330 Journal of Applied Sciences Research, 9(1): 330-340, 2013 ISSN 1819-544X This is a refereed journal and all articles are professionally screened and reviewed

ORIGINAL ARTICLES

Effect of planting date and spacing on growth, yield and essential oil of Mexican marigold ( lucida L.) cultivated in

1Rasha F. Ismail, 2Awaad M. Kandeel, 2Ayman K. Ibrahim and 1Elsayed A. Omer

1Department of Medicinal and Aromatic Research, National Research Centre, , (12622), , Egypt. 2Department of Horticulture, Faculty of , Ain-Shams University, El-Kheima, , Egypt.

ABSTRACT

In order to enrich the Egyptian cultivation with new species of Tagetes, the seeds of Tagetes lucida (Mexican marigold) were imported and cultivated under green house conditions at peat moss media. After 45 days, the seedlings were transplanted in new reclaimed land (sand soil) at the Experimental Station of National Research Centre (Nubaria, Behira Governorate) during two successive seasons (2009 and 2010) to investigate the effect of planting date and plant spacing on growth and yield of Tagetes lucida plant. The experiment included 9 treatments (the combination between 3 planting dates; February, March and April and three plant spacing; (40, 50 and 60 cm). The growth parameters (plant height, plant diameter, number of branches,as well as fresh and dry yield of plant herb) were significantly responded to planting date and plant spacing in both seasons. Essential oil percentage of the plant fresh herb did not show dramatic changes according neither to planting date nor to plant spacing and ranged from 0.4 to 0.6 %. Essential oil yield ml/plant and L/ feddan significantly responded to planting date and plant spacing in both seasons. The best treatment for the maximum production of fresh and dry herb and essential oil of T. lucida was the cultivation in April at 50 cm plant spacing. Thus, we recommend the cultivation of Tagetes lucida plant at 50cm of plant spacing in April for the highest yield of herb and essential oil yield under the sand soil conditions in the new reclaimed lands in Egypt.

Key words: Tagetes lucida, Mexican Marigold, Planting date, Plant spacing, Growth, Essential oil

Introduction

The genus Tagetes belongs to family and consists of approximately 40-50 species (Strother, 1977; Lawrence, 1985). Tagetes lucida (synonymous: T. florida Sweet, T. schiedeana Less) is a native herb widely used in this area and has an important economic potential (Giron et al., 1991; Damian-Badillo et al., 2008a). Tagetes lucida is an aromatic herb distributed naturally from to , at altitude between 1000 and 2000 m. It has sessile, glabrous, oblong-lanceolate and opposite leaves (5-10 cm long) with terminal flowers. It is known that T. lucida is an important source of antioxidant and antifungal compounds (Aquino et al., 2002; Damian-Badillo et al., 2008b). It has several traditional uses, such as food, antiseptic, control infectious, emotional and spirituals. T. lucida is one of the plants most used by the Latin-American population for the treatment of gastrointestinal disorders (Giron et al., 1991; Damian Badillo et al., 2008a). In Mexico, it is used medicinally an infusion, insecticide and ornamental plant. The infusion is used as a tonic, a remedy for coughs, headaches, fevers, colic, , gastrointestinal ailments,body ache and to speed birth (emmenagogue) (Browner, 1985; Bye, 1986). In Guatemala, extracts of this plant are sold as infusion, tincture and elixir (Cáceres, 1996). These products are used for stomach pains, gastritis, menstrual pains (Girón et al., 1991), to treat infections (Cáceres et al., 1993a) and (Cáceres et al., 1990 and 1993b). This plant is cultivated in the United States, France and England as a flowering herb (Morton, 1981). In Nottinghamshire (England), at the beginning of the twentieth century, the plant was used in place of tarragon in soup (Morton, 1981). Today, this plant is used in the Southern states of USA, as well as in Costa Rica as a spice for food. Nutritional aspect of T. lucida is a pharmacologically and nutraceutiacally important if it is used as spice; because this plant could contributes to maintenance of health human being. For all this, T. lucida is proposal as an alternative plant culture new with advantages with regard to other regional and conventional crop. It promoted as crop and use as herbal remedy, nutraceutical and food reinforcement in accord to official standard (Mejia-Barajas et al., 2012).

Corresponding Author: Elasyed A. Omer, Department of Medicinal and Aromatic Plants Research, National Research Centre, Dokki, (12622), Giza, Egypt. E-mail: [email protected] 331 J. Appl. Sci. Res., 9(1): 330-340, 2013

