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New Zealand Journal of Crop and Horticultural Science

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Effect of plant population on calendula (Calendula officinalis L.) flower production

R. J. Martin & B. Deo
a a b

New Zealand Institute for Crop & Food Research Limited, Private Bag 4704, Christchurch, New Zealand E-mail: martind@crop.cri.nz
b

New Zealand Institute for Crop & Food Research Limited, Private Bag 4704, Christchurch, New Zealand Available online: 22 Mar 2010

To cite this article: R. J. Martin & B. Deo (2000): Effect of plant population on calendula (Calendula officinalis L.) flower production, New Zealand Journal of Crop and Horticultural Science, 28:1, 37-44 To link to this article: http://dx.doi.org/10.1080/01140671.2000.9514120

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New Zealand Journal of Crop and Horticultural Science, 2000, Vol. 28: 37-44 0014-0671/00/2801-0037 $7.00 The Royal Society of New Zealand 2000

37

Effect of plant population on calendula (Calendula officinalis L.) flower production

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R. J. MARTIN B.DEO New Zealand Institute for Crop & Food Research Limited Private Bag 4704 Christchurch, New Zealand email: martind@crop.cri.nz Abstract Calendula (Calendula officinalis L.) was drilled at 3, 6, 12, 24, 48, and 96 kg seed/ha in a trial at Lincoln, New Zealand, in November 1997. Plots were divided into five equally sized sections. Fully open flowers were harvested from Sections 2 and 4 of all plots twice or three times weekly from early February to mid March 1998, after which plant population and crop biomass were determined from the same sections and seed yield/flower determined from Sections 1, 3, and 5. Plant counts increased from 9 plants/m2 at the 3 kg seed/ha rate, to 26 at 6, 46 at 12, 101 at 24, 179 at 48, and 332 plant/m2 at 96 kg seed/ha. Total flower yield was not significantly different at populations over 46 plants/m2, but declined with lower plant populations. The total fresh weight (and number) of fully opened flower heads collected off each plot increased from 0.5 (164) to 1.4 kg/m2 (650/m2) with increasing plant population, but did not differ at the four highest populations. Dried petals made up between 7 and 9% of flower fresh weight. Yields of dry petals/m2 were 36 g at 9 plants/m2 compared to an average of 89 g over the four highest plant populations. Seed weight/head was 0.78 g at 9 plants/m2 compared to c. 0.5 g for the other plant populations. Seed yield increased with increasing plant population from 128 g/m2 at

9 plants/m2 to 300 g/m2 at the three highest populations. Keywords Calendula officinalis L.; calendula; marigold; seed yield; flower yield

INTRODUCTION Calendula (Calendula officinalis L.), also known as pot marigold, is an annual or short-lived perennial herb native to southern Europe that produces orange to yellow flowers. It is commonly grown as an ornamental throughout the world and is cultivated in southern and eastern Europe. It is grown for a drug, calendulae flos, obtained from the flowers, particularly the petals (Bissett 1994). Calendulae flos is a complex mixture including essential oils, glycosides, saponins, triols, and xanthophylls, with the triterpenoids being the most important anti-inflammatory principles (Delia Loggia et al. 1994). Calendulae flos has been used historically as a traditional medicine and food dye (Khodzhaeva & Turakhozhaev 1993), but is currently being increasingly used as an external anti-inflammatory and wound healer (Bissett 1994; Akihisa et al. 1996; Patrick et al. 1996). This has prompted investigation of the potential for calendula flower production and processing in New Zealand. There is also interest in growing calendula seed in New Zealand. Seed has an oil content of 5-20% (Meier zu Beerentrup & Robbelen 1987; Angelini et al. 1997; Cromack & Smith 1998), of which up to 60% is calendic acid (Angelini et al. 1997; Cromack & Smith 1998). This seed oil has similar properties to rung oil and, therefore, could be used as a binder in paints, coatings, and cosmetics (Muuse etal. 1992). Calendula seed is very expensive (c. NZ$700/kg). Therefore, as a first stage to evaluating calendula as a field crop in New Zealand, a trial was carried out to determine whether seeding rate affected the production and/or size of calendula flowers, and hence seed yield.

