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หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Isolation and Culture of Protoplasts from Different Explants of Cotton (Gossypium hirsutum L.)) ผู้เขียน:ดร.ประพนธ์ บุญรำพรรณ, Catherine Pannetier, Yves Chupeau สื่อสิ่งพิมพ์:pdf AbstractProtoplasts were isolated from callus, cotyledons, hypocotyls and leaves of cotton (Gossypium hirsutum L. var. Coker 310) using different digestion media. Protoplast yield per gram of fresh mass of leaves, callus cotyledons and hypocotyls was 22.2, 5.5, 1.7 and 1.1 million respectively. Protoplasts were most viable in basic medium in presence of thidiazuro 0.663 gm/l and 2, 4, 5-trichlorophenoxy acetic acid 7 mg/l. Hypocotyl and leaf-derived protoplasts formed microcolonies. Addition of CaCl2 and glutamine to the basic medium improved protoplast survival, and division of hypocotyl and leaf-derived protoplasts. The growth conditions, such as the light and carbon source of donor plant were the important factors for protoplast isolation and culture. |
หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Construction of High Ethanol Fermenting Halotolerant Hybrid by Intergeneric Protoplast Fusion of Saccharomyces cerevisiae and Zygosaccharomyces rouxii) ผู้เขียน:ดร.สาวิตรี ลิ่มทอง, ศาสตราจารย์, สมคิด ดีจริง, ดร.วิเชียร ยงมานิตชัย, อาจารย์, Wilia Santisopasri สื่อสิ่งพิมพ์:pdf AbstractSaccharomyces cerevisiae M30, a high ethanol producing flocculent strain, was hybridized with Zygosaccharomyces rouxii TISTR1750, a halotolerant non-flocculent strain, by polyethylene glycol induced protoplast fusion. The protoplast fusion of these two yeast genera occurred with frequency of 5.89X10-5. Three fusants, namely RM11, RM37 and RM58, were selected based on their high ethanol fermentation efficiency in glucose broth containing 1.5 and 3% sodium chloride. Ethanol fermentation performance in salted medium of the three fusants revealed that in glucose broth containing 1.5, 3, 5 and 7% sodium chloride RM11 produced the highest ethanol concentrations and yields when compared to the other two fusants and both parental strains. Ethanol production in semisynthetic medium consisted of 18% glucose and 3% sodium chloride in 5 l fermentor with controlling of pH not to be lower than pH 4.5, temperature at 30 C and 300 rpm agitation speed also showed that the efficiency of ethanol production of the fusant RM11 was the highest comparing with both parental strains. It revealed maximal ethanol as 6.85% by weight with 87% of theoretical yield at 60 h of cultivation. S. cerevisiae M30 produced ethanol 6.5% by weight with 83% of theoretical yield, while Zygosac. rouxii TISTR1750 produced 6.36% by weight with 79% of theoretical yield at the same cultivation period. The fusant RM11 exhibited flocculation the same as that of S. cerevisiae M30. |
หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Intraspecific Hybridization Between Color Mutants of Trichoderma harzianum by Protoplast Fusion ) ผู้เขียน:Chiradej Chamswarngt, Kihashiro Ogawa, Wanwilai Gesnarat, Nuanwan Faroongsang สื่อสิ่งพิมพ์:pdf AbstractProtoplasts of two color mutants of Trichoderma harzianwn derived from ultraviolet treatment were prepared by using lytic enzyme system (10 mg Zymolvase. 150 mg Uzukizyme, 10 mg Chitinase and 150 mg Funcelase in 3 ml water, pH 5.0) containing 0.6 M NaC 1 as an osmotic stabilizer. Fusion between protoplasts nt two color mutants was observed in the solution containing 0.05 M glycine-NaOH buffer, pH 7.5, 0.05 M CaCI . 2H-,0 and 35% polyethylene glycol ( MW. 6,000). The fused protoplasts were successfully detected at the rate of '2 % as green colonies developed on a hypersonic solid culture medium (complete medium) overlaid with hypertonic soft medium (MM with 0.5% agar). Production of well defined white and yellow sectors on tear surface of minimal medium supplemented with 0.1U (W/V) d-camphor represented the heterokaryotic usants. Green conidia were constantly obtained after transferring the green part from green-sectored colonies :.into MM agar. This indicated that protoplast fusants were heterozygous diploid. |
หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Protoplast Isolation and Culture of Dendrobium Sonia “Bom 17”) ผู้เขียน:ยุพิน เคนตรี, Ampaiwan Paradornuvat, สุรียา ตันติวิวัฒน์, ดร.ศาลักษณ์ พรรณศิริ, ดร.นิพนธ์ ทวีชัย, ศาสตราจารย์ สื่อสิ่งพิมพ์:pdf AbstractThe optimization of Dendrobium Sonia “Bom 17” protoplast isolation and culture conditions were investigated. Protoplasts were successfully isolated from one-month-old leaves of in vitro-grown plantlets using an enzyme solution comprising 1% Cellulase Onozuka R-10, 0.2% Macerozyme, 0.3 M mannitol, 10 mM CaCl2.2H2O and 10 mM 2 (N-morpholino)-ethanesulfonic acid (MES) at pH 5.8. Approximately 3.97?0.63 ? 105 protoplasts per gram fresh weight were obtained when digested in enzyme solution for 4 h on a shaker under dark conditions, then washed with 0.3 M mannitol solution supplemented with 10 mM CaCl2.2H2O and 10 mM MES, and purified with 0.3 M sucrose solution. Freshly isolated protoplasts were then cultured at final density of 2 ? 105 protoplasts/ml in Kao & Michayluk medium (KM8P) by agarose-bead cultures under dark conditions at 25?C. First cell division was observed after culturing for 2 days and multicellular colonies (15-20 cells) were formed after 2 weeks. |