Planting date and plant spacing are considered as important factors that affect plant growth, yield and oil production. Mohanty et al. (1993) found that planting T. erecta in May produced a greater number of secondary branches and increased plant height and spread. Planting in Sep. produced a greater number of larger flowers per plant and per plot. Mishra (1997) transplanted French marigold seedlings to the field (at Samastipur, Bihar, India)at monthly intervals between September and August. They found that planting in May increased plant height and plant spread and extended the cropping and flowering periods compared to the other treatments. The planting in April significantly increased length of flower stalk and weight per flower. Mohanty et al. (1997) reported that rooted cuttings of T. erecta planted on May and July at Bhubaneswar (Orissa), India gave the greatest number of branches per plant, plant height and stem thickness. Flower buds appeared earlier when cuttings were planted in November or January, but flower yield was highest in the September planting, followed by July and November.The highest flower yield per plant was achieved by planting on 1 September. Samantaray et al. (1999) stated that African Yellow marigold was planted in Orissa on 1 May (D1), 1 July (D2), 1 September (D3), 1 November (D4) or 1 January (D5).The optimum planting date was 1 September, closely followed by July planting. January and November plantings hastened the appearance of flower bud. Acosta de la Luz et al. (2010) mentioned that, t h e best planting date for T. lucida was on May. Acosta de la Luz et al. (2011) r eport ed that it is possible to set the optimal date of cultivation of T. lucida in the period from the beginning of May to the beginning of June. Chanda and Roychoudhury (1991) mentioned that T. erecta plant hight, plant spread, number of primary branches, number of flowers and flower yield/plant were higher at the wider spacing but yield/ha was highest at the closer spacing. Yadav et al. (2004) stated that Tagetes erecta plant height was greatest under 40x30 cm spacing (67.13cm). The greatest stem diameter (1.64 cm), plant spread (43.82 cm), number of leaves (147.92), number of primary (14.54) and secondary (64.11) branches, fresh weight (306.54 g) number of flowers (22.89) and flower yield per plant (231.75 g) were obtained under 60x45 cm spacing. Karuppaiah and Krishna (2005) on French marigold ( L.) reveled that the spacing of 30x30 cm recorded the maximum value of growth characters viz., plant height, number of primary and secondary branches, number of leaves, leaf area and dry matter production. Sreekanth et al. (2006) mentioned that the closer spacing of 40x30 cm recorded the maximum flower diameter, yield per plant and yield/ha of African marigold (Tagetes erecta). Anju and Pandey (2007) reported that the growth parameters of African marigold like maximum number of branches per plant, number of leaves per branch, diameter of stem and canopy of plant were recorded higher in wider spacing 40x30 cm, while plant height was highest (28.46 cm) in closer spacing 20x10 cm. Sunitha et al. (2007) revealed that T. erecta cultivated at the wider spacing (60x60 cm) recorded more number of branches , flowers , seed yield per plant and per hectare than closer spacing of 60x40 cm. Ghosh and Pal (2008) on African marigold (Tagetes erecta) reported that the highest number of primary branches, greatest average spread of the plant, size and weight of individual flower were recorded at the widest spacing (40x40 cm). Singh et al. (2008) mentioned that T. minuta plant rows spaced at 30 cm distance produced 20% and 47% more oil than 45 cm and 60 cm spacing, respectively. Essential oil biosynthesis was adversely affected in plants that were either spaced closer than 60 cm in rows. Chaturvedi et al. (2010) stated that maximum marigold growth parameters in terms of plant height, spread of plant, number of leaves and length of flower stalk were exhibited in treatment 40x30cm. The minimum values in all parameters were manifested mostly in treatment of 20x30 cm. Mohanty et al. (1993) on Tagetes erecta found that the most profitable crop was obtained from a Septemper planting at 40 x 30 cm spacing. Ram et al. (1998) on Tagetes minuta at Pantnagar in India showed that mid- October planting and closer spacing (45cm x 45 cm) resulted in maximum increase in plant height, primary branches number and oil yield. Samantaray et al. (1999) stated that in Orissa, the highest yield of flowers/plant of African yellow marigold was obtained with September planting at the spacing of 40cmx30 cm. Serrato-Cruz and Rivera-Mendez (2002) found that in all planting dates for T. erecta (10 March, 12 April, 11 May, 17 June and 26 July), the shortest plant height and the highest flower bud number per plant were recorded at plant density of 30x30 cm. Sreekanth et al. (2006) found that T. erecta cultivated in October in Hyderabad, Andhra Pradesh, india recorded maximum flower diameter, yield per plant and yield/ha and the closer spacing of 40x30 cm recorded maximum flower diameter, yield/plant and yield/ha, but the maximum colour intensity of T.erecta was recorded in December planting. A wider spacing of 60x30 cm recorded maximum colour intensity. Ghosh and Pal (2008) reported that T. erecta cultivated on12 October at 40x20 cm spacing recorded higher yield of flowers per plot in comparison with all the other treatments. This study aimed to investigate the effect of planting date and plant spacing on growth, yield and essential oil of Tagets lucida plant cultivated in Egypt for the first time.

Materials and Methods

This experiment was carried out at the Agricultural Experimental Station of National Research Centre (Nubaria, Behira Governorate, 150 km North West of Cairo) during the two successive seasons of 2009 and

332 J. Appl. Sci. Res., 9(1): 330-340, 2013

2010. The soil of the experiment site was sandy soil and the irrigation system was drip irrigation with water coming from Elkhatattba canal. The physical and chemical analyses of the soil used for this experiment revealed that, the soil is sand in texture. The physical and chemical characters of the soil were determined before cultivation according to Jackson (1973) and are presented in Table (1). The soil was prepared before cultivation and divided into plots (9m2, 4X 2.25m). 3000kg compost, 200kg calcium super phosphate and 200 kg sulphur were added per feddan (4200m2) during soil preparation. All treatments were fertilized with ammonium nitrate (33.5% N) at the rate of 200kg/fed. at three portions; one month later from transplanting and after the first and second cuts.

