Ghavam, Journal of Herbal Drugs, Vol. 6, No. 4: 187-194, 2016 Journal of Herbal Drug journal homepage: www.jhd.iaushk.ac.ir Genetic improvement of quantity/quality yield of black cumin (Nigella sativa L.) ecotypes cultivated in Iran climatic conditions Saeed Salehi1*, Asad Rokhzadi2, Ghorban Noormohammadi3, Sayyed Mohammadjavad Mirhadi3, Ahmad Reza Golparvar4 1Young Researchers and Elite Club, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran; *Email: [email protected] 2Department of Agronomy, College of Agriculture, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran; 3Department of Agronomy, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran; 4Department of Agronomy and plant Breeding, College of Agriculture, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran; ARTICLE INFO ABSTRACT Background & Aim: Black seed or black cumin (Nigella sativa L.) Type: Short Original Research belongs the family Ranunculaceae. Black seed is one of these species, Topic: Medicinal Plants which is naturally distributed in different parts of the country. It is Received January 5th 2014 extensively cultivated in various regions of Iran. Experimental: The experiment was achieved using randomized Accepted April 5th 2014 complete block design with three replications in the research field of Key words: Islamic Azad University of Isfahan Branch during 2013. Seed of ten ecotypes namely; Semirom, Zawareh, Golpayegan, Fereydan, Meymeh, Nigella sativa L. Kashan, Khansar, Daran, Ardestan and Isfahan were cultivated in the plots Genetic variation comprising four rows. The essential oil was extracted by a Clevenger type Selection criteria apparatus and analyzed by using GC/MS. Results & Discussion: Statistical analysis indicated that there was Regression significant difference among all traits. The results of regression and path Path analysis analysis indicated that the number of branches, grain filling rate and days to blooming traits were the best indirect selection criteria to improve yield in black cumin. According to the results of this research, Fereydan and Daran ecotypes showed the optimum amount of essential oil content and morphological traits. Furthermore, cross between these ecotypes has suitable advantage to increase seed and oil yield in black cumin breeding programs. Industrial and practical recommendations: There are few researches about breeding of black cumin. The results of present study could be applied in research centers as well by farmers to cultivate commercially and production black cumin especially for seed. 1. Introduction is 20-40 cm height, straight, and lightly feathered, with finely divided. The flowers are delicate and usually Black cumin (Nigella sativa L.) is plant belongs the coloured pale blue and white, with five to ten petals. family Ranunculaceae. The body of this annual plants Seeds flattened, oblong, small, black in colour and 187 Ghavam, Journal of Herbal Drugs, Vol. 6, No. 4: 187-194, 2016 inflated capsule composed of three to seven united traits on the quantity/quality ecotypes yield of black follicles, each containing numerous seeds and about cumin (Nigella sativa L.) cultivated in Iran climatic eight species in Iran (Mozaffarian, 1998). Black cumin conditions. is native in Europe, Middle East and Western Asia (Iqbal et al., 2010). It also grows in most parts of Iran 2. Materials and Methods such as Arak, Kermanshah, Isfahan and other parts of the country. Black cumin originally grown in arid and 2.1. Plant Material semi-arid regions (Moody and Mohasel, 1998; This experiment has been conducted in the research D’Antuno et al., 2002). The basic chromosomes field of Islamic Azad University of Isfahan, in center number of X = 6 have been found of Nigella sativa L. Iran (32◦, 38_ N and 51◦, 47_ E, 1550 m above sea The diploid chromosome complement was 2n =12 level), during 2013. The experiment was achieved (Jabeen et al., 2012). The seeds of Nigella sativa using randomized complete block design with three contain thymoquinone, monoterpens such as p-cymene replications. This location is arid and warm area and pinene, nigellidine, nigellimine and saponin (according to the Koppen climate classification) (Ansari and Sadiy, 1989). The seeds have also been characterized by warm and dry summers. The long- found to contain fats, crude fiber, minerals; e.g. Fe, Na, term (30 years) mean annual rainfall and temperature Cu, Zn, P, Ca and vitamins like ascorbic acid, thiamine, of area was 121.1 mm and 33.4 centigrade degrees, niacin, pyridoxine and folic acid (Takruri et al., 1998). respectively. The soil texture of the experimental field Nickavar et al. (2003) reported the chemical was clay loam type with pH 7.37, contains total N (0.75 composition of the extracted fixed oil and volatile oil of %), total P2O5 (35 ppm) and total K2O (452 ppm) with Nigella sativa L. seeds grown in Iran were eight fatty an EC of 4.69(ds/m). Weeds were removed by hand acids (99.5%) and thirty- two