Search results for: Chockpisit Thepsithar

Authors: Aree Thongpukdee, Ekasit Nisayan, Chockpisit Thepsithar

Abstract:

Shoots, with three leaves, of Paphiopedilum 'Delrosi' were used as explants for multiple shoot induction. Modified Hyponex medium was supplemented with thidiazuron (TDZ), N6- benzyladenine (BA) or kinetin (Kn) alone and in combinations with 2,4-dichlorophenoxyacetic acid (2,4-D). All explants were cultured for 15 weeks. It was found that TDZ alone at the concentration of 0.45μM or in combination with 4.52μM 2,4-D and 8.88μM BA in combination with 13.56μM 2,4-D promoted multiple shoots. The highest shoot sprouting efficiencies (80.0, 90.0 and 80.0%) and new shoot numbers (1.5, 1.3 and 1.1) were obtained, respectively. Fresh weight, height, numbers of leaf and root of new shoots and initial explants were discussed.

Keywords: Paphiopedilum, terrestrial orchids, in vitro culture, micropropagation, multiple shoot induction

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Authors: Chockpisit Thepsithar, Aree Thongpukdee

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Two species of Physalis, P.angulataL. and P. peruviana L. were used as models for comparative study to understand the values of micro-morphological, -anatomical and architectural characteristics of leaf for taxonomic purposes and possibly breeding and commercial applications. Both speciespossess amphistomaticleaves with 1-layer epidermis, 3-4-layer spongy mesophyll andbicollateral bundle midrib. Palisade parenchyma cells of P. angulatawere almost twice longer (65-75 μm) than the other one. Type of stomata was similar as anomocyticbut stomatal index(SI) at adaxial surface and abaxial surface of P. angulata were less than of P. peruvianaas 3.57, 4.00 and6.25, 6.66 respectively. Some leaf architectural characteristics such as leaf shape, order of venationalsoprovided information of taxonomic significance

Keywords: Physalis, Solanaceae, micromorphology, anatomy, leaf architecture.

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Authors: Aree Thongpukdee, Chockpisit Thepsithar, Sujitra Timchookul

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The study aimed to investigate characteristics of vegetative tissue for taxonomic purpose and possibly trend of waste application in industry. Stems and branches of 15 species in Solanum found in Thailand were prepared for fiber and examined by light microscopy. Microstructural characteristic data of fiber i.e. fiber length and width, fiber lumen diameter and fiber cell wall thickness were recorded. The longest average fiber cell length (>3.9 mm.) were obtained in S. lycopersicum L. and S. tuberosum L. Fiber cells from S. lycopersicum also revealed the widest average diameter of whole cell and its lumen at >45.5 μm and >29 μm respectively. However fiber cells with thickest wall of > 9.6 μm were belonged to the ornamental tree species, S. wrightii Benth. The results showed that the slenderness ratio, Runkel ratio, and flexibility coefficient, with potentially suitable for feedstock in paper industry fell in 4 exotic species, i.e. Solanumamericanum L., S. lycopersicum, S. seaforthianum Andr., and S. tuberosum L

Keywords: Fiber, microstructure, Solanaceae, Solanum.

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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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Authors: Chockpisit Thepsithar, Aree Thongpukdee

Abstract:

For sterilization of Phalaenopsis culture medium without autoclaving, selected single sterilizing agents and in combinations were added to a 25ml Hyponex medium in a 120ml glass container. Treated liquid and solid media, supplemented with sterilizing agents, were compared to a control medium, autoclaved at 121°C for 15min. It was found that 90(L of 10% povidone-iodine, 150(L of 5.25% sodium hypochlorite, 150(L of 2% mercurochrome, 90(L of 2.5% iodine + 2.5% potassium iodine in combination with 10% providone-iodine (1:3) and 30(L of 2.5% iodine + 2.5% potassium iodide in combination with 2% mercurochrome showed 100% sterile conditions in liquid medium but provided 75, 100, 50, 75 and 80% sterile conditions, respectively, in solid medium. For growth of Phalaenopsis protocorms, 90(L of 10% povidone-iodine in liquid Hyponex medium gave the comparable growth of protocorms to control medium while 150(L of 5.25% sodium hypochlorite in solid medium provided the promising growth of protocorms. Growth of protocorms, whole fresh weight, numbers of leaf and root, root length and number of protocorm-like bodies, was discussed.

Keywords: Phalaenopsis, sterilizing agents, Hyponex medium, sterile medium without autoclaving.

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Authors: Chockpisit Thepsithar, Aree Thongpukdee, Apichya Daorat

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The alternative technique for sterilization of culture medium to replace autoclaving was carried out. For sterilization of culture medium without autoclaving, some commercial pure essential oils, bergamot oil, betel oil, cinnamon oil, lavender oil and turmeric oil, were tested alone or in combinations with some disinfectants, 10% povidone-iodine and 2% iodine + 2.4% potassium iodide. Each essential oil or combination was added to 25-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, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. In vitro sterile conditions were found 20 – 100% from these treated media compared to 100% sterile condition from autoclaved medium. Treated media obtained 100% sterile conditions were chosen for culturing chrysanthemum shoots. It was found that 10% povidoneiodine in combination with cinnamon oil (3:1) and 2% iodine + 2.4% potassium iodide in combination with lavender oil (1:3) at the concentration of 36 3L/25 mL medium provided the promising growth of shoot explants.