Table 1: The physical and chemical properties of the experimental soil during 2009 and 2010 seasons. Physical properties Very coarse Coarse Medium Fine Very fine Silt+Clay Texture Year sand sand sand sand sand

2009 34.77 35.31 0.48 15.98 12.45 1.01 Sandy 2010 37.16 34.71 0.42 14.15 12.43 1.13 Sandy Chemical properties pH E.C. (meq/l) (dSm-1) Cations Anions ++ ++ + + -- - - (2.5:1) (1:1) Ca Mg Na K CO3 HCO3 Cl So4 2009 7.96 0.4 1.5 1.0 0.8 0.2 - 0.8 1.0 0.9 2010 8.27 0.32 1.5 1.0 0.5 0.3 0.7 1.8 0.7

The layout of the experiment was split plot design. The main plots were assigned to planting date; February (D1), March (D2) and April (D3), while the sub plots were assigned to plant spacing; 40 cm (S1), 50 cm (S2) and 60 cm (S3) between each two plants. The experiment included nine treatments represented the combinations between three planting dates and three plant spacing. Each treatment was replicated four times. The seeds of T. lucida were imported from Canada and cultivated in the nursery in 1st January for the first date, 28th January for the second date and 1st March for the third date in both seasons in seed trays at Elmyzan Company greenhouse, (SEKM, Bilbase, Sharkya Governorate). Seedlings of Tagetes lucida were transplanted in the permanent field at 40, 50 and 60 cm of planting spacing in rows of 75 cm apart in 15th February for the first date, 15th March for the second date and 15th April for the third date in both seasons. The agricultural treatments normally done for other Tagetes species were applied for Tagetes lucida in all treatments. Plants were watered with drip irrigation system.

Table 2: Monthly average of metrological data of the experimental farm of National Research Centre, Nubaria, Behira,, Egypt, in two seasons Metrological data Air temperature oC R.H. (%) Month Max. Min. Average Feb. 2009 19.4 11.7 15.6 59.5 Mar. 2009 20.8 12.3 16.6 60.3 Apr. 2009 23.9 12.6 18.3 61.8 May 2009 24.8 13.4 19.1 62.8 June 2009 28.8 14.9 21.9 63.9 July 2009 28.0 14.3 21.1 71.0 Aug. 2009 28.7 16.6 22.7 69.6 Sept.2009 28.7 19.4 24.1 66.8 Feb. 2010 22.1 13.8 17.9 60.1 Mar. 2010 23.1 13.9 18.5 62.5 Apr. 2010 24.9 14.2 19.6 60.8 May 2010 27.2 15.0 21.1 61.9 June 2010 28.6 15.0 21.8 66.0 July 2010 28.3 15.5 21.9 71.3 Aug. 2010 29.5 16.9 23.2 72.1 Sept. 2010 29.0 19.3 24.1 67.8 R.H.: Relative humidity Source: Central Laboratory for Agricultural Climate, Ministry of Agriculture, Cairo, Egypt

Plant height (cm), plant diameter (cm) and number of branches/plant were recorded for five times during the growing season on15th May, 15th June, 15th July, 15th August and 15th September in both seasons. Fresh and dry weight (g/plant) and essential oil percentage (%) of herbage were recorded for three times in the growing season on 15th June (1st cut), 1st August (2nd cut), and 15th September (3rd cut) in both seasons. Essential oil percentage (ml/100g) of fresh herb for each treatment was determined by mixing represented samples from the three replicates of each treatment. A weight of 30 mg fresh herb of each treatment was

333 J. Appl. Sci. Res., 9(1): 330-340, 2013 subjected for hydro-distillation for 3 hours at Clevenger-type apparatus according to Egyptian Pharmacopoeia (1984). The resulted essential oil of each treatment was separately dehydrated with anhydrous sodium sulphate and kept in the deep freezer. The data recorded were analyzed as a split plot design by analysis of variance (ANOVA) using the General Linear Models procedure of CoStat (Snedecor and Cochran, 1967). Least significant difference (LSD) test was applied at 0.05 probability level to compare the mean treatments.

