran, 2018), much higher than green tea leaves. In
the efficiency of cutting propagation. Adventitious rooting is a a golden camellia population there is much difference of flower
complex developmental process that can be affected by internal productivity among plants; a 1.2 m tall tree of C. impressinervis
and external factors (Leakey, 2004). Auxins play a critical role can bloom up to 130 flowers in a year, equaling 0.3 kg dry flowers
in the formation of adventitious root by increasing initiation of (Tran, 2018).
the root primordium and growth via cell division (Ren et al.,
In plantations derived by sexual propagation, seedlings
2019). Auxins promote starch hydrolysis and mobilize sugars and
are genetically variable, which affects growth uniformity,
nutrients to the cutting base (Das et al., 1997). During cell division
physiological characteristics, yield, and early bearing (Mohassea
and auxin transport, auxins act primarily through selective
et al., 2009). On other hand, vegetative propagations (e.g.,
proteolysis and cell wall loosening (da Costa et al., 2013).
cutting) provide genetically uniform plant materials from mother
Camellia impressinervis Hung T. Chang & S. Ye Liang is plants, which can fruit early. The objectives of this study were
known as a golden camellia (Hung and Le, 1979; Manh et to evaluate the effects of exogenous hormones (auxin types and
al., 2019), which has natural distribution in Southern China concentrations) and medium on rooting efficiency to establish an
and North Vietnam and is classified as a critically endangered optimum cutting propagation scheme for Camellia impressinervis.
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160 Effect of exogenous hormone and rooting medium on cutting propagation of golden Camellia

Materials and method replications and each replication contained 30 cuttings. Totally,
270 cuttings were used.
Experimental site, plant material, rooting pot, and cutting
management: The experiment was conducted in a nursery at Exogenous hormone type and concentration: Two hormone
Silviculture Research Institute, Vietnamese Academy of Forest types were tested including IBA and IAA (Indole-3-acetic
Sciences in Hanoi capital, Vietnam (21o04’20.8’’, 105o46’34.7’’). acid). In each hormone type, six concentrations were tested
including 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 % by weight in powder
The cuttings of C. impressinervis were collected from garden of type. There was one control (no exogenous hormone treatment),
local people in Cao Bang province (22o24’56.8’’, 106o25’23.9’’), leading to total 13 treatments. Rooting medium was 100 % sand.
300 km far from Hanoi capital. Trees were dug up from natural A randomized complete block design was set up with three
forest and transplanted in garden in 2013-2014. After planting replications and each replication contained 30 cuttings. Totally,
trees were generally not tended. Trees were 1-1.4 m tall, 2-7 1,170 cuttings were used.
cm diameter at stump, and 0.8-1.4 m crown diameter at a time
of cutting collection in October 2018. Most of the trees started Survival rate and rooting measurements: Survival rates of
blooming in 2017. When cuttings were collected, there appeared cuttings were calculated at 15 days, 1 month, 2 months, 3 months,
some flower buds. The climate conditions, where cuttings were and 4 months of growth by counting alive cuttings for each
collected, include annual precipitation of 1,500-1,700 mm, treatment replication and experiment. Measurements of rooting
temperature of 21-23oC, and air humidity of 85-90 % (Tuan et al., efficiency were assessed at 4 months of growth. The measured
2019). The cuttings were collected from the first branches, which indices included the number of roots (NR, roots >0.3 cm long),
were 8-12 months old. After cutting, branches were protected in length of three longest roots, and mean root length (MRL) for
buckets with ordinary water covering 2-3 cm branch bases. It was each plant. Rooting success was expressed as the percentage of
then transported to nursery in Hanoi. Total time required from cuttings that rooted in each replication, treatment, and experiment.
cutting branches to fishing work in nursery was with in 24 hours. Statistical analysis: Differences in rooting percentage, mean RN
Each branch was cut into two segments from top to create two (MNR), and MRL in each experiment were assessed by univariate
cuttings. Cutting was cut at leaf base perpendicularly. Cutting analysis of variance (ANOVA) and post-hoc test at P = 0.05.
was 8-12 cm long and contained 2-4 leaves. Only 15-20 % area All analyses were conducted using SAS 9.2 (SAS Institute Inc.,
of each leaves was remained (Fig. 1a). Cuttings were then treated Cary, NC, USA).
by soaking in fungicide for 10 minutes and treated again every
seven days by spraying to seedbed until 4 months of growth. Results
Rooting pots were 4 cm in diameter and 8 cm long, made of Rooting medium experiment: Rooting media significantly
non-woven plastic (bottom open), and set on the seedbed in the affected alive percentage at 2 and 3 months and rooting percentage
nursery (Fig. 1a). at 4 months of growth (Table 1). The highest alive percentage at
2 months of growth belonged to medium of 100 % sand (96.4
Seedbed was covered by transparent nylon to avoid water
%), followed by medium of 50 % sand + 50 % soil (93.6 %),
evaporation and then a layer of nursery shade cloth with shading
and by medium of 100 % soil (91.4 %). The similar pattern was
level of 47-53 % (Fig. 1b). A meter was set inside seedbed for
observed at 3 months of growth (Table 1). Rooting percentage at
observing humidity and temperature. The cuttings were watered
4 months of growth was highest in medium of 100 % sand (91.4
twice a day in the morning and afternoon by spraying system
%), followed by medium of 50 % sand + 50 % soil (82.9 %), and
(Fig. 1a). Generally, the seedbed had controlled humidity of >
by medium of 100 % soil (78.6 %).
95 % and temperature of < 27 oC by additional watering and/or
openning of the cover. Rooting media significantly affected MRN and MRL at 4 months
of growth (Fig. 2). Both MRN (4.2 roots/cutting) and MRL (5.4
Experimental design-Rooting medium experiment: Three
cm) were highest in medium of 100 % sand, followed by medium
media were tested including (1) 100 % sand, (2) 100 % soil
of 50 % sand + 50 % soil (3.8 roots/cutting and 5.1 cm), and by
collected from 0-40 cm depth of forested soil, and (3) 50 %
medium of 100 % soil (3.6 roots/cutting and 4.7 cm).
sand + 50 % % soil by volume. Exogenous hormone IBA
(Indole-3-butyric acid) 2 % by weight in powder type was used. The medium of 100 % sand was best for cutting propagation of C.
A randomized complete block design was set up with three impressinervis, which can result up to 91.4 % rotting percentage