H99025 Received 12 July 1999; accepted 3 December 1999

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New Zealand Journal of Crop and Horticultural Science, 2000, Vol. 28

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MATERIALS AND METHODS A field experiment was conducted in the 1997-98 season on a Templeton silt loam overlying sand (New Zealand Soil Bureau 1968) at Lincoln, New Zealand. The experiment followed a crop of oca (Oxalis tuber-osa) or a 1-year fallow, both of which followed Echinacea purpurea and Valeriana officinalis trials. The soil fertility status of the top 300 mm of the whole area, taken just before sowing, was pH 5.9, Ca 10 mg/g, K 14 mg/g, P 22 mg/g, S 6 mg/g, NO3-N 5 mg/g, and total N 0.21%. S.oil physical characteristics on an adjacent area have been given by Martin et al. (1992). The crop was sown on 27 November 1997 with a 9-row Oyjord Drill at six seeding rates: 3,6, 12,24, 48, and 96 kg/ha. A randomised block design was used with four replicates, three on the area following oca, and one on the area following fallow. Plots were 10 m long by 1.35 m wide. Seeding depth was 12 cm, and the crop was covered using light harrows attached to the drill. The crop was sprinkler irrigated at 2-4-week intervals from mid December to the end of February, with c. 25 mm applied at each irrigation. On 29 January 100 kg/ha N was applied as urea (46% N), just at the start of flowering. It was irrigated in with 25 mm water on the next day. The crop was hand weeded twice, at 1 and 3 months after drilling. The crop was sprayed with carbendazim (0.5 kg a.i./ha) and benzimidazole (0.25 kg a.i./ha) on 20 March to control an unidentified white fungal growth on the leaves. Each plot was divided into five 2 m lengths. The second and fourth lengths were sampled 2-3 times/ week from 2 February until 19 March 1998. On each occasion all fully opened flowers in the sampling length were plucked by hand, weighed and counted, then dried at 30C and reweighed. On 5 March height to the tallest flower was measured at four places in each plot. On 19 March each plot was visually assessed for lodging using a score of 1 for no lodging to 5 for completely lodged. On 2 and 3 April all plant material in the second and fourth sampling lengths was cut at ground level. The cut plants were weighed and a 1.2 to 2.2 kg subsample was dried at 80C in a forced-draft oven. On 16 April c. 50 dry flowers heads were taken from the first, third, and fifth lengths of the plots, dried at 35C, and the seed removed and weighed. Thousand seed weights were determined on triplicate samples from a seed counter.

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Martin & DeoEffect of plant population on calendula flower production

Fig 1 Calendula (Calendula offwinalis) cumulative flower number/m2 at six plant densities from early February to mid March 1998. = 9 plants/m2, O = 26 plants/m2, T = 46 plants/m2, V = 101 plants/m2, = 179 plants/m2, and = 332 plants/m2. Vertical bars represent LSDs between treatment means (P < 0.05) at each harvest.
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Data were analysed with analysis of variance using the Genstat statistical package (Genstat 5 Committee 1993). The lodging scores were analysed after a (log 10)+1 transformation.

RESULTS The season was one of the warmest and driest on record at Lincoln with mean temperatures of 15.2, 17.9, 19.6, and 16.8C for December 1997 and January, February, and March 1998 respectively. These temperatures were, on average, 1.3C higher than the long-term mean. Rainfalls for these months were 42.5, 17.2, 13.6, and 31.4 mm respectively, compared to the long-term mean of 55.5 mm/month over this period. Approximately 100 mm of irrigation water was applied during the growth of the crop, but, because of the high temperatures and low humidities during application, this may have been less effective than an equivalent amount of rainfall. Plant number, height, lodging, and biomass Plant counts increased from 9 plants/m2 at 3 kg seed/ ha to 332 plant/m2 at the 96 kg seed/ha rate (Table 1). Establishment based on a seed weight of 8 mg was not significantly different among all sowing rates; it averaged 30%. Plant height at 544 mm for 9 plants/m2 was significantly shorter than the 697 mm for all other plant populations. We also observed that as plant density increased the height of the flowers in the canopy became more even. The log-transformed lodging score data showed increased lodging at the two highest plant