Keywords: Sterilizing agents, essential oils, disinfectants, MS medium, in vitro culture, chrysanthemum, sterilization of medium without autoclaving

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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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Authors: Kullanart Obsuwan, Chockpisit Thepsithar

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This study aimed to investigate effect of different organic supplements on growth of Vanda and Mokara seedlings. Vanda and Mokara seedlings approximately 0.2 and 0.3 cm. in height were sub-cultured onto VW supplemented with 150 ml/L coconut water, 100 g/L potato extract, 100 g/L ‘Gros Michel’ banana (AAA group) and 100 g/L ‘Namwa’ banana (ABB group). The explants were sub-cultured onto the same medium every month for 3 months. The best medium increased stem height to 0.52 and 0.44 Cm. in Vanda and Mokara respectively was supplemented with coconut water. The maximum fresh weight of Vanda (0.59 g) was found on medium supplemented with ‘Gros Michel’ banana while Mokara cultured on medium supplemented with Potato extract had the maximum fresh weight (0.27 g) and number of roots (5.20 roots/shoot) statistically different (p≤ 0.05) to other treatments. However, Vanda cultured on medium supplemented with ‘Namwa’ banana had the maximum number of roots (3.80 roots/shoot). Our results suggested that growth of different orchid genera was responded diversely to different organic supplements. 

 

Keywords: Fresh weight, in vitro propagation, orchid, plant height.

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Authors: Aree Thongpukdee, Chockpisit Thepsithar, Chuthalak Thammaso

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Use of plants grown in local area for edible has a long tradition in different culture. The indigenous knowledge such as usage of plants as vegetables by local people is risk to disappear when no records are done. In order to conserve and transfer this valuable heritage to the new generation, ethnobotanical study should be investigated and documented. The survey of vegetable plants traditionally used was carried out in the year 2012. Information was accumulated via questionnaires and oral interviewing from 100 people living in 36 villages of 9 districts in Amphoe Huai Mek, Kalasin, Thailand. Local plant names, utilized parts and preparation methods of the plants were recorded. Each mentioned plant species were collected and voucher specimens were prepared. A total of 55 vegetable plant species belonging to 34 families and 54 genera were identified. The plant habits were tree, shrub, herb, climber, and shrubby fern at 21.82%, 18.18%, 38.18%, 20.00% and 1.82% respectively. The most encountered vegetable plant families were Leguminosae (20%), Cucurbitaceae (7.27%), Apiaceae (5.45%), whereas families with 3.64% uses were Araceae, Bignoniaceae, Lamiaceae, Passifloraceae, Piperaceae and Solanaceae. The most common consumptions were fresh or brief boiled young shoot or young leaf as side dishes of ‘jaeo, laab, namprik, pon’ or curries. Most locally known vegetables included 45% of the studied plants which grow along road side, backyard garden, hedgerow, open forest and rice field.

Keywords: Ethnobotanical survey, Thailand, vegetable plants.

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Authors: Sunthari Tharapan, Chockpisit Thepsithar, Kullanart Obsuwan

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This study was aimed to investigate the effect of various organic supplements on growth and development of Dendrobium discolor’s protocorms and seedlings growth of Dendrobium Judy Rutz. Protocorms of Dendrobium discolor with 2.0 cm. in diameter and seedlings of Dendrobium Judy Rutz at the same size (0.5 cm. height) were sub-cultured on Hyponex medium supplemented with cow milk (CM), soy milk (SM), potato extract (PE) and peptone (P) for 2 months. The protocorms were developed to seedlings in all treatments after cultured for 2 months. However, the best results were found on Hyponex medium supplemented with P was the best in which the maximum fresh and dry weight and maximum shoot height were obtained in this treatment statistically different (p ≤ 0.05) to other treatments. Moreover, Hyponex medium supplemented with P also stimulated the maximum mean number of 5.7 shoots per explant which also showed statistically different (p ≤ 0.05) when compared to other treatments. The results of growth of Dendrobium Judy Rutz seedlings indicated the medium supplemented with 100 mL/L PE enhanced the maximum fresh and dry weigh per explants with significantly different (p ≤ 0.05) in fresh weight from other treatments including the control medium without any organic supplementation. However, the dry weight was not significantly different (p ≤ 0.05) from medium supplemented with SM and P. There was multiple shoots induction in all media with or without organic supplementation ranging from 2.6 to 3 shoots per explants. The maximum shoot height was also obtained in the seedlings cultured on medium supplemented with PE while the longest root length was found in medium supplemented with SM.

Keywords: Fresh weight, in vitro propagation, orchid, plant height.

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Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im

Abstract:

Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by Zona pellucida with layer of cumulus cells ranging between 59.29-202.14 μm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 μg/mL porcine follicle-stimulating hormone, 1 μg/mL LH, 1μg/mL estradiol with ethanol, and 50 μg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into Zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: Cumulus cells, electron microscopy (SEM and TEM), in vitro, porcine oocyte.

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