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Somatic embryogenesis of Robusta coffee, strain FRT17, was employed using leaf as explants. The experiment was conducted using 2 kinds of media, 23A8 and Peirson et al. (1983) supplemented with various concentrations of auxin and cytokinin. The explants showed better response on Peirson et al. (1983) media + 5 mg/l IBA + 1 mg/l 2ip (20 % response). In addition, temporary Immersion Bioreactor (TIB) was used in in vitro pre-germination stage of mass propagation of Robusta Coffee via Somatic Embryogenesis. Advantage of TIB are to reduced labor cost and media can be changed easily. Two experiments of TIB were used: (1) TIB are glass made using with air pressure pump, (2) TIB are polyethylene made using without air pressure pump. TIB (1) were renewed media once a month, while TIB (2) were filled 0.5 L of liquid media. Cotyledon embryos were produced on the average 1,400 embryos per system by TIB (1). TIB (2) produced on the average 1,000 embryos per system.
The study of plant tissue culture for micropropagation was employed for good oil palm planting material. Tissue culture of oil palm (tenera type),Surat-thani 3 had been done using immature embryo, immature inflorescence and young leaf primordia surrounding the cabbage as explants. Explants had been cultured on MS and Y3 media supplemented with various concentrations of dicamba for callus induction. Immature embryo showed the highest callus percentage at 83.30% on MS media supplemented with 10 ?M dicamba .Immature influrescense and young leaf primordia showed the highest callus percentage at 15.8% on Y3 media supplemented with 15 ?M dicamba, and 24.63% on MS media supplemented with 15 ?M dicamba, respectively. Callus proliferation ability of immature embryo, immature inflorescence and young leaf primordia were 7.02 x, 3.87x ,and 9.13x, on MS media supplemented with 1 ?M, 3 ?M and 1 ?M of dicamba, respectively. The percentage of embryogenic callus induction from the original callus were 50.01% 20.04% and 46.76% from immature embryo , immature inflorescence and young leaf primordia respectively on Y3 media supplemented with NAA 10 ?M and abscisic acid 2 ?M. In addition, the development of somatic embryo from embryogenic callus of each explant parts were 40.08%, 13.36%, and 33.34%, respectively on the same media of embryogenic callus induction. Development of somatic embryo can be observed in diverse types, normal plantlets, polyembryony with 2-3 shoots and normal root, or embryo derived shoot without root. However, these shoots can be rooted on root induction media, ? MS supplemented with NAA 15-45 ?M, percentage of rooting can be obtained at 28-31 %.
The study of the effect of polyamine and silver nitrate for callus proliferation was employ using embryogenic callus as starting material. Callus was culture on MS media supplemented with silver nitrate and 2ip (2-isopentyladenine) at 0.1 mg/l. MS media without silver nitrate showed the higher callus proliferation compare to the media with silver nitrate. The other experiment was done using polyamine, putrescine, spermidine, and spermine at 0, 1, 10, 100 uM each. After 3 months of culture, MS media supplemented with spermidine 10 uM displayed the highest callus proliferation (1.19 gm.). Yellow and green friable callus was emerged from the starting material. In contrast, MS media without polyamine showed the lowest callus proliferation.
Callus proliferation in liquid media was done using opaque embryogenic callus as starting material (0.15-0.2 gm.). After 4 months of culture, MS media supplemented with 50 mg/l 2,4-D showed the highest callus proliferation (3.84 times compare to the starting fresh weight). Callus was then transferred on MS media without plant growth regulator. However, development of callus proliferated in liquid media was lower than that of callus proliferated on solid media. Somatic embryo, obtained from liquid culture callus, has haustorium shape and unable to develop to normal embryo.
Plant tissue culture of oil palm has been applied to the other type of oil palm, fertile pisifera type. Immature embryo and immature female inflorescence were used as explants. The highest callus induction was obtained on MS media supplemented with 5-10 uM and 10-15 uM dicamba from immature embryo (62.4-69.6 %) and immature female inflorescence (16.0-18.4 %), respectively. The highest proliferation of embryogenic callus was obtained on MS media with 2-4 uM dicamba, 6.9-7.2 times from immature embryo and 3.8 times from immature female inflorescence compare to the starting fresh weight, respectively.