Results and Discussion

1- Plant height (cm):

Table (3) represents the results of Tagetes lucida plant height as affected by planting date and/or plant spacing during two growing seasons (2009 and 2010). It is clear that the effect of plant spacing on plant height was significant in both seasons in all dates of measurements, while the effect of planting date on plant height was significant during the first three dates of measurements in the first season and in all dates of measurements in the second season. Except the third date of measurements in the first season, the effect of interaction on plant height was insignificant. Although no significant differences were observed in plant height between S3 (60 cm) and S2 (50 cm) in the first and second dates of measurements in both seasons, the maximum plant height was observed with S3 in most of planting dates. In the beginning of plant age, the differences in plant height between the three planting dates were higher, while the three values decreased with the advance of plant age. During the fifth date of measurements, plant height values were almost the same. Similar results were obtained by Acosta de la Luz et al. (2010) and Acosta de la Luz et al. (2011) on Tagetes lucida, Chanda and Roychoudhury (1991), Belorkar et al. (1992), Mohanty et al. (1993 and 1997), , Natarajan and Vijayakumar (2002), Serrato-Cruz and Rivera-Mendez (2002), Yadav et al. (2004), Anju and Pandey (2007) and Tiwari et al.(2010) on Tagetes erecta, Ram et al. (1998) and Ramesh and Singh (2008) on Tagetes minuta, Karuppaiah and Krishna (2005) on Tagetes patula, Chaturvedi et al. (2010) and Mor et al. (2006) on marigold and Mishra (1997) on French marigold.

Table 3: Effect of planting date and plant spacing on plant height (cm) of Tagetes lucida plants during two seasons. First Season (2009) Second season (2010) Plant height (cm) Planting date Planting date nts Plant spacing spacing

Dates of Mean Mean

Measureme D1 D2 D3 D1 D2 D3

S1 9.2 5.3 6.2 6.9 8.3 6.4 5.4 6.7 15th May S2 10.2 5.9 7.2 7.7 10.0 6.7 6.7 7.8 S3 10.6 6.3 7.6 8.2 10.9 7 6.5 8.1 Mean 10.0 5.8 7.0 9.7 6.7 6.2 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.69 0.84 n.s. 0.8 0.57 n.s. S1 11.7 7.3 8.9 9.3 10.7 7.8 7.3 8.6 15th June S2 12.3 8.2 9.9 10.1 12.3 8.7 8.5 9.8 S3 12.6 8.4 10.2 10.4 12.9 8.7 8.1 9.9 Mean 12.2 8.0 9.6 12.0 8.4 8.0 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.555 1.24 n.s. 0.479 0.863 n.s. S1 35.0 30.2 31.5 32.2 34.8 29.6 34.7 33.0 15th July S2 39.2 36.5 36.7 37.5 42.1 36.8 40.0 39.6 S3 41.2 39.2 38.7 39.7 44.5 38.4 42.7 41.9 Mean 38.4 35.3 35.6 40.5 34.9 39.1 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.619 1.965 1.074 0.683 1.037 n.s. S1 40.7 34.3 39.3 38.1 42.4 37.0 40.6 40.0 15th Aug. S2 47.0 41.0 46.0 44.7 49.1 45.3 47.0 47.1 S3 50.0 46.4 48.1 48.2 51.2 48.1 49.2 49.5 Mean 45.9 40.6 44.5 47.5 43.5 45.6 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 2.959 n.s. n.s. 1.931 1.309 n.s. S1 45.2 40.3 44.5 43.3 49.2 43.3 46.3 46.3 15th Sep. S2 54.1 48.2 52.1 51.4 55.2 51.0 53.1 53.1 S3 56.3 52.3 54.8 54.5 58.7 54.7 56.3 56.6 Mean 51.8 47.0 50.5 54.4 49.7 51.9 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 3.487 n.s. n.s. 2.3 1.378 n.s.

334 J. Appl. Sci. Res., 9(1): 330-340, 2013

On the other hand, Vrsek and Zidovec (2002) mentioned that, plant spacing had no significant effect on Tagetes erecta.

2-Plant diameter (cm):

The results showed that, the effect of planting date or plant spacing on plant diameter was significant in both seasons in all dates of measurements, while the effect of interaction was insignificant in all dates of measurements in both seasons (Table 4). Transplanting Tagetes lucida in February increased plant diameter comparing to March and April planting dates during both seasons in all dates of measurements, but these differences decreased with increasing in plant age. At the end of plant life the plant diameters were almost the same in the three planting measurements. Increasing plant spacing significantly increased plant diameter in most dates of measurements in both seasons which may attributed to the non-competition between plants on the light, water and nutritional elements available in the growth zone. Transplanting Tagetes lucida in Feb. at 60 cm between two hills resulted in the tallest plant and the maximum plant diameter in both seasons. These results are agree with Chanda and Roychoudhury (1991), Mohanty et al. (1993), Yadav et al. (2004), Ghosh and Pal (2008) and Tiwari et al. (2010) on Tagetes erecta, Mishra (1997) on French marigold and Chaturvedi et al. (2010) on marigold. On the other hand, plant spacing did not have a significant effect on Tagetes erecta (Vrsek and Zidovec, 2002).

