Investigating effective factors in in vitro regeneration of sage (Salvia sclarea L.)

Dissertation for Master's Degree in Horticultural Sciences

Medicinal Plants

Abstract

           Salvia belongs to the Lamiaceae family, which includes several species. Sage (Salvia sclarea L.) is used in traditional medicine and in trade, especially in the production of essential oil and as a seasoning and flavoring for food. The distribution area of ??this plant is the Mediterranean area and Iran. Essential oil or substances extracted from the aerial parts of this plant have many effects in the treatment of diseases, for example pain reliever, anti-inflammatory, antioxidant, antifungal and antibacterial. In this research, bud induction and branch regeneration were studied under in vitro conditions using different explants (node, branch tip, hypocotyl and cotyledon) in Marigold plant. Induction of shoot and shoot regeneration in MS medium supplemented with 6-benzylaminopurine (BAP), thidiazoron (TDZ) and kinetin (Kin) in different concentrations (0, 2.2, 4.4 and 8.8 ?M), in combination with indole-3-acetic acid (IAA) (0 and 1.1 ?M) was obtained. In the study of the effect of explant type on the induced bud and regenerated branch, it was observed that the highest number of induced bud and regenerated branch with an average of 25.531 and 17.934 (bud and branch in each treatment), respectively, was obtained in the node explant in the medium containing TDZ, and the lowest average of induced bud (4.625 buds in each treatment) and regenerated branch (3.312 branches in each treatment) was obtained in the tip explant in The medium containing Kin was obtained. In investigating the effect of the type and concentration of plant growth regulators on bud induction and branch regeneration, the highest average induced bud (37.187 buds per treatment) was produced in the concentration of 4.4 ?M TDZ and the highest average regenerated branch (26.053 branches per treatment) in the concentration of 8.8 ?M TDZ was produced in the node explant. The lowest mean of induced bud (1) and regenerated branch (1) was determined in the control medium and in the branch tip explant. The regenerated shoots were rooted in ½ MS medium and MS supplemented with different concentrations of IAA and IBA (0, 0.5 and 1 mg/L) during 4 weeks. The comparison of the averages showed that IAA hormone with the amount of 0.5 mg/liter produced the maximum average number of roots with an average of 2.55 roots, and the minimum average number of roots was observed in the IBA hormone with the amount of 0.5 mg/liter with an average of 1.52. Comparing the average effects of hormone treatment on root length, it was found that control treatment had the highest average root length with an average of 5.65 cm and IAA treatment (1 mg/liter) had the lowest average root length with an average of 3.08 cm. Rooted seedlings were adapted to a mixture of peat and perlite in the ratio (3:1) and 100% humidity and 90% survival in greenhouse conditions.

Key words: sage, direct regeneration, explant, TDZ, root induction

Chapter 1

Introduction and Generalities

1-1- Importance of Medicinal Plants

The history of treating diseases with medicinal plants is as old as the history of human existence on the planet and humans always, according to experience, science and thought and according to the requirements, during their life on the planet The earth has treated itself with the help of medicinal plants. Medicinal plants are an important source of compounds for the pharmaceutical industry and traditional medicine. About 80% of people in developing countries traditionally use medicines derived from plants for their primary health care needs (Cunningham, 1993; Desilva, 1997). There is hardly a day when we do not read about the properties of medicinal plants in scientific journals and articles and do not realize the importance of medicinal plants compared to the side effects and disorders of using chemical drugs. In today's world, the tendency of people to use herbal substances in the treatment and prevention of diseases is more than in the past. Today, the use of medicinal plants in the production of medicinal, food, cosmetic and health products is increasing day by day and has increased the importance and status of these plants. 25% of modern medicines are made from medicinal plants. In 2000, about 60 billion dollars was the income of the countries from the sale of medicinal plants.(Mawaoni, 2008).

Also, the preparation of some effective active ingredients that are very important in the pharmaceutical industry is not possible artificially and can only be extracted naturally from the desired plants. These substances either have an unknown chemical structure in general, or because of their very complex chemical structure, it is difficult and expensive to prepare them artificially in the pharmaceutical industry, such as the cardiac glycosides found in digitalis purpurea, the alkaloids found in catharanthus rosea, and the alkaloids found in ergot mushroom. And this is one of the reasons for the desire of pharmaceutical companies to use plant-based medicines (Omidbigi, 1388).

         2-1- Distribution, history and uses of salvia species. has been  It grows in temperate and subtropical regions, and approximately 70 species have been reported in the flora of Iran. The genus Salvia has been introduced as one of the oldest genera of the mint family, with the highest number of primary chromosome bases (11). Although in some sources Ajuga is often introduced as the oldest genus (Masoud et al., 2010).

The distribution center is in the Mediterranean areas. It prefers warm weather conditions and full sun. Salvia sclarea L. has biotypes with one-year and two-year growth habits (Mihalik et al., 2005). Dweck (2000) introduced the place where this plant grows in the Mediterranean area and Iran. Since the beginning of the 20th century, this type of sage was cultivated only for the use of its flowers. These flowers contain fragrant essential oils that are widely used in health-cosmetic, food and beverage industries. The essential oil of the flowers of this plant along with the essential oil of flowers such as lavender and jasmine have many and different uses in the household chemicals industry (such as perfume, cologne, cream, shampoo, soap, lotion and air freshener sprays and the like). The seed of this plant contains 25 to 32% of oil, which is used in ceramic and porcelain industries.

Every year, large arable lands in the countries of Russia, Bulgaria, Italy, France, Spain and Hungary are allocated for the cultivation of this plant. The origin of this plant has been reported in the sandy and dry regions of the Caucasus, Iran, and the shores of the Mediterranean Europe (Omidbigi B, 2008). It can be seen: one is Pyramidalis [2] which has fast growth and the other is Hirsuta [3] which is covered with thick hairs. The main root of this plant is long and straight and its length is between 100 and 130 cm, which has many branches. The stem is straight, thick, square, covered with hair and its height is different and between 100 and 150 cm. This plant has large collared leaves with an uneven and raised surface. Its width is 10 to 20 cm. The leaves have short petioles. The leaves of the lower part of the stem are large and gradually become smaller towards the top of the stem. The color of the leaves is dark silver. The flowers are pink, purple, or white, and they are placed on the flowering stalks, which are between 40 and 60 cm long. Each cycle contains 3 to 6 flowers. The flowers of this plant are also nectar. The three-sided capsule-type fruit is egg-shaped, 2 to 3 mm long, and its color is brown. The weight of a thousand seeds is 4 to 5 grams (Omidbigi B, 1388).

4-1- Ecological needs of Maryam Glikbir

Marymglikbir is a drought-loving plant, it needs enough light, warm air and little water during its growth. In hot and dry weather, the amount of essential oil of flowers reaches 0.1 to 0.2 percent. Abundant water and cool air increase the vegetative growth of this plant, but it will have a negative effect on the amount of essential oil of flowers and cause it to decrease.