concentrations, plant height, plant growth regulators, leaf number, Ornamental Plants, daminozide, plant growth, treatments, cycocel, growth regulators, flowering, maximum effect, paclobutrazol, Analysis of variance, Agricultural sciences, J. Agric, Calendula officinalis L., Calendula officinalis, Acta Hort, Ornamental Plant, ornamental cabbage and kale, Cricket Ball, HortSci, Growth regulator, North Carolina Flower Growers Bulletin, Cooperative Extension Service Institute of Food and Agricultural Science
Effect of Cycocel and Daminozide on Vegetative Growth, Flowering and the Content of Essence of Pot Marigold (Calendula officinalis) Shahram Shoa Kazemi1, Davood Hashemabadi2*, Ali Mohammadi Torkashvand2 and Behzad Kaviani2 1MSc. Student, Department of Horticultural Science, Rasht Branch, Islamic Azad University
, Rasht, Iran 2Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran SID *Corresponding author,s email: [email protected]
f Pot marigold (Calendula officinalis L.) is a medicinal and ornamental o plant. The effect of different concentrations of chlormequat (cycocel), and daminozide, two plant growth
retardant, on plant height
, flowering, the e content of essence and some other traits in pot marigold (Calendula officinalis) was assessed. Plant growth retardants are commonly applied to iv limit stem elongation and produce a more compact plant. The experiment was done as a factorial in randomized completely blocks design (R.C.B.D.) with 16 treatments and 3 replications in Rasht. Cycocel at 4 concentrations h (0, 500, 1000 and 1500 mg/L) and daminozide at 4 concentrations (0, 1500, c 3000 and 4500 mg/L) were used. Investigated characteristics were plant height, leaf number, flower number, flowering time, fresh weight, dry r matter, the content of essence and carotenoid in flowers. Based on analysis A of variance (ANOVA), the effect of different treatments and their interaction on most traits was significant at 0.05 level of probability. The minimum height (24 cm/plant) in treatment of 500 mg/L cycocel + 3000 mg/L daminozide, the largest number of flowers (4.66 flowers/plant) in treatment of 1000 mg/L cycocel + 4500 mg/L daminozide and most essence content (0.154 mg/100 g) in treatment of 4500 mg/L daminozide without cycocel were obtained. Keywords: Chlormequat, Drench, Plant height, ornamental plants
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INTRODUCTION Calendula officinalis (pot marigold) from the family Asteraceae is probably native to south- ern Europe. It is a short-lived aromatic perennial plant, growing to 80 cm tall, with sparsely branched lax or erect stems. The inflorescences are yellow, comprising a thick capitulum or flower head 4-7 cm diameter surrounded by two rows of hairy bracts. Calendula are considered by many gardening experts as one of the most versatile flowers to grow in a garden, especially since they are easy to grow, and tolerate most soils. Calendula officinalis L. is an annual, aromatic, medicinal and ornamental herb (Gazim et al., 2008). The composite flowers blossom in the spring-summer season
s for 3 times per year (Gilman and Howe, 1999; Omidbaighi, 2005). The leaves and flowers of marigold are applied in horticulture, medicine, cosmetics, perfume, pharmaceutical preparation, food and other industries (Van Wyk and Wink, 2004; Gazim et al., 2008). One of the main components of active ingredient in Calendula officinalis L. are essential oils which the most of them synthesize in its orange petals (Omidbaighi, 2005). Flower essence is using for food and medicine (Hamburger et al., 2003; Janke, 2004; Jimenez-Medina et al., 2006). Plant growth retardants are commonly applied to limit stem elongation and produce a more compact plant. Production of high quality, compact pot plants may be achieved through the use of plant growth retardants including cycocel (Tayama et al., 1992). Effectiveness of plant growth retardants depends on time and method D of application, concentration, type of species and cultivar, and type of target organ as well physi- ological and environmental conditions
(Pobudkiewicz and Nowak, 1994; James et al., 1999). The I most common methods of application of growth retardants are foliar sprays and media drenches S (Al-Khassawneh et al., 2006). Plant growth retardants can delay cell division and elongation of plant aerial parts as well restrict gibberellins biosynthesis, resulted in reduces internodes length f and vegetative growth (Magnitskiy et al., 2006). Adding cycocel has also proven to be effective in controlling growth of some other plants (Al-Khassawneh et al., 2006; Rossini Pinto et al., 2005; o Leclerc et al., 2006). Proper doses of cycocel and daminozide foliar spray and drench rate need to be assessed because they can either inhibit or promote growth and development