Micropropagation of two medicinal plants, Zingiber cassumunar Roxb. and Pueraria candollei Grah. Ex Benth. var mirifica have been done for pathogen-free planting material production. For Zingiber cassumunar Roxb., rhizome has been used as explant. The highest multiple shoot formation was obtained from LS media supplemented with 1 mg/l BA (4.32 shoots/explant). Survival percentage of microshoot in field condition was 98.33 %. Tilling of tissue culture shoots (14.2 shoots) was higher than that of tilling of conventional shoot (shoots emerged from rhizome, 8.5 shoots). For Pueraria candollei micropropagation, immature leaves have been used as explants. The highest callus induction was obtained from MS media + 1.25 mg/l NAA + 0.125 mg/l BA (65%) and MS media + 2.21 mg/l 2,4-D + 0.48 mg/l kinetin (55%). Callus was friable and unable to develop to shoot on various type of media (WPM + NAA + BA + kinetin).
Paphiopedilum is a wild Thai orchid that is very popular as a ornamental plant. Seven local species of this genus have been studies for micropropagation. The appropriate seed propagation media of Lueng Pra-geen was Jitraphan II and Jitraphan III. For Inthanon, Inthanon Lao, Far Hoi, and Doi Tung, seed germination media was ? Jitraphan II followed by transferred to growth promotion media including ? macronutrients of VW (1949) and ? micronutrients of MS (1962) + 75 ml/l coconut milk + 50 gm/l potato mix + 12.5 gm/l mushroom mix and 25 gm/l banana mix. In addition, tissue culture of Paphiopedilum godefroyae was employed using 8 month-old root as explants. The root explants were culture on induction media, MS media supplemented with 2 mg/l BA and 0.5 mg/l NAA. For growth promotion, the microshoots were moved on MS media supplemented with 2 mg/l BA, 0.5 mg/l NAA, and 2 gm/l activated charcoal.
In Vitro propagation of endangered plant has been done in 3 types of wild plants, Aerides krabiensis Seidenf., Vandopsis lissochiloides (Gaud.) Pfitz), and Impatiens Psittacina Hook.f.). The appropriate media for seed propagation of Aerides krabiensis composed of VW media supplemented with 150 ml/l coconut milk, 50 gm/l banana mix, and 5 gm/l sucrose or VW media supplemented with 150 ml/l coconut milk, 100 gm/l potato mix, 10 gm/l sucrose, and 100 gm/l banana mix. For Vandopsis lissochiloides seed propagation, the media composed of VW basal media supplemented with 100 ml/l coconut milk and 50 gm/l banana mix. Seed germination media and axillary bud culture media of Impatiens Psittacina was MS basal media supplemented with 1 and 5 mg/l BA, respectively.
Induction of genetic variation using mutagen or colchicine has been done via plant tissue culture. The treated materials were selected for new characteristics. Multiple shoot propagation of three clones of torch ginger (Etlingera elatior (Jack) R.M. Smith), Bua Daeng Yai, Daeng Indo, and Barn Yen was done using emerged sprouts cultured on Murashige and Skoog (MS) media supplemented with 6-benzylaminopurine (BA) 20-30 uM. Within 2 months, 3.5-4.3 microshoots/sprout was obtained. Each microshoot was cultured for further multiplication and chromosome doubling induction by colchicine. The microshoots were treated with colchicine at 0.03, 0.06 and 0.09%(w/v) for 1, 2 and 3 days in liquid MS media supplemented with BA 10 uM. The best chromosome doubling treatment were 0.06% (w/v) colchocine at 2-3 days. At 0.09% (w/v) colchicine resulted in lethal tissue. Ploidy level of the treated microshoots was investigated by flow cytometry technique using the M1V2 microshoots. Mixoploid microshoots were found in all clones. Treated microshoots (M1V2) displayed 4n peak area above 60%(w/v) were selected for further multiplication. M1V4 microshoots were reinvestigated for ploidy level and 5 microshoots from Bua Daeng Yai clone showed only tetraploid (4n) peak whereas the others two clones displayed mixoploid.
Elephant yam were irradiated by gamma ray at 4.3 and 2.9 krad, callus displayed survival percentage at 35 and 43 %, respectively and microshoots can survive at 38 and 52 %, respectively. However, the microshoots develop from treated callus and treated shoots showed abnormal growth and malformed organs. High tilling and dwarf plantlets were found frequently after subculture. Sun tolerant plantlet was obtained from the preliminary screening. However, the plant was unable to survive in field condition. |