Table 4: Effect of planting date and plant spacing on plant diameter (cm) of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Planting date Planting date

Dates of Mean Mean Plant spacing Plant spacing Measurements Measurements D1 D2 D3 D1 D2 D3

S1 7.0 5.7 3.8 5.5 7.5 5.7 3.9 5.7 15th May S2 7.7 6.2 4.3 6.1 8.5 7.0 5.5 7.0 S3 7.9 6.4 4.5 6.3 9.2 8.0 6.5 7.9 Mean 7.5 6.1 4.2 8.4 6.9 5.3 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.41 0.728 n.s. 0.602 0.611 n.s. S1 9.6 6.9 5.2 7.2 9.8 8.0 6.5 8.1 15th June S2 10.4 7.9 5.8 8.0 11.5 9.6 7.8 9.6 S3 10.6 8.3 6.4 8.5 12.3 10.1 8.5 10.3 Mean 10.2 7.7 5.8 11.2 9.3 7.6 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.781 0.42 n.s. 0.708 0.971 n.s. S1 32.8 27.4 26.1 28.8 36.2 28.8 30.2 31.7 15th July S2 39.8 31.4 32.8 34.7 42.9 37.3 37.0 39.1 S3 41.6 34.8 36.7 37.7 46.5 41.6 39.7 42.6 Mean 38.1 31.2 31.9 41.9 35.9 35.6 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.92 1.58 n.s. 1.55 2.225 n.s. S1 39.6 33.1 33.3 35.3 41.6 35.5 37.4 38.2 15th Aug. S2 43.3 39.5 40.2 41.0 47.0 43.6 43.7 44.7 S3 45.1 42.6 44.2 44.0 50.7 47.7 46.7 48.4 Mean 42.7 38.4 39.2 46.4 42.2 42.6 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 2.397 1.89 n.s. 1.453 1.82 n.s. S1 42.3 34.8 36.5 37.9 42.0 38.9 41.6 40.8 15th Sep. S2 48.4 42.3 44.4 45.0 50.8 47.4 48.2 48.8 S3 51.0 44.9 48.2 48.0 53.4 50.9 51.5 51.9 Mean 47.2 40.7 43.0 48.7 45.7 47.1 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.584 3.101 n.s. 1.695 1.786 n.s.

3-Number of branches/plant:

The results showed that the effect of planting date and plant spacing on the number of branches of Tagetes lucida was significant in the first measurement dates in both seasons, In the next three dates, the effect of plant spacing was significant and the planting date was insignificant, while the interaction effect was insignificant in both seasons (Table 5). The variation in the number of branches according to the dates of planting were higher in the beginning of plant life and decreased with the progress of plant age to be almost the same at the end of

335 J. Appl. Sci. Res., 9(1): 330-340, 2013 plant life. It is interesting to mention that the number of branches tended to be stable in the last three measurements which mean that the branching stopped in these plant stages. Increasing plant spacing significantly increased plant branching in all planting dates in both seasons. The maximum number of branches in Tagetes lucida was resulted from 60cm spacing in all dates of planting. These results are in accordance with Chanda and Roychoudhury (1991) , Mohanty et al. (1993 and 1997), Vrsek and Zidovec (2002), Yadav et al. (2004), Sunitha et al. (2007), Ghosh and Pal (2008) and Anju and Pandey (2007) on Tagetes erecta, Ram et al. (1998) o n Tagetes minuta, Karuppaiah and Krishna (2005) on Tagetes patula, , on Tagetes erecta and Acosta de la Luz et al. (2011) on Tagetes lucida.

Table 5: Effect of planting date and plant spacing on number of branches/plant of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Planting date Planting date

Dates of Mean Mean Plant spacing Plant spacing Measurements Measurements D1 D2 D3 D1 D2 D3

S1 3.3 2.7 1.3 2.4 4.0 2.3 1.0 2.4 15th May S2 4.0 3.3 1.7 3.0 6.0 4.3 1.7 4.0 S3 4.7 4.3 1.7 3.6 7.7 6.0 1.7 5.1 Mean 4.0 3.4 1.6 5.9 4.2 2.0 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.609 0.308 n.s. 0.872 0.959 n.s. S1 10.3 8.7 6.0 8.3 8.7 8.0 6.7 7.8 15th June S2 12.3 9.0 7.3 9.6 12.0 12.0 9.7 11.2 S3 14.3 12.7 8.0 11.7 15.7 13.7 11.7 13.7 Mean 12.3 10.1 7.1 12.1 11.2 9.3 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.575 0.666 n.s. 1.751 1.661 n.s. S1 13.3 11.0 13.3 12.6 11.3 10.0 11.7 11.0 15th July S2 15.3 13.0 15.0 14.4 15.3 14.3 16.7 15.4 S3 18.0 15.7 17.0 16.9 19.3 15.3 20.0 18.2 Mean 15.6 13.2 15.1 15.3 13.2 16.1 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.472 1.332 n.s. 1.611 n.s. n.s. S1 13.3 11.0 13.3 12.6 11.7 10.7 11.7 11.3 15th Aug. S2 15.3 13.0 15.0 14.4 16.3 15.3 17.0 16.2 S3 18.0 15.7 17.0 16.9 20.0 16.7 20.0 18.9 Mean 15.6 13.2 15.1 16.0 14.2 16.2 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.472 1.332 n.s. 1.498 n.s. n.s. S1 13.3 11.7 13.3 12.8 12.0 10.7 11.7 11.4 15th Sep. S2 15.7 13.0 15.0 14.6 16.3 15.3 17.3 16.3 S3 18.0 15.7 17.3 17.0 20.0 16.7 20.0 18.9 Mean 15.7 13.4 15.2 16.1 14.2 16.3 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.518 n.s. n.s. 1.397 n.s. n.s.