Fig. 1. Cuttings in seedbed with spraying system (a), shaded seedbed (b), and rooted cuttings at 4 months of growth in different media (c) [100 %
sand (c-1), 50 % sand + 50 % soil (c-2), and 100 % soil (c-3)].
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 Effect of exogenous hormone and rooting medium on cutting propagation of golden Camellia 161

Table 1. Effect of rooting media on alive cutting percentage and rooting percentage (±SE)
Time of growth
Rooting media 15 days 1 month 2 months 3 months 4 months
Alive cuttings (%) Rooting (%)
100 % sand 100 100 96.4 ±1.01a 93.2 ±1.15a 91.4 ±1.21a
50 % sand + 50 % soil 100 100 93.6 ±0.8b 88.7±1.09b 82.9 ±1.18b
100 % soil 100 100 91.4 ±0.91c 84.5±0.98c 78.6 ±1.05c
Means followed by different alphabets in the same column indicate significant differences by Tukey’s test (P < 0.05).

at 0.5 % IBA and lowest at 3.0 % IBA (Table 2). At 4 months

of growth, rooting percentage was highest at 0.5 % IBA (91.4
%) and lowest at 3.0 % IBA (82.9 %). While, MRN and MRL
were not significantly different among IBA concentrations,
which ranged from 4.0 to 4.2 roots/cutting and 5.1 to 5.3 cm,
respectively (Table 2).
At 1 month of growth, there were no dead cuttings appeared in
IAA treatment (Table 3). At 2 months of growth, the dead cutting
Rooting media Rooting media appeared in treatments of 1.0, 1.5, 2.0, 2.5 and 3.0 % IBA, except
Fig. 2. Effects of rooting media on mean root number (MRN) per 0.5 %. Alive cutting percentages were significantly different
cutting and mean root length (MRL) at 4 months of growth. Letters
indicate significant differences by Tukey’s test (P < 0.05) and vertical among IAA concentrations at 2 and 3 months of growth, which
bars indicate SE. was highest at 0.5 % IAA (Table 3). At 4 months of growth,
rooting percentage was highest at 0.5 % IAA (88.6 %) and lowest
at 4 months of growth with highest MRN and MRL by pretreating
at 3.0 % IAA (71.4 %). While MRN and MRL were not different
the cuttings with 2 % IBA. among IAA concentrations, which ranged from 4.0 to 4.2 roots/
Exogenous hormone type and concentration: Both exogenous cutting and 5.2 to 5.4 cm/root (Table 3).
hormones IBA and IAA had higher rooting percentage than
control (54.2 %; Fig. 3). Between two hormones, IBA showed Discussion
significantly better effect on rotting percentage than IAA. Many plants belong to the callus rooting species as their roots
Except at concentration of 2.5 %, rooting percentage in other initiate from calluses (Yang et al., 2015). Rooting in such
IBA concentrations was significantly higher than that in IAA species is relatively more difficult than species that root from
concentrations (Fig. 3). The difference was highest (11.5 %) the phloem side of the fascicular cambium (Izhaki et al., 2018).
at hormone concentration of 3 % and lowest (1 %) at hormone As C. impressinervis roots from both callus and the phloem
concentration of 2 %. There were no significant differences of side of the fascicular cambium (Fig. 1c and 4), it could be
MRN and MRL between IBA and IAA at 4 months of growth considered as an easy rooting plant, similar to other species
(Table 2 and 3; Fig. 4). in genus Camellia (Yongjun and Ming, 2012). In this study,
cuttings were collected from five unknown-age individuals of C.
At 1 month of growth, there were no dead cuttings in IBA impressinervis, which originated from seeds, and they were not
treatment (Table 2). At 2 months of growth, the dead cuttings tended properly. Thissuggest that genetic variability (Nasri et al.,
appeared only at treatments of 1.0, 2.0, and 3.0 % IBA. Alive 2019) and difference of internal and external conditions among
cutting percentages were significantly different among IBA individuals may cause variation in rooting potential of cuttings.
concentrations at 2 and 3 months of growth, which was highest However, rooting efficiency was high (Table 1, 2, 3), indicating

Exogenous hormones concentrations

Fig. 3. Effects of exogenous hormones and their concentrations on rooting percentage at 4 months of growth.
Different letters indicate significant differences by Tukey’s test (P < 0.05)

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162 Effect of exogenous hormone and rooting medium on cutting propagation of golden Camellia