compounds (86.7%) have weeding during growth stages. Seed of ten black seed been identified in the fixed and volatile oils, ecotypes (Semirom, Zawareh, Golpayegan, Fereydan, respectively. The main fatty acids of the fixed oil were Meymeh, Kashan, Khansar, Daran, Ardestan and linoleic acid (55.6%), oleic acid (23.4%), and palmitic Isfahan) were cultivated the plots comprising four acid (12.5%). The major compounds of the volatile oil rows. Seeds were planted in rows 30 cm apart with were trans-anethole (38.3%), p-cymene (14.8%), inter-row spacing 30 cm. The measured characters limonene (4.3%), and carvone (4.0%). The seeds have involved; plant height (cm), number of branches, the anthelmintic, insecticidal, antimalarial, number of capsules, days to beginning of blooming, antibacterial, antifungal and antitumor activates. The days to full blooming, 1000-seed weight (g), grain seeds are also reported to have antispasmodia, diuretic, filling rate (g/m2/day), essential oil percent (%), carminative, digestive and antiseptic properties (Ali et essential oil yield (g/m2), thymoquinone content (%), al., 2008). Salamati and Zeinali (2013) reported that thymoquinone yield (g/m2), γ-terpinene content (%) investigate genetic diversity and relationships among and γ-terpinene yield (g/m2). morphological traits in 21 genotypes of (Nigella sativa L.) in Iran, seed yield varied from 63.34 g in genotype 2.2. Extraction of essential oil of Shiraz to (147.36 g) in genotype of Zabol. The essential oil was extracted from 100 g of Talebi Kouyokhi et al. (2008) reported that powdered seeds in 1 L of water contained in a 2 L flask phytochemical variations were not only found among and heated by heating jacket at 100 °C for 4 h in a samples of different regions but also among samples of Clevenger–type apparatus, according to producers the same region with different altitude reflecting the outlined British Pharmacopoeia. The collected essential effect of environment on essential oil components. It is oil was dried over anhydrous sodium sulphate and well known that yield and yield components of plants stored at 4 °C until analyzed. are determined by a series of factors including plant genetic (Shafieet al., 2009), climate, edaphic, elevation, 2.3. Composition and identification of the constituents topography and also an interaction of various factors of the essential oils (Rahimmalek et al., 2009). Therefore, the aim of this GC/MS analysis was carried out with an Agilent study was to the role of genetic distance and effective 5975 GC-MSD system. HP-5MS column (30 m x 0.25 188 Ghavam, Journal of Herbal Drugs, Vol. 6, No. 4: 187-194, 2016 mm, 0.25 μm film thickness) was used with helium as such as the seed yield as selection criteria more care carrier gas with flow rate of 1.0 mL/min. The oven must be taken. Because, if the plant is too dwarf, it temperature was kept 20 °C at 50 °C for 4 min and might lead to reduction of the competition of plant to programmed to 280 °C at a rate of 5 °C /min, and kept get light and other environmental parameters, thus the 20 °C constant at 280 °C for 5 min, at split mode. The seed yield would diminish. Therefore, it is necessary injector temperature was at 20°C at 280 °C. Transfer 20 that the best morphologic traits is studied more line temperatures 280 °C. MS were taken at 70 eV. carefully (Golparvar and Hejazi Dehaghani, 2012). Mass range was from m/z 35 to 450. Faravani et al. (2006) reported that significant differences were observed in plant height, weight of 2.4. Statistical analysis 1000 kernel, seed yield, number of stem branches, Analysis of variance of the collected data was vascular bundles, weight of capsule and xylem and conducted based on randomized complete blocks phloem diameter on the 28 landraces of Nigella sativa design model. Then existence of significant difference were collected from Khorasan province in Iran. Black between means of different parameters were evaluated cumin seeds are important in herbal medicines using Duncan's new multiple range test (DNMRT) at commonly employed in Iran. The environmental 5% probability level. SPSS16 program was used to factors like climate and location might be a possible statistical analysis. reason for these variations. Moreover, difference in genetic makeup could also be a contributing factor as 3. Results and discussion indigenous variety was tested in the trial. Nigella sativa L., is a new crop in the country and the first step for 3.1. Growth characteristics successful introduction is evaluation of diverse Analysis of variance (Table 1) indicated the germplasm that not only helps to identify the superior significant difference between ecotypes for plant genotypes, but selected genotypes could also be used in height, number of branches, number of capsules, days agronomic experiments (Shadia et al., 1998). to beginning of blooming, days to full blooming, 1000- 3.2. Essential oil yield seed weight and grain filling rate.