4-Fresh and dry weight of herb (g/plant):

Tagetes lucida fresh and dry weight were significantly affected by planting date and/or plant spacing (Table 6), while the interaction effect was significant in the first season only for fresh weight and in the first two measearing dates for dry weight in both seasons. Although the effect of planting date was significant between the dates in the beginning of plant life in both seasons, these differences between February and April planting were insignificant at the end of plant life which means that plants transplanted in April compensate the delay in transplanting and reached the same weight of plants transplanted in February. This may be attributed to the high temperature in April comparing to the other two dates as shown in Table (2), which stimulate the metabolic processes. This has been reflected on plant height, plant diameter, number of branches and fresh and dry weight of herb. Increasing plant spacing significantly increased plant fresh weight in both seasons in all plant cuts. The maximum fresh weight of Tagetes lucida was obtained from transplanting in February at 60 cm plant spacing in the both seasons but the differences between plants which transplanted in April and those transplanted in February were insignificant. These results are in harmony with Mohanty et al. (1993), Rao and Reddy (2002) and Yadav et al. (2004) on Tagetes erecta, Karuppaiah and Krishna (2005) on Tagetes patula, Mor et al. (2006) on marigold and Singh et al. (2008) on Tagetes minuta.

336 J. Appl. Sci. Res., 9(1): 330-340, 2013

Table 6: Effect of planting date and plant spacing on herb fresh and dry weight (g/plant) of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Fresh weight (g/plant)

Cuts Planting date Planting date Mean Mean Plant spacings spacings Plant D1 D2 D3 D1 D2 D3

S1 79.8 45.4 32.9 52.7 84.4 42.2 33.7 53.4 1st cut S2 123.2 76.7 47.1 82.3 131.1 77.0 49.3 85.8 S3 154.6 88.7 59.3 100.8 143.7 89.1 55.0 95.9 Mean 119.2 70.2 46.4 119.8 69.5 46.0 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 7.265 11.04 12.58 10.34 9.41 n.s. S1 144.7 128.8 135.3 136.3 202.6 172.7 188.5 187.9 2nd cut S2 300.1 221.1 293.1 271.4 289.8 242.5 296.9 276.4 S3 333.2 244.5 324.3 300.7 314.1 278.8 310.3 301.1 Mean 259.3 198.1 198.1 268.8 231.3 265.3 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 15.03 20.03 26.04 14.533 10.173 n.s. S1 189.0 179.2 188.1 185.4 237.6 229.4 245.0 237.3 3rd cut S2 351.8 271.0 334.1 319.0 337.4 317.7 340.7 331.9 S3 373.5 298.4 359.1 343.7 356.5 337.4 356.1 350.0 Mean 304.7 249.5 293.8 310.5 294.8 313.9 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 16.67 23.55 28.88 17.31 5.66 n.s. Dry weight (g/plant) S1 26.2 15.0 11.0 17.4 28.3 13.3 11.0 17.5 1st cut S2 40.9 24.3 15.6 26.9 41.9 24.6 16.7 27.7 S3 50.7 31.5 18.5 33.5 47.5 28.9 17.9 31.4 Mean 39.3 23.6 15.0 39.2 22.3 15.2 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 2.72 4.52 4.71 3.125 3.27 5.41 S1 50.9 42.7 45.6 46.4 50.9 42.7 45.6 46.4 2nd cut S2 102.9 69.1 97.6 89.9 102.9 69.1 97.6 89.9 S3 112.7 79.8 111.5 101.4 112.7 79.8 111.5 101.4 Mean 88.9 63.9 84.9 88.9 63.9 84.9 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 7.28 12.69 12.62 7.28 12.69 12.62 S1 61.8 52.5 61.9 58.7 78.8 75.7 78.1 77.6 3rd cut S2 107.7 86.9 106.1 100.3 120.0 97.9 108.3 108.7 S3 121.1 93.9 114.7 109.9 128.0 110.9 121.1 120.0 Mean 96.9 77.8 94.2 108.9 94.9 102.5 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 4.924 13.344 n.s. 6.412 6.444 n.s.

5-Fresh and dry Yield (ton/Fed.):

Plant spacing and planting date showed significant effects on both fresh and dry yield in both seasons (Table, 7), while the interaction effect was insignificant in the second season for fresh yield and in both seasons for dry yield. The fresh and dry yield significantly increased with increasing plant spacing from 40 to 50 cm and then significantly decreased when plant spacing increased from 50 to 60 cm. Transplanting Tagetes lucida in April was superior in fresh and dry yield in the second season comparing to February and March, while the differences in fresh and dry yield between April and February transplantation were insignificant. The fresh and dry yield of Tagetes lucida plants which transplanted in March showed the lower values comparing to the other two dates in both seasons in the last two dates of cuts. The maximum fresh and dry yield of Tagetes lucida was resulted from Plants transplanted in February and plant spacing of 50 cm in both season, but the differences between plants transplanted in April and those transplanted in February were insignificant which encourage us to recommend its transplanting in April in order to save the soil two months for other use. These results are in agreement with Mohanty et al. (1993) and Natarajan and Vijayakumar (2002) on Tagetes erecta, also Ramesh and Singh (2008) on Tagetes minuta, Acosta de la Luz et al. (2010) on Tagetes lucida and Acosta de la Luz et al. (2011) on Tagetes lucida.

