| Summary: | 碩士 === 國立屏東科技大學 === 熱帶農業研究所 === 90 === Abstract
The purposes of this study are to develop the techniques of callus induction, suspension cell culture, embryogenesis of tomato, to establish a economical, effective and safety system for somatic embryogenesis of tomato and to serve as the references for micropropagation of virus-free seedling and the selection and breeding of disease-resistant strains. Using the stem tissue and leaf disc of 45~50-day-old of Lycopersicon esculentum var. Santa., as the materials to evaluate the effects of the compositions of medium, plant growth regulator, amino acid and vitamin, sucrose concentration, light intensity and explants on callus induction of tomato. The results indicated that the upper and middle sections of the stem and mesophyll cultured on CS-1 medium supplemented with 10~12 mg/l NAA, 6 vitamins (0.5mg/l thiamine-HCl, 0.5 mg/l pyridoxine-HCl, 0.5 mg/l nicotinic acid, 0.1mg/l biotin, 0.4 mg/l folic acid and 0.5 mg/l ascorbic acid), 2 amino acids (1 mg/l glutamic acid and 1 mg/l aspartic acid) and 3% sucrose at 3200~4200 lux light intensity for 10 days were best in callus formation, inducing a high yield, friable, low-browned of callus as well as shorten the induction time. In order to induce the vital and highly regenerable callus, the effects of the compositions of medium, sources and contents of carbon, pH values, light intensity, sources of the culture cell and density of the culture cell on the proliferation of culture cell were evaluated. The results showed that 2.5×105 cells/ml culture cells, isolated from the stem tissue of L. esculentum var. Santa., shaking at 100 rpm in CSW-1 liquid medium supplemented with 10 mg/l NAA, 3% sucrose, 0.03% glucose and 0.03% fructose (pH 6.0~6.5) in the dark for 7 days had the highest proliferation, which increased callus formation for 1.18 times and decreased the rupture of the culture cells. The tomato cells cultured under the same conditions produced, the growth curves with phases identified as the lag phase occurred in 0~1 days, the exponential phase in 1~4 days, the linear phase in 4~5 days, the progressive deceleration in 5~9 days, and the stationary phase in 9~10 days, Consequently the time requested for subculture of cells was 4~5 days. Using the technique of somatic embryogenesis of tomato, the compositions of the plant growth regulator, sucrose concentration, light intensity, sources of the culture cell and number of days requested for of the cell cluster induction on the single cells differentiate to the somatic embryos were evaluated. The culture cells, isolated from the middle section of the stem tissue, which were first cultured in CSW-1 liquid medium supplemented with 10 mg/l NAA in the dark condition for 7 days for cell differentiate and then subcultured in CSW-1 liquid medium supplemented with 1 mg/l NAA and 5~7 mg/l BA in the dark condition for 14 days (replaced with fresh medium at intervals of one week) resulted in the best somatic embryogenesis, about 14.9%.
|
|---|