Fig. 4. Rooted cuttings at 4 months of growth in different exogenous hormones and their concentrations, IBA (a) and IAA (b).
Table 2. Effect of IBA concentrations on alive cutting percentage and rooting percentage (±SE)
Concentration Time of growth
(%) 15 days 1 month 2 months 3 months 4 months
Alive cuttings (%) Rooting (%) MRN/ cutting MRL (cm)
0.5 100 100 100a 95.8 ±0.9a 91.4 ±0.9a 4.0 ±0.7 5.1 ±1.3
1.0 100 100 94.3 ±0.9b 90.1 ±0.8b 88.6 ±1.1b 4.1 ±0.8 5.3 ±1.3
1.5 100 100 100a 87.8 ±1.1c 82.9 ±1.2c 4.2 ±0.7 5.2 ±1.2
2.0 100 100 98.1 ±0.8c 87.8 ±1.0c 86.7 ±1.1d 4.0 ±0.8 5.1 ±1.1
2.5 100 100 100a 86.5 ±1.0d 85.7 ±1.2e 4.1 ±0.8 5.3 ±1.2
3.0 100 100 97.1 ±0.8d 86.9 ±1.1d 82.9 ±1.2c 4.2 ±0.7 5.2 ±1.2
Means followed by different alphabets in the same column indicate significant differences by Tukey’s test (P < 0.05). MRN is mean root number
and MRL is mean root length.
Table 3. Effect of IAA concentrations on alive cutting percentage and rooting percentage (±SE)
Time of growth
Concentration
15 days 1 month 2 months 3 months 4 months
(%)
Alive cuttings (%) Rooting (%) MRN/ cutting MRL (cm)
0.5 100 100 100a 94.6 ±1.0a 88.6 ±0.6a 4.2 ±0.9 5.4 ±1.2
1.0 100 100 97.1 ±1.0 b
85.7 ±0.8 b
80.0 ±0.6 b
4.1 ±0.8 5.3 ±1.1
1.5 100 100 92.0 ±0.9c 88.1 ±0.8c 72.0 ±0.5c 4.1 ±0.7 5.4 ±1.3
2.0 100 100 94.3 ±0.9d 89.6 ±0.7c 85.7 ±0.5d 4.2 ±0.8 5.2 ±1.3
2.5 100 100 97.0 ±1.1 b
91.2 ±0.9 d
87.9 ±0.6 e
4.0 ±0.9 5.3 ±1.2
3.0 100 100 94.3 ±1.0d 90.1 ±0.9d 71.4 ±0.5c 4.1 ±0.8 5.2 ±1.2
Means followed by different alphabets in the same column indicate significant differences by Tukey’s test (P < 0.05). MRN is mean root number
and MRL is mean root length.
high rootability of C. impressinervis in cutting propagation. If forming calluses and roots. Meanwhile, medium of 100 % sand
cuttings are collected from a superior genotype and plants are contains 100 % particles >0.02 mm. Therefore, air content and
tended properly, rooting percentage could be higher. oxygen diffusion rate are higher and suitable for rooting of C.
impressinervis. It is reported that C. impressinervis grows well in
Applying IBA may have an indirect influence through enhancing high moisture but drainage soil (Tran, 2018). Soil with high ratio
translocation and movement of carbohydrates to the cutting of loam and silt particles has high water holding capacity and is
base and consequently stimulate rooting (Aminah et al., 1995). not drainage soil. Therefore, medium of 100 % soil should not
Therefore, rooting efficiency was higher in IBA treatments than be used in cutting propagation for C. impressinervis.
that in IAA treatments (Fig. 3). Similar findings were reported in
other species (Eed and Burgoyne, 2014). Such result may also be C. impressinervis is known as a shrub or small-sized tree, which
related to total phenolic content and peroxidase activity, which are is shorter than 7 m at maturity. This is a shade-tolerant species,