populations (Table 1). Plant height and lodging differed considerably between replicates. Taller plants and more severe lodging occurred at higher plant populations in the replicates following the oca crop. Little lodging occurred in the replicate in areas of low residual fertility, which followed fallow. The lodging reduced flower production and increased rotting of plants. Lodging started in some plots after high winds in late February. Crop biomass doubled over the range of plant populations established, but biomass/plant decreased 15-fold from 45 g for the multi-branched bushy plants at 9 plants/m2 to 3 g for the singlestemmed, spindly plants at 332 plants/m2. Flower number Cumulative flower numbers were not significantly different for plant populations over 46/ha up to the end of February, or over 101/ha during March, but were significantly lower at lower plant populations (Fig. 1). For the three highest plant populations c. 600 flowers/m2 were collected over the 45 days of flower sampling (Table 2), and flower production rates averaged c. 15 flowers/m2 per day from mid February (Fig. 1). However, only 2-6 flowers were produced by each plant at these high plant populations, compared to over 18 at 9 plants/m2 (Table 2). There was no consistent replicate effect on flower number. Flower yield The total weight of fresh flower heads collected off each plot increased nearly 3-fold from 488 g/m2 at 9 plants/m2 to a mean of 1330 g/m2 at the four

Table 2 Total number of calendula (Calendula officinalis) flowers harvested per m2 and per plant, flower fresh weight (fwt) and dry weight (dwt) per m2, flower fresh weight per plant, fresh weight per flower, and flower dry matter (DM)%. (NS = not significant.)
Plant no./m2 9 26 46 101 179 332 LSD (5%) (d.f. 15) F-test probability Flowers/m2 164 334 451 576 583 650 111.4 O.001 Flowers/plant 18.6 13.3 10.3 6.1 3.4 2.1 3.01 <0.01 Flower fwt (g/tir) 488 927 1197 1411 1349 1361 241.2 O.001 Flower dwt (g/nr) 61 110 144 171 159 163 26.8 <0.001 Flower fwt /plant (g) 55.0 37.1 27.3 15.1 7.9 4.3 8.19 <0.001 Fwt/flower (g) 2.97 2.78 2.66 2.46 2.31 2.10 0.165 <0.001 Flower DM (%) 12.6 11.9 12.1 12.1 11.9 12.0 0.67 NS ^ g

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oo c o O Table 3 Ratio of calendula (Calendula officinalis) petal dry weight (dwt) to flower fresh weight (fwt) and to flower dry weight (dwt), petal dry matter (DM) %, calculated petal dwt/m2, and petal harvest index (HI). (NS = not significant.) Plant no./m2 9 26 46 101 179 332 LSD (5%) Petal dwt: flower fwt 0.0860 0.0812 0.0850 0.0782 0.0725 0.0765 0.00871 <0.05 Petal dwt: flower dwt 0.588 0.611 0.599 0.550 0.533 0.533 0.0406 <0.01 Petal DM (%) 14.2 12.7 13.6 13.4 12.4 13.5 1.29 NS Petal dwt/m2 (g) 36.0 67.2 86.3 93.9 85.4 89.8 13.81 <0.001 Petal HI 0.091 0.113 0.138 0.135 0.122 0.109 0.0293 <0.05
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Martin & DeoEffect of plant population on calendula flower production Fig 2 Calendula (Calendula officinalis) cumulative flower fresh weight/m2 at six plant densities from early February to mid March 1998. = 9 plants/m2, O = 26 plants/m2, T 2= 46 plants/m2, V = 101 plants/m , = 1792plants/m2, and = 332 plants/m . Vertical bars represent LSDs between treatment means (P < 0.05) at each harvest.