6-Oil percentage (%), content (ml/plant) and yield (L/fed.):

The percentage of the essential oil of Tagetes lucida did not show dramatic change due to planting date or plant spacing along plant life in both season and ranged from 0.4 to 0.6% (Table 8). The differences in the mean

337 J. Appl. Sci. Res., 9(1): 330-340, 2013 values of oil percent due to planting dates reached 4.2 %, 8.5% and 8.3 % for 1st, 2nd and 3rd plant sampling date, respectively in the first season. The corresponding values in the second season were 3.8, 6.1 and 12.5 %, respectively. The differences in the mean values of the essential oil percent due to plant spacing were 15.2, 20.5 and 12.5 % for the 1st, 2nd and 3rd plant sampling date in the first season, respectively. The corresponding values were 3.8, 25.5 and 13.3 %, respectively. Essential oil content and oil yield of Tagetes lucida significantly responded to planting dates and plant spacing in both seasons during the three cuts (Table 9). The interaction between planting date and plant spacing was significant on both parameters in all cuts dates in both seasons except the first cut. In the beginning of Tagetes lucida plant age, essential oil content and yield were superior in plants transplanted in February in both seasons. But by the progress of plant life, plants transplanted in April showed the higher essential oil content and yield comparing to those transplanted either in February or in March in both seasons. Increasing plant spacing from 40 cm to 50 cm and to 60cm significantly increased oil content up to 60 cm in both seasons to show the highest content of essential oil with plants cultivated at 60 cm between hills. Regarding essential oil yield of Tagetes lucida plants cultivated at 50 cm showed the highest oil yield in both seasons comparing to those cultivated at 40 and 60cm. This significant increment was the result of increasing herb yield of these plants and also to the number of plants in the same area comparing to 60 cm plant spacing. The best treatment for essential oil content was the cultivation in April at 60 cm plant spacing in both seasons, while cultivation of Tagetes lucida in April at 50 cm plant spacing was the best treatment for production of essential oil yield in both seasons.

Table 7: Effect of planting date and plant spacing on herb fresh and dry yield (Ton/Feddan) of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Fresh yield (ton/fed.)

Cuts Planting date Planting date

Plant spacing spacing Plant Mean Mean D1 D2 D3 D1 D2 D3

S1 1.12 0.64 0.46 0.74 1.18 0.59 0.47 0.75 1st cut S2 1.38 0.86 0.53 0.92 1.47 0.86 0.55 0.96 S3 1.44 0.83 0.55 0.94 1.34 0.83 0.51 0.90 Mean 1.31 0.77 0.51 1.33 0.76 0.51 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.077 0.122 0.134 0.117 0.125 n.s. S1 2.03 1.80 1.89 1.91 2.84 2.42 2.64 2.63 2nd cut S2 3.36 2.48 3.28 3.04 3.25 2.72 3.33 3.10 S3 3.11 2.28 3.03 2.81 2.93 2.60 2.90 2.81 Mean 2.8 2.2 2.7 3.00 2.58 2.95 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.179 0.199 0.311 0.174 0.094 n.s. S1 2.65 2.51 2.63 2.60 3.33 3.21 3.43 3.32 3rd cut S2 3.94 3.04 3.74 3.57 3.78 3.56 3.82 3.72 S3 3.49 2.79 3.35 3.21 3.33 3.15 3.32 3.27 Mean 3.36 2.78 3.24 3.48 3.31 3.52 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.178 0.219 0.31 0.197 0.045 n.s. Dry yield (Ton/Fed.) S1 0.37 0.21 0.15 0.24 0.40 0.19 0.15 0.25 1st cut S2 0.46 0.27 0.18 0.30 0.47 0.28 0.19 0.31 S3 0.47 0.29 0.17 0.31 0.44 0.27 0.17 0.29 Mean 0.43 0.26 0.17 0.44 0.24 0.17 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.027 0.05 n.s. 0.035 0.043 n.s. S1 0.71 0.60 0.64 0.65 0.90 0.78 0.89 0.86 2nd cut S2 1.15 0.77 1.09 1.01 1.18 0.90 1.03 1.04 S3 1.05 0.75 1.04 0.95 1.09 0.82 0.91 0.94 Mean 0.97 0.71 0.92 1.06 0.83 0.94 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.076 0.129 0.133 0.072 0.042 n.s. S1 0.86 0.74 0.87 0.82 1.10 1.06 1.09 1.09 3rd cut S2 1.21 0.97 1.19 1.12 1.34 1.10 1.21 1.22 S3 1.13 0.88 1.07 1.03 1.19 1.04 1.13 1.12 Mean 1.07 0.86 0.86 1.21 1.06 1.15 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.053 0.158 n.s. 0.065 0.067 n.s.