หัวเรื่อง:การพัฒนาไปเป็นต้นอ่อนจากโปรโตพลาสของพืชตระกูลกระหล่ำในสภาพ in-vitro ผู้เขียน:Ariyaporn Pongrat สื่อสิ่งพิมพ์:pdf AbstractNine species of Brassica family were studied for the genetic control of in vitro regenerability from protoplast. In Brassica family, the protoplast culture should be kept in the dark after the protoplast isolation for the first three days to reduce the phenols as they affected cell division and growth of cells during the culture. The callus formation did not depend on the genotype since the protoplasts of all examined species showed a high microcallus and callus formation in V-KM and on MS-media, respectively. The protoplast regeneration of Brassicaceae was strongly dependent on the genotype. The examined species with the Agenome, namely Brassica campestris (AA), B. juncea (AABB) and B. napus (AACC) showed low capability of regeneration. Probably, the A-genome has an inhibitory effect on the regeneration in Brassica species. The genomes of B. nigra (BB) and B. oleracea (CC) had high frequency of protoplast regeneration. Their interaction enhanced the ability to regenerate since B. carinata (BBCC) showed the best regenerability from protoplasts. Moreover, Erucia sativa, Raphanus sativus and Sinapis alba could not form shoot from protoplasts. |
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หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Protoplast Isolation and Culture of Aquatic Plant Cryptocoryne wendtii De Wit) ผู้เขียน:กาญจนรี พงษ์ฉวี, ดร.อุทัยรัตน์ ณ นคร, ศาสตราจารย์, นางสิรนุช ลามศรีจันทร์, ศาสตราจารย์, ดร.สุภาวดี พุ่มพวง, ศาสตราจารย์ สื่อสิ่งพิมพ์:pdf AbstractThe optimum conditions for protoplast isolation and culture of Cryptocoryne wendtii De Wit were investigated. Protoplasts were successfully isolated from in vitro four-week-old leaves using an enzyme mixture comprising 2% Cellulase Onozuka R-10, 0.2% Pectolyase Y-23, 0.5 M mannitol, 2.5 mM CaCl2.2H2O and 5 mM 2 (N-morpholino)-ethanesulfonic acid (MES), pH 5.6. Approximately 1.04?0.06 ? 107 protoplasts per gram fresh weight with 90.79?4.80% viability were obtained after incubating in enzyme solution for 4 hours in the dark and purified with 16 % sucrose gradient centrifugation. Protoplasts were cultured on modified MS medium supplemented with 0.2 mg/l 2,4- dicholorophenoxyacetic acid (2,4-D), 1 mg/l ?-naphthalene acetic acid (NAA), 0.5 mg/l zeatin, 0.15 M sucrose and 0.3 M mannitol by agarose-bead with thin layer liquid culture. The protoplasts regenerated cell walls within 24 hours. First cell division was observed after culturing for 2-3 days, and microcolonies were formed within 4 weeks. Enzyme mixture, osmotic solution, incubation time, age of leaves, and sucrose solution concentration were found to influence both yield and viability of protoplasts. Culture media, plant growth regulators and method of culture affected protoplast division. |
หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Functional Golden Brown Rice Fermentation by Monascus sp. Yellow, White Parental Strains and Their Protoplast Mutants and Fusants) ผู้เขียน:Panida Thongpradis, ดร.สาวิตร ตระกูลน่าเลื่อมใส, ผู้ช่วยศาสตราจารย์, ดร.เจษฎา โพธิรัตน์, ผู้ช่วยศาสตราจารย์, ดร.บุษบา ยงสมิทธ์, ศาสตราจารย์ สื่อสิ่งพิมพ์:pdf AbstractThis report points out the possibility of producing value-added fermented products from brokenmilled rice using yellow pigment-producing molds: Monascus kaoliang TISTR 3642 and its developed strains, 2 protoplast mutants (U2.3 and U2.5) and 3 fusants (F7, F10 and F43). All strains could produce yellow pigments with a single absorption peak at 370 nm, and the amount of pigment produced in depended on the pH and medium formula. Various factors affecting the growth rate, glucoamylase and yellow pigment activities of Monascus in golden-brown rice fermentation were investigated and compared among the parental strains and the five developed strains. Conditions of 38% weight per weight initial moisture content with an initial pH 5 of rice substrate, and room temperature (28–32 ?C) incubation were found optimal for golden brown rice fermentation of all strains. Furthermore, their noteworthy functional properties of antioxidant activities, total phenolic compounds as well as antichoresterol agents (monacolin K) were observed and discussed. |
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หัวเรื่อง:เทคนิคการเตรียมโปรโตพลาสต์ของ osmotolerant yeast ผู้เขียน:ดร.มณี ตันติรุ่งกิจ, Orawan Anuvongpathom สื่อสิ่งพิมพ์:pdf Abstract Protoplasts of osmotolerant yeast strain AM-06 and AM-35 were isolated by digesting cell wall with lytic emzyme mixture containing 10 mg zymolyase-5000, 0.1 M 2-mercaptoethanol and 2/15 M phosphate buffer ( pH 7.5 ) in the presence of 0.6 M and 1.0 M KCl, respectively. The higher yield of protoplast formation was obtained from cells in log phase. In term of osmotic stabilizer, sorbital and mannitol can be used instead of KCl. |
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