. Therefore, the objective of the present study was to evaluate the effect of different concentrations of cycocel and e daminozide on some growth characters especially plant height and the content of essence in Caliv endula officinalis L. MATERIALS AND METHODS h Seeds of Calendula officinalis L. were obtained from Bazr va Nahal Company, Tehran, c Iran. Investigation was carried out in an experimental field in Rasht city located in the northern part of Iran. Seeds were sown in pots filled with 60% sand, 20% cocopeat and 20% animal fertilizer r on November 2011. Plants were treated with a drench application at the rate of 0, 500, 1000 and A 1500 mg/L of cycocel and 0, 1500, 3000 and 4500 mg/L of daminozide 5-6 weeks after potting. control plants
were drenched with water. Investigated characteristics were the plant height, leaf number, flower number, flowering time, fresh weight, dry matter, the content of essence (essential oil) and carotenoid in flowers. Data were calculated at 60-70 days after transplanting. Plant height was measured by a ruler. Leaf number was obtained by counting leaves from center of each plot and their mean was calculated. Fresh weight of plants was weighted by a digital balance. To obtain the plant dry matter, they were cut from crown and dried at 105°C for 24 h. For determination of the essential oil, the plant materials
(flowers) were dried in 45°C. The essential oil was obtained in a Clevenger apparatus by steam distillation. Thus, the 50 g of dried plant materials was extracted with 1000 ml of water. The water collected was re-extracted with 0.5 ml hexane. The essence and hexane was separated from water physically and weighted until the plant essence obtained. Leaf carotenoids were determined using acetone as extracting solvent and the absorbance was measured at 440, 645 and 663 nm. Sample extract was prepared as follows: 0.5 g of dry sample was thoroughly crushed and homogenized in a mortar with a pestle using 20 ml of 80% acetone. Filtered
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extract was reached to 50 ml by adding of 80% acetone. Concentration of carotenoids was calculated by following formula after spectrophotometry: The amount of carotenoids = 4.69 Ч A440 0.268 Ч (20.2) A645 + (8.02) A663 Where; A is wave length. Data processing of the results was carried out by an EXCEL. analysis of variance
(ANOVA) was done using SPSS statistical software and means were compared using Duncan's test.
RESULTS The overall results of the effects of different concentrations of cycocel and daminozide on the plant height, leaf number, flower number, flowering time, fresh weight, dry matter, the content of essence and carotenoid in flowers are summarized in Tables 2 and 3.
Plant height Based on analysis of variance (Table 2), the effect of different treatments and their in- teraction on the plant height was significant at 0.05 level of probability. Plant height did not decrease linearly with increasing the cycocel concentration (Table 3). Calendula officinalis L. plants treated with all concentrations of cycocel and daminozide were shorter than the control plants D (Table 3). 500 mg/L cycocel and 4500 mg/L daminozide produced the shortest plants, individually (25.43 and 24.16 cm/plant, respectively). Among all treatments, plants treated with 500 mg/L cy- I cocel along with 1500 mg/L daminozide (C2D2) had the least plant height (23.33 cm/plant) (Table S 3). C2D3 treatment was good for control of plant height (24.00 cm/plant), too. The highest plant height (38.33 cm/plant) was obtained in C1D1 treatment (control). Comparison between different f concentrations of cycocel and daminozide shows that daminozide is more suitable plant retardant than cycocel for reduction of plant height (Table 3). o Leaf number Analysis of variance (Table 2) showed that the effect of different treatments and their in- e teraction had no significant effect on leaf number. Table 3 shows that the largest number of leaf iv (5.00/plant) was obtained in C1D2 and C2D3. The smallest number of leaf (2.75 and 2.83/plant) was calculated in C1 and D1. h Flower number c Mean comparison obtained from the data showed that the maximum (4.66/plant) and min- imum (1.41/plant) number of flower were obtained from plants treated with 1000 mg/L cycocel r along with 4500 mg/L daminozide (C3D4) and C1 (control), respectively (Table 3). Plants treated A with highest concentration of cycocel (C4) had 1.58 flower/plant. Treatments of C3D1, C2D3 and C2D2 by induction of 4.33 flowers/plant were proper, too. Analysis of variance presented in Table 2 showed that the effect of cycocel and interaction effect of cycocel and daminozide on the number Table 1. Treatments used in the present study and their symbols.
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Table 2. Analysis of variance (ANOVA) for the effect of different concentrations of cycocel and daminozide on plant height, leaf number, fresh weight, dry matter, flower number, time to flowering, essence and carotenoid con- tents of Calendula officinalis L.
C D CЧD Error CV (%)
ns: Non significant, *: Significant at 5%
Fresh weight 33.05* 9.38* 7.88* 69.27 26.21
Dry Flower matter number
0.27ns 1.03ns 0.05ns 0.08 18.92
0.07* 0.18ns 0.50* 1.33 45.8
Time to flowering 155.96* 14.85* 39.55* 85.16 7.85
Essence content 45.91* 5.32* 2.55* 32.12 28.61
Carotenoid content 12.59* 6.11* 52.55* 9.12 35.91
of flower per plant were significant at 0.05 probability. The effect of daminozide on the number of flower per plant was no significant.
