Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, signific...Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences （P〈0.05） were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences （P〈0.05） in the total number of colony forming units （CFUs） of bacteria, actinomycetes and fungi between soils amended with RP-transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher （P〈0.05） CFUs of bacteria, actinomycetes and fungi resistant to kanamycin （Km） were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Km＇ （kanamycin resistant） bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Km＇ bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase （5.4-5.9x）, polyphenol oxidase （0.7-1.4x）, invertase （0.5-0.79x）, cellulase （0.23-0.35x） and phosphodiesterase （0.16-0.2x）. The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities.展开更多
为筛选出黄籽油菜遗传转化中卡拉霉素或卡拉霉素（kanamycin ，Kan or km ）最佳质量浓度，首先将油菜子叶接种在不同质量浓度Kan（0～180 mg／L）的分化培养基上，结果发现Kan质量浓度小于60 mg／L有利于绿苗分化。然后在不同质量浓度...为筛选出黄籽油菜遗传转化中卡拉霉素或卡拉霉素（kanamycin ，Kan or km ）最佳质量浓度，首先将油菜子叶接种在不同质量浓度Kan（0～180 mg／L）的分化培养基上，结果发现Kan质量浓度小于60 mg／L有利于绿苗分化。然后在不同质量浓度Kan的分化培养基上对农杆菌侵染的下胚轴进行筛选，结果表明初代培养Kan质量浓度以20～40 mg／L为宜，同样质量浓度继代培养后可获得抗性愈伤和转基因阳性苗。最后利用Kan抗性对转基因黄籽油菜后代进行研究，苗期到薹花期生长的植株进行叶片涂抹法筛选，Kan最适质量浓度为4 g／L ，时间为4～10 d；T1代抗性分析、GUS染色分析、T1代籽粒的PCR检测说明转基因在T1代、T2代可遗传并出现分离。β葡萄糖苷酸酶基因（GUS）和聚合酶链式反应（PCR）验证叶片涂抹进行Kan抗性筛选的方法准确性高，可达100％。展开更多
A 4.34kb EcoR I fragment of kanamycin resistance plasmid from pET - 9a was purified by a DNA purification kit. The fragment was labeled with digoxigenin-dUTP with a commercial kit. A dot-blot hybridization and a colon...A 4.34kb EcoR I fragment of kanamycin resistance plasmid from pET - 9a was purified by a DNA purification kit. The fragment was labeled with digoxigenin-dUTP with a commercial kit. A dot-blot hybridization and a colony hybridization test with the probe were successfully developed for the surveillance of Kanamycin resistance to E.coli from swine. It was shown that the methods obtained 100% concordance in a positive tate. It was indicated that the method was available for the surveillance of kanamycin resistance to ? coli from swine.展开更多
文摘Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences （P〈0.05） were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences （P〈0.05） in the total number of colony forming units （CFUs） of bacteria, actinomycetes and fungi between soils amended with RP-transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher （P〈0.05） CFUs of bacteria, actinomycetes and fungi resistant to kanamycin （Km） were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Km＇ （kanamycin resistant） bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Km＇ bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase （5.4-5.9x）, polyphenol oxidase （0.7-1.4x）, invertase （0.5-0.79x）, cellulase （0.23-0.35x） and phosphodiesterase （0.16-0.2x）. The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities.
文摘A 4.34kb EcoR I fragment of kanamycin resistance plasmid from pET - 9a was purified by a DNA purification kit. The fragment was labeled with digoxigenin-dUTP with a commercial kit. A dot-blot hybridization and a colony hybridization test with the probe were successfully developed for the surveillance of Kanamycin resistance to E.coli from swine. It was shown that the methods obtained 100% concordance in a positive tate. It was indicated that the method was available for the surveillance of kanamycin resistance to ? coli from swine.
