Dissertation
Dissertation > Agricultural Sciences > Gardening > Fruit trees gardening > Miscellaneous fruit > Pomegranate

Preliminary Study on the Regeneration System and the Reporter Gene GFP Instantaneous Expression of Soft-seed Pomegranate Variety ’Tunisia’

Author LianHongKe
Tutor ChenYanHui
School Henan Agricultural University
Course Pomology
Keywords Soft-seed pomegranate variety‘Tunisia’ Leaf regeneration Stem regeneration GFP Genetic transformation
CLC S665.4
Type Master's thesis
Year 2011
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Using the leaves and stems of soft-seed pomegranate variety‘Tunisia’as the explants, the regeneration system, the effect of different concentrations antibiotics on the explants and reporter gene GFP instantaneous expression were discussed. The results of the research were indicated as follows:1. The leaf regeneration system of soft-seed pomegranate variety‘Tunisia’was established. The results indicated that: the vitro leaves from climate chamber were soaking 4min in 0.1% HgCl2; the most suitable medium of leaves differentiation into callus was MS+BA1.0mg/L+NAA1.0mg/L; the most suitable medium of adventitious buds formation was MS+NAA0.3mg/L+TDZ1.5mg/L+KT1.0mg/L; the most suitable medium of stem formation was MS+NAA1.0mg/L+BA0.2mg/L+KT1.0mg/L rooting medium was MS+1/2NAA0.5mg/L, the new roots were stronger and the transplant survival percent could reach more than 95%.2. The stem segments regeneration system of soft-seed pomegranate variety‘Tunisia’were established.The results indicated that: the vitro stems from climate chamber were soaking 5min in 0.1% HgCl2; the most suitable medium of stems differentiation callus was MS+ NAA0.1mg/L+TDZ1.0mg/L;the most suitable medium of adventitious buds formation was MS+ NAA 0.3mg/L +TDZ 1.5mg/L + KT1.0mg/L; the most suitable medium of stem formation was MS+NAA 1.0mg/L+ BA0.2mg/L + KT 1.0mg/L; rooting medium was MS + 1/2NAA 0.5mg/L, the new roots were stronger and the transplant survival percent could reach more than 95%.3. The superiorities of 2 regeneration systems of soft-seed pomegranate variety‘Tunisia’were compared. The research showed that: the comprehensive effect of leaves regeneration was better than that of stems regeneration.4. The PVP concentration for soft-seed pomegranate variety‘Tunisia’was sifted. The results indicated that: the optimal PVP concentration was 1.0g/L.5. The antibiotic concentrations for soft-seed pomegranate variety‘Tunisia’were sifted. The results indicated that: callus was significantly inhibited when the Kan concentration was 50mg/L, the genetically engineered materials could be sifted; the strains growing could be effectively restrained when the Cef concentration was 500mg/L.6. The instantaneous expression state of reporter GFP gene was researched. The green fluorescent was all found in genetically modified materials and controls and there were not obvious difference.

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