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highest plant populations, which did not have significantly different flower weights (Table 2). Similarly, the mean dry weight of flowers increased to 159 g/m2 for the four highest plant populations. Flower fresh weight per plant decreased 13-fold from 55 g/plant at 9 plants/m2 to 4 g/plant at 332 plants/m2 (Table 2), and the weight per flower also decreased by 30% from 3 g to just over 2 g. As a result, there was a considerable range in flower number and flower yield accumulation among plant populations (Fig. 1 and 2). Flower dry matter percentage varied from 11.6 to 14.1% over the season with no consistent trend (data not presented); at final harvest there was no significant difference between plant populations (Table 2). There was no consistent replicate effect on flower yield, indicating that any potential increased flower production associated with increased fertility was offset by the increased lodging. Petal yield The dried orange petals made up between 7 and 9% of the fresh weight and 53 and 61% of the dry weight of the flowers; the higher proportion was at the four lower plant populations (Table 3). There was no effect of plant population on petal dry matter percentage. Yields of dry petals/m2 were significantly lower for 9 plant/m2 (36 g) compared to the four highest plant populations (89 g on average). Petal harvest index (dry weight of petals/biomass dry weight) ranged from 9 to 14%, but, apart from the 9 plants/m2 density, did not differ with increasing plant population (Table 3).

Seed yield Seed yield, calculated from flower numbers and seed yield per head, increased with increasing plant population from 128 g/m2 at 9 plants/m2 to 335 g/ m2 at 332 plants/m2, but did not differ significantly at the three highest plant populations (Table 4). Seed weight/head was significantly higher at 9 plants/m2 (0.78 g) than the other plant populations, which averaged 0.51 g. The dry weights of heads excluding seeds followed a similar pattern. Seed made up 8590% of the head dry weight. There was no significant effect of plant population on individual seed weight. The estimated harvest index for seed was c. 29%, with no significant differences among treatments (Table 4).

DISCUSSION Seed yields increased at a decreasing rate with increasing plant density up to 332 plants/m2, with 85% of maximum seed yield produced at 101 plants/m2. This latter density is somewhat higher than the 4060 plants/m2 suggested as optimum by Van Dijk & Borm (1992) and Cromack & Smith (1998). Flower numbers and weights showed a similar response, but with maxima at 101 plants/m2, indicating that the optimum plant density for flower production is similar to that for seed production. Our establishment rate was about half that of Cromack & Smith (1998), but probably reflects the difficult conditions under which this crop established. Crusting soils and hot drying winds made

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New Zealand Journal of Crop and Horticultural Science, 2000, Vol. 28 emergence through a capped soil difficult. Our establishment rate was also affected by weed competition. Although there are no herbicide label recommendations for calendula in New Zealand, herbicides have been successfully used overseas to control weeds in this crop (Lamont & O'Connell 1986; Pank & Ennet 1988). Therefore, the optimum seeding rate of 24 kg/ha (101 plants/m2) in this trial may be reduced under more favourable establishment conditions towards the 10 kg/ha optimum found by Van Dijk & Borm (1992). Zhukova et al. (1996) found that dense populations delayed development of plants and diminished their size. In our trial any initial delay in flower production at higher plant populations would soon have been compensated for by the increased numbers of flowers produced at the higher seeding rates. Plant size decreased with increasing plant population, but the flowers appeared to be at a more even height near the top of the canopy, which made hand harvesting easier and may make mechanical harvesting easier because less vegetative bulk will have to be handled. Another approach may be to use growth regulators to compensate for delayed development in dense populations. Growth regulators have been shown to increase flowers per plant (Pal et al. 1986) and the volume of some of the chemical constituents of calendulae flos (Abdalla et al. 1986). Growth regulators may also reduce lodging, especially under higher fertility situations, as we observed that the higher density plants in the replicates in the area which had oca as the previous crop grew taller and lodged more than those in the replicate in the area which had been left fallow the previous year. Our seed was a mixture producing singles, semi doubles and doubles, mainly with orange petals but some lemon. Seed catalogues show a range of calendula flower types in terms of size, appearance, colour, and time to flowering. Whether the different flower types or colours have any effect on chemical composition and yield of calendulae flos is unknown, although Angelini et al. (1997) found a wide range in seed oil and calendulic acid contents between calendula types. In Italy, time from planting to flowering ranged from 17 to 59 days, depending on cultivar and season (Angelini et al. 1997). There may therefore be considerable scope to increase yields by selecting the best type suited to Canterbury conditions. Consistency of quality needs to be examined as Piccaglia et al. (1997) found that pigment levels in calendula flower heads varied considerably between years.

