الفهرس 
Material and methodsOnly 14 pages are availabe for public view
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Abstract
Echinacea plant, ”American coneflower or purpul coneflower” (Fain Asteraceae), is native to Kansas and Nebraska, states U.S.A. It was widely used in folk medicine by the American Indians for the treatment of many illnesses. The plant has recently found a renewed interest due to its
immtmostimulatory activity.
The most important species are E. purpurea, Ii. angus*lia, and E. pallida. Due to difficulty of germination of the two other species, only E.purpurea was used in this study.
The present study aimed to investigate the suitable methods tbr propagation and cultivation of this plant, as well as, its agricultural requinnents, i.e. spacing and fertilization, for optimal growth and yield of
herb and active principles.
The final goal of this study was to aclimatize and introduce Echinacea plant into cultivation in Egypt as one of the economically
important medicinal plants.
The achieved results from these investigations could be summarized
as follows :
1- Cultivation of Echinacea plants :
- The untreated seeds of E. purpurea showed 70% germination, however, those of Langustifolia and E. pallida failed to germinate. Stratification of the dry or wet seeds (at 5°C in refrigarator for one month) before sowing increased the germination percentage of E. purpurea to 83 and 85%, while only 1% of E. angusq/blia and E. pallida seeds germinated due to such treatments.
Seeds of Is. purpurea sown in May-June gave 92% germination, while those sown in September or October gave 87 and 89% respectively compared with 77 and 64% for January and Agust sowings respectively.
Echinacea plants raised from seedlings gave higher yields of dried aerial parts (100.00 g), roots (23.20 g), and flowering heads (46.60g) per plant, in August of the first planting season, compared with 54.70g, 15.60 g and 36.60 g respectively for the plants raised from suckers.Comparing the annual cultivation of Echinacea with duration for another growing season, the results showed that duration significantly increased the growth and biomass production. As a mean value of two successive years of investigations, duration increased the dry weight of the aerial parts to 160.10 g, the roots to 52.41g, the flowering heads to 142.85 g and the number of produced suckers to 9.37, as well as, 24.56% for the root/plant ratio. The recorded values in case of annual cultivation were 62.90 g, 12.30 g, 50.50 g, 6.60 and 16.37%, respectively. Accordingly, the gained plant material after two growing seasons, amounted to 1223.00 g, 52.41 g and 193.3 g for the aerial parts, roots and flowering heads respectively.
2- Growth and yield parameters of E. purpurea plant as influenced 1), planting season and developing stages :
Spring cultivation produced higher yield of flowers (54.319 g) compared with autumn cultivation (45.00 g) however, the later rather favoured the root production (40.49 g) which led to higher yield of the whole plant (139.66 g). The cost and period of seed bedding in greenhouse, in case of autumn cultivation, as well as, the duration of the growing season, are factors that should be considered when comparing between the advantages of each method.
In case of autumn cultivation, the growing season lasted for 140 days after transplantion, while in spring cultivation, the period increased to
195 days.In spring cultivation, the plants were transplanted in the first week of May, while that took place in first week of March, in case of autumn cultivation. In the first case the plants attained the fruiting stage at the end of November, while in the second, they attained that stage of growth in the first week of August. In both cases, the highest rate of growth of the aerial pails took place in the flowering stage, however, the roots showed the highest rate of growth in the fruiting stage. In general, the plants showed the highest weight for aerial parts, roots and flowring heads at the fruiting
stage.3- Effects of plant spacing on growth and yield of E. purpurea plant:
- Echinacea purpurea plants were spaced at 20, 40 or 60 cm distances. The obtained results revealed that the plant growth positively responded to the increased planting distance, that was reflected in higher values for the plant height, weight of the aerial parts, roots and flowering heads, at all the growing stages. At the fruiting stage, however, a considerable difference could be noticed between the plants spaced at 20 cm on one side, and the plants spaced at 40 or 60 cm on the other side. At this stage of growth, the differences between the plants spaced at 40 cm or 60 cm were not significant in the first season, while significant in the second one- Although the increased spacing increased the growth and yield indices per plant, however, the contrary was true when the studied characteres were measured for the cultivated unit area. For example, at the fruiting stage of the first season, the plants cultivated at 20 cm distance gave the values of 920.00, 253.00, 489.00 g/m2 for the aerial parts, roots, and flowers respectively versus, 291.90, 87.60 and 198.00 g/m2 respectively for the plants cultivated at 60 cm distance.
