Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8

Search results for: MS medium

8 Adaptive Responses of Carum copticum to in vitro Salt Stress

Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

Abstract:

Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: Antioxidant enzymes, Carum copticum, organic solutes, salt stress.

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7 In vitro Environmental Factors Controlling Root Morphological Traits of Pineapple (Ananas comosus L. Merr)

Authors: S. Mohajer, R. M. Taha, M. Adel

Abstract:

Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Keywords: Environmental factors, in vitro rooting, pineapple, tissue culture.

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6 Indirect Regeneration and Somatic Embryogenesis from Leaf and Stem Explants of Crassula ovata (Mill.) Druce – An Ornamental Medicinal Plant

Authors: A. B. A. Ahmed, Amar, D. I., R. M. Taha

Abstract:

This research aims to investigate callus induction, somatic embryogenesis and indirect plant regeneration of Crassula ovata (Mill.) Druce – the famous ornamental plant. Experiment no.1: Callus induction was obtained from leaf and stem explants on Murashige and Skoog (MS) medium supplemented with various plant growth regulators (PGRs). Effects of different PGRs, plant regeneration and subsequent plantlet conversion were also assessed. Indirect plant regeneration was achieved from the callus of stem explants by the addition of 1.5 mg/L Kinetin (KN) alone. Best shoot induction was achieved (6.5 shoots/per explant) after 60 days. For successful rooting, regenerated plantlets were sub-cultured on the same MS media supplemented with 1.5 mg/L KN alone. The rooted plantlets were acclimatized and the survival rate was 90%. Experiment no.2: Results revealed that 0.5 mg/L 2,4-D alone and in combination with 1.0 mg/L 6-Benzyladenine (BA) gave 89.8% callus from the stem explants as compared to leaf explants. Callus proliferation and somatic embryo formation were also evaluated by ‘Double Staining Method’ and different stages of somatic embryogenesis were revealed by scanning electron microscope. Full Strength MS medium produced the highest number (49.6%) of cotyledonary stage somatic embryos (SEs). Mature cotyledonary stage SEs developed into plantlets after 12 weeks of culture. Wellrooted plantlets were successfully acclimatized at the survival rate of 85%. Indirectly regenerated plants did not show any detectable variation in morphological and growth characteristics when compared with the donor plant.

Keywords: Callus induction, Crassula ovata, Double Staining, Indirect plant regeneration, Somatic embryogenesis.

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5 Enhanced Efficiency for Propagation of Phalaenopsis cornu-cervi (Breda) Blume & Rchb. F. Using Trimmed Leaf Technique

Authors: Suphat Rittirat, Sutha Klaocheed, Kanchit Thammasiri

Abstract:

The effects of thidiazuron (TDZ) and benzyladenine (BA) on protocorm-like bodies (PLBs) induction from leaf explants was investigated. It was found that TDZ was superior to BA. The highest percentage and number of PLBs per leaf explant at 30 and 5.3, respectively were obtained on ½ MS medium supplemented with 9µM TDZ. The regenerated plantlets were potted and acclimatized in the greenhouse. These plants grew well and developed into normal plants after 3 month of transplantation. The 100% survival of plantlets was achieved when planted on pots containing sphagnum moss. 

Keywords: Orchid, PLBs, sphagnum moss, thidiazuron.

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4 In vitro Propagation of Purple Nutsedge (Cyperus rotundus L.) for Useful Chemical Extraction

Authors: Chockpisit Thepsithar, Nongnuch Euawong, Nukul Jonghomkajorn

Abstract:

The in vitro culture procedure of purple nutsedge (Cyperus rotundus L.) for multiple shoot induction and tuber formation was established. Multiple shoots were significantly induced from a single shoot of about 0.5 – 0.8 cm long, on Murashige and Skoog (MS) medium supplemented with 4.44 μM 6- benzyladinine (BA) alone or in combination with 2.85 μM 1- indoleacetic acid (IAA), providing 17.6 and 15.3 shoots per explant with 31.2 and 27.5 leaves per explant, respectively, within 6 weeks of culturing. Moreover, MS medium supplemented with 4.44 μM BA and 2.85 μM IAA was suitable for tuber induction, obtaining 5.9 tubers with 3.4 rhizomes per explant. In combination with ancymidol and higher concentration of sucrose, 11.1 μM BA and 60 g/L sucrose or 11.1 μM BA, 7.8 μM ancymidol and 60 g/L sucrose induced 3.5 tubers with 1.6 rhizomes or 3.5 tubers without rhizome, respectively. However, MS medium containing 3.9 or 7.8 μM ancymidol in combination with either 60 or 80 g/L sucrose enchanced significant root formation at 20.9 – 23.6 roots per explant.

