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Chloramphenicol acetyl transferase (CAT) gene was used as a reporter gene to assess the conditions for polyethylene glycol (PEG)-mediated transfection of kiwifruit protoplasts. The effect of plasmid concentration and the presence of carrier DNA were each assessed by analysing CAT activity in transfected protoplasts using thin-layer chromatography (TLC) autoradiographic detection of acetylated chloramphenicol. A gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) non-radioactive method was developed for monitoring CAT gene activity. This method provides a high speed of analysis (30 min) and precise means of detecting acetylated products at the nanomolar level, enabling quantification at very low transfection rates. Using this method we optimized plasmid and PEG concentration and also assessed the effect of heat shock on transfection. The best CAT activity was obtained using 30% polyethylene glycol 4000 and by submitting protoplasts to heat shock (45 °C, 5 min) prior to transfection. 相似文献
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栽培中华猕猴桃的染色体观察 总被引:5,自引:0,他引:5
对原产地位于赣鄂边界幕阜山地区的十二个大果型中华猕猴桃优株或株系的染色体数目观察表明,这些栽培类型全部为四倍体,2n=4x=116。讨论了染色体倍性与果实大小的关系及几种可能的育种方法。 相似文献
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David H. Lewis Garry K. Burge David M. Schmierer Paula E. Jameson 《Physiologia plantarum》1996,98(1):179-186
The cytokinin content in fruit tissue of the kiwifruit ( Actinidia deliciosa [A. Chev.] C. F. Liang et A. R. Ferguson var. deliciosa cv. Hayward) was monitored during fruit development to identify which cytokinins were present and if they were linked with specific stages of fruit growth. Cytokinins were isolated and purified by column chromatography and high-performance liquid chromatography and quantified by radioimmunoassay. A novel HPLC step utilising an amine column was successfully introduced as a preparative step in the separation of the O - and 9-glucosides from the free bases and ribosides. The radioimmunoassay results were validated, and the different cytokinins identified, by gas chromatography-mass spectrometry. Cytokinins detected in fruit included the cytokinin free bases, zeatin and isopentenyladenine, their ribosides, nucleotides and both O - and 9-glucosides. Both qualitative and quantitative changes of the cytokinins occurred during fruit development. A decrease in cytokinin concentration occurred after anthesis (from 342 pmol g−1 fresh weight at anthesis to 41 pmol g−1 fresh weight 27 days after anthesis). A large increase in cytokinin concentration and content per fruit occurred as the fruit reached commercial maturity (to 1900 pmol g−1 fresh weight). Individual cytokinins showed quite different patterns. Zeatin, in particular, showed a peak in concentration (13 pmol g−1 fresh weight) 11 days after anthesis that correlated with the beginning of the cell division phase of fruit growth. The accumulation of cytokinin (mostly zeatin riboside or zeatin nucleotide) in mature fruit may be of significance for the postharvest storage of kiwifruit fruit. 相似文献
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为了选育耐贮藏且风味品质优异的猕猴桃(Actinidia)新品种,对美味猕猴桃(A.deliciosa)的3个野生品种‘kvf54’、‘kvf6’、‘鑫美’和3个栽培种‘秦美’、‘华美’、‘徐香’的品质和贮藏特性等进行比较分析。结果表明,常温和1℃贮藏下‘kvf54’和‘徐香’的硬度下降趋势相近,优于其他品种,且‘kvf54’的维生素C (VC)含量最高。而在相同贮藏条件下,‘鑫美’可溶性固形物含量积累速度最快,淀粉和可滴定酸含量最少,贮藏性能不佳。‘kvf54’相比于其他品种的贮藏性更好,VC、糖类物质等含量高,是可用于培育耐贮藏新品种的优质材料。 相似文献