Time to flowering variance analysis
of data (Table 2) showed that the impact of cycocel, daminozide and their D interaction effect was significant at 0.05 probability on time to flowering. The mean comparison of data in different treatments (Table 3) showed that the lowest time of flowering is related to C4D2 I treatment (98.00 days). The longest time of flowering was obtained in treatments of C1D1 (122.60 S Table 3. Mean comparison of the effect of different concentrations of cycocel and daminozide on plant height, leaf number, fresh weight, dry matter, flower number, time to flowering, essence and f carotenoid contents of Calendula officinalis L.
Treatments Plant Leaf
C1 e C2 iv C3 C4 D1 h D2 D3 c D4 C1D1 r C1D2 C1D3 A C1D4
35.83a 25.43b 34.25a 34.08a 35.41a 24.58b 24.25b 24.16b 38.33a 35.00ab 32.23bc 35.66ab
2.75a 4.25a 4.33a 3.83a 2.83aa 4.08a 4.33a 3.91a 4.33a 5.00a 4.66a 3.33a
Fresh weight 24.41b 32.00a 32.25a 33.33a 25.50b 30.91a 31.58a 33.00a 21.33c 23.33c 29.66b 26.33b
Dry Flower Time to matter number flowering
4.33a 5.66a 5.58a 5.41a 4.41a 5.41a 5.91a 5.41a 5.33a 5.00a 6.00a 5.00a
1.41b 2.58a 2.50a 1.58b 2.58a 2.66a 2.41a 2.41a 2.00d 3.33b 3.00c 2.33cd
119.66a 114.16a 104.58b 121.33a 118.00a 121.33a 106.91b 118.66a 122.60a 112.66b 115.00b 115.33ab
Essence content 0.12c 0.13b 0.14a 0.14a 0.12c 0.13b 0.14a 0.14a 0.12c 0.13b 0.12c 0.15a
Carotenoi d content 5.21b 5.87b 6.51a 6.91a 5.11b 6.14a 6.53a 6.64a 5.11c 5.16bc 6.68b 7.02ab
In each column means followed by the same letters are not significantly different at 5 % level of probability using Duncan's test.
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days). Time to flowering in treatments of C3 (104.58 days) and D2 (106.91 days) were also good (Table 3). On the other hand, time to flowering in treatments of C3D2, C3D3 and C4 (about 120.00 days for all) was not suitable (Table 3). Fresh weight Based on Table 2, the effect of different concentrations cycocel, daminozide and the inter- action effect was significant at 0.05 probability on plant fresh weight. Among concentrations of cycocel, C4 treatment had maximum effect on fresh weight (33.33 g/plant), also, C1 (control) treatments with 24.41 g fresh weight was minimum (Table 3). Plant fresh weight increased as the concentration of cycocel and daminozide increased. Among concentrations of daminozide, D4 treatment had maximum effect on fresh weight (33.00 g/plant), also, D1 (control) treatments with 25.50 g fresh weight was minimum (Table 3). Among all treatments, maximum (34.66 and 34.33 g/plant) and minimum (21.33 g/plant) of plant fresh weight were obtained in C3D4, C4D4 and C1D1 (control), respectively (Table 3). Dry matter Table 2 shows that the effects of cycocel, daminozide and the reciprocal effect were no sig- D nificant effect on plant dry matter. Table 3 shows that dry matter in plants treated with 500 mg/L cycocel along with 3000 mg/L daminozide (6.33 g/plant) was higher than that of other treatments. I The 6.00 g dry matter was obtained in 5 treatments including C1D3, C2D1, C3D2, C3D4 and C4D4 S (Table 3). Mean comparison obtained from the data showed that the least dry matter (4.33 and 4.41 g/plant) was calculated from C1 and D1 treatments. f Essential oil contents o The effect of cycocel, daminozide and interaction effect of these two factors were significant (p0.05) on the content of petal essential oil (Table 2). Maximum essential oil (0.15 ml/100 g FW) was obtained when pot marigold was planted in treatment of C1D4 (Table 3). Minimum essential e oil (0.12 ml/100 g FW) was obtained when pot marigold was planted in C1, D1, C1D1, C1D3 and iv C4D1 (Table 3). Carotenoid content h ANOVA showed that the effect of cycocel, daminozide and interaction effect of these two c factors were significant (p0.05) on the content of petal carotenoid (Table 2). Maximum carotenoid (8.22 mg/L) was obtained when pot marigold plants were planted in treatment of C3D4 (Table 3). r Minimum essential oil (5.11 mg/L) was obtained when pot marigold plants were planted in D1, A C1D1 (Table 3). DISCUSSION One of the most important applications of plant growth retardant is elevation of plant qual- ity, especially ornamental plants by reduction of vegetative growth. Plant growth retardants decrease the internodes length and eliminate the apical dominance (Lee et al., 1999). Plant growth retardants increase cytokinins which