Keywords: Purple nutsedge, Cyperus rotundus, multiple shoot induction, tuber formation

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3 In vitro Culture Medium Sterilization by Chemicals and Essential Oils without Autoclaving and Growth of Chrysanthemum Nodes

Authors: Wittaya Deein, Chockpisit Thepsithar, Aree Thongpukdee

Abstract:

Plant tissue culture is an important in vitro technology applied for agricultural and industrial production. A sterile condition of culture medium is one of the main aspects. The alternative technique for medium sterilization to replace autoclaving was carried out. For sterilization of plant tissue culture medium without autoclaving, ten commercial pure essential oils and 5 disinfectants were tested. Each essential oil or disinfectant was added to a 20-mL Murashige and Skoog (MS) medium before medium was solidified in a 120-mL container, kept for 2 weeks before evaluating sterile conditions. Treated media, supplemented with essential oils or disinfectants, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. Sterile conditions of MS medium were found 100% from betel oil or clove oil (18 mL/20 mL medium), cinnamon oil (36 mL/20 mL medium), lavender oil or holy basil oil (108 mL/20 mL medium), and lemon oil or tea tree oil or turmeric oil (252 mL/20 mL medium), compared to 100% sterile condition from autoclaved medium. For disinfectants, 2% iodine + 2.4% potassium iodide, 2% merbromine solution, 10% povidone-iodine, 6% sodium hypochlorite or 0.1% thimerosal at 36 mL/20 mL medium provided 100% sterile conditions. Furthermore, growth of new shoots from chrysanthemum node explants on treated media (fresh weight, shoot length, root length and number of node) were also reported and discussed in the comparison of those on autoclaved medium.

Keywords: Chrysanthemum, disinfectants, essential oils, MS medium, sterilizing agents, sterilization of medium without autoclaving.

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2 Leaf Pigments Help Almond Explants Tolerating Osmotic Stress

Authors: Soheil Karimi, Abbas Yadollahi, Kazem Arzani, Ali Imani

Abstract:

This study was conducted to evaluate the response of almond genotypes to osmotic stress in vitro in order to screen drought tolerance. Explants subjected to polyethyleneglycol osmotic stress (0, 3.5, and 7.0% WV) on the MS medium. Concentrations of photosynthesis pigments, anthocyanins, and carothenoids were significantly reduced under osmotic stress. Under osmotic stress, leaf water content, cellular membrane stability and pigments concentrations were significantly higher in the leaves of drought tolerant genotypes. The results revealed that carotenoids and anthocyanins may act as photoprotectant compounds in almond leaves and involved in drought tolerance system of the plant.

Keywords: Almond, Anthocianins, Carotenoids, in vitro; Leaf Osmotic Stress, Leaf Pigments, Polyethylene Glycol.

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1 Direct and Indirect Somatic Embryogenesis from Petiole and Leaf Explants of Purple Fan Flower (Scaevola aemula R. Br. cv. 'Purple Fanfare')

Authors: Shyama Ranjani Weerakoon

Abstract:

Direct and indirect somatic embryogenesis (SE) from petiole and leaf explants of Scaevola aemula R. Br. cv. 'Purple Fanfare' was achieved. High frequency of somatic embryos was obtained directly from petiole and leaf explants using an inductive plant growth regulator signal thidiazuron (TDZ). Petiole explants were more responsive to SE than leaves. Plants derived from somatic embryos of petiole explants germinated more readily into plants. SE occurred more efficiently in half-strength Murashige and Skoog (MS) medium than in full-strength MS medium. Non-embryogenic callus induced by 2, 4-dichlorophenoxyacetic acid was used to investigate the feasibility of obtaining SE with TDZ as a secondary inductive plant growth regulator (PGR) signal. Non-embryogenic callus of S. aemula was able to convert into an “embryogenic competent mode" with PGR signal. Protocol developed for induction of direct and indirect somatic embryogenesis in S. aemula can improve the large scale propagation system of the plant in future.

Keywords: Petiole and leaf explants, Scaevola aemula, Somaticembryogenesis

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