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Martin & DeoEffect of plant population on calendula flower production The seed yields obtained here were similar to those of Cromack & Smith (1998) who harvested yields of 2-2.51 seed/ha with an oil yield up to 19%. Oil content was not measured in this trial. Angelini et al. (1997) stated that, for calendula to be economic, cultivars with high seed oil content and a reduced tendency to shed seed should be selected. In common with overseas studies (Meier zu Beerentrup & Robbelen 1987; Baraynk et al. 1995) we found shedding of seed a problem when sampling our crop. There is scope to select for improved fruit shape and seed retention (Theobald & Robbelen 1989). Our harvested seed weights were at the low end of the range obtained by Angelini et al. (1997), and were lighter than the seed sown for this trial. The reasons for this are not known but may be because of the shorter time to maturity under the high temperatures experienced by our crop in the warm 1997-98 season. Seed yields have been increased with nitrogen fertiliser applications (Nordestgaard 1988; Barman & Pal 1994), although in this trial lodging was worse in those parts of the trial where more fertiliser had been applied to previous crops. The major problem with the commercialisation of calendula will be the harvesting of the flowers and the seed (Baraynk et al. 1995). The calendula plant is indeterminate, and unharvested plants in our trial continued to flower throughout the winter. An estimate will have to be made about the best time to harvest flowers or seed. For seed production in the Netherlands, Breemhaar & Bouman (1995) suggested that the optional harvest time was when 5070% of the seeds were mature. Semi-mechanical methods for harvesting flowers are being developed but still result in considerable yield losses (van der Mheen 1993). For seed production, the crop has to be swathed or desiccated (Nordestgaard 1988; Breemhaar & Bouman 1995). The seed is curly and needs multistage cleaning (Breemhaar & Bouman 1995), although an experimental dresser has been developed to remove appendages from seed (Bilsland & Berlage 1986). This preliminary study has shown that calendula can be easily grown in New Zealand for flower or seed production. Small areas are already grown to supply hand-picked flowers to the local pharmaceutical industry. Whether the crop will be grown on a larger scale will depend on markets, profitability, and the development of suitable harvesting technology. Further agronomic research is needed to ensure that validated crop management guidelines are available to growers if and when large-scale calendula production occurs.
ACKNOWLEDGMENTS

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We thank Natures Kiss (New Zealand) Ltd for supplying the seed and Julian Hodgkinson for assistance with maintaining and sampling the crop. The trial was funded through the Public Good Science Fund administered by the New Zealand Foundation for Research, Science and Technology.

REFERENCES Abdalla, N. M.; El-Gengaihi, S.; Sadrak, I. 1986: A study on the influence of Cycocel and Alar 85 on growth, flowering and active ingredients of Calendula officinalis L. Acta Agronomica Hungarica 35: 41-45. Akihisa, T.; Yasukawa, K.; Oinuma, H.; Kasahara, Y.; Yamanouchi, S.; Takido, M.; Kumaki, K.; Tamura, T. 1996: Triterpene alcohols from the flowers of Compositae and their anti-inflammatory effects. Phytochemistry 43: 1255-1260. Angelini, L. G.; Moscheni, E.; Colonna, G.; Belloni, P.; Bonari, E. 1997: Variation in agronomic characteristics and seed oil composition of new oilseed crops in central Italy. Industrial Crops and Products 6: 313-323. Baranyk, P.; Zeleny, V.; Zukalova, H.; Horejs, P. 1995: Oil content of some species of alternative oil plants. Rostlinna Vyroba 41: 433-438. Barman, D.; Pal, P. 1994: Effect of nitrogen and phosphorus on seed yield in calendula {Calendula officinalis L.). Orissa Journal of Agricultural Research 7: 17-21. Bilsland, D. M.; Berlage, A. G. 1986: A filament thresher for seed. American Society of Agricultural Engineers Paper 86-3061. 9 p. Bisset, N. J. 1994: Herbal drugs andphytopharmaceuticals. Medpharm, Stuttgart and CRC, Bota Raton, Ann Arbor, London and Tokyo. 566 p. Breemhaar, H. G.; Bouman, A. 1995: Harvesting and cleaning Calendula officinalis, a new arable oilseed crop for industrial application. Industrial Crops and Products 4: 255-260. Cromack, H. T. H.; Smith, J. M. 1998: Calendula officinalisproduction potential and crop agronomy in southern England. Industrial Crops and Products 7: 223-229. Delia Loggia, R.; Tubaro, A.; Sosa, S.; Becker, H.; Saar, St.; Isaac, O. 1994: The role of triterpenoids in the topical anti-inflammatory activity of Calendula officinalis flowers. Planta Medica 60: 516520. Genstat 5 Committee 1993: Genstat 5 Release 3 Reference Manual. Oxford University Press, Oxford. 796 p.