higher in IBA treated cuttings, particularly during the initiation growing well under shade of others in evergreen broadleaved
and expression phases (Rout, 2006). forests (Tuan et al., 2019). Therefore, shading seedbed in cutting
propagation is required, which is not a case in other species (Yang
Effects of media on rooting efficiency were reported in many et al., 2015), especially for light demanding plants. Effects of light
plants (Eed and Burgoyne, 2014). Air content and oxygen intensity on rooting were reported (Fuerncranz et al., 1990). The
diffusion rate in media are important for rooting (Ercisli et al., shading level of 47-53 % was used in the present study, which
2002). In the present study, medium of 100 % soil with high ratio resulted in 91.4 % rooting percentage of cuttings treated with
of loam and silt particles indicate low air soil, which inhibited 0.5 % IBA in medium of 100 % sand. The rooting efficiency
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 Effect of exogenous hormone and rooting medium on cutting propagation of golden Camellia 163

could be improved if different shading levels are considered. Ercisli, S., O. Anapali, A. Esisken and U. Sahin, 2002. The effects of IBA,
Therefore, further study should be conducted by testing the effects rooting media and cutting collection time on rooting of kiwifruit.
of different shadings on rootability. Gartenbauwissenschaft., 67: 34-38.
Fuernkranz, H.A., C.A. Nowak and C.A. Maynard, 1990. Light effects
Effects of rooting media, and exogenous hormone types and on in vitro adventitious root formation in axillary shoots of mature
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air content and oxygen diffusion rate compared to that in 100 % Izhaki, A., Y. Yitzhakab, T. Blaua, I. Davida, A. Rotbauma, J. Riovb
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hormone IBA promoted rooting better than IAA. In addition, IBA Rout, G.R. 2006. Effect of auxins on adventitious root development from
concentrations significantly affected rooting efficiency. The best single node cuttings of Camellia sinensis (L.) Kuntze and associated
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recommended the use of medium of 100 % sand and 0.5 % IBA Seth, S. and J. Panagrahi, 2019. In vitro organogenesis of Abutilon
in cutting propagation of C. impressinervis with the control of indicum (L.) Sweet from leaf derived callus and assessment of genetic
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Acknowledgements Report. Silviculture Research Institute. Hanoi, Vietnam.
Tuan, N.V, P.D. Trung, T.D. Manh, N.T. Thang, D.V. Thuyet, D.T. Duc,
This research was funded by Vietnam Ministry of Science and M.T. Linh, N.T.T. Phuong, N.V. Khuong, V.T. Lam, N.H. Thinh, H.T.
Technology under grant number 17/18/ĐTĐL.CN-ĐP. Son, T.N. Bon, H.T. Luong, V.V. Thuan and T.V. Do, 2019. Possible
planting areas for Golden Camellia - Camellia impressinervis in
Vietnam. Asian J. Agric. Hort. Res., 3: 1-7.
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