enhance the amount of leaf chlorophyll (Rossini Pinto et al., 2005). Some of the most important factors concerning plant growth retardants are type, time, number, application method and concentration of growth retardant (Cramer and Bridgen, 1998). Cycocel and daminozide are two important plant growth retardant. Several studies revealed effectiveness of cycocel in decreasing plant height (Rossini Pinto et al., 2005; Olivera and Browing, 1993; Garner, 2004; Karlovic et al., 2004; Hashemabadi and Zarchini, 2010). Studies of Al-Khassawneh et al. (2006) on growth and flowering of Iris nigricans showed that cycocel reduced plant height only at the highest drench concentration. These researchers revealed that cycocel spray at Journal of Ornamental Plants, Volume 4, Number 2: 107-114w, Jwunwe,.2S0I1D4 .1i1r1
the higher concentrations (1000-1500 mg/L) reduced plant height. In current study, cycocel caused decreasing of plant height in Calendula officinalis L. Karlovic et al. (2004) reported decreasing height in Chrysanthemum by 2000, 3000 and 4000 mg/L cycocel. Hashemabadi and Zarchini (2010) showed that the least stem length (29.93 cm) was obtained by using 1500 mg/L cycocel in rose. Saffari et al. (2004) sprayed Rosa damascena with cycocel and found that 3000 mg/L cycocel decreased stem length about 5 cm relative to the control. Increasing application rates
did not positively influence plant development when compared to the lower rates used in the study. Cycocel (1000 and 2000 mg/L) decreased Zinnia plant height (Hojjati et al., 2009). Current study confirms these studies. Cycocel, also, reduced plant height in Euphorbia and Bougeinvillia (Shekari et al., 2004), Rosa (Saffari et al., 2004) and Pelargonium (Latimer and Beden, 1994). Other plant growth retardants such as prohexadione-Ca, uniconazole, paclobutrazol and bayleton are applied for decreasing the plants growth as spray or drench (Gibson and Whipker, 2000; Bazzocchi and Giorgioni, 2003). Gholampour et al. (2012) showed that the 1500 mg/L of cycocel resulted in about 50 and 20% shorter Brassica oleracea cultivar `Kamome White' and `Nagoya Red' plants than the control plants, 60 and 90 days after transplant, respectively. The growth of these cultivars decreased with increasing the concentration of cycocel. Some other researches demonstrated the positive effect of cycocel and daminozide on reduction of stem length in some species such as Euphorbia, D Rosa, Pelargonium and Bougainvillea (Saffari et al., 2004; Joyce et al., 2004; Shekari, 2006). Studies of Al-Khassawneh et al. (2006) on growth and flowering of Iris nigricans showed I that maximum number of leaves (average of 12.2-13.6) was obtained when the plants were unS treated with cycocel and paclobutrazol, sprayed with 250 mg/L paclobutrazol, or drenched with 0.25 mg/L. Our finding is consistent with these results. Study of Agrawal and Dikshit (2008) on f Achras sapota L. demonstrated positive effect of cycocel on leaf number. Current study showed that the highest dry matter percentage was obtained from untreated plants with cycocel. In agree- o ment with our finding, Al-Khassawneh et al. (2006) also showed that untreated plants had the highest leaf dry weight
. Study of Garib Sahi (2009) on Zinnia elegans revealed that spraying plants with 2000 mg/L cycocel and 1 mg/L CaCl2 increased leaves and roots dry weight. Our study e showed the positive effect of plant growth retardants on increasing the content of essential oil and iv carotenoid in pot marigold petals. Some studies confirm these findings (Khalid and El-Ghorab, 2006; Gliozeris, 2007). Optimum concentrations of cycocel in these studies were 500 and 1000 mg/L. Plant growth retardants increase cytokinins synthesis and this hormones enhances the content h of pigments in plants (Fletcher et al., 2000). c In conclusion, some concentrations of cycocel and daminozide increased morphological and physiological characteristics in Calendula officinalis L. The 500 mg/L of cycocel along with r 1500 mg/L daminozide resulted in shorter plants than the other concentrations and control plants. A Maximum flower number, fresh weight, dry matter and carotenoid content were obtained in plants treated with 1000 mg/L cycocel along with 4500 mg/L daminozide. The 4500 mg/L caused the highest amount of essential oil.
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SS Kazemi, D Hashemabadi