4- Effects of fertilization treatments on the growth and yield indices of E. purpurea.
Four levels of nitrogen., NO, N I, N2 and N3, as well as, three levels of potasium fertilizers, KO, K1 and K2, were applied to Echinacea plants, either separately or in combination. The collected data from this study showed the following :
All the applied levels of nitrogen increased .the growth and yield characters of this plant. The third level, treatment (N3K0) gave the greatest effect on the studied characters, the recorded values due to this treatment in the first season were., 80.00 cm, 153.20 g. 38.00 g and 88.00 g per plant for the plant height, aerial parts dry weights, roots dry weight and flower dry weight respectively, compared with 36.30 cm, 52.00 g, 14.00 g and 36.00 g respectively for the No Ko treatment. On the contrary, nitrogen fertilization diversely affected the root/plant ratio. Results of the second season followed the same trend.
- The two levels of potassium fertilizers possitively affected the studied characters over the control. The first level (K 1) gave the best results, however, the second level (K2) almost gave lower values than K 1. The recorded values in the first season due to K1 NO treatment were 80.00 cm 209.20 g, 24.00 cm, 49.30 g, 36.60 and 109.00 g for the plant height, aerial partS dry weight, root lengh, root dry weight, number and weight of the flowering heads respectively, compared with 36.30 cm, 52.00 g, 17.30 cm, 14.00 g, 8.60 and 36.00 g, for the same characters respectively, in case of the NO KO treatment.
Regardaing the effects of the combined treatments consisted of nitrogen and potassium, it was clearly noticed that addation of potassium at its lower level (K I ), together with the two lower levels of nitrogen (N I and N2), almost gave better results than the use of nitrogen alone, further addition of potassium at K2 level namely., N I K2 and N2 K2 treatments, decreased the recorded values compared with K I , and sometimes the values became lower than those corresponding to nitrogen alone. Acorrdingly, the NO K I treatment remained the best treatment followed by N3 KO., 5- Chemical investigasions of Epurpurea plants : 5.1. The alkamides :
The alkamide content in the roots was relatively low at the beginning of the growth cycle. They ammounted to 0.041 and 0.053% at the seedling and vegetative stages respectively, after which they increased to 0.073% at the flowering stage, and reached the maximum of 0.084% at the fruiting stage.
- The aerial parts, however contained.0.080°/0 and 0.071% of alkamides at the seedling and vegetative stages respectively. Then the content sharply decreased to 0.031 and 0.029% at the flowering and fruiting
stages respectively.
The fertilization treatments N3 KO and NOK 1, which proved to be the best among the nitrogen and potassium treatments regarding their effect on the agronomic characters, they also remarkably increased the alkamide content in all the plant organs. The NOK 1 was superior in this concern, it increased the alkamide content to 0.073, 0.070 and 0.031”/o in the roots, flowers and aerial parts respectively, versus 0.019, 0.016 and 0.011% respectively in the unfertilized plants.
- Regardless of the fertilization treatments, the plants at the flowering stage accumulated the highest alkamide percentage in the roots (0.051 %), followed by the flowers (0.043%), and aerial parts (0.022%).
- The HPLC profiles of the alkamides in the different plant organs were almost superimposable, however, some quantitative variations were experienced. Eleven alkamides were identified, dodeca- 2E, 4E, 8Z, 10 (E) tetraenoic acid isobutylamide and its isomer (Z) constituted from 46 to 75% of the alkamide mixture, followed by trideca - 2E, 7Z - dies -10, 12 - diynoic acid isobutylamide.
The proportional composition of the alkamide mixture was characteristic for the plant organs of E. purpurea and almost stable against changes in the plant developement or the fertilization treatments. The observed fluctuation in the proportional composition of the alkamides was rather attributed to the accumulation of the whole alkamides, the process which aliready enhanced towards the fruiting stage, as well as, due to the N3 KO and NO K 1 fertilization treatments.