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New Zealand Journal of Crop and Horticultural Science, 2000, Vol. 28

Pank, F.; Ennet, D. 1988: Chemical weed control in medicinal plant crops. Part 10: marigold (Calendula officinalis L.). Pharmazie 43: 503-506. Patrick, K. F. M.; Kumar, S.; Edwardson, P. A. D.; Hutchinson, J. J. 1996: Induction of vascularization by an aqueous extract of the flowers of Calendula officinalis L., the European marigold. Phytomedicine 3: 11-18. Piccaglia, R.; Marotti, M.; Chiavari G.; Gandini, N. 1997: Effects of harvesting date and climate on the flavonoid and carotenoid contents of marigold (Calendula officinalis L.). Flavour and Fragrance Journal 12: 85-90. Theobald, D.; Rbbelen, G. 1989: Flowering biology and fruit formation of Calendula officinalis as a basis for continued breeding success. Angewandte Botanik 63: 313-322. Van der Mheen, H. J. 1993: Research on herb cultivation for flower production. Publikatie Proefstation voor de Akkerbouw en de Groenteteelt in de Vollegrond, Lelystad No. 70A: 154-156. Van Dijk, N.; Borm, G. E. L. 1992: Crop management research on Calendula officinalis (marigold) in 1992. Publikatie Proefstation voor de Akkerbouw en de Groenteteelt in de Vollegrond, Lelystad No. 73A: 108-109. Zhukova, L. A.; Voskresneskaya, O. L.; Grosheva, N. P. 1996: Morphological and physiological characteristics of ontogenesis in pot marigold (Calendula officinalis L.) plants grown at different densities. Russian Journal of Ecology 27: 100 106.

Khodzhaeva, M. A.; Turakhozhaev, M. T. 1993: Carbohydrates of Calendula officinalis. Chemistry of Natural Compounds 29: 533-534. Lamont, G.; O'Connell, M. A. 1986: An evaluation of pre-emergent herbicides in field grown cut flowers. Plant Protection Quarterly 1: 95-100. Martin, R. J.; Jamieson, P. D.; Wilson, D. R.; Francis, G. S. 1992: Effects of soil moisture deficits on the yield and quality of 'Russet Burbank' potatoes. New Zealand Journal of Crop and Horticultural Science 20: 1-9. Meier zu Beerentrup, H.; Robbelen, G. 1987: Calendula and coriandrumnew potential oilcrops for industrial uses. Zeitschrift fr Fett Wissenschaft und Technologie der Fette, le und Wachse 89: 227-230. Muuse, B. G.; Cuperus, F. P.; Derksen, J. T. P. 1992: Composition and physical properties of oils from new oilseed crops. Industrial Crops and Products 1: 57-65. New Zealand Soil Bureau 1968: General survey of the soils of the South Island. New Zealand Soil Bureau Bulletin No. 27. 404 p. Nordestgaard, A. 1988: Trials with calendulas for seed production. Tidsskrift for Planteavl 92: 213-215. Pal, P.; Hore, J.; Poi, A. K. 1986: Effect of growth regulating chemicals on growth and yield of flowers of Calendula officinalis. Environment and Ecology 4: 541-543.

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