338 J. Appl. Sci. Res., 9(1): 330-340, 2013

Table 8: Effect of planting date and plant spacing on oil percentage (%) of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Planting date Planting date Cuts Mean Mean Plant spacing Plant spacing D1 D2 D3 D1 D2 D3

S1 0.52 0.52 0.55 0.53 0.60 0.58 0.60 0.59 1st cut S2 0.51 0.52 0.54 0.52 0.52 0.51 0.53 0.52 S3 0.45 0.49 0.44 0.46 0.46 0.48 0.48 0.47 Mean 0.49 0.51 0.51 0.53 0.52 0.54 S1 0.46 0.46 0.40 0.44 0.46 0.48 0.42 0.45 2nd cut S2 0.47 0.54 0.57 0.53 0.48 0.54 0.54 0.52 S3 0.5 0.52 0.51 0.51 0.54 0.54 0.50 0.53 Mean 0.47 0.51 0.49 0.49 0.52 0.49 S1 0.45 0.51 0.52 0.49 0.40 0.50 0.50 0.47 3rd cut S2 0.50 0.45 0.50 0.48 0.48 0.4 0.48 0.45 S3 0.50 0.57 0.55 0.54 0.48 0.54 0.52 0.51 Mean 0.48 0.51 0.52 0.54 0.48 0.50

Table 9: Effect of planting date and plant spacing on oil content (ml/plant) and yield (L/Feddan.) of Tagetes lucida plants during two seasons.

First Season (2009) Second season (2010)

Oil content (ml/plant)

Cuts Planting date Planting date

Plant spacing Plant spacing Mean Mean D1 D2 D3 D1 D2 D3

S1 0.41 0.24 0.18 0.28 0.51 0.24 0.20 0.32 1st cut S2 0.63 0.53 0.25 0.47 0.68 0.39 0.26 0.45 S3 0.70 0.43 0.26 0.46 0.66 0.43 0.26 0.45 Mean 0.58 0.40 0.23 0.62 0.36 0.24 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.093 0.128 n.s. 0.06 0.05 n.s. S1 0.67 0.59 0.54 0.60 0.93 0.83 0.79 0.85 2nd cut S2 1.41 1.19 1.67 1.42 1.39 1.31 1.60 1.43 S3 1.67 1.27 1.65 1.53 1.70 1.51 1.55 1.58 Mean 1.25 1.02 1.29 1.34 1.21 1.32 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.072 0.101 0.126 0.07 0.05 0.12 S1 0.85 0.91 0.98 0.91 0.95 1.15 1.23 1.11 3rd cut S2 1.76 1.22 1.67 1.55 1.62 1.27 1.64 1.51 S3 1.87 1.70 1.98 1.85 1.71 1.82 1.85 1.79 Mean 1.49 1.28 1.54 1.43 1.41 1.57 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.086 0.126 0.149 0.09 0.07 0.15 Oil yield (L/Fed.) S1 5.81 3.30 2.53 3.88 7.09 3.43 2.83 4.45 1st cut S2 7.04 5.97 2.85 5.29 7.64 4.40 2.93 4.99 S3 6.49 4.05 2.43 4.33 6.17 3.99 2.46 4.21 Mean 6.45 4.44 2.60 6.97 3.94 2.74 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 1.033 1.44 n.s. n.s. 0.56 n.s. S1 9.32 8.29 7.58 8.40 13.05 11.60 11.09 11.91 2nd cut S2 15.80 13.37 18.71 15.96 15.58 14.67 17.96 16.07 S3 15.55 11.87 15.44 14.28 15.83 14.05 14.48 14.79 Mean 13.55 11.18 13.91 14.82 13.44 14.51 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.845 1.026 1.464 0.84 0.47 1.45 S1 11.91 12.79 13.69 12.80 13.30 16.06 17.15 15.50 3rd cut S2 19.70 13.66 18.71 17.36 18.14 14.23 18.31 16.90 S3 17.43 15.88 18.43 17.25 15.97 17.00 17.28 16.75 Mean 16.34 14.11 16.95 15.81 15.76 17.58 LSD at P. spacing P. date Interaction P. spacing P. date Interaction 5% level 0.964 1.252 1.671 0.93 0.23 1.61

339 J. Appl. Sci. Res., 9(1): 330-340, 2013

Similar results were obtained by Ram et al. (1998), Singh et al. (2008) and Ramesh and Singh (2008) on Tagetes minuta and also Acosta de la Luz et al. (2010) and Acosta de la Luz et al. (2011) on Tagetes lucida. In conclusion, the recommend treatment for the best fresh and dry yield as well as essential oil yield of Tagetes lucida under the Egyptian environmental conditions in the new reclaimed soil (sand soil) Nubaria, Behira Governorate, with drip irrigation system is transplanting the plants in April at 50 cm of plant spacing.

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