5.2. The essential oil :
- The GC/MS investigation of the essential oil of E. purpurea flowers revealed the presence of 56 peaks, from which 21 constituents were identified. Germacrene - D was the major constituent (55.592%), followed by methyl chavicol (estragol) (11.923%), and anethol (5.486%), then lower concentrations of the other constituents were detected. The oil of E. purpurea flowers consisted mainly of sesquiterpenoids, (about 69%) and phenolic ethers (about 16%).The flowering heads of E. purpurea plant possessed the highest essential oil content (0.108%), followed by the aerial parts (0.090%) and the
minimum in the roots (0.010%).
- Both the nitrogen and potassium fertilization significantly increased the oil content in the plant organs, however, the second level of nitrogen (N2K0) and the first level of potassium (NOK ) were the best treatments in this regard. 5.3. The anthocyanins :
- It was noticed that the ray flowers of E.purpurea plants under investigation could be classified in two distinguished colors, i.e. dark purple and light purple, the dark one contained 28.51 mg/100g of total
anthocyanins, compared with 14.37 mg/100g in the light one.
- The chromatographic investigation revealed that the anthocyanins of E.purpurea flowers were, (I) cyanidin 3, 5 diglucoside, (11) cyanidin gluco-pyrano - malonyl glycoside, and (III) malvidine glycoside. Both the two types of flowers contained the same anthocyanins, however, the dark purple flowers were characterized with relatively higher contents of anthocyanins I and III, but lower content of anthocyanin 11.
compared with the light purple flowers.
-The N3 KO and NOK 1 treatments resulted in the highest increase in the anthocyanin content of the flowers, the recorded values were 23.90 and 30.30 mg/100g respectively, compared with 17.50 mg/100g in the
untreated plants.Stratification of E. purpurea seeds at 5°C for one month, before sowing increased their germination percentage to about 83-85%, compared
with 70% for the untreated seeds.
Seeds of E. purpurea best germinated in May - June fi)llowed by September - October, compared with sowing in january and August.
E. purpurea could be propagated by seeds and also by suckers, the first method gave better yields of aerial parts, roots and flowers than the
second one.
- Is. purpurea cultivated in Egypt grew successfully and completed their growth cycle in one year. However, in Europe the plant is considered as a biennial plant, attaining the flowering stage in the second season.
-When duration was practiced with E.purpurea plants, the gained biomass in the second season of the growth was much more than the gained yield from the annual cultivation. The economic factors should be considered in the choise between the two methods.
- E. purpurea could be cultivated in either spring or autumn. For autumn cultivation the seeds was sown first in the greenhouse before transplanting to the open field where they completed the growth cycle in 140 days, while in spring cultivation the growth lasted for 195 days. The spring - cultivated plants attained the fruiting stage at the end of November, while that took place at the beginning of August when the plants were cultivated in autum.
-In general, the highest yields of aerial parts, roots and flowering heads were attained at the fruiting stage.
-For the highest biomass production per unit area, E. purpurea plants should be cultivated at 20 cm distance between plants, on rows 50 cm
inbetween (10 plants /m2).purpurea plants responded well to nitrogen or potassium fertilization. The best response was due to the third level of nitrogen (200 kg/fed. of ammonum sulphate), and due to the lower level of potassium (50 kg/fed. of potassium sulphate). Combined treatments of the two fertilizers were less effective.
The alkamide content in the roots increased alongwith the plant growth to reach the maximum at the fruiting stage, while the contrary took place in the aerial parts. At the flowering stage, the highest alkamide content was recorded in the roots, followed by the flowers, and the lowest in the aerial parts.The two isomers E / Z of the amide dodeca - 2E, 4E, 8Z, 10 E / Z-tetraenoic acid isobutylamide, were the main alkamides (46-75%). The alkamide profile was almost the same in all the plant organs with slight quantitative variations characterizing the different plant organs.
Twenty one compounds were identified in the essential oil of E. purpurea flowers. Germacrene - D was the major constituent, followed by methyle chavicol. The oil consisted mainly of sesquiterpenes (65%) and phenolic ethers (16%).
- The flowering heads were the richest in the oil, followed by the aerial parts, then the roots. The fertilization treatments NOK I and N2K2 remarkably increased the oil content in all the plant organs.
-The ray flowers of Epurpurea were characterized with either dark purple or light purple color. Three types of anthocyanins were identified in both the two types. However, the two types of flowers differed in the proportional composition of their anthocyanins, as well as, higher content of the total anthocyanins in the dark purple type.
- The anthocyanin content responded positively to the applied fertilizers, the N3 KO and NO K 1 treatments were superior in this regard.